U.S. patent application number 17/085096 was filed with the patent office on 2021-06-17 for pseudopolymorphic forms of a hiv protease inhibitor.
The applicant listed for this patent is Janssen Sciences Ireland Unlimited Company. Invention is credited to Luc Donne Marie-Louise Janssens, Daniel Joseph Christiaan Thone, Hans Wim Pieter Vermeersch, Piet Tom Bert Paul Wigerinck.
Application Number | 20210179631 17/085096 |
Document ID | / |
Family ID | 1000005432626 |
Filed Date | 2021-06-17 |
United States Patent
Application |
20210179631 |
Kind Code |
A1 |
Vermeersch; Hans Wim Pieter ;
et al. |
June 17, 2021 |
Pseudopolymorphic Forms of a HIV Protease Inhibitor
Abstract
New pseudopolymorphic forms of (3R,3aS,6aR)-hexahydrofuro
[2,3-b] furan-3-yl (1S,2R)-3-[[(4-aminophenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate and processes for
producing them are disclosed.
Inventors: |
Vermeersch; Hans Wim Pieter;
(Gent, BE) ; Thone; Daniel Joseph Christiaan;
(Beerse, BE) ; Janssens; Luc Donne Marie-Louise;
(Malle, BE) ; Wigerinck; Piet Tom Bert Paul;
(Terhagen, BE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Janssen Sciences Ireland Unlimited Company |
Ringaskiddy |
|
IE |
|
|
Family ID: |
1000005432626 |
Appl. No.: |
17/085096 |
Filed: |
October 30, 2020 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
15980993 |
May 16, 2018 |
10858369 |
|
|
17085096 |
|
|
|
|
15600932 |
May 22, 2017 |
10000504 |
|
|
15980993 |
|
|
|
|
14817827 |
Aug 4, 2015 |
|
|
|
15600932 |
|
|
|
|
14183712 |
Feb 19, 2014 |
|
|
|
14817827 |
|
|
|
|
13939494 |
Jul 11, 2013 |
|
|
|
14183712 |
|
|
|
|
12536807 |
Aug 6, 2009 |
8518987 |
|
|
13939494 |
|
|
|
|
10514352 |
Nov 12, 2004 |
7700645 |
|
|
PCT/EP2003/050176 |
May 16, 2003 |
|
|
|
12536807 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 493/04
20130101 |
International
Class: |
C07D 493/04 20060101
C07D493/04 |
Foreign Application Data
Date |
Code |
Application Number |
May 16, 2002 |
EP |
02076929.5 |
Claims
1. A pseudopolymorph of (3R,3aS,6aR)-hexahydrofuro [2,3-b]
furan-3-yl (1S,2R)-3 -[[(4-aminophenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate, wherein the
pseudopolymorph is a C.sub.1-C.sub.4 alcohol solvate.
2. The pseudopolymorph according to claim 1, wherein the
C.sub.1-C.sub.4 alcohol is a straight chained saturated
alcohol.
3. The pseudopolymorph according to claim 1, in which the ratio of
the compound to the alcohol ranges between (5:1) and (1:5).
4. The pseudopolymorph according to claim 1, in which the ratio of
the compound to the alcohol ranges between (1:0.2) and (1:3).
5. The pseudopolymorph according to claim 1, in which the ratio of
the compound to the alcohol ranges between (1:1) and (1:2).
6. The pseudopolymorph according to claim 1, in which the ratio of
the compound to the alcohol is about 1:1.
7. The pseudopolymorph according to claim 1, additionally
comprising water molecules.
8. A pharmaceutical formulation comprising a therapeutically
effective amount of a pseudopolymorph of claim 1 and a
pharamceutically acceptable carrier or diluent.
9. A method of treating a retroviral infection in a human
comprising administering to the human a pseudopolymorph according
to claim 1.
Description
TECHNICAL FIELD
[0001] This invention relates to novel pseudopolymorphic forms of
(3R,3aS,6aR)-hexahydro-furo [2,3-b] furan-3-yl
(1S,2R)-3-[[(4-aminophenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate, a method for their
preparation as well as their use as a medicament.
BACKGROUND OF THE INVENTION
[0002] Virus-encoded proteases, which are essential for viral
replication, are required for the processing of viral protein
precursors. Interference with the processing of protein precursors
inhibits the formation of infectious virions. Accordingly,
inhibitors of viral proteases may be used to prevent or treat
chronic and acute viral infections. (3R,3aS,6aR)-hexahydrofuro
[2,3-b] furan-3-yl (1S,2R)-3-[[(4-aminophenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate has HIV protease
inhibitory activity and is particularly well suited for inhibiting
HIV-1 and HIV-2 viruses.
[0003] The structure of (3R,3aS,6aR)-hexahydrofuro [2,3-b]
furan-3-yl (1S,2R)-3-[[(4-amino-phenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate, is shown below:
##STR00001##
[0004] Compound of formula (X) and processes for its preparation
are disclosed in EP 715618, WO 99/67417, U.S. Pat. No. 6,248,775,
and in Bioorganic and Chemistry Letters, Vol. 8, pp. 687-690, 1998,
"Potent HIV protease inhibitors incorporating high-affinity
P.sub.2-igands and (R)-(hydroxyethylamino)sulfonamide isostere",
all of which are incorporated herein by reference.
[0005] Drugs utilized in the preparation of pharmaceutical
formulations for commercial use must meet certain standards,
including GMP (Good Manufacturing Practices) and ICH (International
Conference on Harmonization) guidelines. Such standards include
technical requirements that encompass a heterogeneous and wide
range of physical, chemical and pharmaceutical parameters. It is
this variety of parameters to consider, which make pharmaceutical
formulations a complex technical discipline.
[0006] For instance, and as example, a drug utilized for the
preparation of pharmaceutical formulations should meet an
acceptable purity. There are established guidelines that define the
limits and qualification of impurities in new drug substances
produced by chemical synthesis, i.e. actual and potential
impurities most likely to arise during the synthesis, purification,
and storage of the new drug substance. Guidelines are instituted
for the amount of allowed degradation products of the drug
substance, or reaction products of the drug substance with an
excipient and/or immediate container/closure system.
[0007] Stability is also a parameter considered in creating
pharmaceutical formulations. A good stability will ensure that the
desired chemical integrity of drug substances is maintained during
the shelf-life of the pharmaceutical formulation, which is the time
frame over which a product can be relied upon to retain its quality
characteristics when stored under expected or directed storage
conditions. During this period the drug may be administered with
little or no risk, as the presence of potentially dangerous
degradation products does not pose prejudicial consequences to the
health of the receiver, nor the lower content of the active
ingredient could cause under-medication.
[0008] Different factors, such as light radiation, temperature,
oxygen, humidity, pH sensitivity in solutions, may influence
stability and may determine shelf-life and storage conditions.
[0009] Bioavailability is also a parameter to consider in drug
delivery design of pharmaceutically acceptable formulations.
Bioavailability is concerned with the quantity and rate at which
the intact form of a particular drug appears in the systemic
circulation following administration of the drug. The
bioavailability exhibited by a drug is thus of relevance in
determining whether a therapeutically effective concentration is
achieved at the site(s) of action of the drug.
[0010] Physico-chemical factors and the pharmaco-technical
formulation can have repercussions in the bioavailability of the
drug. As such, several properties of the drug such as dissociation
constant, dissolution rate, solubility, polymorphic form, particle
size, are to be considered when improving the bioavailability.
[0011] It is also relevant to establish that the selected
pharmaceutical formulation is capable of manufacture, more
suitably, of large-scale manufacture.
[0012] In view of the various and many technical requirements, and
its influencing parameters, it is not obvious to foresee which
pharmaceutical formulations will be acceptable. As such, it was
unexpectedly found that certain modifications of the solid state of
compound of formula (X) positively influenced its applicability in
pharmaceutical formulations.
SUMMARY OF THE INVENTION
[0013] Present invention concerns pseudopolymorphic forms of
compound of formula (X) for the preparation of pharmaceutical
formulations. Such pseudopolymorphic forms contribute to
pharmaceutical formulations in improved stability and
bioavailability. They can be manufactured in sufficient high purity
to be acceptable for pharmaceutical use, more particularly in the
manufacture of a medicament for inhibiting HIV protease activity in
mammals.
[0014] In a first aspect, the present invention provides
pseudopolymorphs of (3R,3aS,6aR)-hexahydrofuro [2,3-b] furan-3-yl
(1S,2R)-3-[[(4-aminophenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate.
[0015] Pseudopolymorphs provided include alcohol solvates, more in
particular, C1-C4 alcohol solvates; hydrate solvates; alkane
solvates, more in particular, C1-C4 chloroalkane solvates; ketone
solvates, more in particular, C1-C5 ketone solvates; ether
solvates, more in particular, C1-C4 ether solvates; cycloether
solvates; ester solvates, more in particular, C1-C5 ester solvates;
and sulfonic solvates, more in particular, C1-4 sulfonic solvates,
of the compound of formula (X). Preferred pseudopolymorphs are
pharmaceutically acceptable solvates, such as hydrate and
ethanolate. Particular pseudopolymorphs are Form A (ethanolate),
Form B (hydrate), Form C (methanolate), Form D (acetonate), Form E
(dichloromethanate), Form F (ethylacetate solvate), Form G
(1-methoxy-2-propanolate), Form H (anisolate), Form I
(tetrahydrofuranate), Form J (isopropanolate) of compound of
formula (X). Another particular pseudopolymorph is Form K
(mesylate) of compound of formula (X).
[0016] In a second aspect, present invention relates to processes
for preparing pseudopolymorphs. Pseudopolymorphs of compound of
formula (X) are prepared by combining compound of formula (X) with
an organic solvent, water, or mixtures of water and water miscible
organic solvents, and applying any suitable technique to induce
crystallization, to obtain the desired pseudopolymorphs.
[0017] In a third aspect, the invention relates to the use of the
present pseudopolymorphs, in the manufacture of pharmaceutical
formulations for inhibiting HIV protease activity in mammals. In
relation to the therapeutic field, a preferred embodiment of this
invention relates to the use of pharmaceutically acceptable
pseudopolymorphic forms of compound of formula (X) for the
treatment of an HIV viral disease in a mammal in need thereof,
which method comprises administering to said mammal an effective
amount of a pharmaceutically acceptable pseudopolymorphic form of
compound of formula (X).
[0018] The following drawings provide additional information on the
characteristics of the pseudopolymorphs according to present
invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0019] FIG. 1, FIG. 2 and FIG. 3 are the powder X-ray diffraction
patterns of the Form A (1:1).
[0020] FIG. 4 depicts Form A (1:1) in three dimensions with the
atoms identified.
[0021] FIG. 5 is a comparison of the Raman spectra of Forms A, B,
D, E, F, H, (1:1) and the amorphous form at the carbonyl stretching
region of 1800-100 cm.sup.-1 and the region 3300-2000
cm.sup.-1.
[0022] FIG. 6 is a comparison of the expanded Raman spectra of
Forms A, B, D, E, F, H, (1:1) and the amorphous form at the
carbonyl stretching region of 600-0 cm.sup.-1.
[0023] FIG. 7 is a comparison of the expanded Raman spectra of
Forms A, B, D, E, F, H, (1:1) and the amorphous form at the
carbonyl stretching region of 1400-800 cm.sup.-1.
[0024] In FIGS. 5, 6, and 7, P1 corresponds to Form A, P18
corresponds to Form B, P19 corresponds to amorphous form, P25
corresponds to Form E, P27 corresponds to Form F, P50 corresponds
to Form D, P68 corresponds to Form H, P69 corresponds to Form C,
P72 corresponds to Form I, and P81 corresponds to Form G.
[0025] FIG. 8 is the Differential Scanning calorimetric (DSC)
thermograph of Form A (1:1).
[0026] FIG. 9 is the Infrared (IR) spectrum that reflects the
vibrational modes of the molecular structure of Form A as a
crystalline product.
[0027] FIG. 10 is the IR spectrum that reflects the vibrational
modes of the molecular structure of Form B as a crystalline
product.
[0028] FIG. 11: IR spectrum of forms A, B, and amorphous form, at
spectral range 4000 to 400 cm.sup.-1.
[0029] FIG. 12: IR spectrum of forms A, B, and amorphous form, at
spectral range 3750 to 2650 cm.sup.-1.
[0030] FIG. 13: IR spectrum of forms A, B, and amorphous form, at
spectral range 1760 to 1580 cm.sup.-1.
[0031] FIG. 14: IR spectrum of forms A, B, and amorphous form, at
spectral range 980 to 720 cm.sup.-1.
[0032] In FIGS. 11, 12, 13 and 14, curve A corresponds to Form A,
curve B corresponds to Form B, and curve C corresponds to the
amorphous form.
[0033] FIG. 15: DSC Thermograph curves of Form A (curve D), Form A
after Adsorption/Desorption (ADS/DES) (curve E), and Form A after
ADS/DES hydratation tests (curve F).
[0034] FIG. 16: Thermogravimetric (TG) curves of Form A (curve D),
Form A after ADS/DES (curve E), and Form A after ADS/DES
hydratation tests (curve F).
[0035] FIG. 17: TG curve of Form A at 25.degree. C. under dry
nitrogen atmosphere in function of time.
[0036] FIG. 18: ADS/DES curves of Form A.
[0037] FIG. 19: ADS/DES curves of the hydratation test of Form
A.
[0038] FIG. 20: ADS/DES curves of Form B.
[0039] FIG. 21: IR spectrum of Form K.
[0040] FIG. 22: Raman spectrum of Form K.
[0041] FIG. 23: DSC curve of Form K.
[0042] FIG. 24: TG curve of Form K.
[0043] FIG. 25: ADS/DES isotherm of Form K, batch 1.
[0044] FIG. 26: ADS/DES isotherm of Form K, batch 2.
DETAILED DESCRIPTION
[0045] The term "polymorphism" refers to the capacity of a chemical
structure to occur in different forms and is known to occur in many
organic compounds including drugs. As such, "polymorphic forms" or
"polymorphs" include drug substances that appear in amorphous form,
in crystalline form, in anhydrous form, at various degrees of
hydration or solvation, with entrapped solvent molecules, as well
as substances varying in crystal hardness, shape and size. The
different polymorphs vary in physical properties such as
solubility, dissolution, solid-state stability as well as
processing behaviour in terms of powder flow and compaction during
tabletting.
[0046] The term "amorphous form" is defined as a form in which a
three-dimensional long-range order does not exist. In the amorphous
form the position of the molecules relative to one another are
essentially random, i.e. without regular arrangement of the
molecules on a lattice structure.
[0047] The term "crystalline" is defined as a form in which the
position of the molecules relative to one another is organised
according to a three-dimensional lattice structure.
[0048] The term "anhydrous form" refers to a particular form
essentially free of water. "Hydration" refers to the process of
adding water molecules to a substance that occurs in a particular
form and "hydrates" are substances that are formed by adding water
molecules. "Solvating" refers to the process of incorporating
molecules of a solvent into a substance occurring in a crystalline
form. Therefore, the term "solvate" is defined as a crystal form
that contains either stoichiometric or non-stoichiometric amounts
of solvent. Since water is a solvent, solvates also include
hydrates. The term "pseudopolymorph" is applied to polymorphic
crystalline forms that have solvent molecules incorporated in their
lattice structures. The term pseudopolymorphism is used frequently
to designate solvates (Byrn, Pfeiffer, Stowell, (1999) Solid-state
Chemistry of Drugs, 2nd Ed., published by SSCI, Inc).
[0049] The present invention provides pseudopolymorphs of
(3R,3aS,6aR)-hexahydrofuro [2,3-b] furan-3-yl
(1S,2R)-3-[[(4-aminophenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate.
[0050] In one embodiment pseudopolymorphs are alcohol solvates,
more in particular, C.sub.1-C.sub.4 alcohol solvates; hydrate
solvates; alkane solvates, more in particular, C.sub.1-C.sub.4
chloroalkane solvates; ketone solvates, more in particular,
C.sub.1-C.sub.5 ketone solvates; ether solvates, more in particular
C.sub.1-C.sub.4 ether solvates; cycloether solvates; ester
solvates, more in particular C.sub.1-C.sub.5 ester solvates; or
sulfonic solvates, more in particular, C.sub.1-C.sub.4 sulfonic
solvates, of the compound of formula (X). The term "C.sub.1-C.sub.4
alcohol" defines straight and/or branched chained saturated and
unsaturated hydrocarbons having from 1 to 4 carbon atoms
substituted with at least a hydroxyl group, and optionally
substituted with an alkyloxy group, such as, for example, methanol,
ethanol, isopropanol, butanol, 1-methoxy-2-propanol and the like.
The term "C.sub.1-C.sub.4 chloroalkane" defines straight and/or
branched chained saturated and unsaturated hydrocarbons having from
1 to 4 carbon atoms substituted with at least one chloro atom, such
as, for example, dichloromethane. The term "C.sub.1-C.sub.5 ketone"
defines solvents of the general formula R'-C(.dbd.O)--R wherein R
and R' can be the same or different and are methyl or ethyl, such
as, acetone and the like. The term "C.sub.1-C.sub.4 ether" defines
solvents of the general formula R'-O-R wherein R and R' can be the
same or different and are a phenyl group, methyl or ethyl, such as,
anisole and the like. The term "cycloether" defines a 4- to
6-membered monocyclic hydrocarbons containing one or two oxygen
ring atoms, such as tetrahydrofuran and the like. The term
"C.sub.1-C.sub.5 ester" defines solvents of the general formula
R'--O--C(.dbd.O)--R wherein R and R' can be the same or different
and are methyl or ethyl, such as ethylacetate and the like. The
term "C.sub.1-C.sub.4 sulfonic solvent" defines solvents of the
general formula R--SO.sub.3H wherein R can be a straight or
branched chained saturated hydrocarbon having from 1 to 4 carbon
atoms, such as mesylate, ethanesulfonate, butanesulfonate,
2-methyl-1-propanesulfonate, and the like.
[0051] Pseudopolymorphs of the present invention, which are
pharmaceutically acceptable, for instance hydrates, alcohol
solvates, such as, ethanolate, are preferred forms.
[0052] Several pseudopolymorphs are exemplified in this application
and include Form A (ethanolate), Form B (hydrate), Form C
(methanolate), Form D (acetonate), Form E (dichloromethanate), Form
F (ethylacetate solvate), Form G (1-methoxy-2-propanolate), Form H
(anisolate), Form I (tetrahydrofuranate), Form J (isopropanolate),
or Form K (mesylate) of compound of formula (X).
[0053] Solvates can occur in different ratios of solvation. Solvent
content of the crystal may vary in different ratios depending on
the conditions applied. Solvate crystal forms of compound of
formula (X) may comprise up to 5 molecules of solvent per molecule
of compound of formula (X), appearing in different solvated states
including, amongst others, hemisolvate, monosolvate, disolvate,
trisolvate crystals, intermediate solvates crystals, and mixtures
thereof. Conveniently, the ratio of compound of formula (X) to the
solvent may range between (5:1) and (1:5). In particular, the ratio
may range from about 0.2 to about 3 molecules of solvent per 1
molecule of compound of formula (X), more in particular, the ratio
may range from about 1 to about 2 molecules of solvent per 1
molecule of compound of formula (X), preferably the ratio is 1
molecule of solvent per 1 molecule of compound of formula (X).
[0054] Solvates may also occur at different levels of hydration. As
such, solvate crystal forms of compound of formula (X) may in
addition comprise under certain circumstances, water molecules
partially or fully in the crystal structures. Consequently, the
term
[0055] "Form A" will be used herein to refer to the ethanolate
forms of compound of formula (X) comprising up to 5 molecules of
solvent per 1 molecule of compound of formula (X), intermediate
solvates crystals, and the mixtures thereof; and optionally
comprising additional water molecules, partially or fully in the
crystal structures. The same applies for Form B through Form K. In
case a particular "Form A" needs to be denoted, the ratio of
solvation will follow the "Form A", for instance, one molecule of
ethanol per one molecule of compound (X) is denoted as Form A
(1:1).
[0056] The X-ray powder diffraction is a technique to characterise
polymorphic forms including pseudopolymorphs of compound of formula
(X) and to differentiate solvate crystal forms from other crystal
and non-crystal forms of compound of formula (X). As such, X-ray
powder diffraction spectra were collected on a Phillips PW 1050/80
powder diffractometer, model Bragg-Brentano. Powders of Form A
(1:1), around 200 mg each sample, were packed in 0.5 mm glass
capillary tubes and were analysed according to a standard method in
the art. The X-ray generator was operated at 45 Kv and 32 mA, using
the copper K.alpha. line as the radiation source. There was no
rotation of the sample along the chi axis and data was collected
between 4 and 60.degree. 2-theta step size. Form A (1:1) has the
characteristic two-theta angle positions of peaks as shown in FIGS.
1, 2 and 3 at: 7,04.degree..+-.0,5.degree.,
9,24.degree..+-.0,5.degree., 9,96.degree..+-.0,5.degree.,
10,66.degree..+-.0,5.degree., 11,30.degree..+-.0,5.degree.,
12,82.degree..+-.0,5.degree., 13,80.degree..+-.0,5.degree.,
14,56.degree..+-.0,5.degree., 16,66.degree..+-.0,5.degree.,
17,30.degree..+-.0,5.degree., 18,28.degree..+-.0,5.degree.,
19,10.degree..+-.0,5.degree., 20,00.degree..+-.0,5.degree.,
20,50.degree..+-.0,5.degree., 21,22.degree..+-.0,5.degree.,
22,68.degree..+-.0,5.degree., 23,08.degree..+-.0,5.degree.,
23,66.degree..+-.0,5.degree., 25,08.degree..+-.0,5.degree.,
25,58.degree..+-.0,5.degree., 26,28.degree..+-.0,5.degree.,
27,18.degree..+-.0,5.degree., 28,22.degree..+-.0,5.degree.,
30,20.degree..+-.0,5.degree., 31,34.degree..+-.0,5.degree.,
32,68.degree..+-.0,5.degree., 33,82.degree..+-.0,5.degree.,
39,18.degree..+-.0,5.degree., 41,20.degree..+-.0,5.degree.,
42,06.degree..+-.0,5.degree., and 48,74.degree..+-.0,5.degree..
[0057] In another set of analytical experiments, X-ray single
diffraction was applied to Form A (1:1), which resulted in the
following crystal configuration, listed in the table below.
TABLE-US-00001 TABLE 1 Crystal Data Crystal shape Prism Crystal
dimensions 0.56 .times. 0.38 .times. 0.24 mm Crystal color
Colorless Space Group P 2.sub.1 2.sub.1 2.sub.1 orthorhombic
Temperature 293 K Cell constants a = 9.9882(6) .ANG. b = 16.1697(8)
.ANG. c = 19.0284(9) .ANG. alpha (.alpha.) = 90.degree. beta
(.beta.) = 90.degree. gamma (.gamma.) = 90.degree. Volume 3158.7(3)
.ANG..sup.3 Molecules/unit cell (Z) 4 Density, in Mg/m.sup.3 1.248
.mu. (linear absorption coefficient) 1.340 mm.sup.-1 F(000) 1272
Intensity Measurements Diffractometer Siemens P4 Radiation Cu
K.alpha. (.lamda. = 1.54184 .ANG.) Temperature ambient 2.theta.max
138.14.degree. Correction Empirical via .PSI.-scans Number of
Reflections Measured Total: 3912 Structure Solution and Refinement
Number of Observations 3467 [F.sup.2>2.sigma.(F.sup.2)] Residual
(R) 0.0446
[0058] The resulting three-dimensional structure of Form A (1:1) is
depicted in FIG. 4.
[0059] Table 2 shows the atomic coordinates (x 10.sup.4) and
equivalent isotropic displacement parameters
(.ANG..sup.2.times.10.sup.3) for Form A (1:1). Atoms are numbered
as exhibited in FIG. 4. The x, y and z fractional coordinates
indicate the position of atoms relative to the origin of the unit
cell. U(eq) is defined as one third of the trace of the
orthogonalized U.sub.ij tensor.
TABLE-US-00002 x y z U(eq) O1 7778(3) 2944(2) 9946(1) 70(1) C2
7171(4) 3513(2) 9487(2) 64(1) C3 6831(3) 3046(2) 8823(2) 52(1) C3A
7953(3) 2411(2) 8793(2) 55(1) C4 7527(4) 1533(2) 8708(2) 65(1) C5
7425(5) 1241(2) 9457(2) 70(1) O6 8501(3) 1642(2) 9809(1) 76(1) C6A
8582(4) 2416(2) 9534(2) 62(1) O7 5533(2) 2702(1) 8945(1) 51(1) O8
5168(2) 2636(1) 7768(1) 53(1) C9 4791(3) 2534(1) 8368(1) 42(1) N10
3590(2) 2256(1) 8562(1) 43(1) C11 2638(3) 1916(2) 8068(2) 44(1) C12
2223(3) 1071(2) 8310(2) 58(1) C13 3381(3) 501(2) 8387(2) 56(1) C14
3937(4) 340(2) 9038(2) 67(1) C15 4989(5) -200(2) 9111(3) 80(1) C16
5494(5) -581(3) 8530(3) 96(2) C17 4975(6) -413(3) 7881(3) 98(2) C18
3926(5) 126(2) 7810(2) 78(1) C19 1423(3) 2464(2) 7976(2) 45(1) O20
494(2) 2112(1) 7502(1) 61(1) C21 1829(3) 3307(2) 7740(2) 48(1) N22
699(3) 3880(1) 7721(1) 49(1) C23 521(4) 4312(2) 7048(2) 58(1) C24
-61(4) 3785(2) 6473(2) 67(1) C25 -1453(5) 3497(3) 6654(2) 86(2) C26
-47(7) 4247(3) 5779(2) 102(2) S27 510(1) 4414(1) 8440(1) 50(1) O28
572(3) 3860(1) 9015(1) 61(1) O29 -693(2) 4873(1) 8345(1) 65(1) C30
1854(3) 5080(2) 8509(2) 50(1) C31 1803(3) 5825(2) 8159(2) 54(1) C32
2871(4) 6341(2) 8195(2) 56(1) C33 4033(4) 6133(2) 8564(2) 55(1) C34
4063(4) 5385(2) 8909(2) 59(1) C35 2998(4) 4869(2) 8883(2) 56(1) N36
5076(3) 6667(2) 8596(2) 72(1) C37 1920(10) 2231(7) 5258(4) 232(6)
C38 1310(10) 1590(6) 5564(4) 191(5) O39 1768(4) 1393(2) 6249(2)
94(1)
[0060] Table 3 shows the anisotropic displacement parameters
(.ANG..sup.2.times.10.sup.3) for Form A (1:1). The anisotropic
displacement factor exponent takes the formula:
-2.pi..sup.2[h.sup.2a*.sup.2U.sub.11+ . . . +2 h k a*
b*U.sub.12]
TABLE-US-00003 U.sub.11 U.sub.22 U.sub.33 U.sub.23 U.sub.13
U.sub.12 O1 65 (2) 89 (2) 55 (1) -4 (1) -12 (1) -3 (1) C2 53 (2) 68
(2) 71 (2) -7 (2) -8 (2) -11 (2) C3 38 (2) 63 (2) 55 (2) 4 (1) -2
(1) -12 (1) C3A 37 (2) 78 (2) 49 (1) 9 (1) 1 (1) -3 (2) C4 61 (2)
74 (2) 61 (2) -4 (2) -6 (2) 10 (2) C5 72 (3) 67 (2) 71 (2) 8 (2)
-11 (2) -7 (2) O6 78 (2) 80 (2) 70 (1) 16 (1) -21 (1) -8 (2) C6A 47
(2) 80 (2) 59 (2) 5 (2) -6 (2) -7 (2) O7 34 (1) 69 (1) 50 (1) 0 (1)
-1 (1) -9 (1) O8 42 (1) 68 (1) 50 (1) 3 (1) 2 (1) -12 (1) C9 35 (2)
41 (1) 49 (1) 1 (1) -3 (1) 3 (1) N10 31 (1) 50 (1) 49 (1) -1 (1) 1
(1) -2 (1) C11 32 (2) 41 (1) 57 (1) -4 (1) 0 (1) -2 (1) C12 44 (2)
42 (1) 87 (2) 2 (1) 2 (2) -4 (1) C13 50 (2) 39 (1) 78 (2) 0 (1) 8
(2) 0 (1) C14 64 (2) 56 (2) 80 (2) 0 (2) 5 (2) 9 (2) C15 68 (3) 72
(2) 100 (3) 18 (2) 7 (2) 12 (2) C16 77 (3) 68 (2) 143 (4) 26 (3) 34
(3) 28 (2) C17 114 (4) 72 (2) 109 (3) -6 (2) 32 (3) 38 (3) C18 89
(3) 60 (2) 85 (2) -4 (2) 10 (2) 10 (2) C19 30 (2) 44 (1) 61 (1) -3
(1) -5 (1) -5 (1) O20 44 (1) 56 (1) 83 (1) -6 (1) -18 (1) -6 (1)
C21 36 (2) 42 (1) 64 (2) 2 (1) -4 (1) -1 (1) N22 42 (1) 47 (1) 57
(1) 1 (1) 0 (1) 3 (1) C23 59 (2) 50 (1) 64 (2) 7 (1) -8 (2) 1 (2)
C24 79 (3) 59 (2) 62 (2) 1 (1) -11 (2) 6 (2) C25 75 (3) 83 (2) 101
(3) 6 (2) -30 (3) -5 (2) C26 143 (5) 99 (3) 65 (2) 14 (2) -15 (3)
-6 (3) S27 44 (1) 47 (1) 61 (1) 2 (1) 2 (1) 1 (1) O28 64 (2) 58 (1)
61 (1) 9 (1) 3 (1) -7 (1) O29 46 (1) 58 (1) 92 (2) -4 (1) 6 (1) 10
(1) C30 50 (2) 46 (1) 54 (1) 2 (1) 1 (1) 1 (1) C31 50 (2) 48 (1) 64
(2) 6 (1) -4 (2) 6 (1) C32 59 (2) 45 (1) 65 (2) 4 (1) 2 (2) 1 (1)
C33 57 (2) 55 (2) 52 (1) -4 (1) 1 (1) -3 (1) C34 56 (2) 63 (2) 59
(2) 6 (1) -13 (2) -3 (2) C35 63 (2) 52 (1) 53 (1) 5 (1) -8 (2) -2
(2) N36 67 (2) 70 (2) 80 (2) 4 (2) -5 (2) -19 (2) C37 290 (10) 260
(10) 145 (7) 68 (7) 67 (8) 120 (10) C38 280 (10) 187 (7) 104 (4) 1
(5) -53 (6) -80 (10) O39 99 (2) 91 (2) 93 (2) 1 (2) -13 (2) -28
(2)
[0061] Raman spectroscopy has been widely used to elucidate
molecular structures, crystallinity and polymorphism. The
low-frequency Raman modes are particularly useful in distinguishing
different molecular packings in crystal. As such, Raman spectra
were recorded on a Bruker FT-Raman RFS100 spectrometer equipped
with a photomultiplier tube and optical multichannel detectors.
Samples placed in quartz capillary tubes were excited by an argon
ion laser. The laser power at the samples was adjusted to about 100
mW and the spectral resolution was about 2 cm.sup.-1. It was found
that Forms A, B, D, E, F, and H, (1:1) and the amorphous form have
the Raman spectra which appear in FIGS. 5, 6, and 7.
[0062] In addition, Forms A and B were characterized using a
.mu.ATR (Micro-Attenuated Total Reflectance) accessory (Harrick
Split-Pea with Si crystal). The infrared spectra were obtained with
a Nicolet Magna 560 FTIR spectrophotometer, a Ge on KBr
beamsplitter, and a DTGS with KBr windows detector. Spectra were
measured at 1 cm.sup.-1 resolution and 32 scans each, in a
wavelength range of from 4000 to 400 cm.sup.-1, and application of
baseline correction. The wavenumbers for Form A obtained are
exhibited in the following Table 4.
TABLE-US-00004 TABLE 4 Wavenumbers (cm.sup.-1) and relative
intensities of absorption bands (.sup.1) 3454w, 3429w, 3354w,
3301w, 3255w, 3089w, 3060w, 3041w, 3028w 2964w, 2905w, 2875w,
2856w, 2722vw, 2684vw, 2644vw, 2603vw, 2234vw 1704s, 1646w, 1595s,
1550m, 1503m, 1466w, 1453w, 1444w, 1413w 1373w, 1367w, 1340w,
1324m, 1314m, 1306m, 1290w, 1266m, 1244m, 1229m 1187w, 1146s,
1124m, 1104m, 1090m, 1076m, 1052m, 1042s, 1038m, 1024s 987s, 971m,
944m, 909w, 890w, 876w, 841m, 792w, 768s, 742s, 732w, 697m, 674s,
645w, 630m 598w, 593w, 574m, 564s, 553vs, 538m, 533m, 531m, 526m,
508m, 501m, 491m, 471m, 458w, 445w, 442w, 436w, 428w, 418w vs =
very strong, s = strong, m = medium, w = weak, vw = very weak, br =
broad
[0063] The IR spectrum in FIG. 9 reflects the vibrational modes of
the molecular structure as a crystalline product.
[0064] The wavenumbers obtained for Form B are exhibited in the
following Table 5.
TABLE-US-00005 TABLE 5 Wavenumbers (cm.sup.-1) and relative
intensities of absorption bands (.sup.1) 3614w, 3361m, 3291m,
3088w, 3061w, 3043w, 3028w 2967w, 2905w, 2872w, 2222vw 1703s,
1631w, 1595s, 1553m, 1502w, 1467w, 1453w, 1444w, 1436w 1388vw,
1374vw, 1366w, 1355vw, 1340w, 1308m, 1291w, 1267m, 1245m 1187w,
1148s, 1125m, 1105m, 1091m, 1077m, 1052m, 1044m, 1025s 990m, 972w,
944m, 912w, 891w, 876vw, 862w, 843w, 836w, 792w, 769m, 757w, 743m,
717w, 699m, 672m 598w, 591w, 585w, 576m, 566m, 553vs, 536m, 509w,
502m, 484w, 471w, 432vw, 425w, 418w (.sup.1) vs = very strong, s =
strong, m = medium, w = weak, vw = very weak, br = broad
[0065] The IR spectrum in FIG. 10 reflects the vibrational modes of
the molecular structure of Form B as a crystalline product.
[0066] Following the same analytical IR method, Form B and the
amorphous form were also characterised and compared with Form A, as
shown in FIGS. 11 to 14. IR spectra of the different physical forms
showed distinct spectral differences, most relevant are those in
Table 6:
TABLE-US-00006 TABLE 6 Wavenumbers (cm.sup.-1) and relative
intensities of absorption bands (.sup.1) Form A Form B Amorphous
form 3454m, 3429m, 3353m, 3615m, 3356m, 3291m, 3462m, 3362m, 3249m,
3255m, 3089w, 3060m, 3089m, 3061m, 3043w, 3062m, 3026m 3041w, 3028w
3027w 2963m, 2905m, 2869m, 2966m, 2905m, 2873m 2959m, 2871m 2856m
1704s, 1646m, 1596s, 1703s, 1630m, 1595s, 1704s, 1628s, 1596s,
1549s, 1503s 1552s, 1502m 1525s, 1502s 1306s, 1266s, 1244s 1308s,
1267s, 1245s 1312s, 1259s 1146s, 1104s, 1090s, 1148s, 1105s, 1090s,
1143s, 1090s, 1014s 1076s, 1052s, 1042s, 1077s, 1052s, 1044s,
1038s, 1023s 1024s 987s, 971s, 954s, 945s, 989s, 972s, 944s, 925m,
960s, 953s, 950s, 944s, 912m, 909m, 891s, 915m, 912s, 891s, 862s,
937s, 922s, 832s 876s, 841s, 827s 843s 792m, 768s, 742s, 697s,
792s, 769s, 744s, 699s, 750br, 702s, 672s 674s 672s (.sup.1) s =
strong, m = medium, w = weak, vw = very weak, br = broad
[0067] The physical Forms A, B, and amorphous form are identified
through spectral interpretation, focused on absorption bands
specific for each form. Unique and specific spectral differences
between forms are noticed in 3 spectral ranges: from 3750 to 2650
cm.sup.-1 (range 1), from 1760 to 1580 cm.sup.-1 (range 2) and from
980 to 720 cm.sup.-1 (range 3).
Range 1 (From 3750 to 2650 cm.sup.-1)
[0068] FIG. 11: Form A shows a double band with absorption maxima
at 3454 cm.sup.-1 and 3429 cm.sup.-1. Form B shows a single
absorption band at 3615 cm.sup.-1 and amorphous form shows a single
absorption band at 3362 cm.sup.-1.
Range 2 (From 1760 to 1580 cm.sup.-1)
[0069] FIG. 12: Form A shows a single absorption band at 1646
cm.sup.-1, Form B shows a single absorption band at 1630 cm.sup.-1
and amorphous form shows a single absorption band at 1628 cm.sup.-1
with a clearly higher intensity compared to the Form B band.
Additionally, amorphous form shows a less intense, broad band at
1704 cm.sup.-1 compared to both forms A and B bands at about 1704
cm.sup.-1.
[0070] Range 3 (From 980 to 720 cm.sup.-1)
[0071] FIG. 13: Form A shows a distinct set of 5 absorption bands
at 911, 890, 876, 862 and 841 cm.sup.-1. Form B shows a similar set
but the 876 cm.sup.-1 band is missing. Amorphous form shows a
single broad band at about 750 cm.sup.-1, both forms A and B show
two maxima at about 768 cm.sup.-1 and 743 cm.sup.-1.
[0072] Thermomicroscopy is another useful technique in the study of
solid-state kinetics. The kinetics of nucleation processes from
solutions or melts, including the analysis of the nucleation speed,
can be quantified. The simplest and most widely used method is the
melting point determination. As such, a Mettler model FP 82
controller with heating stage was used on a Leitz microscope. A few
particles of Form A were placed on a glass slide and observed while
heating at 10.degree. C. per minute. The melting range for Form A
(1:1) was found to be between 90.degree. and 110.degree. C.
[0073] On another means of characterization, the solubility of Form
A (1:1) was also a matter subject to study. Its solubility in
different solvents at approximate 23.degree. C. was determined to
be as follows:
TABLE-US-00007 TABLE 7 Approximate solubility for Form A (1:1), in
mg/ml Approximate solubility Solvent Form A (mg/ml) Acetone 106-211
Dichloromethane 105-209 1-Methoxy-2-propanol 160-213
Ethylmethylketone 102-204 Ethylacetate 71-107 Ethanol absolute
<3.4 Heptane <3.4 Water <3.5 Isopropylether <3.4
Methacyanate >200 Methanol <3.4 2-Propanol <3.4
Tetrahydrofurane 102-203 Toluene <3.5
[0074] Further solubility investigations were performed in function
of pH. As such, the aqueous solubilities of Form A (1:1) were
measured in solvents with different pH. An excess of the solute was
equilibrated with the solvent at 20.degree. C. for at least 24
hours. After removing the undissolved compound, the concentration
in solution was determined using UV spectrometry.
TABLE-US-00008 TABLE 8 Solubility for Form A (1:1) in function of
pH Solvent Solubility (mg/100 ml solution) Water 16 (pH 5.9) Buffer
pH 2 (citrate/HCl) 18 (pH 2.0) Buffer pH 3 (citrate/HCl) 10 (pH
3.0) Buffer pH 4 (citrate/HCl) 9 (pH 4.0) 0.01N HCl 18 (pH 2.1)
0.1N HCl 83 (pH 1.1) 1.0N HCl 620 (pH 0.2)
[0075] Solubility of Form A (1:1) in function of HP.beta.CD
(hydroxypropyl-.beta.-cyclodextrin) was measured. An excess of
product was equilibrated with the solvent during 2 days at
20.degree. C. After removing the undissolved compound, the
concentration in solution was determined using UV spectrometry.
TABLE-US-00009 TABLE 9 Solubility for Form A (1:1) in function of
HP.beta.CD solvent Solubility in mg/ml solution Water 0.16 (pH =
5.9) 5% HP.beta.CD in water 2.4 (pH = 5.8) 10% HP.beta.CD in water
6.5 (pH = 6.0) 20% HP.beta.CD in water 17 (pH = 6.0) 40% HP.beta.CD
in water 40 (pH = 5.9)
[0076] In a second aspect, the present invention relates to
processes for preparing pseudopolymorphs. Pseudopolymorphs of
compound of formula (X) are prepared by combining compound of
formula (X) with an organic solvent, or water, or mixtures of water
and water miscible organic solvents, applying any suitable
technique to induce crystallization, and isolating the desired
pseudopolymorphs.
[0077] By techniques for inducing crystallization are to be
understood those processes for the production of crystals, which
include amongst others, dissolving or dispersing compound of
formula (X) in a solvent medium, bringing the solution or
dispersion of compound of formula (X) and the solvent(s) to a
desired concentration, bringing the said solution or dispersion to
a desired temperature, effecting any suitable pressure, removing
and/or separating any undesired material or impurities, drying the
formed crystals to obtain the pseudopolymorphs in a solid state, if
such state is desired.
[0078] Bringing the solution or dispersion of compound of formula
(X) and solvents to a desired concentration does not necessarily
imply an increase in the concentration of compound of formula (X).
In certain cases, a decrease or no change in concentration could be
preferable. By bringing the said solution or dispersion to a
desired temperature, one will understand the acts of heating,
cooling or leaving at ambient temperature.
[0079] The techniques used for obtaining a desired concentration
are those common in the art, for instance, evaporation by
atmospheric distillation, vacuum distillation, fractioned
distillation, azeotropic distillation, film evaporation, other
techniques well known in the art and combinations thereof. An
optional process for obtaining a desired concentration could as
well involve the saturation of the solution of compound of formula
(X) and solvent, for example, by adding a sufficient volume of a
non-solvent to the solution to reach the saturation point. Other
suitable techniques for saturating the solution include, by way of
example, the introduction of additional compound of formula (X) to
the solution and/or evaporation of a portion of the solvent from
the solution. As referred to herein, saturated solution encompasses
solutions at their saturation points or exceeding their saturation
points, i.e. supersaturated.
[0080] Removing and/or separating any undesired material or
impurities may be performed by purification, filtering, washing,
precipitation or similar techniques. Separation, for example, can
be conducted by known solid-liquid separation techniques. Filtering
procedures known to those skilled in the art can as well be used in
the present process. The filtrations can be performed, amongst
other methods, by centrifugation, or using Buchner style filter,
Rosenmund filter or plates, or frame press. Preferably, in-line
filtration or safety filtration may be advantageously intercalated
in the processes disclosed above, in order to increase the purity
of the resulting pseudopolymorphic form. Additionally, filtering
agents such as silica gel, Arbocel.RTM., dicalite diatomite, or the
like, may also be employed to separate impurities from the crystals
of interest.
[0081] Crystals obtained may be also dried, and such drying process
may optionally be used in the different crystallization passages,
if more than one crystallization passage is applied. Drying
procedures include all techniques known to those skilled in the
art, such as heating, applying vacuum, circulating air or gas,
adding a desiccant, freeze-drying, spray-drying, evaporating, or
the like, or any combination thereof.
[0082] Processes for crystallization of pseudopolymorphs of
compound of formula (X) embrace multiple combinations of techniques
and variations thereof. As such, and by way of example,
crystallization of pseudopolymorphs of compound of formula (X) may
be executed by dissolving or dispersing compound of formula (X) at
a suitable temperature in the solvent whereby portion of the said
solvent evaporates increasing the concentration of the compound of
formula (X) in the said solution or dispersion, cooling the said
mixture, and optionally washing and/or filtering and drying
resulting solvate crystals of compound of formula (X). Optionally,
pseudopolymorphs of compound of formula (X) may be prepared by
dissolving or dispersing compound of formula (X) in a solvent
medium, cooling said solution or dispersion and subsequently
filtering and drying the obtained pseudopolymorph. Another example
of preparation of solvates of compound of formula (X) could be by
saturating compound of formula (X) in the solvent medium, and
optionally filtering, washing and drying obtained crystals.
[0083] Crystal formation may as well involve more than one
crystallization process. In certain cases, one, two or more extra
crystallization steps may be advantageously performed for different
reasons, such as, to increase the quality of the resulting solvate.
For instance, pseudopolymorphs of the present invention could also
be prepared by adding a solvent to an initial starting base
material of compound of formula (X), stirring the solution at a
fixed temperature until the substances would be fully solved,
concentrating the solution by vacuum distillation, and cooling. A
first crystallization would take place and the formed crystals
would be newly washed with a solvent, and followed by dissolution
of compound of formula (X) with the solvent to form the desired
pseudopolymorph. Recrystallization of the reaction mixture would
occur, followed by a cooling step from reflux. The formed
pseudopolymorph would optionally be filtered and allowed to
dry.
[0084] By dissolving or dispersing compound of formula (X) in the
organic solvent, water or a mixture of water and water miscible
organic solvents, one may obtain different degrees of dispersion,
such as suspensions, emulsions, slurries or mixtures; or preferably
obtain homogeneous one-phase solutions.
[0085] Optionally, the solvent medium may contain additives, for
example one or more dispersing agents, surfactants or other
additives, or mixtures thereof of the type normally used in the
preparation of crystalline suspensions and which are well
documented in the literature. The additives may be advantageously
used in modifying the shape of crystal by increasing the leniency
and decreasing the surface area.
[0086] The solvent medium containing the solution may optionally be
stirred for a certain period of time, or vigorously agitated using,
for example, a high shear mixer or homogeniser or a combination of
these, to generate the desired droplet size for the organic
compound.
[0087] Examples of organic solvents useful for the present
invention include C.sub.1-C.sub.4 alcohols such as methanol,
ethanol, isopropanol, butanol, 1-methoxy-2-propanol, and the like;
C.sub.1-C.sub.4 chloroalkanes such as dichloromethane;
C.sub.1-C.sub.4 ketones such as acetone; C.sub.1-C.sub.4 ethers
such as anisole, and the like; cycloethers such as tetrahydrofuran;
C.sub.1-C.sub.4 esters such as ethylacetate; C.sub.1-C.sub.4
sulfonates such as mesylate, ethanesulfonate, butanesulfonate,
2-methyl-1-propanesulfonate; and the like.
[0088] Examples of mixtures of water and water miscible organic
solvents include, mixtures of water with all organic solvents
listed above provided they are miscible in water, e.g.
ethanol/water, for instance in a 50/50 ratio.
[0089] Preferred solvents are those pharmaceutically acceptable
solvents. However, pharmaceutically non-acceptable solvents may
also find their use in the preparation of pharmaceutically
acceptable pseudopolymorphs.
[0090] In a preferred method, the solvent is a pharmaceutically
acceptable solvent since it results in a pharmaceutically
acceptable pseudopolymorph. In a more preferred method, the solvent
is ethanol.
[0091] In a particular embodiment, pharmaceutically acceptable
pseudopolymorphs of compound of formula (X) can be prepared
starting from pseudopolymorphic forms of compound of formula (X),
which may not be necessarily pharmaceutically acceptable. For
instance, Form A may be prepared starting from Form J.
Pseudopolymorphs may also be prepared starting from the amorphous
form.
[0092] In the mixtures of water and water miscible organic
solvents, the amount of water can vary from about 5% by volume to
about 95% by volume, preferably from about 25% to about 75% by
volume, more preferably from about 40% to about 60% by volume.
[0093] It should also be noted that the quality of selected organic
solvent (absolute, denaturated, or other) also influences the
resulting quality of the pseudopolymorph.
[0094] Control of precipitation temperature and seeding may be
additionally used to improve the reproducibility of the
crystallization process, the particle size distribution and form of
the product. As such, the crystallization can be effected without
seeding with crystals of the compound of the formula (X) or
preferably in the presence of crystals of the compound of the
formula (X), which are introduced into the solution by seeding.
Seeding can also be effected several times at various temperatures.
The amount of the seed material depends on the amount of the
solution and can readily be determined by a person skilled in the
art.
[0095] The time for crystallization in each crystallization step
will depend on the conditions applied, the techniques employed
and/or solvents used.
[0096] Breaking up the large particles or aggregates of particles
after crystal conversion may additionally be performed in order to
obtain a desired and homogeneous particle size. Accordingly, the
solvate crystal forms of compound of formula (X) are optionally
milled after undergoing conversion. Milling or grinding refers to
physically breaking up the large particles or aggregates of
particles using methods and apparatus well known in the art for
particle size reduction of powders. Resulting particle sizes may
range from millimeters to nanometers, yielding i.e. nanocrystals,
microcrystals.
[0097] The yield of the preparation process of the pseudopolymorphs
of compound of formula (X) may be 10% or more, a more preferred
yield would vary from 40% to 100%.
[0098] Interestingly, the yield varies between 70% and 100%.
[0099] Suitably, pseudopolymorphs of the present invention have a
purity greater than 90 percent. More suitably, the present
pseudopolymorphs have a purity greater than 95 percent. Even more
suitably, the present pseudopolymorphs have a purity greater than
99 percent.
[0100] In a third aspect, the present invention relates to a
pharmaceutical formulation comprising a therapeutically effective
amount of a pseudopolymorph of compound of formula (X), and a
pharmaceutically acceptable carrier or diluent thereof.
[0101] In one embodiment, present invention relates to the use of
pharmaceutically acceptable pseudopolymorphic forms of compound of
formula (X), preferably Form A, in the manufacture of a medicament
for treating diseases caused by retroviruses, such as HIV
infections, for example, Acquired Immune Deficiency Syndrome (AIDS)
and AIDS-related complex (ARC).
[0102] In another embodiment, present invention provides a method
for the treatment of a retroviral infection, for example an HIV
infection, in a mammal such as a human, which comprises
administering to the mammal in need thereof an effective
antiretroviral amount of a pharmaceutically acceptable
pseudopolymorphic form of compound of formula (X), preferably Form
A.
[0103] Present invention also relates to a method in which the
treatment of a HIV viral infection comprises the reduction of HIV
load. Present invention also relates to a method in which the
treatment of said HIV viral infection comprises the increase of
CD4+ cell count. Present invention relates as well to a method in
which the treatment of said HIV viral infection comprises
inhibiting HIV protease activity in a mammal.
[0104] Pharmaceutically acceptable pseudopolymorphic forms of
compound of formula (X), preferably Form A, also referred to herein
as the active pharmaceutical ingredients, may be administered by
any route appropriate to the condition to be treated, preferably
orally. It will be appreciated however, that the preferred route
may vary with, for example, the condition of the recipient.
[0105] For each of the above-indicated utilities and indications
the amount required of the active ingredient will depend upon a
number of factors including the severity of the condition to be
treated and the identity of the recipient and will ultimately be at
the discretion of the attendant physician or veterinarian. The
desired dose preferably may be presented as one, two, three or four
or more subdoses administered at appropriate intervals throughout
the day.
[0106] For an oral administration form, pseudopolymorphs of the
present invention are mixed with suitable additives, such as
excipients, stabilizers or inert diluents, and brought by means of
the customary methods into the suitable administration forms, such
as tablets, coated tablets, hard capsules, aqueous, alcoholic, or
oily solutions. Examples of suitable inert carriers are gum arabic,
magnesia, magnesium carbonate, potassium phosphate, lactose,
glucose, or starch, in particular, corn starch. In this case the
preparation can be carried out both as dry and as moist granules.
Suitable oily excipients or solvents are vegetable or animal oils,
such as sunflower oil or cod liver oil. Suitable solvents for
aqueous or alcoholic solutions are water, ethanol, sugar solutions,
or mixtures thereof. Polyethylene glycols and polypropylene glycols
are also useful as further auxiliaries for other administration
forms.
[0107] For subcutaneous or intravenous administration, the
pseudopolymorphs of compound of formula (X), if desired with the
substances customary therefor such as solubilizers, emulsifiers or
further auxiliaries, are brought into solution, suspension, or
emulsion. The pseudopolymorphs of compound of formula (X) can also
be lyophilized and the lyophilizates obtained used, for example,
for the production of injection or infusion preparations. Suitable
solvents are, for example, water, physiological saline solution or
alcohols, e.g. ethanol, propanol, glycerol, in addition also sugar
solutions such as glucose or mannitol solutions, or alternatively
mixtures of the various solvents mentioned.
[0108] Suitable pharmaceutical formulations for administration in
the form of aerosols or sprays are, for example, solutions,
suspensions or emulsions of the pseudopolymorphs of compound of
formula (X) in a pharmaceutically acceptable solvent, such as
ethanol or water, or a mixture of such solvents. If required, the
formulation can also additionally contain other pharmaceutical
auxiliaries such as surfactants, emulsifiers and stabilizers as
well as a propellant. Such a preparation customarily contains the
active compound in a concentration from approximately 0.1 to 50%,
in particular from approximately 0.3 to 3% by weight.
[0109] Pseudopolymorphs of the present invention may also be
presented in a formulation comprising micrometer-, nanometer- or
picometer-size particles of the pseudopolymorph of compound of
formula (X), which formulation may contain other pharmaceutical
agents and may optionally be converted to solid form.
[0110] It may be convenient to formulate the present
pseudopolymorphs in the form of nanoparticles which have a surface
modifier adsorbed on the surface thereof in an amount sufficient to
maintain an effective average particle size of less than 1000 nm.
Useful surface modifiers are believed to include those that
physically adhere to the surface of the antiretroviral agent but do
not chemically bind to the antiretroviral agent.
[0111] It may be further convenient to store the pseudopolymorphs
of compound of formula (X) in packaging materials which are
protective to mechanical, environmental, biological or chemical
hazards, or degradation. Conditioning drug substances can be
achieved by employing packaging materials impermeable to moisture,
such as sealed vapour lock bags. Conditioning drug products, such
as tablets, capsules, can be achieved by employing for instance,
aluminium blisters.
[0112] It should be understood that in addition to the ingredients
particularly mentioned above, formulations of this invention
includes other agents conventional in the art having regard to the
type of formulation in question, for example those suitable for
oral administration may include flavouring agents or taste masking
agents.
[0113] The following examples are intended for illustration only
and are not intended to limit the scope of the invention in any
way.
EXAMPLE 1
[0114] The industrial scale synthesis of Form A (1:1) was performed
using the following steps. First a solution was prepared with
isopropanol and (3R,3aS,6aR)-hexahydrofuro [2,3-b] furan-3-yl
(1S,2R)-3-[[(4-aminophenyl) sulfonyl] (isobutyl)
amino]-1-benzyl-2-hydroxypropylcarbamate. The solution was
concentrated by vacuum distillation at 70.degree. C. and
200-500mbar pressure and cooled from a T>35.degree. to a T
between 15.degree. and 20.degree. C. for about 10 hours. The
crystals formed were newly washed with 13 liters isopropanol and
filtered. A subsequent recrystallization from ethanol/water (90
liters/90 liters) was performed. This was followed by a new
dissolution step, but with 60 liters ethanol instead.
Recrystallization of the reaction mixture from ethanol occurred,
followed by a cooling step from reflux to -15.degree. C.
approximately and during 10 hours. The ethanolate formed was
filtered and let to dry at about 50.degree. C. and about 7 mbar.
The yield of this process was at least 75%.
Example 2
[0115] In another example a mixture of Form D and Form B were
prepared. Acetone was used as a solvent during the crystallisation
process to form Form D. The crystallisation process then comprised
the step of stirring the initial starting compound (10g) in 70 ml
acetone. The solution was subsequently refluxed until the compound
was completely solved. 40 ml of water were added and the solution
was subsequently cooled slowly until room temperature and stirred
overnight. Formed crystals were filtered and dried in the vacuum
oven at 50.degree. C. 7.6 g of product resulted from the
crystallization, being the yield of this process of about 75%.
EXAMPLE 3
[0116] In another example Form J crystals were prepared.
Isopropanol was used as a solvent during the crystallisation
process to form Form J. The crystallisation process then comprised
the step of solving the initial starting material in the hot
solvent. The solution was subsequently cooled until room
temperature. Formed crystals were filtered and dried in the vacuum
oven at 50.degree. C. The crystals contained about 50 mol %
isopropanol.
EXAMPLE 4
[0117] In this example, the mass losses for different
pseudopolymorphs in thermogravimetric (TG) experiments were
calculated. Thermogravimetry is a technique that measures the
change in mass of a sample as it is heated, cooled or held at
constant temperature. Approximately 2 to 5 mg of sample were placed
on a pan and inserted into the TG furnace, model Netzsch
Thermo-Microbalance TG 209 coupled to a Bruker FTIR Spectrometer
vector 22. The samples were heated in a nitrogen atmosphere at a
rate of 10.degree. C./min, up to a final temperature of 250.degree.
C. The detection limit of residual solvents was in the order of
0.1% for distinct stepwise solvent loss over a narrow temperature
range (few degrees Celsius).
[0118] The following TG data were obtained:
[0119] Form A: a weight loss of 4.2% was observed in the
temperature range of 25-138.degree. C. (ethanol+little water) and
of 6.9% (ethanol+CO.sub.2) in the temperature range of
25-200.degree. C. Ethanol loss rate was maximal at 120.degree. C.
CO.sub.2 loss was due to chemical degradation and was visible at
around 190.degree. C.
[0120] Form B: a weight loss of 3.4% was observed in the
temperature range 25-78.degree. C. (water) and of 5.1% in the
temperature range 25-110.degree. C. (ethanol+water for
T>78.degree. C.). From 110-200.degree. C. further 1.1% weight
was lost (ethanol).
[0121] Form C: a weight loss of 2.1% was observed in the
temperature range 25-83.degree. C. (water+methanol) and of 4.2% in
the temperature range 25-105.degree. C. (methanol for
T>83.degree. C., distinct step). From 105-200.degree. C. further
2.1% weight was lost (methanol). No ethanol was observed in the gas
phase.
[0122] Form D: a weight loss of 0.1% was observed in the
temperature range 25-50.degree. C., of 4.2% in the temperature
range 25-108.degree. C. (acetone+ethanol for T>50.degree. C.),
of 8.2% in the temperature range 25-157.degree. C. (acetone+ethanol
for T>108.degree. C.) and of 10.5% in the temperature range
25-240.degree. C. (acetone+ethanol for T>157.degree. C.).
[0123] Form E: a weight loss of 0.2% was observed in the
temperature range 25-75.degree. C. (water), of 1.8% in the
temperature range 25-108.degree. C. (dichloromethane+ethanol for
T>75.degree. C.), of 6.8% in the temperature range
25-157.degree. C. (dichloromethane+ethanol for T>108.degree. C.)
and of 8.8% in the temperature range 25-240.degree. C.
(dichloromethane+ethanol for T>157.degree. C.).
[0124] Form F: a weight loss of 0.1% was observed in the
temperature range 25-50.degree. C. (probably water), of 1.7% in the
temperature range 25-108.degree. C. (ethylacetate+ethanol for
T>50.degree. C.), of 6.6% in the temperature range
25-157.degree. C. (ethylacetate+ethanol for T>108.degree. C.)
and of 9% in the temperature range 25-240.degree. C.
(ethylacetate+ethanol for T>157.degree. C.).
[0125] Form G: a weight loss of 0.0% was observed in the
temperature range 25-50.degree. C., of 3.7% in the temperature
range 25-108.degree. C. (1-methoxy-2-propanol+ethanol for
T>50.degree. C., distinct step), of 8% in the temperature range
25-157.degree. C. (1-methoxy-2-propanol+ethanol for
T>108.degree. C.) and of 12.5% in the temperature range
25-240.degree. C. (1-methoxy-2-propanol+ethanol for
T>157.degree. C.).
[0126] Form H: a weight loss of 0.8% was observed in the
temperature range 25-100.degree. C. (anisole+little ethanol) and of
8.8% in the temperature range 25-200.degree. C. (anisole+ethanol
for T>100.degree. C.).
[0127] Form I: a weight loss of 0.3% was observed in the
temperature range 25-89.degree. C. (water) and of 11.0% in the
temperature range 25-200.degree. C. (tetrahydrofurane for
T>89.degree. C.). No ethanol was observed in the gas phase.
[0128] Table 10 shows approximate expected mass losses for
different Forms in thermogravimetric (TG) experiments.
[0129] Mass loss in % (M+x.LM=100%)
TABLE-US-00010 Pseudo- BP polymorph [.degree. C.] Hemisolvate
Monosolvate Disolvate Trisolvate Form D 56 5.0 9.6 17.5 24.1 Form H
152 9.0 16.5 28.3 37.2 Form E 40 7.2 13.4 23.7 31.8 Form G 119 7.6
14.1 24.8 33.1 Form F 76 7.4 13.9 24.3 32.6 Form A 78 4.0 7.8 14.4
20.2 Form B 100 1.6 3.2 6.2 9.0 Form C 65 2.8 5.5 10.5 14.9 Form I
66 6.2 11.6 20.8 28.3
[0130] In another set of thermogravimetric methods, Form A, Form A
after Adsorption/Desorption, and Form A after Adsorption/Desorption
hydratation tests, were all transferred into an aluminum sample
pan. The TG curve was recorded on a TA Instrument Hi-Res TGA 2950
thermogravimeter at the following conditions: [0131] initial
temperature: room temperature [0132] heating rate: 20.degree.
C./min [0133] resolution factor: 4 [0134] final condition:
300.degree. C. or <80[(w/w) %]
[0135] The TG curves of the samples are collected in FIG. 16.
[0136] Table 11 shows mass losses for the forms tested:
TABLE-US-00011 TG (% weight change) Form A Up to 80.degree. C.
>80.degree. C. Form A 0.3 7.1 Form A after ADS/DES 2.9 4.0 Form
A after A/D hydratation test 5.4 0.5
[0137] The loss of weight at temperatures up to 80.degree. C. is
mainly due to the evaporation of solvent (water) present in the
sample. The loss of weight at temperatures above 80.degree. C. is
mainly due to the evaporation of solvent (ethanolate) present in
the sample.
[0138] A TG curve of form A at 25.degree. C. under dry nitrogen
atmosphere in function of time is collected in FIG. 17. The loss of
weight at 25.degree. C. after 10 hours was around 0.6%. This was
due to the evaporation of solvent.
EXAMPLE 5
[0139] In another example, measurements of differential scanning
calorimetry (DSC) were also performed. For such purpose, a Perkin
Elmer DSC 204 thermal analysis system was used. From 2 to 5 mg
sample of Form A were accurately weighed into a DSC pan. The
experiments were performed in an open pan. The sample was
equilibrated to approximately 30.degree. C. and then heated at a
rate of 10.degree. C. per minute, up to a final temperature of
200.degree. C. The DSC data was obtained following a standard
method in the art. The Form A was characterized by differential
scanning calorimetry (DSC) in which it showed a sharp endotherm in
the range 80-119.degree. C., showing a peak at about 105.6.degree.
C., with a delta H=-98.33 J/g onset. Accordingly, the ethanol
solvate crystal Form A of compound of formula (X) (1:1) showed the
thermograph pattern, which appears in FIG. 8.
[0140] In another set of DSC measurements, Form A, Form A after
Adsorption/Desorption, and Form A after Adsorption/Desorption
hydratation tests were examined. About 3 mg of the samples were
transferred into a 30 .mu.l perforated aluminum Perkin Elmer sample
pan. The sample pan was closed with the appropriate cover and the
DSC curve recorded on a Perkin Elmer Pyris DSC, at the following
conditions: [0141] initial temperature: 25.degree. C. [0142]
heating rate: 10.degree. C./min [0143] final temperature:
150.degree. C. [0144] nitrogen flow: 30 ml/min
[0145] Form A showed an endothermic signal at about 104.6.degree.
C. and a heat of fusion of 95.8 J/g caused by the evaporation of
the ethanolate and the melting of the product. Form A after ADS/DES
showed a broad endothermic signal due to a mixture of ethanolate
Form A and hydrated Form B. Form A after ADS/DES hydratation test
showed an endothermic signal at about 73.5.degree. C. and a heat of
fusion of 126 J/g caused by the evaporation of water and the
melting of the product. Thermograph curves are depicted in FIG.
15.
EXAMPLE 6
[0146] In another example stability studies of the Form A in three
different conditions were tested out. They included conditions of
25.degree. C. and 60% RH, 40.degree. C. and 75% RH, and 50.degree.
C. These studies revealed that at 25.degree. C. and 60% RH
long-term stability, the amount of ethanol and water is stable.
[0147] Table 12 shows the Stability study for Form A. Long term
stability at 25.degree. C./60% RH (Relative Humidity), with brown
glass bottles as sample container.
TABLE-US-00012 Test Release data 0 month 1 month 3 month Residual
solvent: 7.5 7.6 7.6 7.1 % (w/w) ethanol % (w/w) Water 0.10 0.27
0.26 0.55
EXAMPLE 7
[0148] Adsorption-Desorption tests
[0149] About 23 mg of Form A were transferred into a VTI vapor
sorption analyzer model SGA100 and the weight change with respect
to the atmospheric humidity was recorded at the following
conditions: [0150] drying temperature: 40.degree. C. [0151]
equilibrium: .ltoreq.0.05% in 5min. or 60 min. [0152] data
interval: 0.05% or 2 min. [0153] temperature: 25.degree. C. [0154]
first cycle RH (%) adsorption: 5, 10, 20, 30, 40, 50, 60, 70, 80,
90, 95 [0155] RH (%) desorption: 95, 90, 80, 70, 60, 50, 40, 30,
20, 10, 5 [0156] second cycle RH (%) adsorption: 5, 10, 20, 30, 40,
50, 60, 70, 80, 90, 95 [0157] RH (%) desorption: 95, 90, 80, 70,
60, 50, 40, 30, 20, 10, 5
[0158] At the drying step about 0.6% weight loss was registered.
The obtained dried product was not hygroscopic, it adsorbed up to
0.7% water at high relative humidity. During the desorption cycle a
loss of weight of 1.4% was registered, this indicated that the
product was losing ethanolate. The obtained product after ADS/DES
was a mixture of ethanolate form and hydrated form.
[0159] The ADS/DES curve is collected in FIG. 18.
Adsorption-Desorption Hydratation Tests
[0160] About 23 mg of Form A were transferred into a VTI vapor
sorption analyzer model SGA100 and the weight change with respect
to the atmospheric humidity was recorded at the following
conditions: [0161] equilibrium: .ltoreq.0.0005% in 5 min. or 90
min. [0162] data interval: 0.05% or 2 min [0163] temperature:
25.degree. C. [0164] cycle RH (%) adsorption/desorption: 5.95
[0165] repeat the cycle 11 times
[0166] At the end of this test a loss of weight of 5.2% was
registered. This was comparable with the TG result (TG 5.4% up to
80.degree. C.). The ethanolate form was transferred into a hydrated
form. The ADS/DES hydratation test curves are collected in FIG.
19.
EXAMPLE 8
[0167] The stability of Form A was studied after storage of the
compound in a sample container with an inner cover made of single
LD-PE (string sealed), and and outer cover made of PETP/Alu/PE
(Moplast) heat sealed. A long term stability study at 25.degree.
C./60% RH, and an accelerated stability study at 40.degree.
C./75%RH, were performed for a period of 6 months, and the samples
analysed at different time points as shown in following tables.
TABLE-US-00013 TABLE 13 Long term stability at 25.degree. C./60% RH
Release tests Remark Specification data 0 month 1 month 3 month 6
month Polymorphism .degree. C. (onset) For information only 97.3
97.3 95.5 97.9 97.5 DSC .degree. C. max For information only 104
104.2 103.5 104.2 104 Residual % (w/w) ethanol <=10.0% 6.71 6.31
6.33 6.40 6.33 solvents % (w/w) 2-propanol <=0.5% 0.04 0.04 0.05
0.05 0.05 % (w/w) THF <=0.5% <0.01 <0.01 <0.01 <0.01
<0.01 % (w/w) acetone <=0.5% <0.01 <0.01 <0.01
<0.01 <0.01 % (w/w) CH.sub.2Cl.sub.2 <=0.06% <0.01
<0.01 <0.01 <0.01 <0.01 Water (KF) % (w/w) <=7.0%
0.63 0.23 0.34 0.32 0.46 X-Ray For information only C C -- -- --
powder diffraction C: chrystal
TABLE-US-00014 TABLE 14 Accelerated stability at 40.degree. C./75%
RH Release Tests Remark Specification data 0 month 1 month 3 month
6 month Polymorphism .degree. C. (onset) For information only 97.3
97.3 97.5 98.0 97.8 DSC .degree. C. max For information only 104
104.2 103.4 1039 104.3 Residual % (w/w) ethanol <=10.0% 6.71
6.31 6.73 6.32 6.50 solvents % (w/w) 2-propanol <=0.5% 0.04 0.04
0.05 0.05 0.05 % (w/w) THF <=0.5% <0.01 <0.01 <0.01
<0.01 <0.01 % (w/w) acetone <=0.5% <0.01 <0.01
<0.01 <0.01 <0.01 % (w/w) CH.sub.2Cl.sub.2 <=0.06%
<0.01 <0.01 <0.01 <0.01 <0.01 Water (KF) % (w/w)
<=7.0% 0.63 0.23 0.37 0.34 0.42 X-Ray For information only C C
-- -- -- powder diffraction
[0168] Form A exhibited chemical and crystallographic stability at
the conditions mentioned in tables 13 and 14.
EXAMPLE 9
[0169] The stability of Form A was studied after storage of the
compound in a sample container with an inner cover made of single
LD-PE (string sealed), and and outer cover made of vapor loc bag
(LPS) heat sealed. A long term stability study at 25.degree. C./60%
RH, and an accelerated stability study at 40.degree. C./75% RH,
were performed for a period of 6 months, and the samples analysed
at different time points as shown in following tables.
TABLE-US-00015 TABLE 15 Long term stability at 25.degree. C./60% RH
Release Tests Remark Specification data 0 month 1 month 3 month 6
month Polymorphism .degree. C. (onset) For information only 97.3
97.3 96.3 96.2 98.5 DSC .degree. C. max For information only 104
104.2 103.1 103.8 103.9 Residual % (w/w) ethanol <=10.0% 6.71
6.31 6.42 6.35 6.52 solvents % (w/w) 2-propanol <=0.5% 0.04 0.04
0.06 0.05 0.05 % (w/w) THF <=0.5% <0.01 <0.01 <0.01
<0.01 <0.01 % (w/w) acetone <=0.5% <0.01 <0.01
<0.01 <0.01 <0.01 % (w/w) CH.sub.2Cl.sub.2 <=0.06%
<0.01 <0.01 <0.01 <0.01 <0.01 Water (KF) % (w/w)
<=7.0% 0.63 0.23 0.32 0.38 0.49 X-Ray For information only C C
-- -- -- powder diffraction
TABLE-US-00016 TABLE 16 Accelerated stability at 40.degree. C./75%
RH Release Tests Remark Specification data 0 month 1 month 3 month
6 month Polymorphism .degree. C. (onset) For information only 97.3
97.3 97.8 97.5 97.9 DSC .degree. C. max For information only 104
104.2 103.4 103.7 104.0 Residual % (w/w) ethanol <=10.0% 6.71
6.31 6.35 6.31 6.30 solvents % (w/w) 2-propanol <=0.5% 0.04 0.04
0.06 0.05 0.05 % (w/w) THF <=0.5% <0.01 <0.01 <0.01
<0.01 <0.01 % (w/w) acetone <=0.5% <0.01 <0.01
<0.01 <0.01 <0.01 % (w/w) CH.sub.2Cl.sub.2 <=0.06%
<0.01 <0.01 <0.01 <0.01 <0.01 Water (KF) % (w/w)
<=7.0% 0.63 0.23 0.31 0.36 0.51 X-Ray For information only C C
-- -- -- powder diffraction
[0170] Form A exhibited chemical and crystallographic stability at
the conditions mentioned in tables 15 and 16.
EXAMPLE 10
[0171] For the purpose of chemical stability testing, Form A was
stored for a period of 1, 4 and 8 weeks under different conditions.
These conditions were 40.degree. C./75% RH, 50.degree. C.,
RT/<5% RH, RT/56% RH, RT/75% RH and 0.3 da ICH light. The
compound was analysed after storage by HPLC and by visual
inspection. The HPLC method used in this study was HPLC method 909.
The results of the tests are reported in the following table.
TABLE-US-00017 TABLE 17 HPLC Sum of impurities Appearance
Conditions 1 week 4 week 8 week 1 week 4 weeks 8 weeks Reference
1.07 -- -- slightly-yellow -- -- 0.3 da ICH light 1.01 -- --
slightly-yellow -- -- 40.degree. C./75% RH 1.03 0.98 0.99
slightly-yellow slightly-yellow slightly-yellow 50.degree. C. 1.05
1.08 1.06 slightly-yellow slightly-yellow slightly-yellow RT/<5%
RH -- 1.02 1.04 -- slightly-yellow slightly-yellow RT/56% RH --
1.02 0.99 -- slightly-yellow slightly-yellow RT/75% RH -- 1.00 1.01
-- slightly-yellow slightly-yellow
[0172] It was concluded that Form A is chemically stable after
storage in all investigated conditions.
EXAMPLE 11
[0173] Different fractions of Form B were characterized with
thermogravimetry (TG), differential scanning calorimetry (DSC) and
infrared spectroscopy (IR). The results of the tests are reported
in the following table.
TABLE-US-00018 TABLE 18 TG % weight DSC change Max Extra Fractions
<100.degree. C. IR (.degree. C.) (.degree. C.) Form B fraction 1
5.65 Hydrate, Ref 69.1 -- after ADS/DES 4.30 .+-.Hydrate, Ref, --
-- +amorphous Form B fraction 2 5.91 ~Hydrate, Ref 75.6 -- after 5
d 40.degree. C./75% 3.56 ~Hydrate, Ref 74.1 -- RH Form B fraction 3
3.13 .+-.Hydrate, Ref, 77.0 67.8 +amorphous after 5 d 40.degree.
C./75% 2.33 .+-.Hydrate, Ref, 77.4 62.8 RH +amorphous
[0174] .about.hydrate, Ref: identical with reference
EXAMPLE 12
[0175] The adsorption and desorption of water at 25.degree. C. at
different conditions of relative humidity was investigated on 38 mg
of Form B. The weight change as a function of relative humidity was
registered. The results are displayed in FIG. 20. At the drying
step about 5.6% weight loss was registered for Form B. The obtained
dried product was hygroscopic, it adsorbed up to 6.8% water at high
relative humidity. After the desorption cycle about 1.2% water
remained on the sample. The obtained product after ADS/DES was a
mixture of hydrate and amorphous product.
EXAMPLE 13
[0176] Aqueous solubilities of Form B were measured in solvents
with different pH. An excess of the solute was equilibrated with
the solvent at 20.degree. C. for at least 24 hours. After removing
the undissolved compound, the concentration in solution was
determined using UV spectrometry.
TABLE-US-00019 TABLE 19 Solvent Solubility (mg/100 ml solution)
Water 10 (pH 5.1) Buffer pH 2 (citrate/HCl) 23 (pH 2.0) Buffer pH 3
(citrate/HCl) 13 (pH 3.0) Buffer pH 4 (citrate/HCl) 12 (pH 4.0)
0.01N HC1 18 (pH 2.1) 0.1N HCl 150 (pH 1.1) 1.0N HCl 510 (pH
0.14)
EXAMPLE 14
[0177] The stability of the crystal structure of Form B was studied
after storage of the compound for a period of two weeks at room
temperature (RT) under <5%, 56% and 75% relative humidity (RH),
50.degree. C. and 40.degree. C./75% RH. The samples were analyzed
with thermogravimetry (TG), differential scanning calorimetry
(DSC), infrared spectroscopy (IR) and X-ray diffraction (XRD). The
results of the tests are reported in the following table.
TABLE-US-00020 TABLE 20 TG DSC condition <100.degree. C.
<225.degree. C. IR XRD Max (.degree. C.) Appearance 0 days 5.65
0.16 Ref Ref 69.1 slightly yellow-orange after ADS/DES 4.30 0.18
.noteq.Ref -- -- slightly yellow-orange RT/<5% RH 0.32 0.07
.noteq.Ref .noteq.Ref 71.2 slightly yellow-orange RT/56% RH 5.71
0.25 ~Ref ~Ref 71.0 slightly yellow-orange RT/75% RH 6.20 0.10 ~Ref
~Ref 71.5 slightly yellow-orange 50.degree. C. 0.23 0.06 .noteq.Ref
.noteq.Ref 76.4 slightly yellow-orange 40.degree. C. 75% RH 5.77
0.07 ~Ref .+-.Ref 70.4 slightly yellow-orange ~Ref: identical with
reference .+-.Ref: similar with reference .noteq.Ref: different
with reference
EXAMPLE 15
[0178] In the chemical stability test program Form B was stored for
a period of 1, 4 and 9 weeks under different conditions. These
conditions were 40.degree. C./75% RH, 50.degree. C., RT/<5% RH,
RT/56% RH, RT/75% RH and 0.3 da ICH light. The compound was
analysed after storage by HPLC and by visual inspection. The HPLC
method used in this study was HPLC method 909. The results of the
tests are reported in the following table, from which it was
concluded that Form B is chemically stable.
TABLE-US-00021 TABLE 21 HPLC Sum of impurities Appearance Condition
1 week 4 week 9 week 1 week 4 weeks 9 weeks Reference 1.35 -- --
lightly yellow- -- -- orange 0.3 da ICH light 1.30 -- --
light-orange -- -- 40.degree. C./75% RH 1.43 1.38 1.41 lightly
yellow- Orange light-orange orange 50.degree. C. 1.46 1.50 1.46
lightly yellow- light-orange light-orange orange RT/<50% RH --
1.48 1.37 -- light-orange light-orange RT/56% RH -- 1.11 1.35 --
lightly yellow- light-orange orange RT/75% RH -- 1.34 1.29 --
light-orange light-orange
EXAMPLE 16
[0179] Form K was prepared by adding neat methanesulfonic acid to a
solution of Form A in 4852-4767-2031.1
[0180] THF at r.t. Form K was subsequently mixed with alkali halide
and pressed to a pellet (Ph. Eur.) and analyzed by Infrared
spectrometry (IR) at the following conditions: [0181] apparatus:
Nicolet Magna 560 FTIR spectrophotometer [0182] number of scans: 32
[0183] resolution: 1 cm.sup.-1 [0184] wavelength range: 4000 to 400
cm.sup.-1 [0185] baseline correction: yes [0186] detector: DTGS
with KBr windows [0187] beamsplitter: Ge on KBr [0188] alkali
halide: KBr (potassium bromide)
[0189] The IR spectrum of Form K, as shown in FIG. 21, reflects the
vibrational modes of the molecular structure of the mesylate
solvate as a crystalline product.
TABLE-US-00022 TABLE 22 Wavenumbers (cm.sup.-1) and relative
intensities of absorption bands (.sup.1) 3362m, 3064w 2985m, 2964m,
2906m, 2873m, 2632w, 2585w 1687s, 1627w, 1601w 1554m, 1495m, 1480w,
1470w, 1452w, 1443w, 1421w 1383w, 1373w, 1369w, 1345m, 1324m,
1314m, 1299w, 1268m, 1245m, 1221m, 1202s 1190s, 1166vs, 1122m,
1091m, 1077m, 1051s, 1043s, 1023m, 1002m 992m, 969w, 943w, 912w,
888w, 867vw, 836w, 813vw 773m, 754w, 743m, 711w, 700m, 658m, 634w
581w, 556m, 505w, 472vw, 452vw, 435vw, 417vw (.sup.1) vs = very
strong, s = strong, m = medium, w = weak, vw = very weak, br =
broad
EXAMPLE 17
[0190] Form K was transferred to a glass capillary cell and
analyzed by Raman spectrometry at the following conditions: [0191]
Raman mode: Nondispersive Raman [0192] apparatus: Nicolet FT-Raman
module [0193] number of scans: 64 [0194] resolution: 4 cm.sup.-1
[0195] wavelength range: 3700 to 100 cm.sup.-1 [0196] laser:
Nd:YVO4 [0197] laser frequency: 1064 cm.sup.-1 [0198] detector:
InGaAs [0199] beamsplitter: CaF.sub.2 [0200] sample geometry:
180.degree. reflective [0201] polarization: no
[0202] The Raman spectrum of Form K, as shown in FIG. 22, reflects
the vibrational modes of the molecular structure of the mesylate as
a crystalline product.
TABLE-US-00023 TABLE 23 Wavenumbers (cm.sup.-1) and relative
intensities of absorption bands (.sup.1) 3080m, 3068m, 3059m,
3043w, 3022w, 3006m 2989s, 2978s, 2962s, 2933vs, 2906m, 2871m
1685vw, 1628w, 1603s, 1585w, 1495w, 1479w, 1466w, 1450m, 1423w
1381w, 1346w, 1336w, 1313w, 1290w, 1271w, 1244w, 1230w, 1209m
1190w, 1182m, 1163vs, 1122w, 1105w, 1090m, 1049vs, 1032w, 1003s
968w, 955w, 941w, 914w, 897w, 877w, 866w, 845w, 823m, 814m 783m,
771m, 742w, 658w, 634m, 621w 577w, 561m, 534w, 524w, 497w, 451w,
436w 337w, 308w, 287m, 247w, 206w, 162m, 129m (.sup.1) vs = very
strong, s = strong, m = medium, w = weak, vw = very weak
EXAMPLE 18
[0203] About 3 mg of Form K were transferred into a standard
aluminium TA-Instrument sample pan. The sample pan was closed with
the appropriate cover and the DSC curve recorded on a
TA-Instruments Q1000 MTDSC equipped with a RCS cooling unit, at the
following conditions: [0204] initial temperature: 25.degree. C.
[0205] heating rate: 10.degree. C./min [0206] final temperature:
200.degree. C. [0207] nitrogen flow: 50 ml/min
[0208] The DSC curve as depicted in FIG. 23, shows the melting with
decomposition of a crystalline product. The melting of Form K
occurs at 158.4.degree. C. Due to the decomposition, the heat of
fusion calculation can only be used to indicate the crystalline
property of the product.
EXAMPLE 19
[0209] Form K was transferred into an aluminum sample pan. The TG
curve was recorded on a TA Instruments Hi-Res TGA 2950
thermogravimeter at the following conditions: [0210] initial
temperature: room temperature [0211] heating rate: 20.degree.
C./min [0212] resolution factor: 4 [0213] final condition:
300.degree. C. or <80[(w/w) %]
[0214] The TG curve is exhibited in FIG. 24. The loss of weight of
around 0.2% up to 60.degree. C. was due to the evaporation of
solvent. The loss of weight at temperatures above 140.degree. C.
was due to the evaporation and decomposition of the product.
EXAMPLE 20
Adsorption-Desorption
[0215] About 21 mg of Form K were transferred into a VTI vapor
sorption analyzer model SGA100 and the weight change with respect
to the atmospheric humidity was recorded at the following
conditions: [0216] drying temperature: 40.degree. C. [0217]
equilibrium: 0.05% in 5 min. or 60 min. [0218] data interval: 0.05%
or 2.0 min. [0219] temperature: 25.degree. C. [0220] first cycle RH
(%) adsorption: 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95 [0221] RH
(%) desorption: 95, 90, 80, 70, 60, 50, 40, 30, 20, 10, 5 [0222]
second cycle RH (%) adsorption: 5, 10, 20, 30, 40, 50, 60, 70, 80,
90, 95 [0223] RH (%) desorption: 95, 90, 80, 70, 60, 50, 40, 30,
20, 10, 5
[0224] The Adsorption-Desorption isotherm is shown in FIG. 25. Form
K is hygroscopic. At the initial drying step a loss of weight of
0.3% was registered, comparable to the TG result. Form K adsorbed
up to 1.5% water at high relative humidity. The product dried
completely during the desorption cycle.
[0225] A different study of the adsorption and desorption of water
by Form K at 25.degree. C. at different conditions of relative
humidity was investigated on an amount of about 18 mg of the
mesylate solvate. The weight change as a function of relative
humidity was registered. The result is displayed in FIG. 26.
[0226] At the drying step about 0.6% weight loss is registered for
Form K. The obtained dried product is slightly hygroscopic, it
adsorbed up to 1.7% water at high relative humidity. The product
dried completely during the desorption cycle.
EXAMPLE 21
[0227] Aqueous solubilities of Form K were measured in solvents
with different pH. An excess of the solute was equilibrated with
the solvent at 20.degree. C. for at least 48 hours. After removing
the undissolved compound, the concentration in solution was
determined using UV spectrometry.
TABLE-US-00024 TABLE 24 Solvent Solubility (mg/100 ml solution)
Water 19 (pH 3.3) Buffer pH 2 (citrate/HCl) 21 (pH 2.0) Buffer pH 3
(citrate/HCl) 12 (pH 3.0) Buffer pH 4 (citrate/HCl) 11 (pH 4.0)
0.01N HCl 24 (pH 2.0) 20% HP.beta.CD in water 2100 (pH 1.6)
EXAMPLE 22
[0228] The stability of the crystal structure of Form K batch 1 was
studied after storage of the compound for a period of four weeks at
room temperature (RT) under 75% relative humidity (RH), 50.degree.
C. and 40.degree. C./75% RH. The stability of the crystal structure
of Form K batch 2 was studied after storage of the compound for a
period of four weeks at room temperature (RT) under <5%, 56% and
75% relative humidity (RH), 50.degree. C. and 40.degree. C./75% RH.
The samples were analyzed with thermogravimetry (TG), differential
scanning calorimetry (DSC) and infrared spectroscopy (IR). The
results of the tests are reported in the following table.
TABLE-US-00025 TABLE 25 DSC TG Max Extra compound conditions
<80.degree. C. <125.degree. C. IR (.degree. C.) (.degree. C.)
Appearance Form K 0 days 0.47 0.15 Ref 143.7 -- slightly orange
Batch 1 RT/75% RH 2.87 0.19 .noteq.Ref 146.6 64.3 slightly orange
50.degree. C. 0.32 0.14 ~Ref 140.6 45.6 orange 40.degree. C./75% RH
1.48 3.71 -- -- -- brown oil Form K 0 days 0.16 0.11 Ref 155.8 --
slightly orange Batch 2 RT/<5% RH 0.00 0.03 ~Ref 156.9 --
slightly orange RT/56% RH 0.27 0.03 .+-.Ref 154.6 -- slightly
orange RT/75% RH 1.82 0.07 .noteq.Ref 149.2 67.0 slightly orange
50.degree. C. 0.12 0.12 ~Ref 156.8 -- slightly orange 40.degree.
C./75% RH 3.26 3.08 -- -- -- brown oil ~Ref: identical with
reference .+-.Ref: similar with reference .noteq.Ref: different
with reference
EXAMPLE 23
[0229] In the chemical stability test program Form K batch 1 was
stored for a period of 1 and 4 weeks under different conditions.
These conditions were 40.degree. C./75% RH, 50.degree. C., RT/75%
RH and 0.3 da ICH light. Form K batch 2 was also stored for a
period of 1 and 4 weeks under different conditions. These
conditions were 40.degree. C./75% RH, 50.degree. C., RT/<5% RH,
RT/56% RH, RT/75% RH and 0.3 da ICH light. The compound was
analysed after storage by HPLC and by visual inspection. The HPLC
method used in this study was HPLC method 909. The results of the
tests are reported in the following table.
TABLE-US-00026 TABLE 26 HPLC Sum of impurities 1 4 appearance
compound conditions week weeks 1 week 4 weeks Form K Reference 3.57
-- slightly-orange -- batch 1 0.3 da ICH 2.93 -- slightly-orange --
light 40.degree. C./75% 5.36 >90* slightly-orange brown oil RH
50.degree. C. 3.99 27.53 slightly-orange orange RT/75% -- 3.61 --
slightly-orange RH Form K Reference 1.50 -- slightly-orange --
Batch 2 0.3 da ICH 1.17 -- slightly-orange -- light 40.degree.
C./75% 1.75 >85* slightly-orange brown oil RH 50.degree. C. 1.46
1.25 slightly-orange slightly-orange RT/<5% -- 1.58 --
slightly-orange RH RT/56% -- 1.45 -- slightly-orange RH RT/75% --
1.46 -- slightly-orange RH
EXAMPLE 24
[0230] A randomized, placebo-controlled, double-blind, multiple
dose escalation trial was performed to examine the safety,
tolerability and pharmacokinetics of Form A after oral
administration twice or three times daily, in healthy subjects.
Four dosages of Form A (400 mg b.i.d., 800 mg b.i.d., 800 mg
t.i.d., and 1200 mg t.i.d.) were tested in 4 panels of 9 healthy
subjects. Within each panel, 6 subjects were treated with Form A
and 3 subjects with placebo for 13 days with a single intake in the
morning of day 14. (b.i.d.=twice daily, t.i.d.=three times
daily).
[0231] Form A was readily absorbed and concentration-time profiles
of Form A after repeated dosing were dependent on the dose
administered. Steady-state plasma concentrations were reached
generally within 3 days, although Con (conc. at administration
time) and AUC.sub.24h (area under de curve or bioavailability)
slightly decreased over time at all dose levels. AUC.sub.24h and
C.sub.ss,av (conc. at average steady-state) were dose-proportional
(daily dose) at 400 mg b.i.d., 800 mg t.i.d. and 1200 mg t.i.d.,
but was more than dose-proportional at 800 mg b.i.d. Cmax (maximum
conc.) was dose-proportional with respect to dose per intake. Less
than 2% of unchanged Form A was excreted in the urine at all dose
levels.
* * * * *