U.S. patent application number 13/919518 was filed with the patent office on 2013-10-24 for crystal forms of anamorelin.
This patent application is currently assigned to Helsinn Therapeutics (U.S.), Inc.. The applicant listed for this patent is Benjamin Littler, Keith Lorimer, Bernhard Paul, Marcia Philipson. Invention is credited to Benjamin Littler, Keith Lorimer, Bernhard Paul, Marcia Philipson, Seemon H. Pines.
Application Number | 20130281701 13/919518 |
Document ID | / |
Family ID | 49380715 |
Filed Date | 2013-10-24 |
United States Patent
Application |
20130281701 |
Kind Code |
A1 |
Lorimer; Keith ; et
al. |
October 24, 2013 |
CRYSTAL FORMS OF ANAMORELIN
Abstract
Crystalline forms of anamorelin which are useful as
pharmaceutical agents are disclosed. Methods of production and
isolation of these polymorphs and pharmaceutical compositions which
include these polymorphs and pharmaceutical methods of treatment
are also disclosed. The crystalline polymorphs of the present
invention are useful as they act directly on the pituitary gland
cells to release growth hormone.
Inventors: |
Lorimer; Keith; (West
Lafayette, IN) ; Pines; Seemon H.; (New Providence,
NJ) ; Philipson; Marcia; (Doylestown, PA) ;
Paul; Bernhard; (Slingerlands, NY) ; Littler;
Benjamin; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Lorimer; Keith
Philipson; Marcia
Paul; Bernhard
Littler; Benjamin |
West Lafayette
Doylestown
Slingerlands
San Diego |
IN
PA
NY
CA |
US
US
US
US |
|
|
Assignee: |
Helsinn Therapeutics (U.S.),
Inc.
Bridgewater
NJ
|
Family ID: |
49380715 |
Appl. No.: |
13/919518 |
Filed: |
June 17, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12831009 |
Jul 6, 2010 |
8466173 |
|
|
13919518 |
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|
11165598 |
Jun 22, 2005 |
7825138 |
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12831009 |
|
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60583757 |
Jun 29, 2004 |
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Current U.S.
Class: |
546/201 |
Current CPC
Class: |
C07K 5/0202 20130101;
C07D 401/06 20130101; C07K 5/06026 20130101 |
Class at
Publication: |
546/201 |
International
Class: |
C07D 401/06 20060101
C07D401/06 |
Claims
1. Crystalline anamorelin clathrate having an X-ray powder
diffraction pattern having at least four 2.theta. values measured
using Cu K.sub..alpha. radiation selected from the group consisting
of 10.1, 11.1, 17.6, 20.0, and 20.8.
2. The crystalline anamorelin clathrate of claim 1 having a degree
of hydration of from 0.5 to 2.5.
3. The crystalline anamorelin clathrate of claim 1 having a degree
of hydration of from 1.0 to 2.0.
4. The crystalline anamorelin clathrate of claim 1 comprising
methanol as an organic volatile impurity.
5. A process for preparing the crystalline anamorelin clathrate of
claim 1 comprising: a) producing a mixture of anamorelin and a
solvent comprised of methanol and water, b) precipitating
crystalline anamorelin clathrate hydrate from the solvent; and c)
isolating the crystalline anamorelin clathrate.
6. The process of claim 5, wherein the solvent is a mixture of
water and methanol and the volume percentage of methanol in the
mixture is from about 5% to about 95%.
7. The process of claim 5, wherein the solvent is a mixture of
water and methanol and the volume percentage of methanol in the
mixture is from about 20% to about 80%.
8. The process of claim 5, wherein the solvent is a mixture of
water and methanol and the volume percentage of methanol in the
mixture is from about 40% to about 60%.
9. The process of claim 5, wherein the solvent is at an elevated
temperature while combining with the anamorelin in step a), or the
solvent is heated to an elevated temperature after combining with
the anamorelin in step a).
10. The process of claim 9, wherein said elevated temperature is
from about 40.degree. C. to about 100.degree. C.
11. The process of claim 9, wherein said elevated temperature is
from about 50.degree. C. to about 80.degree. C.
12. The process of claim 9, wherein said elevated temperature is
from about 65.degree. C. to about 75.degree. C.
13. The process of claim 5, wherein the crystals are precipitated
in step b) by cooling the solvent.
14. A process for preparing the crystalline anamorelin clathrate
hydrate of claim 1 comprising: a) combining
{1-[(1R)-2-](3R)-3-Benzyl-3-(N,N',N'-trimethylhydrazinocarbonyl)piperidin-
-1-yl-]-1-(1H-indol-3-ylmethyl)-2-oxo-ethylcarbamoyl]-1-methylethyl}carbam-
ic acid tert-butyl ester (protected anamorelin) with a solvent; b)
combining the mixture from step a) with an acid; c) neutralizing
the mixture formed in step b); d) precipitating crystals of
anamorelin clathrate hydrate from the solvent; and e) isolating the
crystals.
15. The process of claim 14, wherein the solvent in step a) is
methanol, and the mixture is neutralized in step c) with a mixture
of potassium hydroxide and water.
16. The process of claim 14, wherein the acid is methanesulfonic
acid.
17. The process of claim 14, wherein the solvent is at an elevated
temperature while combining with said protected anamorelin in step
a), or the solvent is heated to an elevated temperature after
combining with said protected anamorelin in step a).
18. The process of claim 17, wherein the elevated temperature is
from about 50.degree. C. to about 75.degree. C.
19. The process of claim 14, wherein the crystals are precipitated
in step d) by cooling the solvent.
20. A method of making a pharmaceutically acceptable salt of
anamorelin comprising reacting the crystalline anamorelin clathrate
of claim 1 with hydrochloric acid.
Description
BACKGROUND OF THE INVENTION
[0001] Growth hormone is a major participant in the control of
several complex physiologic processes, including growth and
metabolism. Growth hormone is known to have a number of effects on
metabolic processes, e.g., stimulation of protein synthesis and
free fatty acid mobilization and to cause a switch in energy
metabolism from carbohydrate to fatty acid metabolism. Deficiency
in growth hormone can result in a number of severe medical
disorders, e.g., dwarfism.
[0002] The release of growth hormone from the pituitary is
controlled, directly or indirectly, by number of hormones and
neurotransmitters. Growth hormone release can be stimulated by
growth hormone releasing hormone (GHRH) and inhibited by
somatostatin. In both cases the hormones are released from the
hypothalamus but their action is mediated primarily via specific
receptors located in the pituitary. Other compounds which stimulate
the release of growth hormone from the pituitary have also been
described. For example, arginine, L-3,4-dihydroxyphenylalanine
(1-Dopa), glucagon, vasopressin, PACAP (pituitary adenylyl cyclase
activating peptide), muscarinic receptor agonists and a synthetic
hexapeptide, GHRP (growth hormone releasing peptide) release
endogenous growth hormone either by a direct effect on the
pituitary or by affecting the release of GHRH and/or somatostatin
from the hypothalamus.
[0003] The use of certain compounds for increasing the levels of
growth hormone in mammals has previously been proposed. For
example, U.S. Pat. Nos. 6,303,620 and 6,576,648 (the entire
contents of which are incorporated herein by reference), disclose a
compound:
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide, having the following chemical
structure:
##STR00001##
which acts directly on the pituitary cells under normal
experimental conditions in vitro to release growth hormone
therefrom. This compound is also known under the generic name
"anamorelin." This growth hormone releasing compound can be
utilized in vitro as a unique research tool for understanding,
inter alia, how growth hormone secretion is regulated at the
pituitary level. Moreover, this growth hormone releasing compound
can also be administered in vivo to a mammal to increase endogenous
growth hormone release.
SUMMARY OF THE INVENTION
[0004] It has been found that
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide can be crystallized. At least
four crystalline forms with purities of greater than 99 area
percent have been produced. These crystalline forms of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide are expected to be useful as
pharmaceutical compositions, which can be used to increase
endogenous growth hormone release.
[0005] In one embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide.
[0006] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one or at least two 2.theta.
values measured using Cu K.sub..alpha. radiation are selected from
the group consisting of about: 10.1, 11.1, 17.6, 20.0 and 20.8. In
yet another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group listed above
and at least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group consisting of
about: 9.6, 17.3, 18.2, 22.1 and 23.5.
[0007] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one or at least two 2.theta.
values measured using Cu K.sub..alpha. radiation are selected from
the group consisting of about: 17.0, 19.4, 21.5, 26.2, and 33.3. In
yet another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group listed above
and at least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group consisting
about: 9.3, 23.3, 20.4, 22.9, and 23.5.
[0008] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one or at least two 2.theta.
values measured using Cu K.sub..alpha. radiation are selected from
the group consisting of about 19.2, 20.1, 23.0, 26.2, and 27.0. In
yet another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group listed above
and at least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group consisting
about: 16.3, 21.4, 24.0, 29.8, and 31.5.
[0009] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one or at least two 2.theta.
values measured using Cu K.sub..alpha. radiation are selected from
the group consisting of about 10.2, 11.2, 18.7, 20.6, and 23.4. In
yet another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group listed above
and at least one or at least two 2.theta. values measured using Cu
K.sub..alpha. radiation are selected from the group consisting of
about 9.9, 13.8, 14.3, 16.7, and 19.8.
[0010] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one 2.theta. value measured
using Cu K.sub..alpha. radiation is selected from the group
consisting of: 10.1, 17.0, 20.6, and 23.0, and optionally a second
additional 2.theta. value measured using Cu K.sub..alpha. radiation
is selected from the group consisting of about: 17.6, 19.2, 23.0,
23.4, 26.2, and 33.3 or optionally a second additional 2.theta.
value measured using Cu K.sub..alpha. radiation is selected from
the group consisting of: 9.5, 16.7, 17.5, 17.9, 20.0, 21.5, 23.5,
23.9, and 27.5.
[0011] Further the present invention is directed to a process for
preparing crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide. The method comprises combining
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide with a solvent. Precipitating the
crystals from the solvent and isolating the crystals. In one
embodiment, the solvent is selected from the group consisting of:
water, methanol, ethanol, n-propanol, isopropanol, butanol, and
mixtures thereof. In another embodiment, the solvent is a mixture
of methanol and water. In a further embodiment, the solvent
includes between 40% v/v methanol and 60% v/v methanol. In a still
further embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method described
above.
[0012] Further the present invention is a process for preparing
crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide. The method comprises the steps
of combining
{1-[(1R)-2-](3R)-3-Benzyl-3-(N,N',N'-trimethylhydrazinocarbonyl-
)piperidin-1-yl-]-1-(1H-indol-3-ylmethyl)-2-oxo-ethylcarbamoyl]-1-methylet-
hyl}carbamic acid tert-butyl ester with a solvent and an acid. The
method further comprises neutralizing the mixture, precipitating
the crystals from the solvent, and isolating the crystals. In one
embodiment the solvent is methanol, and the mixture is neutralized
with a mixture of potassium hydroxide and water. In another
embodiment the acid is methanesulfonic acid. In yet another
embodiment the present invention is a crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method described
above.
[0013] Further, the present invention is directed to a
pharmaceutical composition comprising crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide and at least one pharmaceutically
acceptable carrier or diluent.
[0014] Further the present invention is directed to a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide.
[0015] The present invention provides crystalline polymorphic forms
of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide, each of which are more stable,
and have advantageous manufacturing properties relative to
amorphous forms of this compound. Due to the high purity of these
crystalline forms, they can fulfill more exacting pharmaceutical
regulations and specifications often required for pharmaceutical
formulations. Furthermore, the polymorphs of the present invention
are readily suspended in solvents, they are easily dried and
filtered, and as such are amenable to large-scale production.
Additionally, these crystalline forms are stable for extended
periods of time without the need for specialized storage
conditions.
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] FIG. 1 is a characteristic X-Ray Powder Diffraction (XRPD)
pattern for form A
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method described
in Example 1 (Vertical axis: Intensity (CPS); Horizontal axis:
Diffraction Angle, in Two Theta (degrees).
[0017] FIG. 2 is a characteristic X-Ray Powder Diffraction (XRPD)
pattern for form B
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method described
in Example 2 (Vertical axis: Intensity (CPS); Horizontal axis:
Diffraction Angle, in Two Theta (degrees).
[0018] FIG. 3 is a characteristic X-Ray Powder Diffraction (XRPD)
pattern for form D
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method described
in Example 5 (Vertical axis: Intensity (CPS); Horizontal axis:
Diffraction Angle, in Two Theta (degrees).
[0019] FIG. 4 is a characteristic X-Ray Powder Diffraction (XRPD)
pattern for form C
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method described
in Example 8 (Vertical axis: Intensity (CPS); Horizontal axis:
Diffraction Angle, in Two Theta (degrees).
[0020] FIG. 5 is a characteristic Differential Scanning Calorimetry
(DSC) thermogram for form C
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide as described in Example 8.
DETAILED DESCRIPTION OF THE INVENTION
[0021] The foregoing and other objects, features and advantages of
the invention will be apparent from the following more particular
description of preferred embodiments of the invention, as
illustrated in the accompanying drawings in which like reference
characters refer to the same parts throughout the different views.
The drawings are not necessarily to scale, emphasis instead being
placed upon illustrating the principles of the invention.
[0022] The present invention is directed to crystalline compounds
of the following Structural Formula:
##STR00002##
[0023] The chemical name of Formula I is
1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxylic
acid 1,2,2-trimethylhydrazide which is the equivalent of
2-amino-N-[(1R)-2-[-3-benzyl-3-(N,N',N'-trimethylhydrazinocarbonyl)piperi-
-din-1-yl]-1-((1H-indol-3-yl)methyl)-2-oxoethyl]-2-methylpropionamide.
[0024] In a preferred embodiment, the compound of Formula I has the
(R) configuration at the chiral carbon designated by the asterisk
(*) in Formula I. The chemical name of the compound of Formula I
having the (R) configuration at the designated chiral carbon is:
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide:
##STR00003##
This compound is better known as "anamorelin."
[0025] It is to be understood that crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide includes hydrates thereof, unless
specifically excluded. In a preferred embodiment crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is in the form of a dihydrate. As
used herein a "dihydrate" is when each molecule of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is associated with about two
molecules of water. As used herein a "monohydrate" is when each
molecule of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarbo-x-
ylic acid 1,2,2-trimethylhydrazide is associated with about one
molecule of water.
[0026] As used herein a "crystalline form" is a solid substance
having a highly regular chemical structure. When a compound
recrystallizes from a solution or slurry, it may crystallize with
different spatial lattice arrangements, a property referred to as
"polymorphism," with the different crystal forms individually being
referred to as a "polymorph." While polymorphs of a given substance
have the same chemical composition, they may differ from each other
with respect to one or more physical properties, such as solubility
and dissociation, true density, melting point, crystal shape,
compaction behavior, flow properties, and/or solid state
stability.
[0027] The polymorphic behavior of drugs can be of crucial
importance in pharmacy and pharmacology. The differences in
physical properties exhibited by polymorphs affect pharmaceutical
parameters such as storage stability, compressibility and density
(important in formulation and product manufacturing), and
dissolution rates (an important factor in determining
bio-availability). Differences in stability can result from changes
in chemical reactivity (e.g. differential oxidation, such that a
dosage form discolors more rapidly when it is one polymorph than
when it is another polymorph) or mechanical changes (e.g. tablets
crumble on storage as a kinetically favored polymorph converts to
thermodynamically more stable polymorph) or both (e.g. tablets of
one polymorph are more susceptible to breakdown at high humidity).
In addition, the physical properties of the crystal may be
important in processing: for example, one polymorph might be more
likely to form solvates or might be difficult to filter and wash
free of impurities (i.e. particle shape and size distribution might
be different between one polymorph relative to other).
[0028] In certain specific embodiments, the present invention is
directed to crystalline polymorphs of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide. Four such crystalline polymorphs
described herein are designated as crystalline forms A, B, C, and
D, and may be distinguished by their respective X-ray powder
diffraction patterns. Crystalline forms A, B, C, and D of the
present invention may exist in anhydrous forms as well as hydrated
and solvated forms, all of which are intended to be encompassed
within the scope of the present invention.
[0029] In one embodiment, X-ray powder diffraction patterns of the
polymorphs are measured on a Shimadzu XRD-6000 X-ray powder
diffractometer using Cu K.sub..alpha. radiation. The instrument is
equipped with a fine focus X-ray tube. The tube voltage and
amperage ware set to 40 kV and 40 mA, respectively. The divergence
and scattering slits are set at 10 and the receiving slit is set at
0.15 mm. Diffracted radiation is detected by NaI scintillation
detector. A theta-two theta continuous scan at 3.degree./min (0.4
sec/0.02.degree. step) from 2.5 to 40.degree. 2.theta. is used. A
silicon standard is analyzed to check the instrument alignment.
Samples were prepared for analysis by placing them in an aluminium
holder with silicon insert. Some shifting in peak positions can
occur due to differences in sample height. As defined herein, the
2-theta position has an error of +/-0.2.degree.. Further, some
shifting in peak positions can occur due to differences in the
sample, such as, for example, particle size and crystal
density.
[0030] In one specific embodiment, the present invention is
directed to crystalline form A of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide, produced by slow evaporation
from a solution thereof as described in Example 1, characterized by
the following X-ray powder diffraction pattern (also presented in
FIG. 1) expressed in terms of 2.theta., d-spacings and relative
intensities:
TABLE-US-00001 TABLE 1 Peak Data List for crystalline form A of
(3R)-1-(2-methylalanyl-D-
tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxylic acid 1,2,2-
trimethylhydrazide Peak 2Theta d Relative No. (deg) (A) Intensity 1
2.6 33.7 3 2 2.9 30.6 4 3 4.0 22.34 3 4 4.7 18.8 3 5 7.1 12.4 3 6
9.3 9.5 8 7 16.1 5.5 4 8 16.6 5.3 4 9 17.0 5.2 100 10 17.6 5.0 6 11
19.4 4.6 9 12 20.1 4.4 3 13 20.4 4.4 8 14 20.6 4.3 4 15 21.5 4.1 10
16 22.9 3.9 7 17 23.1 3.9 6 18 23.3 3.8 6 19 23.5 3.8 9 20 24.1 3.7
4 21 25.4 3.5 4 22 26.2 3.4 37 23 26.5 3.4 4 24 27.1 3.3 6 25 27.4
3.3 5 26 29.8 3.0 6 27 30.0 3.0 4 28 30.7 2.9 4 29 30.9 2.9 3 30
31.5 2.8 5 31 33.3 2.7 18 32 34.7 2.6 4 33 35.1 2.6 3 34 37.2 2.4 4
35 37.3 2.4 5 36 37.5 2.4 4 37 37.9 2.4 3 38 38.1 2.4 3 39 38.4 2.3
4 40 38.8 2.3 4
[0031] In another embodiment, the present invention is directed to
crystalline form B of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide, produced by slow evaporation
from a solution thereof as described in Example 2, characterized by
the following X-ray powder diffraction pattern (also presented in
FIG. 2) expressed in terms of 2.theta., d-spacings and relative
intensities:
TABLE-US-00002 TABLE 2 Peak Data List for crystalline form B of
(3R)-1-(2-methylalanyl-D-
tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxylic acid 1,2,2-
trimethylhydrazide Peak 2Theta D Relative No. (deg) (A) Intensity 1
8.1 10.9 8 2 9.0 9.8 4 3 9.6 9.2 8 4 10.7 8.3 4 5 13.6 6.5 8 6 14.3
6.2 12 7 16.3 5.4 40 8 17.5 5.1 12 9 18.0 4.9 12 10 19.2 4.6 68 11
20.1 4.4 56 12 21.4 4.2 20 13 23.0 3.9 100 14 24.0 3.7 16 15 24.6
3.6 4 16 25.3 3.5 12 17 26.2 3.4 100 18 27.0 3.3 52 19 29.8 3.0 16
20 31.5 2.8 32 21 33.1 2.7 16 22 34.1 2.6 12 23 35.3 2.5 8 24 37.4
2.4 4 25 38.6 2.3 8
[0032] In another embodiment, the present invention is directed to
crystalline form C of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide, produced by slow evaporation
from a solution thereof as described in Example 3.
[0033] In another embodiment, the present invention is directed to
crystalline form D of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide, produced by slow evaporation
from a solution thereof as described in Example 5, characterized by
the following X-ray powder diffraction pattern (also presented in
FIG. 3) expressed in terms of 2.theta., d-spacings and relative
intensities:
TABLE-US-00003 TABLE 3 Peak Data List for crystalline form D of
(3R)-1-(2-methylalanyl-D-
tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxylic acid 1,2,2-
trimethylhydrazide Peak 2Theta d Relative No. (deg) (A) Intensity 1
9.4 9.4 3 2 9.9 9.0 24 3 10.2 8.7 76 4 10.6 8.3 15 5 11.2 7.9 55 6
11.7 7.6 8 7 13.8 6.4 24 8 14.3 6.2 43 9 14.6 6.1 9 10 15.1 5.9 24
11 16.7 5.3 24 12 17.4 5.1 18 13 17.9 5.0 23 14 18.2 4.9 15 15 18.7
4.7 53 16 19.8 4.5 40 17 20.6 4.3 100 18 21.1 4.2 5 19 21.6 4.1 9
20 21.8 4.1 10 21 22.5 4.0 7 22 23.4 3.8 56 23 23.8 3.7 6 24 24.9
3.6 9 25 25.8 3.5 13 26 26.4 3.4 6 27 27.5 3.2 11 28 27.7 3.2 21 29
28.8 3.1 13 30 30.0 3.0 14 31 30.4 3.0 18 32 31.3 2.9 13 33 31.9
2.8 6 34 32.9 2.7 4 35 33.6 2.7 18 36 34.1 2.6 9 37 34.4 2.6 4 38
35.5 2.5 7 39 36.8 2.4 10 40 37.4 2.4 11
[0034] Further, the present invention is directed to a process for
the preparation of crystalline
(3R)-1-(2-methyalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxyl-
-ic acid 1,2,2-trimethylhydrazide which comprises crystallization
of amorphous
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperid-
inecarboxylic acid 1,2,2-trimethylhydrazide. In one embodiment, the
method broadly includes crystallization of a solute from a solvent
or slurry under conditions which yield crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide.
[0035] Thus, for example, crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide may be prepared by
crystallizastion of the amorphous form, from a solution or from a
slurry thereof in a solvent. In one embodiment, the solution or
slurry of amorphous
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide in the solvent is formed by
combining the
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecar-
-boxylic acid 1,2,2-trimethylhydrazide with the solvent.
[0036] In another preferred embodiment, a solution of amorphous
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide in the solvent is formed by
dissolving the
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecar-
-boxylic acid 1,2,2-trimethylhydrazide in the solvent.
[0037] Suitable solvents are those from which
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide can be crystallized by the
methods described herein. Examples of suitable solvents include
those selected from the group consisting of alcohols (e.g.,
methanol, ethanol, n-propanol, isopropanol, butanol), polar organic
solvents (e.g., dimethyl sulfoxide and ethylene glycol) and
water.
[0038] In a preferred embodiment, the solvent is selected from the
group comprising: methanol, ethanol, n-propanol, isopropanol,
butanol, and mixtures thereof. In a more preferred embodiment, the
solvent is methanol. In a most preferred embodiment, the solvent is
a mixture of methanol and water containing, for example, between 5%
and 95% v/v methanol, between 20% and 80% v/v methanol, preferably
between 40% and 60% v/v methanol.
[0039] In another embodiment, crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide can be prepared from an
aqueous-based solvent that includes, for example, between 5% and
95% v/v ethanol, between 20% and 80% v/v ethanol, and preferably
between 40% and 60% v/v ethanol.
[0040] In a further embodiment, crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide can be prepared from an
aqueous-based solvent that includes, for example, between 5% and
95% v/v isopropanol, between 20% and 80% v/v isopropanol, and
preferably between 40% and 60% v/v isopropanol.
[0041] In one embodiment, the solvent may be heated to an elevated
temperature. As used herein an "elevated temperature" is higher
than ambient temperature, such as for example, between 40.degree.
C. and 100.degree. C., between 50.degree. C. and 80.degree. C., or
preferably between 65.degree. C. and 75.degree. C. In another
embodiment, the solvent is heated to an elevated temperature prior
to combining of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide with the solvent. In another
embodiment, the solvent is heated to an elevated temperature after
combining
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide with the solvent.
[0042] In one embodiment, crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is precipitated from the solvent.
In one embodiment, precipitation is induced by concentration of the
solution of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide in the solvent by reduction of
the volume of the solvent. The solvent may be reduced, for example,
by evaporation of the solvent. In another embodiment, precipitation
may be induced by allowing the solution of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide in the solvent to cool.
[0043] In another embodiment, precipitation may be induced by
rapidly cooling the solution, or by slowly cooling the solution. In
a specific embodiment, precipitation may be induced by slowly
cooling the solution to about 20.degree. C. at a rate ranging from
about 0.2.degree. C./hr to about 20.degree. C./hr, preferably from
about 1.degree. C./hr to about 10.degree. C./hr. In a preferred
embodiment, the solution is cooled from the initial elevated
temperature to 60.degree. C. at a rate of 20.degree. C./h, then
from 60.degree. C. at a rate of 5.degree. C./h, and then from
45.degree. C. to ambient temperature at a rate of 15.degree. C./h.
The temperature and the cooling rate can be appropriately decided
depending on the choice of solvent and the volume thereof. For the
purpose of this invention ambient temperature is from about
20.degree. C. to about 25.degree. C.
[0044] In one embodiment, the precipitated crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is isolated from the solvent. As
used herein the term "isolated" encompasses crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide which is separated away from
other material, such as the materials contained in the medium in
which it was produced. In one embodiment, the isolated crystals are
essentially free of chemicals or contaminating solvents from the
source from which the crystals were derived or produced. In one
embodiment, the crystals are isolated from the solvent by
filtration of the solvent. In a preferred embodiment, the
crystalline form is isolated from the solvent by filtration of the
solvent after precipitation of the crystals.
[0045] In one embodiment, the isolated crystals are washed with,
for example, a methanol/water mixture and dried in an oven. The
crystals are preferably dried in an oven, at a temperature of
between about ambient temperature and about 70.degree. C.,
preferably from about 40.degree. C. to about 60.degree. C.
Preferably, the drying takes place under vacuum.
[0046] In a further embodiment, "seeds" of the desired polymorph
are added to the solvent in order to promote formation of a
particular polymorph in the solvent. In one embodiment, the
amorphous
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is added to the solvent, and
after cooling has begun the solvent is seeded with crystals of the
desired polymorph. Alternatively, the seed crystals can be added
once the solvent is saturated with amorphous of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide.
[0047] In one embodiment the crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is formed from
1-[(1R)-2-](3R)-3-Benzyl-3-(N,N',N'-trimethylhydrazinocarbonyl)piperidin--
1-yl-]-1-(1H-indol-3-ylmethyl)-2-oxo-ethylcarbamoyl]-1-methylethyl}carbami-
c acid tert-butyl ester. The tert-butoxycarbonyl group on
1-[(1R)-2-](3R)-3-Benzyl-3-(N,N',N'-trimethylhydrazinocarbonyl)piperidin--
1-yl-]-1-(1H-indol-3-ylmethyl)-2-oxo-ethylcarbamoyl]-1-methylethyl}carbami-
c acid tert-butyl ester is removed to form the free amine in the
presence of an acid. In one embodiment the acid is trifluoroacetic
acid, hydrofluoric acid, hydrochloric acid, hydrobromic acid or
sulfuric acid. In a preferred embodiment the acid is
methanesulfonic acid. Once the tert-butoxycarbonyl group is
removed, the mixture is neutralized using a base such as, for
example, potassium hydroxide, sodium hydroxide, calcium hydroxide,
ammonium hydroxide or potassium carbonate. In a preferred
embodiment, the mixture is neutralized with a mixture of water and
a base such as, for example, potassium hydroxide, sodium hydroxide,
calcium hydroxide, ammonium hydroxide or potassium carbonate. The
remainder of the crystallization process is carried out as
described above.
[0048] In one embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method described
above. Variations in the methods of crystallization may produce
variations in the crystals, including, for example, variations in
the particle size and crystal density, which may result in
variations in the XRPD patterns between crystal forms such as, for
example, peak splitting, crystals produced by such methods are also
encompassed by the present invention
[0049] In one embodiment, form C of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide can be produced by the methods
described in Example 4 or Example 6, wherein a different XRPD
spectrum is generated than by crystallization of form C
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide by slow evaporation from a
solution thereof as described in Example 3.
[0050] "Slow evaporation" as used herein, means evaporation under
atmospheric conditions or under an inert atmosphere of a solvent or
mixture of solvents from a solution or slurry thereof with the
compound to be crystallized.
[0051] In one embodiment, the present invention is a crystalline
form C of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the methods described
in Example 4.
[0052] In one embodiment, the present invention is a crystalline
form C of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the methods described
in Example 6. In one embodiment crystalline form C of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is a monohydrate. In a preferred
embodiment, crystalline form C of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide is a dihydrate
[0053] In another embodiment, the present invention is directed to
crystalline form C dihydrate of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the methods described
in Example 8 characterized by the following X-ray powder
diffraction pattern (also presented in FIG. 4) measured on a
Shimadzu XRD-6000 X-ray powder diffractometer using Cu
K.sub..alpha. radiation. Samples are placed on Si zero-return micro
samples holders. The tube voltage and amperage are set to 40 kV and
40 mA, respectively. The divergence and scattering slits are set at
10.degree. and the receiving slit is set at 0.30 mm. A theta-two
theta continuous scan at 2.degree./min from 3.0 to 45.degree.
2.theta., with a sampling pitch of 0.02 deg and a preset time of
0.60 seconds. The X-ray powder diffraction pattern is expressed in
terms of 2.theta., d-spacings and relative intensities:
TABLE-US-00004 TABLE 4 Peak Data List for crystalline form C of
(3R)-1-(2-methylalanyl-D-
tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxylic acid 1,2,3-
trimethylhydrazide dihydrate. Peak 2Theta d Relative No. (deg) (A)
Intensity 1 9.6 9.2 8 2 10.1 8.8 100 3 10.8 8.2 4 4 11.1 8.0 25 5
13.8 6.4 7 6 14.6 6.1 4 7 15.2 5.8 6 8 16.7 5.3 6 9 17.3 5.1 9 10
17.6 5.0 18 11 18.2 4.9 12 12 20.0 4.4 18 13 20.5 4.3 7 14 20.8 4.3
12 15 22.1 4.0 8 16 23.5 3.8 13 17 25.8 3.5 4 18 27.5 3.2 5 19 30.5
3.0 4 20 35.0 2.6 3
[0054] In another embodiment, crystalline form C dihydrate may be
characterized by Differential Scanning Calorimetry (DSC) or by any
other method known to persons skilled in the art.
[0055] In one embodiment, the present invention is directed to form
C
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide dihydrate characterized by
differential scanning calorimetry analysis (FIG. 5) which showed a
broad endotherm centered at 68.5. .degree. C., followed by the melt
endotherm with onset of 110.degree. C. and peak of 116.8.degree.
C.
[0056] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about: 10.1, 11.1, 17.6, 20.0 and 20.8. In yet
another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one, at least two, at least three, at least four or at least
five 2.theta. values measured using Cu K.sub..alpha. radiation are
selected from the group listed above and at least one, at least
two, at least three, at least four or at least five 2.theta. values
measured using Cu K.sub..alpha. radiation are selected from the
group consisting of about: 9.6, 17.3, 18.2, 22.1 and 23.5.
[0057] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about: 17.0, 19.4, 21.5, 26.2, and 33.3. In yet
another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one, at least two, at least three, at least four or at least
five 2.theta. values measured using Cu K.sub..alpha. radiation are
selected from the group listed above and at least one, at least
two, at least three, at least four or at least five 2.theta. values
measured using Cu K.sub..alpha. radiation are selected from the
group consisting of about: 9.3, 23.3, 20.4, 22.9, and 23.5.
[0058] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about: 19.2, 20.1, 23.0, 26.2, and 27.0. In yet
another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one, at least two, at least three, at least four or at least
five 2.theta. values measured using Cu K.sub..alpha. radiation are
selected from the group listed above and at least one, at least
two, at least three, at least four or at least five 2.theta. values
measured using Cu K.sub..alpha. radiation are selected from the
group consisting of about: 16.3, 21.4, 24.0, 29.8, and 31.5.
[0059] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about 10.2, 11.2, 18.7, 20.6, and 23.4. In yet
another embodiment, the present invention is a crystalline
composition, having an X-ray powder diffraction pattern wherein at
least one, at least two, at least three, at least four or at least
five 2.theta. values measured using Cu K.sub..alpha. radiation are
selected from the group listed above and at least one, at least
two, at least three, at least four or at least five 2.theta. values
measured using Cu K.sub..alpha. radiation are selected from the
group consisting of about: 9.9, 13.8, 14.3, 16.7, and 19.8.
[0060] In another embodiment, the present invention is crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, or at least four 2.theta. values measured using Cu
K.sub..alpha. radiation is selected from the group consisting of
about: 10.1, 17.0, 20.6, and 23.0, and optionally at least two, at
least three, or at least four additional 2.theta. values measured
using Cu K.sub..alpha. radiation is selected from the group
consisting: 17.6, 19.2, 23.0, 23.4, 26.2, and 33.3, or optionally
at least two, at least three, at least four, at least five, at
least six, at least seven, at least eight, or at least nine
additional 2.theta. value measured using Cu K.sub..alpha. radiation
is selected from the group consisting of about: 9.5, 16.7, 17.5,
17.9, 20.0, 21.5, 23.5, 23.9, and 27.5.
[0061] Further, the crystalline forms of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide of the present invention are
useful as pharmaceutical compositions.
[0062] In one embodiment, the present invention is a pharmaceutical
composition comprising crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide as described herein and at least
one pharmaceutically acceptable carrier or diluent.
[0063] Pharmaceutical compositions containing a polymorph of the
present invention may be prepared by conventional techniques, as
described in U.S. Pat. No. 6,576,648, the entire contents of which
are incorporated herein by reference.
[0064] In another embodiment, the present invention relates to a
pharmaceutical composition comprising, as an active ingredient, a
polymorph of the present invention together with a pharmaceutically
acceptable carrier or diluent.
[0065] The pharmaceutical carrier or diluent employed may be a
conventional solid or liquid carrier. The carriers, diluents,
preparation, composition, dosage and administration of the
pharmaceutical compositions of the present invention are as
described in U.S. Pat. No. 6,576,648, the entire contents of which
are incorporated herein by reference.
[0066] The compound of the present invention is expected to possess
the ability to release endogenous growth hormone in vivo. The
compound may therefore be used in the treatment of conditions which
require increased plasma growth hormone levels such as in growth
hormone deficient humans or in elderly patients or livestock.
[0067] Thus, in one embodiment, the present invention is directed
to a pharmaceutical composition for stimulating the release of
growth hormone from the pituitary of a mammal, the composition
comprising, as an active ingredient, a polymorph of the present
invention together with a pharmaceutically acceptable carrier or
diluent.
[0068] In one embodiment, the mammal, is a human, canine, murine,
feline, bovine, ovine, swine or caprine. In a preferred embodiment,
the mammal is a human.
[0069] Further the present invention is a method of stimulating the
release of growth hormone from the pituitary of a mammal in need
thereof, the method comprising administering to the mammal a
therapeutically effective amount of a polymorph of the present
invention.
[0070] As used herein, "a therapeutically effective amount" refers
to an appropriate amount of active ingredient to obtain therapeutic
or prophylactic effect and can be determined by standard
pharmaceutical procedures in cell cultures or experimental
animals.
[0071] In one embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide.
[0072] In another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about: 10.1, 11.1, 17.6, 20.0 and 20.8. In yet
another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group listed
above and at least one, at least two, at least three, at least four
or at least five 2.theta. values measured using Cu K.sub..alpha.
radiation are selected from the group consisting of about: 9.6,
17.3, 18.2, 22.1 and 23.5.
[0073] In another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about: 17.0, 19.4, 21.5, 26.2, and 33.3. In yet
another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group listed
above and at least one, at least two, at least three, at least four
or at least five 2.theta. values measured using Cu K.sub..alpha.
radiation are selected from the group consisting of about: 9.3,
23.3, 20.4, 22.9, and 23.5.
[0074] In another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about: 19.2, 20.1, 23.0, 26.2, and 27.0. In yet
another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group listed
above and at least one, at least two, at least three, at least four
or at least five 2.theta. values measured using Cu K.sub..alpha.
radiation are selected from the group consisting of about: 16.3,
21.4, 24.0, 29.8, and 31.5.
[0075] In another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group
consisting of about 10.2, 11.2, 18.7, 20.6, and 23.4. In yet
another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, at least four or at least five 2.theta. values measured
using Cu K.sub..alpha. radiation are selected from the group listed
above and at least one, at least two, at least three, at least four
or at least five 2.theta. values measured using Cu K.sub..alpha.
radiation are selected from the group consisting of about: 9.9,
13.8, 14.3, 16.7, and 19.8.
[0076] In another embodiment, the present invention is a method of
stimulating the release of growth hormone from the pituitary of a
mammal, the method comprising administering to said mammal a
therapeutically effective amount of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide having an X-ray powder
diffraction pattern wherein at least one, at least two, at least
three, or at least four 2.theta. values measured using Cu
K.sub..alpha. radiation is selected from the group consisting of
about: 10.1, 17.0, 20.6, and 23.0, and optionally at least two, at
least three, or at least four additional 2.theta. values measured
using Cu K.sub..alpha. radiation is selected from the group
consisting: 17.6, 19.2, 23.0, 23.4, 26.2, and 33.3, or optionally
at least two, at least three, at least four, at least five, at
least six, at least seven, at least eight, or at least nine
additional 2.theta. value measured using Cu K.sub..alpha. radiation
is selected from the group consisting of about: 9.5, 16.7, 17.5,
17.9, 20.0, 21.5, 23.5, 23.9, and 27.5.
[0077] To those skilled in the art, it is well known that the
current and potential uses of growth hormone in humans are varied
and multitudinous. Thus, the polymorph of the present invention can
be administered for purposes stimulating release of growth hormone
from the pituitary and would then have similar effects or uses as
growth hormone itself. The polymorphs of the present invention are
expected to be useful, for example, for stimulation of growth
hormone release in the elderly, prevention of catabolic side
effects of glucocorticoids, prevention and treatment of
osteoporosis, treatment of chronic fatigue syndrome (CFS),
treatment of acute fatigue syndrome and muscle loss following
elective surgery, stimulation of the immune system, acceleration of
wound healing, accelerating bone fracture repair, accelerating
complicated fractures, e.g. distraction osteogenesis, treatment of
wasting secondary to fractures, treatment of growth retardation,
treating growth retardation resulting from renal failure or
insufficiency, treatment of cardiomyopathy, treatment of wasting in
connection with chronic liver disease, treatment of
thrombocytopenia, treatment of growth retardation in connection
with Crohn's disease, treatment of short bowel syndrome, treatment
of wasting in connection with chronic obstructive pulmonary disease
(COPD), treatment of complications associated with transplantation,
treatment of physiological short stature including growth hormone
deficient children and short stature associated with chronic
illness, treatment of obesity and growth retardation associated
with obesity, treatment of anorexia, treating growth retardation
associated with the Prader-Willi syndrome and Turner's syndrome;
increasing the growth rate of a patient having partial growth
hormone insensitive syndrome, accelerating the recovery and
reducing hospitalization of burn patients; treatment of
intrauterine growth retardation, skeletal dysplasia,
hypercortisolism and Cushing's syndrome; induction of pulsatile
growth hormone release; replacement of growth hormone in stressed
patients, treatment of osteochondrodysplasias, Noonan's syndrome,
schizophrenia, depressions, Alzheimer's disease, delayed wound
healing and psychosocial deprivation, treatment of catabolism in
connection with pulmonary dysfunction and ventilator dependency;
treatment of cardiac failure or related vascular dysfunction,
treatment of impaired cardiac function, treatment or prevention of
myocardial infarction, lowering blood pressure, protection against
ventricular dysfunction or prevention of reperfusion events;
treatment of adults in chronic dialysis; attenuation of protein
catabolic responses after major surgery, reducing cachexia and
protein loss due to chronic illness such as cancer or AIDS;
treatment of hyperinsulinemia including nesidioblastosis, adjuvant
treatment for ovulation induction; stimulation of thymic
development and prevention of the age-related decline of thymic
function, treatment of immunosuppressed patients; treatment of
sarcopenia, treatment of wasting in connection with AIDS;
improvement in muscle strength, mobility, maintenance of skin
thickness, metabolic homeostasis and renal homeostasis in the frail
elderly, stimulation of osteoblasts, bone remodelling and cartilage
growth; regulation of food intake; stimulation of the immune system
in companion animals and treatment of disorder of aging in
companion animals, promoting growth in livestock and stimulation of
wool growth in sheep, increasing milk production in livestock,
treatment of metabolic syndrome (syndrome X), treatment of insulin
resistance, including NIDDM, in mammals, e.g. humans, treatment of
insulin resistance in the heart, improvement of sleep quality and
correction of the relative hyposomatotropism of senescence due to
high increase in REM sleep and a decrease in REM latency, treatment
of hypothermia, treatment of frailty associated with ageing,
treatment of congestive heart failure, treatment of hip fractures,
treatment of immune deficiency in individuals with a depressed
T4/T8 cell ratio, treatment of muscular atrophy, treatment of
musculoskeletal impairment in elderly, enhancing the activity of
protein kinase B (PKB), improvement of the overall pulmonary
function, treatment of sleep disorders, treatment of growth
retardation in connection with asthma, treatment of growth
retardation in connection with juvenile rheumatic arthritis, and
treatment of growth retardation in connection with cystic
fibrosis.
[0078] Optionally, the pharmaceutical composition of the invention
may comprise a polymorph of the present invention combined with one
or more compounds exhibiting a different activity, e.g., an
antibiotic or other pharmacologically active material.
[0079] Apart from the pharmaceutical use of a polymorph of the
present invention, it may be useful in vitro tools for
investigating the regulation of growth hormone release.
[0080] The polymorphs of the present invention may also be a useful
in vivo tool for evaluating the growth hormone releasing capability
of the pituitary. For example, serum samples taken before and after
administration of the polymorph to humans can be assayed for growth
hormone. Comparison of the growth hormone in each serum sample
would directly determine the ability of the patient's pituitary to
release growth hormone.
[0081] The polymorph of the present invention may be administered
to commercially important animals to increase their rate and extent
of growth, and to increase milk production.
[0082] A further use of the polymorph of the present invention is
in combination with other secretagogues such as GHRP (2 or 6), GHRH
and its analogues, growth hormone and its analogues or somatomedins
including IGF-1 and IGF-2.
Clathrates
[0083] The crystalline anamorelin of any of the foregoing
embodiments is preferably a clathrate, such that the stoichiometry
of water to anamorelin in the crystalline lattice can vary without
impacting the crystalline structure of the molecule. The degree of
hydration (i.e. stoichiometirc ratio of water to anamorelin) can
range from greater than zero to as much as 3 without changing the
crystalline form of the molecule which, as we have noted, is
preferably defined by an X-ray powder diffraction pattern having at
least four 2.theta. values measured using Cu K.sub..alpha.
radiation selected from the group consisting of 10.1, 11.1, 17.6,
20.0, and 20.8. In one embodiment the crystalline anamorelin
clathrate has a degree of hydration of from 0.5 to 2.5. In another
embodiment the crystalline anamorelin clathrate has a degree of
hydration of from 1.0 to 2.0. Moreover, in any of these embodiment,
the crystalline anamorelin clathrate can further include an organic
volatile impurity without impacting the crystalline structure of
the molecule, such as methanol, ethanol, or isopropanol.
Pharmaceutically Acceptable Salts
[0084] The invention also provides pharmaceutically acceptable
salts made from the compounds of this invention, as well as methods
of making such pharmaceutically acceptable salts. Thus, in one
embodiment the invention provides a method of making a
pharmaceutically acceptable salt of anamorelin by reacting
anemorelin free base of the present invention with a
pharmaceutically acceptable acid, preferably hydrochloric acid. In
a particularly preferred embodiment the anamorelin free base is a
crystalline clathrate defined by an X-ray powder diffraction
pattern having at least four 2.theta. values measured using Cu
K.sub..alpha. radiation selected from the group consisting of 10.1,
11.1, 17.6, 20.0, and 20.8. In one embodiment the crystalline
anamorelin clathrate has a degree of hydration of from 0.5 to 2.5.
In another embodiment the crystalline anamorelin clathrate has a
degree of hydration of from 1.0 to 2.0.
[0085] The invention will now be described more specifically by the
examples.
Example 1
Crystallization of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form A
[0086] 0.0103 g of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was dissolved in methanol (0.1
mL) in a glass vial. The glass vial was then covered with
PARAFILM.RTM. (thermoplastic film) which was perforated with a
single hole. The solvent was then allowed to evaporate under
ambient conditions. An X-ray diffraction pattern showed the
compound was crystalline (FIG. 1).
Example 2
Crystallization of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form B
[0087] 0.0124 g of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was dissolved in methanol:water
(2:1) mixture (0.15 mL) in a glass vial. The glass vial was then
covered with PARAFILM.RTM. (thermoplastic film) which was
perforated with a single hole. The solvent was then allowed to
evaporate under ambient conditions. An X-ray diffraction pattern
showed the compound was crystalline (FIG. 2).
Example 3
Crystallization of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form C
[0088] 0.1003 g of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was added to methanol (2 mL) in a
glass vial. The mixture was filtered into a clean vial. The glass
vial was then covered with PARAFILM.RTM. (thermoplastic film) which
was perforated with a single hole. The solvent was then allowed to
evaporate under ambient conditions. XRPD analysis of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide indicated that the molecule was
crystalline [data not shown].
Example 4
Crystallization of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form C
[0089]
{1-[(1R)-2-](3R)-3-Benzyl-3-(N,N',N'-trimethylhydrazinocarbonyl)pip-
eridin-1-yl-]-1-(1H-indol-3-ylmethyl)-2-oxo-ethylcarbamoyl]-1-methylethyl}-
-carbamic acid tert-butyl ester (4.72 kg) was dissolved in methanol
(5 L/kg) and heated to 55+/-5.degree. C. Methanesulfonic acid (1.5
equiv) was added dropwise over 45 minutes while maintaining a
temperature of 55+/-5..degree. C. After the addition was complete,
the batch temperature was increased and maintained at
60+/-5.degree. C. The progress of the reaction was monitored using
HPLC Upon the completion of the deprotection, the batch was heated
to 70+/-5.degree. C. and a solution of potassium hydroxide (3
equiv) in water (5 L/kg of Boc-protected compound) was added
dropwise over 2 hours while maintaining the temperature. The heat
was then removed from the vessel and the batch was allowed to cool
to 22.degree. C. over 41 hours.
[0090] The batch was filtered and the cake was washed with 50%
aqueous methanol followed by water. The filter cake was then
slurried in water at 50+/-5.degree. C. for 24 hours, filtered, and
washed with water. The filter cake was again slurried in water at
50+/5.degree. C. for 17 hours, filtered, and washed with water. The
product was then loaded onto trays and dried under vacuum at
70+/-5.degree. C. The final yield of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was 3.25 kg (81%) with a purity
of 99.8 area % by HPLC Moisture of this product was determined
using Karl Fischer in two separate measurements to be 2.4% and
3.2%, which indicates that the sample is in a monohydrate form In
an alternative embodiment the steps of slurring the filter cake in
purified water at 50+/-5.degree. C. for 24 hours, refiltering, and
washing with purified water, and repeating the slurry/filter/wash
process can be omitted.
Example 5
Preparation of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form D
[0091] 15 g of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was produced by the method
described in Example 4 except that the steps in which the filter
cake was reslurried in water at 50+/-5.degree. C. were omitted and
the product was dried at room temperature in a vacuum oven. This
sample was charged to a vessel with 76 mL (5 vol) water. The
mixture was heated to 50.degree. C. under moderate stirring
overnight (16 h). The still warm mixture was filtered through
Whatman #1 filter paper, washed with water (15 mL; 1 vol) and dried
in a vacuum oven at 60.degree. C. for 54 hours and then dried at
70. OC for 120 hours. An X-ray diffraction pattern showed the
compound was crystalline (FIG. 3).
Example 6
Crystallization of (3R)-1-(2-methylalanyl-D-tryptophyl)-3
[0092]
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinec-
-arboxylic acid 1,2,2-trimethylhydrazide amorphous was suspended in
methanol (5 L/kg) and the mixture was heated to 65.degree. C. while
being stirred. Water (4 L/kg) was then added slowly while the
temperature was maintained between 65-68.degree. C. After the
addition was complete the mixture was cooled to 60.degree. C. at a
rate of 20.degree. C./h, then from 60.degree. C. to 45.degree. C.
at a rate of 5.degree. C./h, and then from 45.degree. C. to ambient
temperature at a rate of 15.degree. C./h. The crystals were
filtered and washed with methanol-water (5:4 1.5 L/kg) and dried in
vacuo.
[0093] Using this protocol approximately 175 g of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide with a purity of 99.3% area was
isolated under traceable conditions from crude
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide (approximately 90 area %).
[0094] XRPD analysis of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide indicated that the molecule was
crystalline [data not shown]. KF analysis showed that the sample
contained 2.1% water. OVI results showed that the product contained
methanol of 1.9%.
Example 7
Hygroscopicity study of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide (form C)
[0095] The hygroscopicity of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form C produced by the method
disclosed in Example 4 (monohydrate) was evaluated using Dynamic
Vapor Sorption analysis (DVS) and Thermogravimetric analysis
(TGA).
Dynamic Vapor Sorption Analysis (DVS)
[0096] The samples were analyzed using a Hiden IGAsorp vapor
sorption analyzer. Each sample was placed in a sample pan and
exposed to drying at 25.degree. C. in a dry nitrogen stream for two
hours. This assured that the samples being analyzed were at a
constant weight prior to the start of the analysis. The samples
were then analyzed at 25.0.degree. C. The adsorption isotherm was
conducted between: 10-90% RH with a step size of 10%. The
desorption isotherm was conducted between 85-0% RH with a step size
10%. After the isotherms were complete, each of the samples were
heated to 125.degree. C. until the weight loss curve reached an
asymptote or for 4 hours.
Thermogravimetric Analysis (TGA)
[0097] The samples were analyzed using a Mettler 851e
thermogravimetric analyzer. Samples were weighed into a crucible.
The samples were heated from 30.degree. C. to 300.0.degree. C.
ramped at 10.00.degree. C./min with an air purge.
[0098] The
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperid-
-inecarboxylic acid 1,2,2-trimethylhydrazide initially contained
2.5% by weight bound water (shown by TGA analysis). Bound water was
identified by a step transition in the TGA thermogram. DVS analysis
showed that during the adsorption isotherm the sample adsorbed
water steadily until it contained 6% by weight at 35% RH which
indicated the presence of a stable dihydrate. No other significant
additional adsorption was observed up to 90% RH. During the
desorption isotherm, the dihydrate form was observed down to 30% RH
after which desorption of moisture occurred.
[0099] In conclusion DVS results showed that the dihydrate was
formed above a relative humidity of 35%.
Example 8
Preparation of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form C dihydrate and
characterization of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form C monohydrate and
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide form C dihydrate
[0100] One gram of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide produced by the method of Example
4 (monohydrate) was spread evenly on a Petri dish and equilibrated
in a desiccator over a saturated solution of BaCl.sub.22H.sub.2O
for 2.5 days. The relative humidity in this environment was
.about.90%. The conversion to the dihydrate was confirmed by sample
weight measurement before and after equilibration.
[0101] The starting material (monohydrate) and dihydrate were then
analyzed by DSC, and KF. XRPD was performed which investigated if
moisture sorption altered the crystal lattice of the starting
material.
X-Ray Powder Diffraction (XRPD)
[0102] The X-ray powder diffraction patterns were measured on a
Shimadzu XRD-6000 X-ray powder diffractometer using Cu
K.sub..alpha. radiation. Samples were placed on Si zero-return
micro samples holders. The tube voltage and amperage were set to 40
kV and 40 mA, respectively. The divergence and scattering slits
were set at 10 and the receiving slit was set at 0.30 mm. A
theta-two theta continuous scan at 2.degree./min from 3.0 to
45.degree. 2.theta., with a sampling pitch of 0.02 deg and a preset
time of 0.60 seconds.
Differential Scanning Calorimetry Analysis (DSC)
[0103] Samples were analyzed using a Mettler 821e Differential
Scanning Calorimeter DSC Samples were weighed in pan, covered with
a pierced lid and then crimped (sealed). Analysis conditions were
30-300.0.degree. C. ramped at 10.degree. C./min with a nitrogen
purge.
[0104] XRPD analysis of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide monohydrate indicated that the
molecule was crystalline. DSC analysis showed an endothermic
transition with onset of 109.5.degree. C. and peak value of
116.4.degree. C. The extended onset was believed to be due to
dehydration of the material. KF analysis showed that the sample
contained 2.4% water. This corresponded to an average molar ratio
of water/sample of 0.8 or =.about.1, which would indicate the
compound was a monohydrate.
[0105] XRPD results of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide dihydrate indicated that the
molecule was crystalline (FIG. 4) and exhibited a similar pattern
to the monohydrate starting material. DSC analysis showed a broad
endotherm centered at 68.5.degree. C. due to water loss followed by
the melt endotherm with onset of 110.degree. C. and peak of
116.8.degree. C. (FIG. 5). KF analysis showed that the sample
contained 5.6% water. This corresponded to an average molar ratio
of water/sample of 1.7 or .about.2.0, which would indicate the
compound was a dihydrate.
[0106] XRPD results indicated that both the monohydrate and its
dihydrate were crystalline and exhibited similar powder patterns.
DSC and KF results confirmed the formation of the dihydrate from
the monohydrate.
Example 9
Stability of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide
[0107] Crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was placed in double plastic
bags, closed under a nitrogen blanket, and placed inside a small
fiberboard container, the physical appearance, water content,
purity and crystallinity were tested every three months for a
period of 12 months under a variety of conditions. The results of
this study can be seen in Table 5.
[0108] The purity and water content of
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was analyzed using HPLC and KF
respectively before, during, and after the 12 month stability
study. As can be seen from Table 5 the purity of crystalline
(3R)-1-(2-methylalanyl-D-tryptophyl)-3-(phenylmethyl)-3-piperidinecarboxy-
lic acid 1,2,2-trimethylhydrazide was relatively unchanged after 12
months. Karl Fischer Analysis found that the water content
increased from 3.2% to approximately 6% after 12 months.
TABLE-US-00005 TABLE 5 12 month stability study Test Specifi-
Storage Interval (Months) (Test Minded) cations Conditions Initial
1 3 6 9 12 18 24 36 Physical White to 5.degree. C. Off-white
Off-white Off-white Off-white White Off-white Description off-white
solid solid solid solid solid solid (TM-106) solid 25.degree. C.
60% RH Off-white Off-white Off-white Off-white Off-white solid
solid solid solid solid 30.degree. C. 60% RH -- -- -- -- --
40.degree. C. 60% RH Off-white Off-white Off-white -- -- solid
solid solid Water content For 5.degree. C. 3_% 5.3% 6.0% 5.8% 5.6%
6.1% by Karl Fisher Informa- 25.degree. C. 60% RH 5.8% 6.0% 5.9%
5.7% 5.8% (USP <921> tion 30.degree. C. 60% RH -- -- -- -- --
method 1 of Only 40.degree. C. 75% RH 6.1% 6.1% 5.9% -- -- SOP
INS002) HPLC Weight For 5.degree. C. 99.8% 92.0% 93.2% 93.9% 94.5%
94.3% % (TM-808) Informa- 25.degree. C. 60% RH 92.1% 93.2% 94.2%
94.6% 94.2% tion 30.degree. C. 60% RH -- -- -- -- -- Only
40.degree. C. 75% RH 91.8% 93.0% 94.1% -- -- HPLC Area % Not Less
5.degree. C. 99.8% 99.7% 99.7% 99.7 99.8% 99.7% (TM-808).sup.1 Than
25.degree. C. 60% RH 99.7% 99.7% 99.7 99.8% 99.7% 95% 30.degree. C.
60% RH -- -- -- -- -- 40.degree. C. 75% RH 99.7% 99.7% 99.7 -- --
DSC (USP For 5.degree. C. Onset Onset: Onset: Onset: -- --
<891>, SUP Informa- 108.2.degree. C. 81.8.degree. C.
112.1.degree. C. 112.3.degree. C. INS034) tion Peak: Peak: Peak:
Peak: Only 112.7.degree. C. 110.4.degree. C. 117.1.degree. C.
116.9.degree. C. Onset: 116.0.degree. C. Peak: 118.4.degree. C.
25.degree. C. 60% RH Onset: Onset: Onset: -- -- 107.0.degree. C.
112.6.degree. C. 112.5.degree. C. Peak: Peak: Peak: 110.3.degree.
C. 117.7.degree. C. 116.9% Onset: 121.3.degree. C. Peak:
122.2.degree. C. 30.degree. C. 60% RH -- -- -- -- -- 40.degree. C.
75% RH Onset: Onset: Onset: -- -- 89.2.degree. C. 112.2.degree. C.
125.5.degree. C. Peak: Peak: Peak: 110.3.degree. C. 117.5.degree.
C. 117.8.degree. C. Onset: 129.9.degree. C. Peak: 134.9.degree. C.
Onset: 151.1.degree. C. Peak: 165.2.degree. C. Crystallinity For
5.degree. C. Crystal- Crystal- Crystal- Crystal- Crystal- Crystal-
using Optical Informa- lize lize lize lize lize lize Microscopy
tion 25.degree. C. 60% RH Crystal- Crystal- Crystal- Crystal-
Crystal- (USP<695>) only lize lize lize lize lize 30.degree.
C. 60% RH -- -- -- -- -- 40.degree. C. 75% RH Crystal- Crystal-
Crystal- -- -- lize lize lize
[0109] While this invention has been particularly shown and
described with references to preferred embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
scope of the invention encompassed by the appended claims.
* * * * *