U.S. patent application number 15/588845 was filed with the patent office on 2017-08-24 for crystalline pl3 kinase inhibitors.
This patent application is currently assigned to Respivert Limited. The applicant listed for this patent is Respivert Limited. Invention is credited to Rudy Laurent Maria Broeckx, Alex Herman Copmans, Walter Ferdinand Maria Filliers, Carina Leys, Patrick Hubert, J Nieste, Filip Marcel Vanhoutte.
Application Number | 20170239256 15/588845 |
Document ID | / |
Family ID | 49160309 |
Filed Date | 2017-08-24 |
United States Patent
Application |
20170239256 |
Kind Code |
A1 |
Broeckx; Rudy Laurent Maria ;
et al. |
August 24, 2017 |
CRYSTALLINE Pl3 KINASE INHIBITORS
Abstract
There is provided inter alia
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-di hydroquinazolin-5-yl)-N,
N-bis(2-methoxyethyl)hex-5-ynamide in the form of a solid
crystalline hydrate and in solid crystalline anhydrous form. There
are also provided dry powder pharmaceutical compositions for
inhalation containing such solid crystalline forms.
Inventors: |
Broeckx; Rudy Laurent Maria;
(Beerse, BE) ; Filliers; Walter Ferdinand Maria;
(Beerse, BE) ; Nieste; Patrick Hubert, J; (Beerse,
BE) ; Copmans; Alex Herman; (Beerse, BE) ;
Vanhoutte; Filip Marcel; (Beerse, BE) ; Leys;
Carina; (Beerse, BE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Respivert Limited |
Buckinghamshire |
|
GB |
|
|
Assignee: |
Respivert Limited
Buckinghamshire
GB
|
Family ID: |
49160309 |
Appl. No.: |
15/588845 |
Filed: |
May 8, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14381289 |
Aug 27, 2014 |
9642799 |
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PCT/GB2013/050624 |
Mar 13, 2013 |
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15588845 |
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61610012 |
Mar 13, 2012 |
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61610023 |
Mar 13, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 31/519 20130101; A61P 7/00 20180101; A61K 47/26 20130101; A61K
31/52 20130101; A61P 11/06 20180101; C07D 487/04 20130101; A61P
43/00 20180101; A61P 29/00 20180101; A61P 35/04 20180101; A61K
47/12 20130101; A61P 11/00 20180101; A61P 37/00 20180101; A61K
9/0075 20130101; A61K 9/0073 20130101; A61P 11/14 20180101 |
International
Class: |
A61K 31/519 20060101
A61K031/519; A61K 9/00 20060101 A61K009/00; A61K 47/12 20060101
A61K047/12 |
Claims
1.-19. (canceled)
20. A method of treating a condition selected from COPD, asthma,
cystic fibrosis, sarcoidosis, idiopathic pulmonary fibrosis,
cachexia, and lung cancer, said method comprising administering to
a subject an effective amount of a compound of formula (I) in
crystalline form, ##STR00011## wherein the crystalline form is a
hydrate or is anhydrous.
21. The method of claim 20, wherein the condition is COPD.
22. The method of claim 21, wherein the condition is chronic
bronchitis and emphysema.
23. The method of claim 20, wherein the condition is asthma.
24. The method of claim 23, wherein the condition is paediatric
asthma.
25. The method of claim 20, wherein the condition is lung
cancer.
26. The method of claim 25, wherein the condition is non-small cell
lung carcinoma.
27. The method of claim 20, wherein the crystalline form is a
hydrate.
28. The method of claim 20, wherein the crystalline form has an
X-ray powder diffraction pattern comprising one, two, three or four
peaks selected from peaks at (.+-.0.2) 17.6, 18.4, 22.5 and
24.2.degree. 2.theta..+-.0.2.degree..
29. The method of claim 20, wherein the crystalline form is
formulated with one or more pharmaceutically acceptable diluents or
carriers in a pharmaceutical composition.
30. The method of claim 29, wherein the one or more
pharmaceutically acceptable diluents or carriers comprise
lactose.
31. The method of claim 30, wherein the lactose is lactose
monohydrate.
32. The method of claim 20, wherein the crystalline form is further
formulated with a stabilizing agent selected from the group
consisting of metal salts of stearic acid and metal salts of
stearyl fumarate in the pharmaceutical composition.
33. The method of claim 32, wherein the stabilizing agent is
magnesium stearate.
34. The method of claim 29, wherein the crystalline form is
micronized.
Description
FIELD OF THE INVENTION
[0001] The present invention provides novel crystalline forms of a
compound that inhibit phosphoinositide 3-kinases (PI3 kinases), and
their use in therapy, especially in the treatment of inflammatory
diseases such as COPD and asthma. The novel crystalline forms are
suitable for use in dry powder formulations for inhalation.
BACKGROUND OF THE INVENTION
[0002] Lipid kinases catalyse the phosphorylation of lipids to
produce species involved in the regulation of a wide range of
physiological processes, including cellular migration and adhesion.
The PI3-kinases are membrane associated proteins and belong to the
class of enzymes which catalyse the phosphorylation of lipids which
are themselves associated with cell membranes. The PI3-kinase delta
isozyme (PI3 kinase .delta.) is one of four isoforms of type I PI3
kinases responsible for generating various 3'-phosphorylated
phosphoinositides, that mediate cellular signalling and has been
implicated in inflammation, growth factor signalling, malignant
transformation and immunity [See Review by Rameh, L. E. and
Cantley, L. C. J. Biol. Chem., 1999, 274:8347-8350].
[0003] The involvement of PI3 kinases in controlling inflammation
has been confirmed in several models using pan-PI3 kinase
inhibitors, such as LY-294002 and wortmannin [Ito, K. et al., J
Pharmacol. Exp. Ther., 2007, 321:1-8]. Recent studies have been
conducted using either selective PI3 kinase inhibitors or in
knock-out mice lacking a specific enzyme isoform. These studies
have demonstrated the role of pathways controlled by PI3 kinase
enzymes in inflammation. The PI3 kinase .delta. selective inhibitor
IC-87114 was found to inhibit airways hyper-responsiveness, IgE
release, pro-inflammatory cytokine expression, inflammatory cell
accumulation into the lung and vascular permeability in
ovalbumin-sensitized, ovalbumin-challenged mice [Lee, K. S. et al.,
J. Allergy Clin. Immunol., 2006, 118:403-409 and Lee, K. S. et al.,
FASEB J., 2006, 20:455-65]. In addition, IC-87114 lowered
neutrophil accumulation in the lungs of mice and neutrophil
function, stimulated by TNF.alpha. [Sadhu, C. et al., Biochem.
Biophys. Res. Commun., 2003, 308:764-9]. The PI3 kinase .delta.
isoform is activated by insulin and other growth factors, as well
as by G-protein coupled protein signalling and inflammatory
cytokines. Recently the PI3 kinase dual .delta./.gamma. inhibitor
TG100-115 was reported to inhibit pulmonary eosinophilia and
interleukin-13 as well as mucin accumulation and airways
hyperesponsiveness in a murine model, when administered by
aerosolisation. The same authors also reported that the compound
was able to inhibit pulmonary neutrophilia elicited by either LPS
or cigarette smoke [Doukas, J. et al., J Pharmacol. Exp. Ther.,
2009, 328:758-765]
[0004] Since it is also activated by oxidative stress, the PI3
kinase .delta. isoform is likely to be relevant as a target for
therapeutic intervention in those diseases where a high level of
oxidative stress is implicated. Downstream mediators of the PI3
kinase signal transduction pathway include Akt (a serine/threonine
protein kinase) and the mammalian target of rapamycin, the enzyme
mTOR. Recent work has suggested that activation of PI3 kinase
.delta., leading to phosphorylation of Akt, is able to induce a
state of corticosteroid resistance in otherwise
corticosteroid-sensitive cells [To, Y. et al., Am. J. Respir. Crit.
Care Med., 2010, 182:897-904]. These observations have led to the
hypothesis that this signalling cascade could be one mechanism
responsible for the corticosteroid-insensitivity of inflammation
observed in the lungs of patients suffering from COPD, as well as
those asthmatics who smoke, thereby subjecting their lungs to
increased oxidative stress. Indeed, theophylline, a compound used
in the treatment of both COPD and asthma, has been suggested to
reverse steroid insensitivity through mechanisms involving
interaction with pathways controlled by PI3 kinase .delta. [To, Y.
et al., Am. J. Respir. Crit. Care Med., 2010, 182:897-904].
[0005] International patent application WO2011/048111 discloses a
number of compounds which are inhibitors of PI3 kinases,
particularly PI3 kinase .delta., including
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-di
hydroquinazolin-5-yl)-N,N-bis(2-methoxyethyl)hex-5-ynamide in the
free base form which is disclosed therein as Example 83. This
compound is also disclosed in WO2012/052753.
##STR00001##
[0006] The above mentioned compound is referred to herein as
"compound of formula (I)" or "compound of formula (I) free
base".
[0007] Prior to the applicant'searlier disclosure (WO2011/048111),
the PI3 kinase inhibitors described to date have typically been
intended for oral administration. However, an undesired consequence
of this approach is that non-targeted body tissues, especially the
liver and the gut, are likely to be exposed to pharmacologically
active concentrations of the drug. An alternative strategy is to
design treatment regimens in which the drug is dosed directly to
the inflamed organ via topical therapy. In the case of controlling
inflammation (or providing another therapeutic effect) in the
lungs, this may be achieved by inhalation of the drug, which has
the benefit of retaining the drug predominantly in the lungs
thereby minimising the risks of systemic toxicity. In order to
achieve a sustained duration of action an appropriate formulation
which generates a "reservoir" of the active drug may be used.
[0008] The compound of formula (I) has, accordingly, been described
as being useful for topical administration to the lung (see
WO2011/048111).
[0009] As well as providing affinity for the target organ and
sustained efficacy, a drug for topical administration to the lung
via inhalation must also be formulated so as to provide a
predictable dose of the drug, which in turn must have predictable
and reproducible properties. Achieving acceptable and reproducible
chemical and physical stability of the drug in the formulation is a
key goal in the product development of pharmaceutical products for
all types of pharmaceutical dosage forms. Crystalline forms are
preferred, as are forms which are amenable to micronisation.
[0010] For inhalation use, there are 3 main dosage forms--a dry
powder inhaler (DPI), a metered dose inhaler (MDI) and an aqueous
based nebuliser (hand-held or table-top). However the majority of
global sales of inhalation products are DPIs and thus provide a
well-accepted way of delivering drugs by inhalation. There are
numerous commercialised DPI products, such as Flixotide
(fluticasone propionate), Advair (fluticasone
propionate/salmeterol), Symbicort (budesonide/formoterol),
Pulmicort (budesonide), Serevent (salmeterol), Foradil
(formoterol).
[0011] Dry powder inhalation formulations typically consist of a
blend of drug particles (size below 10 microns and normally below 5
microns) with a diluent, typically lactose. Since the usual doses
required for inhaled therapies are in the microgram range, the
diluent facilitates pharmaceutical processing and dispensing of
individual doses e.g. into capsules or blisters or the metering of
doses from a bulk reservoir, for subsequent administration to the
patient. Therefore, typically, the mass of diluent (the most common
being lactose) may be greater than that of the drug substance. In
this environment, acceptable formulations of some products can be
achieved by simply blending the drug product with lactose. Other
products may require other additional excipients or other
processing steps in order for the product to meet the requirements
of regulatory authorities. For example, U.S. Pat. No. 7,186,401 B2
(Jagotec A G et al.) discloses that the addition of magnesium
stearate to dry powder formulations for inhalation improves the
moisture resistance of the formulations and allows a high fine
particle dosage or fine particle fraction to be maintained under
humid conditions. WO00/53157 (Chiesi) describes magnesium stearate
as a lubricant to be employed in dry powder formulations for
inhalation which is capable if increasing the fine particle dose of
certain drugs. US2006/0239932 (Monteith) discloses an inhalable
solid pharmaceutical formulation comprising certain active
ingredient substances susceptible to chemical interaction with
lactose, lactose and magnesium stearate. It is disclosed that
magnesium stearate inhibits lactose induced degradation of the
active ingredient, presumably via the Maillard reaction which
involves the reaction of an amine group on the active ingredient
with lactose. US2012/0082727 (Chiesi) discloses a method of
inhibiting or reducing chemical degradation of an active ingredient
bearing a group susceptible to hydrolysis selected from the group
consisting of a carbonate group, a carbamate group and an ester
group in a powder formulation for inhalation comprising carrier
particles (such as lactose particles) said method comprising
coating at least a portion of the surface of said carrier particles
with magnesium stearate.
[0012] Thus, there remains a need to provide forms of selective PI3
kinase inhibitors for use in inhalation therapy which have the
potential to provide therapeutic efficacy in asthma, COPD and other
inflammatory diseases of the lungs. In particular, it remains an
objective to provide a compound of formula (I) in a crystalline
form which has appropriate physical and chemical stability,
preferably amenable to micronization, and compatible with
pharmaceutical excipients for inhalation therapy, especially
lactose.
SUMMARY OF THE INVENTION
[0013] In a first aspect, the present invention provides a compound
of formula (I)
##STR00002##
that is
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-
-methoxyethyl)hex-5-ynamide in the form of a solid crystalline
hydrate.
[0014] In a second aspect, the Present invention provides a
compound of formula (I)
##STR00003##
that is
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-
-methoxyethyl)hex-5-ynamide in solid crystalline anhydrous
form.
[0015] Such substances are hereinafter referred to as "solid
crystalline forms of the invention". Pharmaceutical formulations
which contain the solid crystalline forms of the invention
(optionally micronized) are hereinafter referred to as
"formulations of the invention".
[0016] As explained in the Examples, the solid crystalline forms of
the invention have high melting point (around 183.degree. C. or
above), appear to have good physical stability (as determined by
XRPD, TGA, DSC, DVS and IR analysis) and have good chemical
stability (as determined by HPLC analysis). The solid crystalline
forms of the invention have good physical stability when combined
with lactose. The solid crystalline hydrate form has good chemical
stability when combined with lactose. The solid crystalline
anhydrous form has good chemical stability when combined with
lactose in the presence of a metal salt of stearic acid such as
magnesium stearate.
[0017] The solid crystalline hydrate form and the solid crystalline
anhydrous form appear to have related (but distinct) crystal
structures.
BRIEF DESCRIPTION OF THE FIGURES
[0018] FIG. 1 shows an XRPD pattern acquired on a sample of
compound of formula (I) in the form of a solid crystalline
hydrate.
[0019] FIG. 2 shows an XRPD pattern acquired on a sample of
compound of formula (I) in solid crystalline anhydrous form.
[0020] FIG. 3 shows a DVS isotherm plot of a sample of compound of
formula (I) in the form of a solid crystalline hydrate.
[0021] FIG. 4 shows a DVS change in mass plot of a sample of
compound of formula (I) in the form of a solid crystalline
hydrate.
[0022] FIG. 5 shows a DVS isotherm plot of a sample of compound of
formula (I) in solid crystalline anhydrous form.
[0023] FIG. 6 shows a DVS change in mass plot of a sample of
compound of formula (I) in solid crystalline anhydrous form.
[0024] FIG. 7 shows an IR spectrum of a sample of compound of
formula (I) in the form of a solid crystalline hydrate.
[0025] FIG. 8 shows an IR spectrum of a sample of compound of
formula (I) in solid crystalline anhydrous form.
[0026] FIG. 9 shows thermal analysis of a sample of compound of
formula (I) in the form of a solid crystalline hydrate by DSC.
[0027] FIG. 10 shows thermal analysis of a sample of compound of
formula (I) in solid crystalline anhydrous form by DSC.
[0028] FIG. 11 shows thermal analysis of a sample of compound of
formula (I) in the form of a solid crystalline hydrate by TGA.
[0029] FIG. 12 shows thermal analysis of a sample of compound of
formula (I) in solid crystalline anhydrous form by TGA.
[0030] FIG. 13 shows a DVS isotherm plot of a sample of micronized
compound of formula (I) in solid crystalline anhydrous form.
[0031] FIG. 14 shows an IR spectrum of a sample of a blend of
compound of formula (I) in solid crystalline hydrate form and
Lactohale200.RTM..
[0032] FIG. 15 shows an XRPD pattern acquired on a sample of a
blend of compound of formula (I) in solid crystalline hydrate form
and Lactohale200.RTM..
[0033] FIG. 16 shows an IR spectrum of a sample of a blend of
compound of formula (I) in solid crystalline anhydrous (micronized)
form and Lactohale200.RTM..
[0034] FIG. 17 shows an XRPD pattern acquired on a sample of a
blend of compound of formula (I) in solid crystalline anhydrous
form (micronized) and Lactohale200.RTM..
[0035] FIG. 18 shows an IR spectrum of a sample of a blend of
compound of formula (I) in solid crystalline anhydrous form
(micronized), Lactohale200.RTM. and magnesium stearate.
[0036] FIG. 19 shows an XRPD pattern acquired on a sample of a
blend of compound of formula (I) in solid crystalline anhydrous
form (micronized), Lactohale200.RTM. and magnesium stearate
DETAILED DESCRIPTION OF THE INVENTION
Compound of Formula (I) as Active Ingredient
[0037] The compound of formula (I) is a dual PI3K delta PI3K gamma
inhibitor, wherein the term inhibitor as employed herein is
intended to refer to a compound that reduces (for example by at
least 50%) or eliminates the biological activity of the target
protein, for example the PI3K delta isozyme, in an in vitro enzyme
assay. The term delta/gamma inhibitor as employed herein is
intended to refer to the fact that the compound inhibits, to some
degree, both enzyme isoforms although not necessarily to the same
extent. Compound of formula (I) is active in cell based screening
systems and thereby demonstrates that it possesses suitable
properties for penetrating cells and thereby exert intracellular
pharmacological effects.
[0038] Generic processes for synthesising the compound of formula
(I) are disclosed in WO2011/048111, the contents of which are
incorporated by reference in their entirety, and a method similar
to that of Example 1 can be employed. See also WO2012/052753, the
contents of which are incorporated by reference in their entirety,
where a specific method for synthesising the compound of formula
(I) is provided in the Example.
[0039] Suitably compound of formula (I) is protected from light
during and after synthesis e.g. by use of amber glassware or light
impervious packaging (e.g. foil packaging).
[0040] The pharmaceutical formulation of the invention comprises
compound of formula (I) as active ingredient in a therapeutically
effective amount. A therapeutically effective amount of compound of
formula (I) is defined as an amount sufficient, for a given dose or
plurality of divided doses, to achieve a therapeutically meaningful
effect in a subject when administered to said subject in a
treatment protocol.
[0041] Pharmaceutical formulations of the invention are suitably
dry powder pharmaceutical formulations for inhalation.
[0042] In one embodiment, the dry powder pharmaceutical formulation
comprises from about 0.004 wt. % to about 50 wt. % of compound of
formula (I) based on weight of the dry powder pharmaceutical
formulation and based on weight of compound of formula (I) as free
base; for example from about 0.02 wt. % to about 50 wt. %, from
about 0.02 wt. % to about 25 wt. %, or from about 0.02 wt. % to
about 15 wt. % or from about 0.02 wt. % to about 20 wt. %.
Preferably, the dry powder pharmaceutical formulation comprises
from about 0.1 wt. % to about 20 wt. % e.g. from about 0.1 wt. % to
about 5 wt. % of compound of formula (I) based on the weight of the
dry powder pharmaceutical formulation.
[0043] A pharmaceutical formulation of the invention may contain
compound of formula (I) as a single active ingredient. However, the
pharmaceutical formulation may contain further active ingredients.
The pharmaceutical formulation may also be co-administered together
with one or more other active ingredients (or one or more
pharmaceutical formulations containing one or more active
ingredients). Exemplary further active ingredients are mentioned
below.
[0044] Compound of formula (I) is suitably prepared in particulate
form such that it is suitable for dry powder inhalation. A
pharmaceutical formulation of the invention may typically contain
drug particles having a volume median diameter (D50) from about 0.5
.mu.m to about 10 .mu.m particularly from about 1 .mu.m to about 5
.mu.m.
[0045] A suitable method for determining particle size is laser
diffraction, e.g. using a Mastersizer 2000S instrument from Malvern
Instruments. Instruments are also available from Sympatec. For
particle size distributions, the median value D50 is the size in
microns that splits the particle size distribution with half above
and half below. The primary result obtained from laser diffraction
is a volume distribution, therefore D50 is actually Dv50 (median
for a volume distribution) and as used herein refers to particle
size distributions obtained using laser diffraction. D10 and D90
values (when used in the context of laser diffraction, taken to
mean Dv10 and Dv90 values) refer to the particle size wherein 10%
of the distribution lies below the D10 value, and 90% of the
distribution lies below the D90 value, respectively.
[0046] Particles of suitable size for use in a dry powder
inhalation formulation may be prepared by any suitable method known
to the person skilled in the art. Drug particles of suitable size
for inhalation may be prepared by particle size reduction methods
including milling or more preferably micronization e.g. using a jet
mill micronization device (eg as manufactured by Hosokawa Alpine).
Alternatively, particulates of suitable size may be produced at the
first instance by spray drying, spray freezing, controlled
crystallisation approaches e.g. controlled precipitation,
super-critical fluid crystallisation, sonocrystallisation or other
suitable crystallisation procedure, for example in a continuous
crystallisation apparatus. Thus one aspect of the invention
provides compound of formula (I) in micronized form.
Solvates--Hydrate Form of Compound of Formula (I)
[0047] In one embodiment, there is provided compound of formula (I)
in the form of a hydrate. In particular, there is provided compound
of formula (I) in the form of a solid crystalline hydrate obtained
by crystallising compound of formula (I) from dichloromethane
optionally in mixture with methanol (e.g. containing up to 20% e.g.
up to 10% e.g. 4.8% v/v methanol) at ambient temperature e.g.
around 22.degree. C. Formation of the hydrate was found not to
require the addition of water to the reaction mixture (i.e. any
residual water in the solvent, or carried over in product from a
previous reaction step and/or moisture in the atmosphere is
sufficient). However water may be added to the solvent, e.g. 0.1 to
5% water may be added. The detailed preparation of such a solid
crystalline hydrate of compound of formula (I) is provided in
Example 1.
[0048] In one embodiment, there is provided a solid crystalline
hydrate form compound of formula (I) having an XRPD pattern
substantially as shown in FIG. 1. The method of obtaining the XRPD
data is described in the General Procedures and the data discussed
in Example 3.
[0049] Thus, there is provided a hydrate form of compound of
formula (I) in a crystalline form having an X-ray powder
diffraction pattern with at least one (for example, one, two,
three, four, five, six, seven, eight, nine, ten, eleven, twelve,
thirteen or all fourteen) peaks at 5.6, 7.6, 9.6, 11.1, 12.2, 12.6,
13.3, 13.9, 15.9, 17.0, 18.9, 20.3, 21.8, 23.1 (.+-.0.2 degrees,
2-theta values), these peaks being characteristic of the
crystalline hydrate form. The peaks at 9.6, 13.3, 13.9, 17.0, 18.9,
20.3 and 23.1 are particularly characteristic of the hydrate from
and therefore it is preferred to see at least one (for example one,
two, three, four, five, six or all seven) of these peaks.
[0050] Without being limited by theory, the solid crystalline
hydrate form of compound of formula (I) may be a channel hydrate
form. Alternatively, water may reside in pores in the crystal or at
the surface of the crystal. In any event, as shown in the Examples,
water does not form an essential part of the crystal lattice and
the crystal form is stable to removal or addition of water.
[0051] The physical and chemical stabilities of the solid
crystalline hydrate form of the compound of formula (I) disclosed
herein were investigated.
[0052] In order to assess physical stability, samples of the
hydrate form of compound of formula (I) were stored in containers
open to the ambient atmosphere at different temperatures and
relative humidities. The physical stability of the samples was
investigated using thermogravimetric analysis (TGA), differential
scanning calorimetry (DSC), dynamic vapour sorption (DVS), infrared
spectroscopy (IR) and X-ray powder diffraction (XRPD). Full
experimental procedures are provided in the General Procedures
section and the results are summarised in Example 4 (Table 3). As
discussed in Example 4, the hydrate form of compound of formula (I)
was found to have good overall physical stability. However, under
DVS analysis a weight loss of 2.2% was registered and the obtained
dry product was found to be hygroscopic. Small differences in the
IR and XRPD data were observed for the samples under drier
conditions, however these differences were attributed to the loss
of water observed in the DVS studies and the integrity of the
crystalline structure was retained after water loss and subsequent
rehydration.
[0053] In order to assess chemical stability, samples of the
hydrate form of compound of formula (I) were prepared in methanol
and analysed by HPLC. The results are summarised in Example 5
(Table 6) where it is indicated that the hydrate form of compound
of formula (I) was found to be chemically stable, although some
sensitivity towards light was detected.
[0054] Dry powder pharmaceutical formulations typically comprise
lactose as a suitable carrier for the active ingredient. Therefore,
the lactose compatibility of the hydrate form of compound of
formula (I) was investigated.
[0055] Both the physical and chemical compatibilities of the solid
crystalline hydrate form of compound of formula (I) with lactose
were investigated.
[0056] In order to assess physical compatibility, high
concentration compositions of the hydrate form of compound of
formula (I) and lactose were prepared, then analysed at various
temperatures and humidities, as summarised in Example 6. It is
evident that the tested mixtures were physically compatible under
all investigated conditions.
[0057] In order to assess chemical compatibility, lower
concentration (relative to those used in the physical compatibility
studies) compositions of the hydrate form of compound of formula
(I) with lactose were prepared in methanol and analysed by HPLC The
results are summarised in Example 7 (Table 9) where it is indicated
that the hydrate form of compound of formula (I) and lactose are
chemically compatible.
[0058] As a result of the inventors' studies, it can be concluded
that the hydrate form of compound of formula (I), has good physical
and chemically stability. The combination of the hydrate form of
compound of formula (I) with lactose has both chemical and physical
stability, indicating suitability for use in a pharmaceutical
formulation.
Anhydrous Form of Compound of Formula (I)
[0059] In one embodiment, there is provided compound of formula (I)
in anhydrous form. In particular, there is provided compound of
formula (I) in solid crystalline anhydrous form, obtained by
crystallizing the hydrate form of compound of formula (I) from
1-propanol. Suitably, the 1-propanol is dry e.g. containing a
maximum of around 0.9% w/w water. In one embodiment, the 1-propanol
has a maximum of 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.05%
w/w of water. Suitably, the 1-propanol has maximum of 0.2% w/w
water. Suitably, crystallisation is performed in the presence of a
metal scavenger. Suitable metal scavengers are materials that
adsorb the metal while being easily separable from the compound of
interest (i.e. compound of formula (I)). For example,
functionalised silicas are particularly useful as metal scavengers,
as once the metal has been adsorbed, the metal-silica complex may
then be easily separated from the compound of interest by
filtration. Functional groups that form stable complexes with metal
ions include groups containing one or more nitrogen and/or sulphur
centres, and are well known to the person skilled in the art.
[0060] An example of a suitable commercially available metal
scavenger is SiliaMetS.RTM. Thiol (a thiol-derivatised silica gel
suitable for scavenging a variety of metals including Pd, Pt, Cu,
Ag and Pb). Suitably, the metal scavenger is present in amount
sufficient to ensure that the resulting metal ion concentration is
below 20 ppm, preferably below 10 ppm. In one embodiment, the metal
scavenger is present at 1-10% w/w, for example 2-8% w/w or 5% w/w
based on the weight of the compound of formula (I). Suitably
crystallisation is performed by cooling the solution of compound of
formula (I) and solvent from elevated temperature, continuously
(i.e. continuous cooling) or in stages (i.e. alternating between
cooling and holding the solution at a particular temperature).
Suitable temperature gradients (continuous or separate) for cooling
include 95-15.degree. C., 95-20.degree. C., 90-20.degree. C.,
80-20.degree. C. 95-90.degree. C., 95-85.degree. C., 95-80.degree.
C. 90-85.degree. C., 80-20.degree. C. In one embodiment, the
solution is cooled from 80-95.degree. C. to ambient temperature
(e.g. around 20-22.degree. C.). The detailed preparation of such a
solid crystalline anhydrous form of compound of formula (I) is
provided in Example 2.
[0061] In one embodiment, there is provided a solid crystalline
anhydrous form of compound of formula (I) having an X-ray powder
diffraction pattern substantially as shown in FIG. 2. The method of
obtaining the XRPD data is described in the General Procedures and
the data discussed in Example 3.
[0062] Thus, there is provided compound of formula (I) in a
crystalline anhydrous form having an XRPD pattern with at least one
(for example, one, two, three, four, five, six, seven, eight, nine
or all ten) peaks at 5.6, 7.9, 11.2, 12.3, 15.6, 17.6, 18.4, 21.4,
22.5, 24.2 (.+-.0.2 degrees, 2-theta values), these peaks being
characteristic of the crystalline anhydrous form. The peaks at
17.6, 18.4, 22.5 and 24.2 are particularly characteristic for the
anhydrous form and therefore it is preferred to see at least one
(for example one, two, three or all four) of these peaks.
[0063] The physical and chemical stabilities of the compound of
formula (I) in solid crystalline anhydrous form were
investigated.
[0064] In order to assess physical stability, samples of the
anhydrous form of compound of formula (I) in unmicronized and in
micronized form were stored in containers open to the ambient
atmosphere at different temperatures and relative humidities.
Physical stability was investigated using TGA, DSC, DVS, IR and
XRPD as described above for the hydrate form of compound of formula
(I). The results are summarised in Example 4.
[0065] As discussed in Example 4, the anhydrous form of compound of
formula (I) (both unmicronized and micronized) was found to be
physically stable in all investigated conditions.
[0066] In order to assess chemical stability, samples of the
anhydrous form of compound of formula (I) (unmicronized and
micronized) were prepared in methanol and analysed by HPLC. The
results are summarised in Example 5 (Tables 7 and 8) where it is
indicated that the anhydrous form of compound of formula (I) (both
unmicronized and micronized) was found to be chemically stable,
although some sensitivity towards light was detected. It is evident
that the chemical stability of the anhydrous form of the compound
of formula (I) is comparable with the chemical stability of the
hydrate form of compound of formula (I).
[0067] The lactose compatibility of the solid crystalline anhydrous
form of compound of formula (I) was investigated.
[0068] Both the physical and chemical compatibility of the
anhydrous form of compound(I) with lactose was investigated.
[0069] In order to assess physical compatibility, high
concentration compositions of the anhydrous form (micronized) of
compound of formula (I) and lactose were prepared, then analysed at
various temperatures and humidities, as summarised in Example 6. It
is evident that the tested mixtures were physically compatible
under all investigated conditions.
[0070] In order to assess chemical compatibility, lower
concentration (relative to those used in the physical compatibility
studies) compositions of the anhydrous form (micronized) of
compound of formula (I) with lactose were analysed by HPLC. The
results are summarised in Example 7 (Table 10) where it is
indicated that under certain conditions the composition of
anhydrous form and lactose underwent degradation. The degradation
products were investigated and the main degradant was identified by
mass spectrometry as being one or both of the two substances shown
as D019328:
##STR00004##
[0071] This degradation product is likely to be the result of the
addition of water across the alkyne triple bond and may exist as
one of two forms (or may exist in both forms) depending on the
orientation of the addition of the water across the triple bond.
The same degradant has been observed during the forced degradation
of the anhydrous form of compound of formula (I) with metal ions.
As a result of further studies, it appears that the degradation of
the anhydrous form of compound of formula (I) requires metal ions
and water and is accelerated by elevated temperature.
[0072] Further investigation involving accelerated stability
testing (i.e. exposure of the drug substance to 80.degree. C. in a
closed vial, see Example 10) has led the inventors to confirm that
at least the degradation product shown as D019492 in Scheme 1
(below) is generated. Moreover the inventors also concluded that a
further degradation product (D019493) can result from the
hydrolytic cleavage of the pyrimidinone ring and subsequent
intramolecular reaction with the alkyne group. D019349 is a
presumed intermediary degradation product which was observed in
certain circumstances of temperature and RH in stability testing
(data not shown).
##STR00005##
[0073] The addition of magnesium stearate to the combination of
anhydrous form of compound of formula (I) and lactose was
investigated. The combination of anhydrous form of compound of
formula (I) with lactose and magnesium stearate was found to be
physically stable (Example 8). However, surprisingly, it was found
that the addition of magnesium stearate caused an increase in the
chemical stability of the combination of anhydrous form of compound
of formula (I) and lactose (Example 9). A similar stabilising
effect was found using other metal salts of stearic acid,
specifically sodium stearate and calcium stearate (Example 10).
[0074] Without wishing to be bound by theory, it appears that the
metal salt of stearic acid such as magnesium stearate can act as a
protecting agent against chemical degradation of the alkyne group
in compound of formula (I) and against chemical degradation of the
pyrimidinone ring in compound of formula (I) which is observed when
the anhydrous form of compound of formula (I) is in a mixture with
lactose.
[0075] In summary, the inventors have discovered that the solid
crystalline anhydrous form of compound of formula (I) has greater
physical stability than the solid crystalline hydrate form of
compound of formula (I) in isolation, but found that the anhydrous
form was less stable with lactose. However, the inventors have
discovered that this problem can be overcome by the addition of a
metal salt of stearate such as magnesium stearate. The inventors
extrapolate these findings with metal salts of stearic acid to
metal salts of stearyl fumarate.
Pharmaceutical Formulations for Inhalation
[0076] The invention provides pharmaceutical compositions
comprising the solid crystalline forms of the invention in
admixture with one or more diluents or carriers. Suitably the
composition contains lactose as a diluent or carrier.
[0077] As used herein, the term "lactose" refers to a
lactose-containing component, including .alpha.-lactose
monohydrate, .beta.-lactose monohydrate, .alpha.-lactose anhydrous,
.beta.-lactose anhydrous and amorphous lactose. Lactose components
may be processed by micronization, sieving, milling, compression,
agglomeration or spray drying. Commercially available forms of
lactose in various forms are also encompassed, for example
Lactohale.RTM. (inhalation grade lactose; Frieslandfoods),
InhaLac.RTM.70 (sieved lactose for dry powder inhaler; Meggle) and
Respitose.RTM. products. In one embodiment, the lactose component
is selected from the group consisting of .alpha.-lactose
monohydrate, .alpha.-lactose anhydrous and amorphous lactose.
Preferably, the lactose is .alpha.-lactose monohydrate.
[0078] In order to penetrate sufficiently far into the lungs, the
particulate active ingredient (in this case compound of formula
(I)) must be a suitable size as described above. These small
particles will have a tendency to agglomerate. The use of a carrier
such as lactose prevents this agglomeration and can improve
flowability. Furthermore, the use of a carrier ensures that a
correct and consistent dosage reaches the lungs. The active
ingredient will usually form a monolayer on the larger lactose
particle, then during inhalation the active ingredient and the
carrier are separated and the active ingredient is inhaled, while
the majority of the carrier is not. As such, the use of particulate
lactose as a carrier for the active ingredient ensures that each
dose of the dry powder pharmaceutical formulation releases the same
amount of the active ingredient.
[0079] Generally, to prevent agglomeration of the small active
particles, a carrier such as lactose with a particle size of
approximately or at least ten times that of the active ingredient
is used (e.g. lactose having a D50 approximately or at least ten
times that of the active ingredient is used).
[0080] In one embodiment, the dry powder formulation of the present
invention comprises particulate lactose having D50 in the range
40-150 .mu.m.
[0081] The dry powder pharmaceutical formulation of the present
invention comprises particulate lactose as carrier in an amount
sufficient to ensure that the correct and consistent dosage of the
active ingredient reaches the lungs. In one embodiment, the dry
powder pharmaceutical formulation comprises from about 40 wt. % to
about 99.88 or 99.98 wt. %, for example from about 50 wt. % to
about 99.88 or 99.98 wt %, from about 65 wt. % to about 99.88 or
99.98 wt. %, or from about 75 wt. % to about 99.88 or 99.98 wt. %
of particulate lactose based on the weight of the dry powder
pharmaceutical formulation. Preferably, the dry powder
pharmaceutical formulation comprises from about 80 wt. % to about
99.98 wt. % or for example from about 80 wt % to about 99.9% wt %,
for example from about 85 wt. % to about 99.88 or 99.98 wt. %, for
example from about 95 wt. % to about 99 wt. % of particulate
lactose based on the weight of the dry powder pharmaceutical
composition.
[0082] Optionally (and especially when using the solid crystalline
anhydrous form) the composition contains a stabilising agent
selected from metals salt of stearic acid such as magnesium
stearate and metal salts of stearyl fumarate.
[0083] An example metal salt of stearic acid is magnesium stearate.
Alternative metal salts of stearic acid that may be employed
include salts of stearic acid formed with Group I and other Group
II metals, such as sodium stearate, calcium stearate and lithium
stearate. Other metal salts of stearic acid that may be mentioned
include zinc stearate and aluminium stearate.
[0084] Metal salts of stearyl fumarate (e.g. sodium stearyl
fumarate) appear to have similar properties to those of metal salts
of stearic acid (see Shah et al, Drug development and Industrial
pharmacy 1986, Vol. 12 No. 8-9, 1329-1346). In the inventors'
opinion they can be employed as an alternative to metal salts of
stearic acid in the present invention.
[0085] As used herein the term "magnesium stearate" includes
magnesium stearate trihydrate, magnesium stearate dihydrate,
magnesium stearate monohydrate and amorphous magnesium stearate.
Magnesium stearate as defined herein includes a tolerance wherein
any material defined as "magnesium stearate" may contain up to 25%
(e.g. up to 10% e.g. up to 5% e.g. up to 1%) of palmitate salt.
[0086] More generally, metal salts of stearic acid or metal salts
of stearyl fumarate may be employed in anhydrous form or as a
hydrate and may contain up to 25% (e.g. up to 10% e.g. up to 5%
e.g. up to 1%) of palmitate salt.
[0087] As used herein the expression "stabilizing agent selected
from metal salts of stearic acid such as magnesium stearate and
metal salts of stearyl fumarate" can include a mixture of metal
salts of stearic acid and/or stearyl fumarate, although use of a
single salt would be preferred.
[0088] The metal salt of stearic acid such as magnesium stearate or
metal salt of stearyl fumarate is typically obtained as a fine
powder which need not be micronized. Suitably the D50 of the metal
salt of stearic acid such as magnesium stearate or metal salt of
stearyl fumarate is greater than 5 .mu.m e.g. around 10 .mu.m or
greater than 10 .mu.m e.g. in the range 5 to 100 .mu.m e.g. 5 to 50
.mu.m e.g. 5 to 20 .mu.m e.g. 10 to 20 .mu.m. Magnesium stearate
may for example be obtained from Avantor (Hyqual 2257 brand) or
Peter Greven. Sodium stearate and calcium stearate may, for
example, be obtained from Sigma-Aldrich. Sodium stearyl fumarate
may, for example, be obtained from ScienceLab.
[0089] The dry powder pharmaceutical formulation of the present
invention optionally comprises particulate stabilising agent
selected from metal salts of stearic acid such as magnesium
stearate and metal salts of stearyl fumarate in an amount
sufficient to ensure the chemical stability of the formulation ("a
stabilising amount"). Chemical stability is, for example,
demonstrated when the production of degradant D019328 (one or both
substances) is at a level of less than 0.2% wt. % following storage
of the composition containing Compound of formula (I) for 4 weeks
at 50.degree. C. Alternatively or in addition, chemical stability
is, for example, demonstrated when the production of degradant
D019493 is at a level of less than 0.5% wt. % following storage of
the composition containing Compound of formula (I) for 2 weeks at
80.degree. C. Alternatively, or in addition, chemical stability is,
for example, demonstrated when the production of degradant D019492
is at a level of less than 0.4% wt. % following storage of the
composition containing Compound of formula (I) for 2 weeks at
80.degree. C. In one embodiment, the dry powder pharmaceutical
formulation comprises from about 0.01 wt. % to about 15 wt. %, for
example 0.1 wt. % to about 10 wt. %, 10 wt. %, 5 wt. %, 2 wt. % or
1 wt. % of particulate metal salt of stearic acid such as magnesium
stearate or metal salt of stearyl fumarate based on the weight of
the dry powder pharmaceutical formulation. Preferably, the dry
powder pharmaceutical formulation comprises from about 0.5 wt. % to
about 5 wt. % e.g. 1-2% w/w of particulate metal salt of stearic
acid such as magnesium stearate or metal salt of stearyl fumarate
based on the weight of the dry powder pharmaceutical composition.
Suitably the metal salt of stearic acid such as magnesium stearate
or metal salt of stearyl fumarate is present in an amount
sufficient to ensure the physical stability of the formulation.
Physical stability is, for example, demonstrated when the IR
spectrum and XRPD pattern of the composition (especially in
relation to characteristics peaks of Compound of formula (I)) are
substantially unaltered following storage of the composition
containing Compound of formula (I) for 4 weeks at 50.degree. C.
[0090] In one embodiment, the dry powder pharmaceutical formulation
for inhalation of the present invention comprises: [0091] (i) From
about 0.02 to 50 wt %
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-me-
thoxyethyl)hex-5-ynamide in solid crystalline anhydrous form in
particulate form as active ingredient; [0092] (ii) from about 40 to
about 99.88 wt. % particulate lactose; and [0093] (iii) from about
0.1 to about 10 wt. % particulate stabilizing agent selected from
metal salts of stearic acid (such as magnesium stearate) and metal
salts of stearyl fumarate.
[0094] In a further embodiment, the dry powder pharmaceutical
formulation for inhalation of the present invention comprises:
[0095] (i) From about 0.02 to 50 wt %
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-me-
thoxyethyl)hex-5-ynamide in the form of a solid crystalline hydrate
in particulate form as active ingredient [0096] (ii) from about 40
wt. % to about 99.98 wt. % particulate lactose; and [0097] (iii)
optionally from about 0.1 wt. % to about 10 wt. % particulate
stabilizing agent selected from metal salts of stearic acid (such
as magnesium stearate) and metal salts of stearyl fumarate.
Pharmaceutical Uses and Methods of Administration
[0098] There is provided according to one aspect of the present
invention use of solid crystalline forms of the invention for use
as a medicament.
[0099] In one embodiment there is provided the use of a
pharmaceutical formulation of the invention for the treatment of
COPD and/or asthma, in particular COPD or severe asthma, by
inhalation i.e. by topical administration to the lung.
Advantageously, administration to the lung allows the beneficial
effects of the compounds to be realised whilst minimising the
side-effects, for patients.
[0100] In one embodiment the pharmaceutical formulation of the
invention is suitable for sensitizing patients to treatment with a
corticosteroid.
[0101] The pharmaceutical formulations may conveniently be
administered in unit dosage form and may be prepared by any of the
methods well-known in the pharmaceutical art, for example as
described in Remington's Pharmaceutical Sciences, 17th ed., Mack
Publishing Company, Easton, Pa., (1985).
[0102] Topical administration to the lung is achieved by use of an
inhalation device.
[0103] Thus, an aspect of the invention includes an inhalation
device comprising one or more doses of a pharmaceutical formulation
according to the invention. Inhalation devices for dry powder
formulations are typically breath operated such that the dose is
withdrawn from the device and administered to the subject using the
power of the subject's lungs by inhaling from a mouthpiece.
However, optionally, external energy may be provided to assist the
administration of the dose. Typically the inhalation device will
comprise a plurality of doses of a pharmaceutical formulation
according to the invention, e.g. 2 or 4 or 8 or 28 or 30 or 60 or
more doses. Thus the inhalation device may comprise a month's
supply of doses. In an embodiment, a dose is metered into a capsule
for use one by one in an inhalation device adapted to deliver the
contents of a capsule to a subject upon inhalation. Optionally the
doses are divided e.g. such that a dose is administered using two
(or more) inhalations from the inhalation device. According to one
embodiment of the invention the doses of formulation are
pre-metered in the inhalation device. For example the pre-metered
doses may be contained in the pouches of a blister strip or disk or
within capsules. According to another embodiment of the invention
the doses are metered in use. Thus the inhalation device contains a
reservoir of dry powder and the device meters a dose of powder
(typically on a fixed volume basis) prior to or at the time of
administration.
[0104] Example dry powder inhalation devices include SPINHALER,
ECLIPSE, ROTAHALER, HANDIHALER, AEROLISER, CYCLOHALER,
BREEZHALER/NEOHALER, FLOWCAPS, TWINCAPS, X-CAPS, TURBOSPIN,
ELPENHALER, DISKHALER, TURBUHALER, MIATHALER, TWISTHALER,
NOVOLIZER, DISKUS, SKYEHALER, ORIEL dry powder inhaler, MICRODOSE,
ACCUHALER, PULVINAL, EASYHALER, ULTRAHALER, TAIFUN, PULMOJET,
OMNIHALER, GYROHALER, TAPER, CONIX, XCELOVAIR, PROHALER and
CLICKHALER. Another example is MONODOSE inhaler.
[0105] Optionally the inhalation device may be over-wrapped for
storage to protect against ingress of moisture. A desiccant may
optionally be employed within an over-wrap or within the device.
Suitably the pharmaceutical formulation according to the invention
in the inhalation device is protected from light.
[0106] The pharmaceutical formulations according to the invention
may also be useful in the treatment of respiratory disorders
including COPD, chronic bronchitis, emphysema), asthma, paediatric
asthma, cystic fibrosis, sarcoidosis and idiopathic pulmonary
fibrosis and especially asthma, chronic bronchitis and COPD.
[0107] The pharmaceutical formulations according to the invention
may comprise compound of formula (I) as the sole active ingredient,
or may comprise additional active ingredients, e.g. active
ingredients suitable for treating the above mentioned conditions.
For example possible combinations for treatment of respiratory
disorders include combinations with steroids (e.g. budesonide,
beclomethasone dipropionate, fluticasone propionate, mometasone
furoate, fluticasone furoate, flunisolide, ciclesonide,
triamcinolone), beta agonists (e.g. terbutaline, bambuterol,
salbutamol, levalbuterol, salmeterol, formoterol, clenbuterol,
fenoterol, broxaterol, indacaterol, reproterol, procaterol,
vilanterol) and/or xanthines (e.g. theophylline), muscarinic
antagonists, (e.g. ipratropium, tiotropium, oxitropium,
glycopyrronium, glycopyrrolate, aclidinium, trospium), leukotriene
antagonists (e.g. zafirlukast, pranlukast, zileuton, montelukast)
and/or a p38 MAP kinase inhibitor. It will be understood that any
of the aforementioned active ingredients may be employed in the
form of a pharmaceutically acceptable salt.
[0108] In one embodiment, the pharmaceutical formulation of the
invention is administered in combination with an antiviral agent,
for example acyclovir, oseltamivir (Tamiflu.RTM.), zanamivir
(Relenza.RTM.) or interferon.
[0109] In one embodiment the combination of compound of formula (I)
and other active ingredient(s) are co-formulated in the
pharmaceutical formulation of the invention. In another embodiment,
the other active ingredient(s) are administered in one or more
separate pharmaceutical formulations.
[0110] In one embodiment compound of formula (I) is co-formulated
in the pharmaceutical formulation of the invention or
co-administered in a separate formulation with a corticosteroid,
for example for use in maintenance therapy of asthma, COPD or lung
cancer including prevention of the latter.
[0111] In one embodiment the pharmaceutical formulation of the
invention is administered by inhalation and a corticosteroid is
administered orally or by inhalation either in combination or
separately.
[0112] The pharmaceutical formulation of the invention may also
re-sensitise the patient's condition to treatment with a
corticosteroid, when previously the patient's condition had become
refractory to the same.
[0113] In one embodiment of the invention a dose of the
pharmaceutical formulation employed is equal to that suitable for
use as a monotherapy but administered in combination with a
corticosteroid. In one embodiment a dose of the pharmaceutical
formulation which would be sub-therapeutic as a single agent is
employed, and is administered in combination with a corticosteroid,
thereby restoring patient responsiveness to the latter, in
instances where the patient had previously become refractory to the
same.
[0114] Additionally, the pharmaceutical formulation of the
invention may exhibit anti-viral activity and prove useful in the
treatment of viral exacerbations of inflammatory conditions such as
asthma and/or COPD.
[0115] The pharmaceutical formulation of the present invention may
also be useful in the prophylaxis, treatment or amelioration of
influenza virus, rhinovirus and/or respiratory syncytial virus.
[0116] In one embodiment the presently disclosed pharmaceutical
formulations are useful in the treatment or prevention of cancer,
in particular lung cancer, especially by topical administration to
the lung.
[0117] Thus, in a further aspect, the present invention provides a
pharmaceutical formulation as described herein for use in the
treatment of one or more of the above mentioned conditions.
[0118] In a further aspect, the present invention provides a
pharmaceutical formulation as described herein for the manufacture
of a medicament for the treatment of one or more of the above
mentioned conditions.
[0119] In a further aspect, the present invention provides a method
of treatment of the above mentioned conditions which comprises
administering to a subject an effective amount of a pharmaceutical
formulation of the invention thereof.
[0120] Pharmaceutical formulations described herein may also be
used in the manufacture of a medicament for the treatment of one or
more of the above-identified diseases.
[0121] The word "treatment" is intended to embrace prophylaxis as
well as therapeutic treatment.
[0122] Unless otherwise specified, % values as used herein are %
values by weight (wt. %).
[0123] The solid crystalline forms of the invention, and
pharmaceutical formulations containing them, may have the advantage
that they have improved crystallinity (e.g. as measured by XRPD),
improved physical stability (e.g. as measured by XRPD, IR, DVS, DSC
or TGA analysis), improved chemical stability (e.g. as measured by
HPLC), improved physical compatibility with lactose (optionally
when combined with other excipients), improved chemical
compatibility with lactose (optionally when combined with other
excipients), improved particle size distribution on administration
(such as evidenced by improved fine particle mass) or may have
other favourable properties as compared with prior art solid forms
of the compound of formula (I).
Abbreviations
[0124] aq aqueous [0125] COPD chronic obstructive pulmonary disease
[0126] d doublet [0127] DCM dichloromethane [0128] DMAP
4-dimethylaminopyridine [0129] DMSO dimethyl sulfoxide [0130] DPI
dry powder inhaler [0131] DSC differential scanning calorimetry
[0132] DVS dynamic vapour sorption [0133] EDC.HCl
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride [0134]
(ES.sup.+) electrospray ionization, positive mode [0135] EtOAc
ethyl acetate [0136] HPLC high performance liquid chromatography
[0137] HPLC-MS high performance liquid chromatography mass
spectrometry [0138] hr hour(s) [0139] IR infrared [0140] LPS
lipopolysaccharide [0141] (M+H).sup.+ protonated molecular ion
[0142] MDI metered dose inhaler [0143] MeOH methanol [0144] MEK
methylethylketone [0145] MHz megahertz [0146] min minute(s) [0147]
mm Millimetre(s) [0148] ms mass spectrometry [0149] mTOR mammalian
target of rapamycin [0150] m/z mass-to-charge ratio [0151]
NH.sub.4OAc ammonium acetate [0152] NMR nuclear magnetic resonance
(spectroscopy) [0153] Pd(dppf)Cl.sub.2
1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) [0154]
ppm parts per million [0155] q quartet [0156] quin quintet [0157]
RH relative humidity [0158] RRT relative retention time [0159]
R.sup.t retention time [0160] RT room temperature [0161] s singlet
[0162] t triplet [0163] TBDMSCl tert-butyldimethylsilyl chloride
[0164] TGA thermogravimetric analysis [0165] TNF.alpha. tumour
necrosis factor alpha [0166] XRPD X-ray powder diffraction
EXAMPLES
General Procedures
HPLC-MS
[0167] Performed on Agilent HP1200 systems using Agilent Extend C18
columns, (1.8 .mu.m, 4.6.times.30 mm) at 40.degree. C. and a flow
rate of 2.5-4.5 mL min.sup.-1, eluting with a H.sub.2O-MeCN
gradient containing 0.1% v/v formic acid over 4 min. Gradient
information: 0-3.00 min, ramped from 95% H.sub.2O-5% MeCN to 5%
H.sub.2O-95% MeCN; 3.00-3.01 min, held at 5% H.sub.2O-95% MeCN,
flow rate increased to 4.5 mL min.sup.-1; 3.01-3.50 min, held at 5%
H.sub.2O-95% MeCN; 3.50-3.60 min, returned to 95% H.sub.2O-5% MeCN;
flow rate reduced to 3.50 mL min.sup.-1; 3.60-3.90 min, held at 95%
H.sub.2O-5% MeCN; 3.90-4.00 min, held at 95% H.sub.2O-5% MeCN, flow
rate reduced to 2.5 mL min.sup.-1. UV detection was performed at
254 nm using an Agilent G1314B variable wavelength detector.
Mass Spectra (MS)
[0168] Obtained using electrospray ionization (ESI) over the range
m/z 60 to 2000 at a sampling rate of 1.6 sec/cycle using an Agilent
G1956B, over m/z 150 to 850 at a sampling rate of 2 Hz using a
Waters ZMD or over m/z 100 to 1000 at a sampling rate of 2 Hz using
a Shimadzu 2010 LC-MS system.
NMR Spectra
[0169] .sup.1H NMR spectra (except those of Example 10) were
acquired on a Bruker Avance III spectrometer at 400 MHz using
residual undeuterated solvent as reference.
[0170] The .sup.1H NMR spectrum for Example 10 was acquired on a
Bruker Avance spectrometer at 600 MHz using residual undeuterated
solvent as reference.
Dynamic Vapour Sorption (DVS)
[0171] Obtained using a Surface Measurement Systems dynamic vapor
sorption model DVS-1. Using about 19 mg of the sample, the weight
change recorded with respect to the atmospheric humidity at
25.degree. C. was determined using the following parameters: [0172]
drying: 60 min. under dry nitrogen [0173] equilibrium: 0.01%/min.
for min:15 min and max: 60 min. [0174] data interval: 0.05% or 2.0
min. [0175] RH (%) measurement points: [0176] first set:
5,10,20,30,40,50,60,70,80,90,95,90,80,70,60,50,40,30,20,10.5 [0177]
second set:
10,20,30,40,50,60,70,80,90,95,90,80,70,60,50,40,30,20,10,5,0.
X-Ray Powder Diffraction (XRPD)
[0178] XRPD patterns were acquired on a PANalytical (Philips)
X'PertPRO MPD diffractometer equipped with a Cu LFF X-ray tube (45
kV; 40 mA; Bragg-Brentano; spinner stage) were acquired using Cu
K.alpha. radiation and the following measurement conditions: [0179]
scan mode: continuous [0180] scan range: 3 to 50.degree. 20 [0181]
step size: 0.02.degree./step [0182] counting time: 30 sec/step
[0183] spinner revolution time: 1 sec [0184] radiation type:
CuK.alpha.
Incident Beam Path
[0184] [0185] program. divergence slit: 15 mm [0186] Soller slit:
0.04 rad [0187] beam mask: 15 mm [0188] anti scatter slit:
1.degree. [0189] beam knife: +
Diffracted Beam Path
[0189] [0190] long anti scatter shield: + [0191] Soller slit: 0.04
rad [0192] Ni filter: + [0193] detector: X'Celerator
[0194] Samples were prepared by spreading on a zero background
sample holder.
Infrared Spectrometry (IR)
[0195] Micro Attenuated Total Reflectance (microATR) was used and
the sample was analyzed using a suitable microATR accessory and the
following measurement conditions: [0196] apparatus: Thermo Nexus
670 FTIR spectrometer [0197] number of scans: 32 [0198] resolution:
1 cm.sup.-1 [0199] wavelength range: 4000 to 400 cm.sup.-1 [0200]
detector: DTGS with KBr windows [0201] beamsplitter: Ge on KBr
[0202] micro ATR accessory: Harrick Split Pea with Si crystal
[0203] Differential Scanning Calorimetry (DSC)
[0204] DSC data were collected on a TA-Instruments Q2000 MTDSC
equipped with RCS cooling unit. Typically 3 mg of each compound, in
a standard aluminium TA-Instrument sample pan, was heated at
10.degree. C./min from 25.degree. C. to 250/300.degree. C. A
nitrogen purge at 50 ml/min was maintained over the sample.
Thermogravimetric Analysis (TGA)
[0205] TGA data were collected on a TA-Instruments Q500
thermogravimeter Typically 10 mg of each sample was transferred
into a pre-weighed aluminium pan and was heated at 20.degree.
C./min from ambient temperature to 300.degree. C. or <80[(w/w)
%] unless otherwise stated.
Chemical Stability--HPLC
[0206] HPLC analysis was carried out using the following operating
conditions: [0207] Column Waters Xbridge C18
(150.times.3.0.times.3.5 mm) or equivalent (a column is considered
equivalent if performance as specified in SST is met and a
comparable separation of all relevant compounds is demonstrated).
[0208] Column temperature 35.degree. C. [0209] Sample temperature
10.degree. C. [0210] Flow rate 0.45 ml/min Injection volume The
injection volume can be adjusted as long as the qualification
limits of the system are not exceeded (detector and injector) and
the peak shape of the main compound is acceptable. As a guide, 30
.mu.l is considered suitable. [0211] Detection UV detection at 255
nm [0212] Mobile phase Preparation and composition: [0213] A 10 mM
ammonium acetate (0.771 g/l)+0.1%, v/v trifluoroacetic acid in
water [0214] B Acetonitrile [0215] Gradient Analytical run time is
41 minutes
TABLE-US-00001 [0215] Time (minutes) Solvent 0 35 36 41 42 48 % A
95 30 0 0 95 95 % B 5 70 100 100 5 5
[0216] With this HPLC method the degradant D019492 elutes at
RRT0.86.
Chemical Stability--Ultra High Pressure Liquid Chromatography
(UPLC)
[0217] UPLC analysis was carried out using the following operating
conditions: [0218] Column Acquity BEH C.sub.18; 2.1.times.150 mm;
1.7 .mu.m or equivalent (a column is considered equivalent if
performance as specified in SST is met and a comparable separation
of all relevant compounds is demonstrated) [0219] Column
temperature 35.degree. C. [0220] Sample temperature 10.degree. C.
[0221] Flow rate 0.40 ml/min [0222] Injection volume The injection
volume can be adjusted as long as the qualification limits of the
system are not exceeded (detector and injector) and the peak shape
of the main compound is acceptable. As a guide, 4 .mu.l is
considered suitable. [0223] Detection UV detection at 255 nm [0224]
Mobile phase Preparation and composition: [0225] A 10 mM ammonium
acetate (0.771 g/l)+0.1%, v/v trifluoroacetic acid in water [0226]
B Acetonitrile [0227] Gradient Analytical run time is 23
minutes
TABLE-US-00002 [0227] Time (minutes) Solvent 0 19 20 23 23.5 28 % A
95 30 0 0 95 95 % B 5 70 100 100 5 5
[0228] With this UPLC method the degradant D019492 elutes at
RRT0.92-0.93 and the degradant D019493 elutes at RRT 0.86-0.87.
Reagents and Suppliers
[0229] Lactohale200.RTM.: Particle size (Sympatec): D10: 5-15
.mu.m; D50: 50-100 .mu.m; D90: 120-160 .mu.m. Magnesium stearate:
Grade Hyqual.RTM. 2257; supplied by Avantor. Particle size: D10:
typically 3 .mu.m; D50: typically 11.5 .mu.m (10.5-16.5 .mu.m);
D90: typically 24 .mu.m (18-28 .mu.m). Supplied as a fine
powder
Example 1--Preparation of
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-me-
thoxyethyl)hex-5-ynamide (Compound of formula (I)) in the form of a
solid crystalline hydrate
5-Bromo-3-(2-chlorobenzyl)-2-(chloromethyl)quinazolin-4(3H)-one
(2)
##STR00006##
[0231] To a stirred solution of 2-amino-6-bromo-benzoic acid (3.06
g, 14.2 mmol) in toluene (75 mL) cooled to 0.degree. C. in an
ice-bath was added pyridine (0.60 mL, 7.10 mmol) followed by a
solution of chloroacetyl chloride (2.26 mL, 28.4 mmol) in toluene
(75 mL) drop-wise over 1 hr. The reaction mixture was allowed to
warm to RT, and was heated at 115.degree. C. for 3 hr and then
allowed to cool to RT. The solvent volume was reduced by half by
evaporation in vacuo. Upon standing overnight, the product
precipitated and was collected by filtration to afford
2-bromo-6-(2-chloroacetamido)benzoic acid (1a, X=Cl) (1.44 g) as a
white solid: m/z 290/292 (M+H).sup.+ (ES.sup.+). The filtrate was
concentrated in vacuo and the residue triturated with
ethanol/heptane to afford 2-bromo-6-(2-hydroxyacetamido) benzoic
acid (1b X=OH) (1.02 g, combined yield, 59%): m/z 274/276
(M+H).sup.+ (ES.sup.+). Both 1a and 1b can be used without further
purification in the next step.
[0232] To a stirred mixture of compound (1a) (7.50 g, 27.4 mmol),
2-chlorobenzylamine (5.00 mL, 41.05 mmol) and triethylamine (5.70
mL, 41.1 mmol) in toluene (250 mL) was added a solution of
phosphorus trichloride (2.60 mL, 30.1 mmol) in toluene (250 mL)
dropwise over 1 hr. The reaction mixture was heated to 110.degree.
C. for 24 hr, whereupon the hot solution was decanted and
concentrated in vacuo. The residue was triturated with propan-2-ol
(50 mL) to afford the title compound (2) (6.41 g, 59%) as a yellow
solid: R.sup.t 2.67 min; m/z 397/399 (M+H).sup.+ (ES.sup.+).
3-(3-(tert-Butyldimethylsilyloxy)phenyl)-1H-pyrazolo[3,4-c]pyrimidin-4-ami-
ne (6)
##STR00007##
[0234] To a stirred suspension of
3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (3) (8.22 g, 31.5 mmol),
3-phenol boronic acid (13.0 g, 94.5 mmol) and potassium phosphate
(10.0 g, 47.3 mmol) in degassed DMF/water (3:2, 140 mL) was added
Pd(dppf)Cl.sub.2 (13.0 g, 15.7 mmol). The reaction mixture was
flushed with nitrogen, heated at 120.degree. C. for 2 hr and then
allowed to cool to RT. The reaction mixture was diluted with EtOAc
(500 mL) and hydrochloric acid (2 M, 500 mL) and the resulting
suspension was filtered. The filtrate was extracted with
hydrochloric acid (2 M, 2.times.500 mL). The combined aqueous
extracts were basified with a saturated aqueous solution of sodium
carbonate to pH 10. The precipitate formed was filtered and the
filtrate was extracted with EtOAc (3.times.1 L). The combined
organic extracts were dried, filtered and the solvent removed in
vacuo to afford a grey solid. All solid materials generated during
the workup procedure were combined and triturated with DCM to
afford 3-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenol (5) (6.04
g, 84%) as a grey solid: m/z 228 (M+H).sup.+ (ES.sup.+).
[0235] To a stirred solution of the phenol (5) (4.69 g, 20.66 mmol)
and imidazole (2.10 g, 30.99 mmol) in dry DMF (100 mL) was added
TBDMSCl (4.70 g, 30.99 mmol). After 16 hr, further aliquots of
imidazole (2.10 g, 30.99 mmol) and TBDMSCl (4.70 g, 30.99 mmol)
were added and the mixture was stirred for 48 hr. The reaction
mixture was diluted with water (120 mL) and extracted with DCM
(2.times.200 mL). The combined organic extracts were washed with
water (2.times.200 mL), dried, filtered and the volume reduced to
approximately 100 mL by evaporation in vacuo. The resulting slurry
was filtered and the solid washed with heptane (50 mL) to afford
the title compound (6) (6.05 g, 85%) as an off-white solid: m/z 343
(M+H).sup.+ (ES.sup.+).
Intermediate A:
2-((4-Amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)--
5-bromo-3-(2-chlorobenzyl)quinazolin-4(3H)-one
##STR00008##
[0237] To a stirred mixture of
5-bromo-3-(2-chlorobenzyl)-2-(chloromethyl)quinazolin-4(3H)-one (2)
(100 mg, 0.25 mmol) and potassium carbonate (42 mg, 0.30 mmol) in
DMF (2.5 mL) was added a solution of
3-(3-(tert-butyldimethylsilyloxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-am-
ine (6) (94 mg, 0.28 mmol) in DMF (2.5 mL) and the reaction mixture
was stirred at RT for 18 hr. Potassium carbonate (3.times.35 mg,
0.75 mmol) was added in three portions over 30 hr. after which the
solvent was removed in vacuo and the crude material was purified by
flash column chromatography, eluting with 4.5% methanol in DCM, to
afford the title compound, Intermediate A, (94 mg, 64%) as a
off-white solid: R.sup.t 2.01 min; m/z 588/590 (M+H).sup.+,
(ES.sup.+).
Intermediate B: N,N-bis(2-Methoxyethyl)hex-5-ynamide
##STR00009##
[0239] To a solution of hex-5-ynoic acid (7.11 g, 63.4 mmol),
EDC.HCl (14.0 g, 72.9 mmol) and DMAP (387 mg, 3.17 mmol) in DCM
(600 mL) at 0.degree. C. was added bis(2-methoxyethyl)amine (9.3
mL, 63 mmol). The resulting mixture was warmed to RT for 20 hr and
was then washed with hydrochloric acid (1 M, 2.times.500 mL) and
with water (500 mL). The organic layer was dried and was evaporated
in vacuo to afford the title compound, Intermediate B, as a yellow
oil (16 g, 97%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.: 1.88
(3H, m), 2.26 (2H, m), 2.49 (2H, m), 3.32 (6H, s), 3.51 (4H, m),
3.55 (4H, m)
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-me-
thoxyethyl)hex-5-ynamide (I)
##STR00010##
[0241] Intermediate A
((2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl-
)-5-bromo-3-(2-chlorobenzyl)quinazolin-4(3H)-one (65.7 g, 1.0
eq.)), copper(I) iodide (1.06 g, 0.05 moles/mol),
bis(triphenylphosphine)palladium(II) chloride (3.92 g, 0.05
moles/mol), Intermediate B (N,N-bis(2-methoxyethyl)hex-5-ynamide
(63.42 g, 2.5 moles/mol) and diethylamine (837.05 mL; 591.21 g, 7.5
L/mol) were added to a 2 L reactor and the mixture degassed with
argon purging. The reaction mixture was warmed to 55.degree. C.
(reflux temperature) over 30 minutes and then stirred at 55.degree.
C. After 2 hours the mixture was cooled to 22.degree. C. before
being concentrated in vacuo to produce a dark brown semi solid
residue (201.0 g). The residue was then dissolved in MEK (781 mL)
and water added (223 mL). After stirring strongly for 5 minutes the
layers were separated and the aqueous layer discarded. The organic
layer was washed with 10% w/v aqueous NH.sub.4OAc (300 mL) and 2%
w/v aqueous NaCl (112 mL) before being partly concentrated in vacuo
to an heterogeneous mixture in MEK (230 g). The mixture was stirred
for 16 hours then filtered, and the precipitate was washed with MEK
(3.times.25 mL). The resulting solid was dried at 50.degree. C. in
vacuo for 18 hours to give "crude"
6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-me-
thoxyethyl)hex-5-ynamide (compound of formula (I)) (54.13 g; 0.66
equiv; 65.97% yield).
[0242] Crude compound of formula (I) (53.5 g; 1.00 equiv), methanol
(7.28 mL, 0.1 L/mol) and dichloromethane (145.53 mL, 2 L/mol) were
stirred in a 250 mL reactor at 22.degree. C. After 4 hours the
solid was filtered and washed with dichloromethane (29 mL) before
being dried in vacuo at 40.degree. C. for 18 hours to obtain
compound of formula (I) (the title compound) in the form of a
hydrate (45.3 g; 0.85 equiv; 84.67% crystallization yield) as an
off-white solid.
Example 2--Preparation of Compound of Formula (I) in Solid
Crystalline Anhydrous Form
[0243] All reactions described within this example were carried out
under a flow of nitrogen gas. Compound of formula (I) in the form
of a hydrate, as prepared in Example 1 (14.0 g) and 1-propanol (210
mL, 15 L/kg) were added to a 400 ml crystallization vessel. The
resulting heterogeneous mixture was stirred and warmed to
90.degree. C. (with the mixture becoming homogeneous at 85.degree.
C.). Once the solution had reached 90.degree. C., a metal scavenger
(SiliaMetS.RTM. Thiol 0.7 g (5% w/w)) was added and the mixture
warmed to 95.degree. C. After stirring for 15 minutes at 95.degree.
C. the mixture was cooled to 90.degree. C. and stirred for a
further 2 hours at 90.degree. C. The metal scavenger was then
filtered and the homogeneous filtrate was again stirred and warmed
to 95.degree. C., before being cooled to 85.degree. C. and stirred
for 8 hours. The filtrate was then cooled over 8 hours to
20.degree. C. and stirred for an additional 6 hours at 20.degree.
C. The product was then filtered and washed with 1-propanol (6 ml)
before being dried in vacuo at 50.degree. C. for 18 hours to afford
compound of formula (I) in anhydrous form (12.6 g, 90%) as a white
solid.
[0244] The above method may optionally be adapted to facilitate
crystallization with seeding.
Example 3--XRPD Analysis of Compound of Formula (I) in Solid
Crystalline Hydrate and Anhydrous Form
[0245] XRPD analysis of the hydrate and anhydrous forms of compound
of formula (I) was undertaken using the method described in General
Procedures. The resulting diffraction patterns are shown in FIGS. 1
and 2. Both XRPD patterns showed diffraction peaks without the
presence of a halo, thereby indicating that both materials are
crystalline. Peaks and intensities of the two forms are given below
in Tables 1 and 2:
TABLE-US-00003 TABLE 1 Characteristic XRPD peaks for compound of
formula (I) in a hydrate form XRPD peak (.+-.0.2 degrees, 2-theta
values) 5.6 7.6 9.6 11.1 12.2 12.6 13.3 13.9 15.9 17.0 18.9 20.3
21.8 23.1
TABLE-US-00004 TABLE 2 Characteristic XRPD peaks for the anhydrous
form of compound (I) XRPD peak (.+-.0.2 degrees, 2-theta values)
5.6 7.9 11.2 12.3 15.6 17.6 18.4 21.4 22.5 24.2
[0246] The two solid crystalline forms have some peaks in common
indicating that they appear to have related (but distinct) crystal
structures.
Example 4--Thermal Analysis of Compound of Formula (I) in Hydrate
and Anhydrous Form (Anhydrous Form with and without
Micronization)
[0247] Thermal analysis of the hydrate form and anhydrous form
(anhydrous form unmicronized and micronized) of compound of formula
(I) was undertaken using TGA, DVS, XRPD, IR and DSC as described in
General Procedures. Where appropriate, a sample at ambient
temperature and relative humidity (reference sample/"0 days") was
compared with samples stored at various temperatures and relative
humidities (comparative samples).
[0248] Micronized anhydrous form of compound of formula (I) was
prepared using a jet mill micronization device (1.5 bar)
(manufactured by Hosokawa Alpine). The Particle Size Distribution
was measured using laser diffraction (Malvern Mastersizer
instrument). Micronized anhydrous form of compound of formula (I)
had the following particle size distribution: D10 of 1.40 .mu.m;
D50 of 2.77 .mu.m and D90 of 5.29 .mu.m.
[0249] The tested storage conditions were 4 weeks at RT<5% RH,
RT 56% RH, RT 75% RH, 50.degree. C. and 40.degree. C. 75% RH. XRPD
and IR data were also acquired after 1 week at 80.degree. C.
Solid Crystalline Hydrate Form
[0250] TGA: The reference sample (t=0) and comparative samples
(exposed to different storage conditions) were heated at 20.degree.
C./min from RT to 300.degree. C. The TGA curve of the reference
sample (t=0) is illustrated in FIG. 11 and the results for all
samples are illustrated in Table 3. From FIG. 11 it is evident
weight loss of 1.1% was observed in the temperature region from RT
up to 45.degree. C. due to the evaporation of free solvent or
hygroscopic water (as evidenced by the first peak between
29.39.degree. C. and 44.82.degree. C.). Weight loss of 1.5% was
observed between 45.degree. C. and 190.degree. C. due to the
evaporation of bound solvent, and weight loss above 190.degree. C.
was due to evaporation and decomposition of the product. Comparing
this weight loss profile with those of the comparative samples in
Table 3, no significant differences were observed.
[0251] DVS: The DVS isotherm plot for the reference sample is
illustrated in FIG. 3 and the DVS change in mass plot is
illustrated in FIG. 4. During the initial drying step, a weight
loss of 2.2% was registered and the obtained dried product was
found to be hygroscopic. The hygroscopic product adsorbed up to
2.1% moisture depending on the atmospheric humidity and dried
completely during the desorption cycle. The obtained product after
adsorption/desorption was investigated with XRPD and IR and was
found to be comparable to the reference sample. These data indicate
that the hydrate form is hygroscopic.
[0252] XRPD and IR: The XPRD diffraction pattern of the reference
sample (t=0) is illustrated in FIG. 1 and the IR trace is
illustrated in FIG. 7. This diffraction pattern and IR trace were
compared with those of the comparative samples (exposed to
different storage conditions) and the results are illustrated in
Table 3. The diffraction patterns and IR traces were identical or
very similar for most samples, however some small differences in
the XRPD diffraction patterns and IR traces were observed after
storage at elevated temperatures of 50.degree. C. and 80.degree. C.
and under dry conditions (RT/<5% RH), when compared with the
reference sample (as evidenced by the ".+-.Ref" entries in Table
3). These same small differences were also observed in the XRPD
pattern and IR trace of the reference sample after DVS, suggesting
that the differences observed in the 50.degree. C., 80.degree. C.
and RT<5% RH samples were due to drying of the product.
[0253] DSC: The reference sample (t=0) and comparative samples
(exposed to different storage conditions) were heated at 10.degree.
C./min from 25.degree. C. to 300.degree. C. The DSC curve of the
reference sample is illustrated in FIG. 9 and the results for all
samples are illustrated in Table 3. From FIG. 9, it is evident that
the reference sample melted with decomposition at about
183.5.degree. C. with an endothermic signal at 64.5.degree. C. due
to solvent evaporation.
[0254] In summary, it is evident that the hydrate has good physical
stability. The product appears to be hygroscopic and experiences
some loss of water under dry conditions. However the crystalline
form appears to be stable to water loss and water gain.
TABLE-US-00005 TABLE 3 stability data for the hydrate form of the
compound of formula (I) DSC TGA Max Extra Product Condition
<45.degree. C. <190.degree. C. XRD IR (.degree. C.) (.degree.
C.) Appearance Hydrate 0 days* 1.1 1.5 Cryst., Ref Cryst., Ref
183.5 64.5 (44 J/g) white RT/<5% RH 0.7 0.9 .+-.Ref .+-.Ref
183.6 62.7 (23 J/g) white RT/56% RH 1.0 1.1 ~Ref ~Ref 183.5 66.0
(40 J/g) white RT/75% RH 1.2 1.3 ~Ref ~Ref 183.5 65.3 (40 J/g)
white 80.degree. C. (1 week) -- -- .+-.Ref .+-.Ref -- -- white
50.degree. C. 1.0 1.0 .+-.Ref .+-.Ref 183.7 64.7 (37 J/g) white
40.degree. C./75% RH 0.9 1.0 ~Ref ~Ref 183.4 66.9 (39 J/g) white
*ambient temperature and relative humidity Cryst., Ref: crystalline
reference ~Ref: identical to crystalline reference +/- Ref:
comparable to crystalline reference
Solid Crystalline Anhydrous Form (not Micronized)
[0255] TGA (Table 4 and FIG. 12): The reference sample and
comparative samples were heated at 20.degree. C./min from room
temperature to 300.degree. C. The TGA curve of the reference sample
(t=0) is illustrated in FIG. 12 and the results for all samples are
illustrated in Table 4. From FIG. 12 it is evident that for the
reference sample weight loss of 0.2% was observed in the
temperature region from room temperature up to 100.degree. C. due
to the evaporation of free solvent and/or hygroscopic water. Weight
loss of 0.4% was observed between 100.degree. C. and 200.degree.
C., probably due to the evaporation and decomposition of the
product. Comparing this weight loss profile with those of the
comparative samples--differences were observed under dry conditions
of RT/<5% RH where lower % weight losses of 0.7% and 0.9%
occurred.
[0256] DVS: The DVS isotherm plot for the reference sample is
illustrated in FIG. 5 and the DVS change in mass plot is
illustrated in FIG. 6. During the initial drying step, a weight
loss of 0.1% was registered. The obtained dried product exhibited
no hygroscopic behavior and remained in the same solid state form
during the test.
[0257] XRPD and IR: The XPRD diffraction pattern of the reference
sample is illustrated in FIG. 2 and the IR trace is illustrated in
FIG. 8. This diffraction pattern and IR trace were compared with
those of the comparative samples and the results are illustrated in
Table 4. The diffraction patterns and IR traces were found to be
identical for all samples, indicating that no solid state changes
occurred after storage under different conditions.
[0258] DSC: The reference sample and comparative samples were
heated at 10.degree. C./min from 25.degree. C. to 250.degree. C.
The DSC curve of the reference sample is illustrated in FIG. 10 and
the results for all samples are illustrated in Table 4. From FIG.
10, it is evident that the reference sample melted (with possible
decomposition) at about 187.0.degree. C. Comparing the DSC data of
the reference sample with the data for the comparative samples it
is evident that the storage conditions have not altered the melting
point of the substance.
[0259] In summary, it is evident that the solid crystalline
anhydrous form of compound of formula (I) was physically stable
under all investigated conditions.
TABLE-US-00006 TABLE 4 stability data for the anhydrous form of the
compound of formula (I) (unmicronized) DSC TGA Max Product
Condition <100 <200.degree. C. XRD IR (.degree. C.)
Appearance Anhydrous Form 0 days 0.2 0.4 Cryst., Ref Cryst., Ref
187.0 white (unmicronized) 80.degree. C. -- -- ~Ref ~Ref -- white
RT/<5% RH 0.3 0.5 ~Ref ~Ref 187.6 white RT/56% RH 0.4 0.5 ~Ref
~Ref 187.4 white RT/75% RH 0.3 0.4 ~Ref ~Ref 187.0 white 50.degree.
C. 0.3 0.4 ~Ref ~Ref 187.2 white 40.degree. C./75% RH 0.4 0.5 ~Ref
~Ref 187.2 white
Solid Crystalline Anhydrous Form (Micronized)
[0260] TGA: The reference sample and comparative samples were
heated at 20.degree. C./min from room temperature to 300.degree. C.
and the results for all samples are illustrated in Table 5.
[0261] DVS: The DVS isotherm plot for the reference sample is
illustrated in FIG. 13. During the initial drying step, a weight
loss of 0.1% was registered. The product was observed to be
slightly hygroscopic, adsorbing up to 0.9% moisture depending on
the atmospheric conditions. During the desorption cycle the product
was found to dry out completely.
[0262] XRPD and IR: The diffraction pattern and IR trace of the
reference sample were compared with those of the comparative
samples and the results are illustrated in Table 5. The diffraction
patterns and IR traces were found to be identical for all samples,
indicating that no solid state changes occurred after storage under
different conditions.
[0263] DSC: The reference sample and comparative samples were
heated at 10.degree. C./min from 25.degree. C. to 250.degree. C.
The results for all samples are illustrated in Table 5. The DSC
curve for the reference sample showed melting of the product (with
possible decomposition) at about 186.degree. C. An extra exothermic
signal was observed at about 124.degree. C. probably due to
recrystallization of amorphous material or crystal artefacts
present in the sample generated during milling.
[0264] In summary, it is evident that the solid crystalline
anhydrous form of compound of formula (I) in micronized form was
physically stable under all investigated conditions, although it is
slightly hygroscopic.
TABLE-US-00007 TABLE 5 stability data for the anhydrous form of the
compound of formula (I) (micronized) DSC TGA Max Extra Product
Condition <100 <200.degree. C. XRD IR (.degree. C.) (.degree.
C.) Appearance Anhydrous Form 0 days 0.2 0.4 Cryst., Ref Cryst.,
Ref 185.6 123.4 (4 J/g) white (micronized) 80.degree. C. -- -- ~Ref
~Ref -- -- white RT/<5% RH 0.3 0.4 ~Ref ~Ref 185.7 123.7 (4 J/g)
white RT/56% RH 0.4 0.4 ~Ref ~Ref 185.5 123.0 (4 J/g) white RT/75%
RH 0.3 0.4 ~Ref ~Ref 185.6 123.3 (4 J/g) white 50.degree. C. 0.5
0.4 ~Ref ~Ref 185.4 124.9 (4 J/g) white 40.degree. C./75% RH 0.2
0.3 ~Ref ~Ref 185.7 124.5 (2 J/g) white
Example 5--HPLC Analysis of Compound of Formula (I) in Solid
Crystalline Hydrate and Solid Crystalline Anhydrous Form (Anhydrous
Form--Unmicronized and Micronized)
[0265] The chemical stability of the solid crystalline hydrate form
and solid crystalline anhydrous form (anhydrous form both
unmicronized and micronized) of the compound of formula (I) was
determined by comparing a sample at ambient temperature and
relative humidity (reference sample) with samples stored at various
temperatures and relative humidities (comparative samples). The
samples were stored under various conditions for 1, 4 or 8 weeks as
shown in Tables 6, 7 and 8. The samples were then analyzed by HPLC
using the method in General Procedures and by visual
inspection.
Hydrate Form
[0266] From the data provided in Table 6 it is evident that the
hydrate form of compound of formula (I) is chemically stable
although some sensitivity to light was observed.
TABLE-US-00008 TABLE 6 stability data for the hydrate form of the
compound of formula (I) HPLC Sum of impurities Appearance Product
Condition 1 week 4 weeks 8 weeks 1 week 4 weeks 8 weeks Hydrate
Reference 1.52 -- -- white -- -- 0.3 days ICH light* 1.61 -- --
white -- -- 80.degree. C. 1.58 -- -- white -- -- 40.degree. C./75%
RH 1.54 1.46 1.54 white white white 50.degree. C. 1.53 1.53 1.59
white white white RT/<5% RH -- 1.50 1.53 -- white white RT/56%
RH -- 1.53 1.54 -- white white RT/75% RH -- 1.48 1.60 -- white
white *stimulated daylight (light cabinet 700 W/m.sup.2)
Anhydrous Form (Unmicronized)
[0267] From the data provided in Table 7 it is evident that the
anhydrous form of compound of formula (I) is sensitive to light.
After storage in ICH light for 0.3 days, a degradation product was
observed at RRT 1.12 and RRT 1.24.
TABLE-US-00009 TABLE 7 stability data for the anhydrous form of the
compound of formula (I) (solid state) HPLC Sum of impurities
Appearance Product Condition 1 week 4 weeks 8 weeks 1 week 4 weeks
8 weeks Anhydrous Form Reference 0.72 -- -- white -- --
(unmicronized) 0.3 da ICH light 1.08 -- -- slightly -- -- yellow
80.degree. C. 0.71 -- -- white -- -- 40.degree. C./75% RH 0.71 0.72
0.72 white white white 50.degree. C. 0.71 0.71 0.73 white white
white RT/<5% RH -- 0.72 0.74 -- white white RT/56% RH -- 0.71
0.77 -- white white RT/75% RH -- 0.71 0.74 -- white white
Anhydrous Form (Micronized)
[0268] From Table 8 it is evident that the anhydrous form of
compound of formula (I) in micronized form is sensitive to light.
After storage in ICH light for 0.3 days, a degradation product was
observed at RRT 1.12.
TABLE-US-00010 TABLE 8 stability data for the anhydrous form of the
compound of formula (I) (micronized) HPLC Sum of impurities
Appearance Product Condition 1 week 4 weeks 8 weeks 1 week 4 weeks
8 weeks Anhydrous Form Reference 0.72 -- -- white -- --
(micronized) 0.3 da ICH light 0.89 -- -- slightly -- -- yellow
80.degree. C. 0.71 -- -- white -- -- 40.degree. C./75% RH 0.72 0.71
0.76 white white white 50.degree. C. 0.71 0.71 0.76 white white
white RT/<5% RH -- 0.70 0.73 -- white white RT/56% RH -- 0.71
0.74 -- white white RT/75% RH -- 0.73 0.75 -- white white
[0269] The HPLC studies indicate that the chemical stabilities of
the hydrate and anhydrous forms (both unmicronized and micronized)
of compound of formula (I) are comparable, although all forms show
some sensitivity to light.
Example 6--XRPD/IR Analysis of Compound of Formula (I) in Hydrate
Form with Lactose, and in Anhydrous Form with Lactose
[0270] Mixtures of the hydrate form of compound of formula (I) with
lactose, and the anhydrous form (micronized) of compound of formula
(I) with lactose (in each case 50%/50%) were prepared, using
LactoHale.RTM. as lactose source (supplied by
DOMO.RTM./Frieslandfoods). The mixtures were stored under different
temperatures and humidities and were analysed by XRPD and IR at
time zero and after 1 week and 4 weeks of storage. The IR spectra
and the XRPD patterns of the 1 and 4 week stability samples were
compared with the IR spectrum and XRPD pattern generated at time
zero.
Solid Crystalline Hydrate
[0271] Blend preparation: about 250 mg of compound of formula (I)
in hydrate form and 250 mg Lactohale200.RTM. were added to an agate
mortar before being mixed using a pestle and plastic blade (Feton)
for 5 minutes. The physical blends were filled in 10 mL brown glass
flasks with screw lid (closed) and without lid (open). The
following storage conditions were used:
80.degree. C.: 1 week closed; 50.degree. C.: 1 and 4 weeks closed;
40.degree. C./75% RH: 1 and 4 weeks open.
[0272] A reference IR spectrum of a sample of a blend of solid
crystalline hydrate form with lactose is shown in FIG. 14. IR
spectra were acquired after the various storage conditions. No
differences were observed between the IR spectra of the 1 and 4
week stability samples and the IR spectrum at time zero. No
interaction between the hydrate form and lactose was observed and
the hydrate form remained stable under all storage conditions.
[0273] A reference XRPD pattern of a sample of a blend of solid
crystalline hydrate form with lactose is shown in FIG. 15. XRPD
patterns were acquired after the various storage conditions. The
generated XRPD patterns of the 1 and 4 week stability samples were
similar to the diffraction pattern at time zero. It was clearly
visible that the typical diffraction peaks of the hydrate form did
not change in the presence of Lactohale200.RTM., indicating that
the hydrate form is physically stable in the presence of
lactose.
[0274] The IR spectra showed no interaction between the hydrate
form and the lactose, and the XRPD results showed that there was no
solid state conversion of the hydrate form. As such, it may be
concluded that the hydrate form is physically compatible with
lactose.
Solid Crystalline Anhydrous Form
[0275] Blend preparation: about 500 mg of anhydrous compound of
formula (I) (micronized) and 500 mg Lactohale200.RTM. were added to
an agate mortar before being mixed using a pestle and plastic blade
(Feton) for 5 minutes. The physical blends were filled in 10 mL
brown glass flasks with screw lid (closed) and without lid (open).
The following storage conditions were used:
80.degree. C.: 1 week closed; 50.degree. C.: 1 and 4 weeks closed;
40.degree. C./75% RH: 1 and 4 weeks open.
[0276] A reference IR spectrum of a sample of a blend of solid
crystalline anhydrous form (micronized) with lactose is shown in
FIG. 16. IR spectra were acquired after the various storage
conditions. No differences were observed between the IR spectra of
the 1 and 4 week stability samples and the IR spectrum at time
zero. No interaction between the anhydrous form and lactose was
observed and the anhydrous form remained stable under all storage
conditions.
[0277] A reference XRPD patterns of a sample of a blend of solid
crystalline anhydrous form (micronized) with lactose is shown in
FIG. 17. XRPD patterns were acquired after the various storage
conditions. The generated XRPD patterns of the 1 and 4 week
stability samples are similar to the diffraction pattern at time
zero. It is clearly visible that the typical diffraction peaks of
the anhydrous form did not change in the presence of
Lactohale200.RTM., indicating that the anhydrous form is physically
stable in the presence of lactose.
[0278] The IR spectra showed no interaction between the anhydrous
form and the lactose, and the XRPD results showed that there was no
solid state conversion of the anhydrous form. As such, it may be
concluded that the anhydrous form is physically compatible with
lactose.
Example 7--HPLC Analysis of Compound of Formula (I) in Solid
Crystalline Hydrate Form with Lactose, and in Solid Crystalline
Anhydrous Form with Lactose
[0279] The chemical compatibility of the hydrate form and anhydrous
form of compound of formula (I) in combination with lactose was
determined by HPLC analysis.
Hydrate
[0280] Blend preparation: 2 mg of hydrate form and 2 mg of
Lactohale200.RTM. were added to an agate mortar before being mixed
using a pestle and plastic blade (Feton) for 5 minutes. Further
aliquots of Lactohale200.RTM. (starting at 4 mg) were mixed into
the blend, doubling the volume of the mixture each time, until the
mixture contained 6000 mg Lactohale200.RTM. in total.
[0281] The mixtures were analysed by HPLC at time zero and after
different conditions of storage. Samples were stored under the
following conditions: (i) 1, 2 and 3 weeks at 50.degree. C. (ii) 1
week 80.degree. C. (iii) 1, 2 and 3 weeks at 40.degree. C./75% RH.
From Table 9 it is evident that the hydrate form of compound of
formula (I) is stable in combination with lactose for up to 3
weeks, indicating their chemical compatibility.
TABLE-US-00011 TABLE 9 stability data for the hydrate form of the
compound of formula (I) with lactose RRT* RRT* RRT* RRT* RRT* RRT*
Conditions 0.80 0.84 0.86 1.11 1.14 1.32 T = zero 0.15 0.13 0.15
0.09 0.15 0.92 1 week 50.degree. C. 0.14 0.31 0.15 0.94 1 week
80.degree. C. 0.15 0.12 0.15 0.91 1 week 0.16 0.13 0.18 0.24 0.93
40.degree. C./ 75% RH 2 weeks 50.degree. C. 0.15 0.13 0.12 0.10
0.16 0.91 2 weeks 0.13 0.13 0.15 0.10 0.17 0.92 40.degree. C./ 75%
RH 3 weeks 50.degree. C. 0.14 0.11 0.17 0.08 0.14 0.99 3 weeks 0.14
0.10 0.13 0.08 0.14 1.00 40.degree. C./ 75% RH *Area % by HPLC at
RRT indicated. Compound of formula (I) has RRT = 1.0
Anhydrous (Micronized)
[0282] Micronized anhydrous form of compound of formula (I) was
prepared as described in Example 4.
[0283] The test batch was taken from stock containing 3.519 mg
anhydrous form of compound of formula (I) (micronized) and 6006.64
mg Lactohale200.RTM..
[0284] The mixtures were analysed by HPLC at time zero and after
different conditions of storage. Samples were stored under the
following conditions: (i) 1, 2, 3 and 4 weeks at 50.degree. C. (ii)
1 week 80.degree. C. (iii) 1, 2, 3 and 4 weeks at 40.degree. C./75%
RH.
[0285] Table 10 indicates that significant degradation was observed
after storage for 1 week at 80.degree. C. and degradation was also
observed after storage at elevated temperatures of 50.degree. C.
These results suggest that the anhydrous form (micronized) of
compound of formula (I) is not chemically stable in combination
with lactose, therefore the two components would not be compatible
in a pharmaceutical formulation.
[0286] The peak at RRT 0.86 has been attributed to the hydrated
derivative(s) D019328 shown above.
TABLE-US-00012 TABLE 10 stability data for the solid crystalline
anhydrous form of the compound of formula (I) (micronized) with
lactose Conditions RRT* 0.80 RRT* 0.86 RRT* 0.97 RRT* 1.14 RRT*
1.32 T = zero 0.21 0.12 0.12 0.13 1 week 50.degree. C. 0.17 0.23
0.10 0.12 1 week 80.degree. C. 0.52 2.53 0.78 0.19 0.12 1 week
40.degree. C./75% RH 0.19 0.12 0.11 0.13 2 weeks 50.degree. C. 0.19
0.30 0.12 0.13 2 weeks 40.degree. C./75% RH 0.17 0.11 0.12 0.13 3
weeks 50.degree. C. 0.19 0.38 0.12 0.14 3 weeks 40.degree. C./75%
RH 0.19 0.08 0.11 0.14 4 w 50.degree. C. 0.19 0.54 0.11 0.13 4 w
40.degree. C./75% RH 0.18 0.20 0.11 0.14 *Area % by HPLC at RRT
indicated. Compound of formula (I) has RRT = 1.0
Example 8--XRPD/IR Analysis of Compound of Formula (I) in Solid
Crystalline Anhydrous Form with Lactose and Magnesium Stearate
[0287] A mixture of the solid crystalline anhydrous form
(micronized) of compound of formula (I) with lactose was prepared
with the addition of 1% magnesium stearate. The mixtures were
stored under different temperatures and humidities and were
analysed by XRPD and IR at time zero and after 1 week and 4 weeks
of storage.
[0288] Blend preparation: about 500 mg of Lactohale200.RTM. and
about 10 mg magnesium stearate were added to an agate mortar before
being mixed using a pestle and plastic blade (Feton) for 5 minutes.
About 500 mg of anhydrous compound of formula (I) (micronized) was
added to the mixture and the blend was mixed for a further 5
minutes. Samples of the blend were then stored under the various
conditions described in Example 6.
[0289] A reference IR spectrum of a sample of a blend of solid
crystalline anhydrous form (micronized) with lactose and magnesium
stearate is shown in FIG. 18. IR spectra were acquired after the
various storage conditions. No differences were observed between
the IR spectra of the 1 and 4 week stability samples and the IR
spectrum at time zero. No interaction between the anhydrous form;
lactose and magnesium stearate was observed and the anhydrous form
remained stable under all storage conditions.
[0290] A reference XRPD pattern of a sample of a blend of solid
crystalline anhydrous form (micronized) with lactose and magnesium
stearate is shown in FIG. 19. XRPD patterns were acquired after the
various storage conditions. The generated XRPD patterns of the 1
and 4 week stability samples were similar to the diffraction
pattern at time zero. It was clearly visible that the typical
diffraction peaks of the anhydrous form did not change in the
presence of Lactohale200.RTM. and magnesium stearate, indicating
that the anhydrous form is physically stable in the presence of
lactose and magnesium stearate.
[0291] The IR spectra showed no interaction between the anhydrous
form, the lactose and the magnesium stearate, and the XRPD results
showed that there was no solid state conversion of the anhydrous
form. As such, it may be concluded that the anhydrous form is
physically compatible with lactose and magnesium stearate.
Example 9--HPLC Analysis of Compound of Formula (I) in Anhydrous
Form with Lactose and Magnesium Stearate
[0292] The chemical compatibility of the solid crystalline
anhydrous form of compound of formula (I) in combination with
lactose and 1% magnesium stearate was determined by HPLC
analysis.
[0293] The test batch was taken from stock containing 3.704 mg
anhydrous form of compound of formula (I) (micronized), 6017.90 mg
Lactohale200 and 67.33 mg magnesium stearate.
[0294] The data shown in Table 11 indicate a significant increase
in chemical stability compared with the same composition with the
absence of magnesium stearate (see Table 2), as evidenced by only a
small amount of degradation observed after storage for 1 week at
80.degree. C. (see e.g. RRT 0.86, 0.28%). These results suggest
that the chemical stability of the anhydrous form (micronized) of
compound of formula (I) with lactose is significantly improved by
the addition of magnesium stearate to the composition. As such, the
addition of magnesium stearate improves the chemical compatibility
of the anhydrous form (micronized) of compound of formula (I) in
combination with lactose such that they could be compatible in a
pharmaceutical formulation.
TABLE-US-00013 TABLE 11 stability data for the anhydrous form of
the compound of formula (I) (micronized) with lactose and magnesium
stearate RRT* Conditions 0.80 RRT* 0.86 RRT* 1.14 RRT* 1.32 T =
zero 0.21 0.10 0.12 0.13 1 week 50.degree. C. 0.20 0.11 0.11 0.13 1
week 80.degree. C. 0.19 0.28 0.11 0.13 1 week 40.degree. C./ 0.20
0.11 0.11 0.13 75% RH 2 weeks 50.degree. C. 0.20 0.08 0.11 0.14 2
weeks 40.degree. C./ 0.21 0.11 0.11 0.13 75% RH 3 weeks 50.degree.
C. 0.20 0.13 0.11 0.13 3 weeks 40.degree. C./ 0.20 0.11 0.11 0.14
75% RH 4 weeks 50.degree. C. 0.19 0.12 0.11 0.14 4 weeks 40.degree.
C./ 0.20 0.10 0.10 0.13 75% RH *Area % by UPLC at RRT indicated.
Compound of formula (I) has RRT = 1.0
Example 10--UPLC Analysis of Compound of Formula (I) in Anhydrous
Form with Lactose and Metal Salts of Stearic Acid
[0295] The chemical compatibility of the solid crystalline
anhydrous form of compound of formula (I) (micronized) in
combination with lactose and 1% metal salt of stearic acid
(magnesium stearate, sodium stearate and calcium stearate) was
determined by UPLC analysis (micronization of compound of formula
(I) as described in Example 4).
[0296] Test samples were prepared as described in Table 12
below:
TABLE-US-00014 TABLE 12 test samples for UPLC analysis after
accelerated stability testing Metal salt of solid crystalline
anhydrous form of stearic acid compound of formula (I) (micronized)
Lactohale 200 sample 1/ Sample sample 1/sample 2 sample 1/sample 2
sample 2 Drug only 0.50 mg/0.47 mg Drug and 0.58 mg/0.47 mg 749.84
mg/750.06 mg lactose Drug, lactose, 0.46 mg/0.51 mg 749.97
mg/751.59 mg 7.40 mg/7.55 mg Mg stearate Drug, lactose, 0.49
mg/0.45 mg 751.08 mg/753.53 mg 7.67 mg/7.80 mg Ca stearate Drug,
lactose, 0.48 mg/0.45 mg 750.20 mg/750.42 mg 7.78 mg/7.59 mg Na
stearate
[0297] Samples were dispensed into vials, sealed with caps and kept
at 80.degree. C. for 1 or 2 weeks. Sample 1 was used for the 1 week
studies and sample 2 was used for the 2 week studies.
[0298] Results are shown in Table 13 below:
TABLE-US-00015 TABLE 13 results of UPLC analysis after accelerated
stability testing 1 week 1 week 2 weeks 2 weeks 80.degree. C.
80.degree. C. 80 .degree. C. 80.degree. C. Sample RRT* 0.87 RRT*
0.92 RRT* 0.87 RRT* 0.92 Drug only 0.00 0.08 0.00 0.08 Drug and
0.58 0.39 1.80 0.77 lactose Drug, lactose, 0.28 0.29 0.06 0.18 Mg
stearate Drug, lactose, 0.11 0.19 0.17 0.19 Ca stearate Drug,
lactose, 0.00 0.09 0.00 0.09 Na stearate *Area % by UPLC at RRT
indicated. Compound of formula (I) has RRT = 1.0
[0299] Mass spectroscopy analysis indicates that the substance with
RRT 0.87 is D019493 and the substance with RRT 0.92 is D019492
(confirmed by NMR) (see Scheme 1). The NMR resonance assignments
for D019492 are given in Table 14:
TABLE-US-00016 TABLE 14 .sup.1H NMR resonance assignments for
D019492 .sup.1H NMR assignments (600 MHz, DMSO-d.sub.6) .delta. ppm
D019492 1.59 (quin, J = 7.30 Hz, 2 H) 2.20 (t, J = 7.55 Hz, 2 H)
2.46-2.49 (m, 2 H) 3.18 (d, J = 7.90 Hz, 6 H) 3.29-3.39 (m, 8 H)
4.23 (s, 2 H) 5.24 (s, 2 H) 5.76 (s, 2 H) 6.08 (d, J = 7.55 Hz, 1
H) 6.75 (t, J = 7.55 Hz, 1 H) 6.83 (dd, J = 8.12, 1.70 Hz, 1 H)
6.90 (d, J = 7.55 Hz, 1 H) 6.91-6.93 (m, 1 H) 7.01 (t, J = 7.55 Hz,
1 H) 7.09 (d, J = 7.55 Hz, 1 H) 7.29 (m, J = 7.93, 7.93 Hz, 1 H)
7.32 (d, J = 7.18 Hz, 1 H) 7.66 (d, J = 7.93 Hz, 1 H) 7.77-7.82 (m,
1 H) 8.17 (s, 1 H) 9.67 (s, 1 H)
[0300] The data shown in Table 13 indicate a significant increase
in chemical stability for formulations containing a metal salt of
stearic acid compared with the same composition in the absence of a
metal salt of stearic acid, as evidenced by a comparatively small
amount of degradation observed after storage for 1 or 2 weeks at
80.degree. C. These results suggest that the chemical stability of
the anhydrous form of compound of formula (I) with lactose is
significantly improved by the addition of metal salts of stearic
acid to the composition. Therefore the addition of metal salts of
stearic acid improves the chemical compatibility of the anhydrous
form of compound of formula (I) in combination with lactose such
that they could be compatible in a pharmaceutical formulation.
Example 11--Preparation of Pharmaceutical Formulations According to
the Invention
[0301] An exemplary pharmaceutical formulation of the invention
consists of 0.5 wt. % of compound of formula (I) (solid crystalline
anhydrous form, micronised), 98.5 wt. % lactose monohydrate
(inhalation grade) and 1.0 wt. % magnesium stearate, wherein the
wt. % of all components is based on the weight of the dry
pharmaceutical formulation.
Summary of the Results Disclosed in the Examples
[0302] Solid crystalline anhydrous and hydrate forms of compound of
formula (I) have been identified.
[0303] From the TGA, DVS, XRPD, IR and DSC studies, it is evident
that the solid crystalline anhydrous form of compound of formula
(I) (in both unmicronized and micronized forms) and the solid
crystalline hydrate form are both stable, although the hydrate form
has a tendency to lose some water under dry conditions, apparently
without impact on its crystalline structure. The chemical
stabilities of the hydrate and anhydrous forms of the compound of
formula (I) are comparable.
[0304] When the solid crystalline hydrate and anhydrous forms were
tested for their chemical compatibility with lactose, although both
forms were found to be physically compatible, chemical degradation
was observed for the solid crystalline anhydrous form in the
presence of lactose.
[0305] However, the addition of magnesium stearate, calcium
stearate or sodium stearate (examples of a metal salt of stearic
acid) to the combination of the solid crystalline anhydrous form of
compound of formula (I) and lactose was surprisingly found to
significantly reduce chemical degradation. As such, a
pharmaceutical formulation comprising compound of formula (I) in
solid crystalline anhydrous form, lactose and a metal salt of
stearic acid such as magnesium stearate has good physical and
chemical stability.
[0306] All references referred to in this application, including
patent and patent applications, are incorporated herein by
reference to the fullest extent possible.
[0307] Throughout the specification and the claims which follow,
unless the context requires otherwise, the word `comprise`, and
variations such as `comprises` and `comprising`, will be understood
to imply the inclusion of a stated integer, step, group of integers
or group of steps but not to the exclusion of any other integer,
step, group of integers or group of steps.
* * * * *