U.S. patent application number 14/381273 was filed with the patent office on 2015-04-09 for novel pharmaceutical formulations.
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 C. Vanhoutte.
Application Number | 20150099768 14/381273 |
Document ID | / |
Family ID | 49160309 |
Filed Date | 2015-04-09 |
United States Patent
Application |
20150099768 |
Kind Code |
A1 |
Broeckx; Rudy Laurent Maria ;
et al. |
April 9, 2015 |
NOVEL PHARMACEUTICAL FORMULATIONS
Abstract
There is provided inter alia a dry powder pharmaceutical
formulation for inhalation comprising: (i)
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 or a pharmaceutically acceptable salt
thereof, including all stereoisomers, tautomers and isotopic
derivatives thereof and solvates thereof in particulate form as
active ingredient; (ii) particulate lactose as carrier; and (iii) a
particulate stabilizing agent selected from metal salts of stearic
acid such as magnesium stearate and metal salts of stearyl
fumarate.
Inventors: |
Broeckx; Rudy Laurent Maria;
(Turnhout, BE) ; Filliers; Walter Ferdinand Maria;
(Vremde, BE) ; Nieste; Patrick Hubert J.;
(Westerlo, BE) ; Copmans; Alex Herman; (Lille,
BE) ; Vanhoutte; Filip Marcel C.; (Bellegem, BE)
; Leys; Carina; (Stabroek, BE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Respivert Limited |
Buckinghamshire |
|
GB |
|
|
Family ID: |
49160309 |
Appl. No.: |
14/381273 |
Filed: |
March 13, 2013 |
PCT Filed: |
March 13, 2013 |
PCT NO: |
PCT/GB2013/050623 |
371 Date: |
August 27, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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61610012 |
Mar 13, 2012 |
|
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61610023 |
Mar 13, 2012 |
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Current U.S.
Class: |
514/263.21 |
Current CPC
Class: |
A61K 9/0073 20130101;
A61K 9/0075 20130101; A61P 7/00 20180101; C07D 487/04 20130101;
A61K 31/519 20130101; A61P 35/04 20180101; A61P 11/06 20180101;
A61P 11/00 20180101; A61K 47/26 20130101; A61P 43/00 20180101; A61K
31/52 20130101; A61P 11/14 20180101; A61P 35/00 20180101; A61K
47/12 20130101; A61P 29/00 20180101; A61P 37/00 20180101 |
Class at
Publication: |
514/263.21 |
International
Class: |
A61K 9/00 20060101
A61K009/00; A61K 47/26 20060101 A61K047/26; A61K 47/12 20060101
A61K047/12; A61K 31/52 20060101 A61K031/52 |
Claims
1. A dry powder pharmaceutical formulation for inhalation
comprising: a compound of formula (I) ##STR00009## 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-me-
thoxyethyl)hex-5-ynamide or a pharmaceutically acceptable salt
thereof, including all stereoisomers, tautomers and isotopic
derivatives thereof and solvates thereof in particulate form as
active ingredient; (ii) particulate lactose as carrier; and (iii) a
particulate stabilizing agent selected from metal salts of stearic
acid and metal salts of stearyl fumarate.
2. A pharmaceutical formulation according to claim 1, wherein the
compound of formula (I) is in its free base form.
3. A pharmaceutical formulation according to claim 1, wherein the
compound of formula (I) is in solid crystalline form.
4. A pharmaceutical formulation according to claim 3, wherein the
compound of formula (I) is in anhydrous form.
5. A pharmaceutical formulation according to claim 4, wherein the
compound of formula (I) is in solid crystalline form having the
X-ray powder diffraction pattern substantially as shown in FIG.
1.
6. A pharmaceutical formulation according to claim 4, wherein the
compound of formula (I) is in solid crystalline form having a X-ray
powder diffraction pattern containing one, two, three or four peaks
selected from (.+-.0.2) 17.6, 18.4, 22.5 and 24.2 degrees
2-theta.
7. A pharmaceutical formulation according to claim 1 wherein the
active ingredient has been micronized.
8. A pharmaceutical formulation according to claim 1 wherein the
stabilizing agent is a metal salt of stearic acid.
9. A pharmaceutical formulation according to claim 8 wherein the
stabilizing agent is magnesium stearate.
10. A pharmaceutical formulation according to claim 1 wherein the
lactose is .alpha.-lactose monohydrate.
11. An inhalation device comprising one or more doses of a
pharmaceutical formulation according to claim 1.
12. (canceled)
13. (canceled)
14. A method of treatment of a condition selected from: COPD
(including chronic bronchitis and emphysema), asthma including
paediatric asthma, cystic fibrosis, sarcoidosis, idiopathic
pulmonary fibrosis, cachexia and inhibition of the growth and
metastasis of lung tumours including non-small cell lung carcinoma
which comprises administering to a subject an effective amount of a
pharmaceutical formulation according to claim 1.
15-20. (canceled)
21. A method of increasing the stability of a pharmaceutical
formulation containing a compound of formula (I) and lactose to
chemical degradation which comprises including in said formulation
a stabilizing amount of a stabilizing agent selected from metal
salts of stearic acid such as magnesium stearate and metal salts of
stearyl fumarate.
22. A method according to claim 21 wherein the compound of formula
(I) is in solid crystalline anhydrous form.
23. A method according to claim 22 wherein the compound of formula
(I) is in solid crystalline form having the X-ray powder
diffraction pattern substantially as shown in FIG. 1.
24. A method according to claim 22 wherein the compound of formula
(I) is in solid crystalline form having the X-ray powder
diffraction pattern containing one, two, three or four peaks
selected from (.+-.0.2) 17.6, 18.4, 22.5 and 24.2 degrees
2-theta.
25. A method according to claim 21 wherein the stabilizing agent is
a metal salt of stearic acid.
26. A method according to claim 25 wherein the stabilizing agent is
magnesium stearate.
Description
FIELD OF THE INVENTION
[0001] The present invention provides novel dry powder
pharmaceutical formulations for inhalation of a compound that
inhibits phosphoinositide 3-kinases (PI3 kinases), and their use in
therapy, especially in the treatment of inflammatory diseases such
as COPD and asthma.
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-dihydroquinazolin-5-yl)-N,N-bis(2-me-
thoxyethyl)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's earlier 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.
[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) and 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 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.
[0012] One such additional excipient is magnesium stearate which is
known for improving certain properties of formulations containing
it. Thus, U.S. Pat. No. 7,186,401B2 (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.
[0013] Thus, there remains a need to provide formulations 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 formulation of the compound of
formula (I) which has appropriate physical and chemical stability
and other necessary properties for inhalation therapy.
SUMMARY OF THE INVENTION
[0014] In a first aspect, the present invention provides a dry
powder pharmaceutical formulation for inhalation comprising: [0015]
(i)
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)) or a
pharmaceutically acceptable salt thereof, including all
stereoisomers, tautomers and isotopic derivatives thereof and
solvates thereof in particulate form as active ingredient; [0016]
(ii) particulate lactose as carrier; and [0017] (iii) a particulate
stabilizing agent selected from metal salts of stearic acid (such
as magnesium stearate) and metal salts of stearyl fumarate
[0018] Such a formulation is hereinafter referred to as "a
formulation of the invention".
[0019] As explained in the Examples, formulations of the invention
appear to have good physical stability (as determined by XRPD and
IR analysis) and good chemical stability (as determined by HPLC
analysis). Without being limited by theory, it appears from the
inventors' discoveries that the alkyne group of the compound of
formula (I) is susceptible to metal catalysed oxidative degradation
involving hydration of the alkyne. It also appears from the
inventors' discoveries that the pyrimidinone ring of the compound
of formula (I) is susceptible to hydrolytic cleavage. Experiments
conducted by the inventors have determined that formulations of the
invention containing lactose and a metal salt of stearic acid such
as magnesium stearate have superior chemical stability than
corresponding formulations not containing a metal salt of stearic
acid such as magnesium stearate. To the inventors' knowledge it has
not been reported before that a metal salt of stearic acid such as
magnesium stearate can act as a protecting agent against chemical
degradation of alkyne containing compounds (especially in respect
of metal catalysed oxidative degradation involving hydration of the
alkyne) in dry powder inhalation formulations. To the inventors'
knowledge it has also not been reported before that a metal salt of
stearic acid such as magnesium stearate can act as a protecting
agent against hydrolytic cleavage of a drug substance containing a
pyrimidinone ring. The inventors extrapolate these findings with
metal salts of stearic acid to metal salts of stearyl fumarate.
BRIEF DESCRIPTION OF THE FIGURES
[0020] FIG. 1 shows an XRPD pattern acquired on a sample of
compound of formula (I) in solid crystalline anhydrous form.
[0021] FIG. 2 shows an IR spectrum of a sample of a blend of
compound of formula (I) in anhydrous form (micronized) with
Lactohale200.RTM. and magnesium stearate.
[0022] FIG. 3 shows an XRPD pattern acquired on a sample of a blend
of compound of formula (I) in anhydrous form (micronized) with
Lactohale200.RTM. and magnesium stearate.
DETAILED DESCRIPTION OF THE INVENTION
Compound of Formula (I) as Active Ingredient
[0023] 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.
[0024] 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.
[0025] 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).
[0026] The dry powder pharmaceutical formulation of the present
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.
[0027] 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. %, from about 0.02 wt. % to about
20 wt. %, or from about 0.02 wt. % to about 15 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 and based on weight of compound of
formula (I) as free base.
[0028] 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.
[0029] Compound of formula (I) is 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.
[0030] 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.
[0031] 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 (e.g. 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.
[0032] In one embodiment, compound of formula (I) is in free base
form, in the form of a pharmaceutically acceptable salt, or in the
form of a solvate of either. Suitably compound of formula (I) is in
free base form, e.g. in anhydrous form.
[0033] Suitably, compound of formula (I), or a pharmaceutically
acceptable salt or solvate thereof, is in solid crystalline
form.
Pharmaceutically Acceptable Salts of Compound of Formula (I)
[0034] In one embodiment there is provided a pharmaceutically
acceptable salt of compound of formula (I).
[0035] The pharmaceutically acceptable salts as mentioned
hereinabove are meant to comprise the therapeutically active
non-toxic acid addition salt forms that the compound of formula (I)
is able to form. These pharmaceutically acceptable acid addition
salts conveniently can be obtained by treating the base form with
such appropriate acid. Appropriate acids comprise, for example,
inorganic acids such as hydrohalic acids, e.g. hydrochloric or
hydrobromic acid, sulfuric, nitric, phosphoric and the like acids;
or organic acids such as, for example, acetic, propanoic,
hydroxyacetic, lactic, pyruvic, oxalic (i.e. ethanedioic), malonic,
succinic (i.e. butanedioic acid), maleic, fumaric, malic, tartaric,
citric, methanesulfonic, ethanesulfonic, benzenesulfonic,
p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic
and the like acids.
[0036] Thus, specific examples of salts of compound of formula (I)
include the acid additional salts formed with HCl, HBr and
p-toluenesulfonic acid.
Solvates
[0037] The invention also extends to solvates of compound of
formula (I). Examples of solvates include hydrates and hygroscopic
products such as channel hydrates.
Anhydrous Form of Compound of Formula (I)
[0038] 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 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% 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.
[0039] 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 an 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 weight of compound of formula (I). Suitably
crystallisation is performed by cooling the solution of compound of
formula (I) and solvent from elevated temperature (e.g.
80-95.degree. C.), continuously (i.e. continuous cooling) or in
stages (i.e. alternating between cooling and holding 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. and
80-20.degree. C. In one embodiment, the solution is cooled from
around 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. Crystals of compound of formula (I) in solid
crystalline form may be collected by usual separation techniques
(e.g. by filtration or centrifugation).
[0040] In one embodiment, there is provided a solid crystalline
anhydrous form of 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.
[0041] Thus, there is provided compound of formula (I) in a
crystalline anhydrous form having an X-ray powder diffraction
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.
[0042] The chemical compatibility of the anhydrous form of
compound(I) with lactose was investigated.
[0043] In order to assess chemical compatibility, compositions of
the anhydrous form of compound of formula (I) with lactose were
analysed by HPLC. The results are summarised in Example 4 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:
##STR00002##
[0044] 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 with identical mass (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.
[0045] Further investigation involving accelerated stability
testing (i.e. exposure of the drug substance to 80.degree. C. in a
closed vial, see Example 7) 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).
##STR00003##
[0046] 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 5). 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 6). A similar stabilising
effect was found using other metal salts of stearic acid,
specifically sodium stearate and calcium stearate (Example 7).
[0047] Without wishing to be bound by theory, it appears that the
metal salt of stearic acid such as magnesium stearate (or, it is
believed, a metal salt of stearyl fumarate) can act as a protecting
agent against chemical degradation of the alkyne group in the
compound of formula (I) and against chemical degradation of the
pyrimidinone ring in the compound of formula (I) which is observed
when the anhydrous form of compound of formula (I) is in a mixture
with lactose.
Particulate Lactose as Carrier
[0048] 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. (sieved inhalation grade lactose; DFE Pharma)
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.
[0049] In order to penetrate sufficiently far into the lungs, the
particulate active ingredient (in this case compound (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.
[0050] Generally, to prevent agglomeration of the small active
particles, 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).
[0051] In one embodiment, the dry powder formulation of the present
invention comprises particulate lactose having D50 in the range
40-150 .mu.m.
[0052] 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 wt. %, for example from about 50 wt. % to about 99.88
wt. %, for example from about 65 wt. % to about 99.88 wt. %, for
example from about 75 wt. % to about 99.99 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 99.98 wt. %, for example from about 95 wt. % to
about 99 wt. % of particulate lactose based on the weight of the
dry powder pharmaceutical composition.
Particulate Metal Salt of Stearic Acid Such as Magnesium Stearate
or Metal Salt of Stearyl Fumarate as Stabilizing Agent
[0053] An example metal salt of stearic acid is magnesium
stearate.
[0054] 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.
[0055] 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.
[0056] 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.
[0057] 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.
[0058] 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.
[0059] 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 the 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.
[0060] The dry powder pharmaceutical formulation of the present
invention comprises particulate stabilizing agent selected from
metal salt 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 stabilizing agent selected from metal salt of stearic
acid such as magnesium stearate and metal salts 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
stabilizing agent selected from metal salt of stearic acid such as
magnesium stearate and metal salts of stearyl fumarate based on the
weight of the dry powder pharmaceutical composition. Suitably, the
stabilizing agent selected from metal salt of stearic acid such as
magnesium stearate and metal salts 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.
[0061] In one embodiment, the dry powder pharmaceutical formulation
for inhalation of the present invention comprises: [0062] (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 or a pharmaceutically acceptable salt
thereof, including all stereoisomers, tautomers and isotopic
derivatives thereof and solvates thereof in particulate form as
active ingredient; [0063] (ii) from about 40 wt. % to about 99.88
wt. % particulate lactose; and [0064] (iii) 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.
[0065] In a further embodiment, the dry powder pharmaceutical
formulation for inhalation of the present invention comprises:
[0066] (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 free base form; [0067] (ii) from about
40 wt. % to about 99.88 wt. % particulate lactose; and [0068] (iii)
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.
[0069] A further aspect of the invention relates to the use of a
stabilizing agent selected from metal salt of stearic acid such as
magnesium stearate and metal salts of stearyl fumarate in a
pharmaceutical formulation containing a compound of formula (I) and
lactose to increase the stability of the compound of formula (I) to
chemical degradation (particularly in respect of metal ion
catalysed addition of water to the alkyne group and/or hydrolysis
of the pyrimidinone ring of the compound of formula (I)) and to a
method of increasing the stability of a pharmaceutical formulation
containing a compound of formula (I) and lactose to chemical
degradation (particularly in respect of metal ion catalysed
addition of water to the alkyne group and/or hydrolysis of the
pyrimidinone ring of the compound of formula (I)) which comprises
including in said formulation a stabilizing amount of a stabilizing
agent selected from metal salts of stearic acid such as magnesium
stearate and metal salts of stearyl fumarate. Suitably the compound
of formula (I) is in solid crystalline anhydrous form.
[0070] The preferred stabilizing agent is magnesium stearate.
Pharmaceutical Uses and Methods of Administration
[0071] There is provided according to one aspect of the present
invention use of pharmaceutical formulation of the invention as a
PI3 kinase inhibitor.
[0072] 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.
[0073] In one embodiment the pharmaceutical formulation of the
invention is suitable for sensitizing patients to treatment with a
corticosteroid.
[0074] 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).
[0075] Topical administration to the lung is achieved by use of an
inhalation device.
[0076] 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. 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. 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. 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.
[0077] 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.
[0078] 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.
[0079] 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.
[0080] 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.
[0081] 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.
[0082] 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.
[0083] 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.
[0084] 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.
[0085] 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.
[0086] 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.
[0087] 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.
[0088] 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.
[0089] 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.
[0090] 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.
[0091] 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.
[0092] 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.
[0093] 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.
[0094] 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.
[0095] The word "treatment" is intended to embrace prophylaxis as
well as therapeutic treatment.
[0096] Unless otherwise specified, % values as used herein are %
values by weight (wt. %).
[0097] Pharmaceutical formulations of the invention may have the
advantage that they have improved physical stability (e.g. as
measured by XRPD and/or IR analysis), improved chemical stability
(e.g. as measured by HPLC), improved physical compatibility with
lactose, improved chemical compatibility with lactose, improved
particle size distribution on administration (such as evidenced by
improved fine particle mass) or may have other favourable
properties as compared with similar formulations that do not
contain a stabilizing agent selected from metal salt of stearic
acid such as magnesium stearate and metal salts of stearyl
fumarate.
Abbreviations
[0098] aq aqueous COPD chronic obstructive pulmonary disease d
doublet DCM dichloromethane DMAP 4-dimethylaminopyridine DMSO
dimethyl sulfoxide DPI dry powder inhaler DSC differential scanning
calorimetry DVS dynamic vapour sorption EDC.HCl
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride
(ES.sup.+) electrospray ionization, positive mode EtOAc ethyl
acetate HPLC high performance liquid chromatography HPLC-MS high
performance liquid chromatography mass spectrometry hr hour(s) IR
infrared LPS lipopolysaccharide (M+H).sup.+ protonated molecular
ion MDI metered dose inhaler MeOH methanol MEK methylethylketone
MHz megahertz min minute(s)
mm Millimetre(s)
[0099] ms mass spectrometry mTOR mammalian target of rapamycin m/z
mass-to-charge ratio NH.sub.4OAc ammonium acetate NMR nuclear
magnetic resonance (spectroscopy) Pd(dppf)Cl.sub.2
1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) ppm
parts per million q quartet quin quintet RH relative humidity RRT
relative retention time R.sup.t retention time RT room temperature
s singlet t triplet TBDMSCI tert-butyldimethylsilyl chloride TGA
thermogravimetric analysis TNF.alpha. tumour necrosis factor alpha
XRPD X-ray powder diffraction
EXAMPLES
General Procedures
HPLC-MS
[0100] 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)
[0101] 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
[0102] .sup.1H NMR spectra (except those of Example 7) were
acquired on a Bruker Avance III spectrometer at 400 MHz using
residual undeuterated solvent as reference.
[0103] The .sup.1H NMR spectrum for Example 7 was acquired on a
Bruker Avance spectrometer at 600 MHz using residual undeuterated
solvent as reference.
X-Ray Powder Diffraction (XRPD)
[0104] 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:
scan mode: continuous scan range: 3 to 50.degree. 2.theta. step
size: 0.02.degree./step counting time: 30 sec/step spinner
revolution time: 1 sec radiation type: CuK.alpha.
Incident Beam Path
[0105] program. divergence slit: 15 mm Soller slit: 0.04 rad beam
mask: 15 mm anti scatter slit: 1.degree. beam knife: +
Diffracted Beam Path
[0106] long anti scatter shield: + Soller slit: 0.04 rad Ni filter:
+ detector: X'Celerator
[0107] Samples were prepared by spreading on a zero background
sample holder.
Infrared Spectrometry (IR)
[0108] Micro Attenuated Total Reflectance (microATR) was used and
the sample was analyzed using a suitable microATR accessory and the
following measurement conditions:
apparatus: Thermo Nexus 670 FTIR spectrometer number of scans: 32
resolution: 1 cm.sup.-1 wavelength range: 4000 to 400 cm.sup.-1
detector: DTGS with KBr windows beamsplitter: Ge on KBr micro ATR
accessory: Harrick Split Pea with Si crystal
Chemical Stability--High Performance Liquid Chromatography
(HPLC)
[0109] HPLC analysis was carried out using the following operating
conditions: [0110] 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).
[0111] Column temperature 35.degree. C. [0112] Sample temperature
10.degree. C. [0113] Flow rate 0.45 ml/min [0114] 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. [0115] Detection UV detection at 255
nm [0116] Mobile phase Preparation and composition: [0117] A 10 mM
ammonium acetate (0.771 g/l)+0.1%, v/v trifluoroacetic acid in
water [0118] B Acetonitrile [0119] Gradient Analytical run time is
41 minutes
TABLE-US-00001 [0119] Time (minutes) Solvent 0 35 36 41 42 48 % A
95 30 0 0 95 95 % B 5 70 100 100 5 5
[0120] With this HPLC method the degradant D019492 elutes at
RRT0.86.
[0121] Chemical Stability--Ultra High Pressure Liquid
Chromatography (UPLC)
[0122] UPLC analysis was carried out using the following operating
conditions: [0123] 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) [0124] Column
temperature 35.degree. C. [0125] Sample temperature 10.degree. C.
[0126] Flow rate 0.40 ml/min [0127] 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. [0128] Detection UV detection at 255 nm [0129]
Mobile phase Preparation and composition: [0130] A 10 mM ammonium
acetate (0.771 g/l)+0.1%, v/v trifluoroacetic acid in water [0131]
B Acetonitrile [0132] Gradient Analytical run time is 23
minutes
TABLE-US-00002 [0132] Time (minutes) Solvent 0 19 20 23 23.5 28 % A
95 30 0 0 95 95 % B 5 70 100 100 5 5
[0133] With this UPLC method the degradant D019492 elutes at
RRT=0.92-0.93 and the degradant D019493 elutes at
RRT=0.86-0.87.
Reagents and Suppliers
[0134] Lactohale200.RTM.: supplied by Frieslandfoods. 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
5-Bromo-3-(2-chlorobenzyl)-2-(chloromethyl)quinazolin-4(3H)-one
(2)
##STR00004##
[0136] 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.dbd.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.dbd.OH) (1.02 g, combined yield, 59%): m/z 274/276
(M+H).sup.+ (ES.sup.+). Both 1a and 1 b can be used without further
purification in the next step.
[0137] 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-d]pyrimidin-4-ami-
ne (6)
##STR00005##
[0139] 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.+).
[0140] 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
TBDMSCI (4.70 g, 30.99 mmol). After 16 hr, further aliquots of
imidazole (2.10 g, 30.99 mmol) and TBDMSCI (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
##STR00006##
[0142] 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
##STR00007##
[0144] 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)
##STR00008##
[0146] 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).
[0147] 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) (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
[0148] All reactions described within this example were carried out
under a flow of nitrogen gas. Compound of formula (I) 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.
[0149] The above method may optionally be adapted to facilitate
crystallization with seeding.
Example 3
XRPD Analysis of Compound of Formula (I) in Solid Crystalline
Anhydrous Form
[0150] XRPD analysis of the anhydrous form of compound of formula
(I) (Example 2) was undertaken using the method described in
General Procedures. The resulting diffraction pattern is shown in
FIG. 1. The XRPD pattern showed diffraction peaks without the
presence of a halo, thereby indicating that both materials are
crystalline. Characteristic peaks of the forms are given below in
Table 1:
TABLE-US-00003 TABLE 1 Characteristic XRPD peaks for the anhydrous
form of compound of formula (I) XRPD peaks (.+-.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
Example 4
HPLC Analysis of Compound of Formula (I) in Solid Crystalline
Anhydrous Form (Micronized) with Lactose
[0151] The chemical compatibility of the solid crystalline
anhydrous form of compound of formula (I) (micronized) in
combination with lactose was determined by HPLC analysis.
[0152] Micronized anhydrous form of compound of formula (I) was
prepared using a jet mill micronization device (1.5 bar)
(manufactured by Hosokawa Alpine) to produce the following particle
size distribution: D10=1.40 .mu.m; D50=2.77 .mu.m and D90=5.29
.mu.m (the particle size distribution was determined using laser
diffraction (Malvern Mastersizer instrument).
[0153] The test batch was taken from stock containing 3.519 mg
anhydrous form of compound of formula (I) (micronized) and 6006.64
mg Lactohale200.
[0154] 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 at 80.degree. C. (iii) 1, 2, 3 and 4 weeks at 40.degree.
C./75% RH.
[0155] The data shown in Table 2 indicate that significant
degradation was observed after storage for 1 week at 80.degree. C.
and degradation was also observed after storage for 4 weeks at
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.
[0156] The peak at RRT 0.86 has been attributed to the hydrated
derivative(s) D019328 shown above.
TABLE-US-00004 TABLE 2 stability data for the anhydrous form of the
compound of formula (I) (micronized) with lactose RRT* RRT* RRT*
RRT* RRT* Conditions 0.80 0.86 0.97 1.14 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 weeks 50.degree. C. 0.19 0.54 0.11 0.13 4 weeks 40.degree.
C./75% RH 0.18 0.20 0.11 0.14 *Area % by HPLC at the RRT indicated.
Compound of formula (I) has RRT = 1.0
Example 5
XRPD/IR Analysis of Compound of Formula (I) in Solid Crystalline
Anhydrous Form with Lactose and Magnesium Stearate
[0157] 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 (micronization of
compound of formula (I) as described in Example 4).
[0158] 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.
[0159] 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 conditions for 1 week storage
were: 40.degree. C./75% RH open; 1 week 50.degree. C. closed; and 1
week 80.degree. C. closed. The conditions for 4 week stability
storage were: 4 weeks 50.degree. C. closed; 4 weeks 40.degree.
C./75% RH open.
[0160] The IR spectrum acquired at time zero is shown in FIG. 2. IR
spectra were prepared for samples in the stability studies. 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.
[0161] The XRPD spectrum acquired at time zero is shown in FIG. 3.
XRPD spectra were prepared for samples in the stability studies.
The generated XRPD patterns of the 1 and 4 week stability samples
were similar to the diffraction pattern at time zero. It was
clearly evident 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.
[0162] 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 a result, it may be concluded that the anhydrous form is
physically compatible with lactose and magnesium stearate.
Example 6
HPLC Analysis of Compound of Formula (I) in Anhydrous Form with
Lactose and Magnesium Stearate
[0163] The chemical compatibility of the solid crystalline
anhydrous form (micronized) of compound of formula (I) in
combination with lactose and 1% magnesium stearate was determined
by HPLC analysis (micronization of compound of formula (I) as
described in Example 4).
[0164] 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.
[0165] The data shown in Table 3 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-00005 TABLE 3 stability data for the anhydrous form of the
compound of formula (I) (micronized) with lactose and magnesium
stearate RRT* RRT* RRT* RRT* Conditions 0.80 0.86 1.14 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./75% RH
0.20 0.11 0.11 0.13 2 weeks 50.degree. C. 0.20 0.08 0.11 0.14 2
weeks 40.degree. C./75% RH 0.21 0.11 0.11 0.13 3 weeks 50.degree.
C. 0.20 0.13 0.11 0.13 3 weeks 40.degree. C./75% RH 0.20 0.11 0.11
0.14 4 weeks 50.degree. C. 0.19 0.12 0.11 0.14 4 w 40.degree.
C./75% RH 0.20 0.10 0.10 0.13 *Area % by HPLC at the RRT indicated.
Compound of formula (I) has RRT = 1.0
Example 7
UPLC Analysis of Compound of Formula (I) in Anhydrous Form with
Lactose and Metal Salts of Stearic Acid
[0166] The chemical compatibility of the solid crystalline
anhydrous form (micronized) of compound of formula (I) 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).
[0167] Test samples were prepared as described in Table 4
below:
TABLE-US-00006 TABLE 4 test samples for UPLC analysis after
accelerated stability testing solid crystalline anhydrous form
Metal salt of (micronized) Lactohale 200 stearic acid of compound
(I) sample 1/ sample 1/ Sample sample 1/sample 2 sample 2 sample 2
Drug only 0.50 mg/ 0.47 mg Drug and 0.58 mg/ 749.84 mg/ lactose
0.47 mg 750.06 mg Drug, lactose, 0.46 mg/ 749.97 mg/ 7.40 mg/ Mg
stearate 0.51 mg 751.59 mg 7.55 mg Drug, lactose, 0.49 mg/ 751.08
mg/ 7.67 mg/ Ca stearate 0.45 mg 753.53 mg 7.80 mg Drug, lactose,
0.48 mg/ 750.20 mg/ 7.78 mg/ Na stearate 0.45 mg 750.42 mg 7.59
mg
[0168] 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.
[0169] Results are shown in Table 5 below:
TABLE-US-00007 TABLE 5 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
[0170] 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 6:
TABLE-US-00008 TABLE 6 .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)
[0171] The data shown in Table 5 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 8
Preparation of Pharmaceutical Formulations According to the
Invention
[0172] 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.
[0173] 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.
* * * * *