U.S. patent application number 15/345234 was filed with the patent office on 2017-10-05 for crystalline and amorphous forms of a beta-arrestin effector.
The applicant listed for this patent is Trevena, Inc.. Invention is credited to Gregory Alcorn, Yuriy B. Kalyan, Graham Richard Lawton, Matthew Ronsheim, Ritesh Sanghvi, Jiaher Tian, Meiki Yu, Shao Hong Zhou.
Application Number | 20170283464 15/345234 |
Document ID | / |
Family ID | 53774370 |
Filed Date | 2017-10-05 |
United States Patent
Application |
20170283464 |
Kind Code |
A1 |
Sanghvi; Ritesh ; et
al. |
October 5, 2017 |
Crystalline And Amorphous Forms Of A Beta-Arrestin Effector
Abstract
The present disclosure provides novel crystalline forms of a
compound that acts as a .beta.-arrestin effector, processes for
preparing novel crystalline and amorphous forms of a compound that
acts as a .beta.-arrestin effector and uses thereof.
Inventors: |
Sanghvi; Ritesh; (East
Northport, NY) ; Alcorn; Gregory; (Valley Forge,
PA) ; Lawton; Graham Richard; (Smithtown, NY)
; Yu; Meiki; (Jamaica, NY) ; Ronsheim;
Matthew; (Port Jefferson, NY) ; Tian; Jiaher;
(Syosset, NY) ; Zhou; Shao Hong; (Commack, NY)
; Kalyan; Yuriy B.; (Staten Island, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Trevena, Inc. |
King of Prussia |
PA |
US |
|
|
Family ID: |
53774370 |
Appl. No.: |
15/345234 |
Filed: |
November 7, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14616487 |
Feb 6, 2015 |
9518086 |
|
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15345234 |
|
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61936914 |
Feb 7, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07K 7/64 20130101; A61K
38/12 20130101; A61P 9/04 20180101; A61P 43/00 20180101; C07K 7/06
20130101; A61K 38/00 20130101; A61P 13/12 20180101; A61P 9/00
20180101; A61P 9/12 20180101 |
International
Class: |
C07K 7/64 20060101
C07K007/64; C07K 7/06 20060101 C07K007/06; A61K 38/12 20060101
A61K038/12 |
Claims
1-42. (canceled)
43. A process for preparing a crystalline form of a peptide having
a sequence of SEQ. ID. NO.1, comprising crystallizing SEQ. ID. NO.
1 to form a crystalline form of a peptide having a sequence of SEQ.
ID. NO. 1.
44. The process of claim 43, wherein the crystallizing comprises
dissolving peptide in an aqueous solution and crystallizing the
peptide to form the crystalline form therefrom.
45. The process of claim 44, wherein the aqueous solution is
deionized water.
46. The process of claim 44, wherein the aqueous solution consists
of deionized water.
47. The process of claim 44, wherein the process further comprises
precipitating the peptide after dissolving the peptide in the
aqueous solution.
48. The process of claim 47, wherein the precipitating comprises
heating the aqueous solution and cooling the aqueous solution to
ambient temperature to precipitate the peptide in the crystalline
form.
49. The process of claim 48, wherein the process further comprises
cooling the aqueous solution to a temperature of about 0 to about
5.degree. C. to precipitate the peptide in the crystalline
form.
50. The process of claim 43, further comprising lyophilizing the
crystalline form.
51. A pharmaceutical composition comprising a crystalline form of a
peptide having a sequence of SEQ. ID. NO.1.
52. A method of treating a cardiovascular or a cardiorenal disorder
comprising administering to a patient in need thereof the
pharmaceutical composition of claim 51.
53. A crystalline form of a peptide having a sequence of SEQ. ID.
NO. 1.
54. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising a peak at about 18.5.+-.0.5
degrees 2.theta..
55. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising a peak at about 10.1.+-.0.5
degrees 2.theta..
56. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising a peak at about 8.2.+-.0.5
degrees 2.theta..
57. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising a peak at about 20.2.+-.0.5
degrees 2.theta..
58. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising a peak at about 24.4.+-.0.5
degrees 2.theta..
59. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising peaks at about 18.5, and at
about 10.1.+-.0.5 degrees 2.theta..
60. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising peaks at about 10.1, and at
about 8.2.+-.0.5 degrees 2.theta..
61. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2, and at
about 20.2.+-.0.5 degrees 2.theta..
62. The crystalline form of claim 53, characterized by an X-ray
powder diffraction pattern comprising peaks at about 20.2, and at
about 10.1.+-.0.5 degrees 2.theta..
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to and the benefit of U.S.
Provisional Application No. 61/936,914, filed Feb. 7, 2014, which
is hereby incorporated by reference in its entirety.
FIELD
[0002] The present disclosure describes novel crystalline forms of
a compound that acts as .beta.-arrestin effector, processes for
preparing and precipitating amorphous and crystalline forms of the
compound, and uses thereof.
BACKGROUND
[0003] U.S. Pat. No. 8,486,885 discloses peptides that act as GPCR
agonist of GPCR receptors (e.g., angiotensin II). GPCR agonist
causes activation of a heterotrimeric "G protein". Such activation
leads to second messenger/down-stream signaling (e.g., via
diacylglycerol, inositol-triphosphate, calcium, etc.) causing
changes in physiological function (e.g., blood pressure and fluid
homeostasis). One particular peptide disclosed in U.S. Pat. No.
8,486,885 is referred to therein as "SEQ ID NO. 27", which has the
following amino acid sequence: NH.sub.2-Sarcosine L-Arginine
L-Valine L-Tyrosine L-Isoleucine L-Histidine L-Proline D-Alanine
--OH referred to as NH.sub.2-Sar Arg Val Tyr Ile His Pro
D-Ala-OH.
[0004] SEQ ID NO. 27 referred to in U.S. Pat. No. 8,486,885
(hereinafter referred as SEQ. ID. NO. 1) is an agonist of
.beta.-arrestin/GRK-mediated signal transduction via AT1
angiotensin receptor. The amino acid sequence, including, but not
limited to, formula, variables, derivatives, of the peptide or
peptide mimetic of SEQ ID NO. 1, the ability of the compound to
effect G protein-mediated signaling or GPCR activity, or the
absence of such signaling/activity, methods for preparation of SEQ.
ID. NO. 1, and other related peptides are disclosed in U.S. Pat.
No. 8,486,885, the contents of which are incorporated herein by
reference in their entirety.
[0005] There remains a need in the art for improved forms of SEQ.
ID. NO. 1 with improved properties. There also remains a need in
the art for improved processes for preparing the peptide of SEQ.
ID. NO. 1.
SUMMARY
[0006] The present disclosure provides novel crystalline
modifications of the peptide of SEQ. ID. NO. 1, processes for
preparing SEQ. ID. NO. 1, and optionally isolating such forms.
[0007] Surprisingly, the peptide of SEQ ID NO. 1 can be
crystallized and is superior in properties. Surprisingly, amorphous
SEQ ID NO. 1 can be prepared, by precipitating SEQ. ID. NO. 1.
Crystalline forms of SEQ ID NO. 1 are distinguished from prior art
by improved stability, processability and can also be used in for
pharmaceutical formulations.
[0008] In some embodiments, crystalline forms of SEQ. ID. NO. 1 are
provided. In some embodiments, the crystalline form is Form I of
SEQ. ID. NO. 1 (hereinafter, Form I).
[0009] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 18.5.+-.0.5
degrees 2.theta..
[0010] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 10.1.+-.0.5
degrees 2.theta..
[0011] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 8.2.+-.0.5
degrees 2.theta..
[0012] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 20.2.+-.0.5
degrees 2.theta..
[0013] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 24.4.+-.0.5
degrees 2.theta..
[0014] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 18.5, and at
about 10.1.+-.0.5 degrees 2.theta..
[0015] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 10.1, and at
about 8.2.+-.0.5 degrees 2.theta..
[0016] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2, and at
about 20.2.+-.0.5 degrees 2.theta..
[0017] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 20.2, and at
about 10.1.+-.0.5 degrees 2.theta..
[0018] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 20.2, and at
about 24.4.+-.0.5 degrees 2.theta..
[0019] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 20.2, at about
10.1, and at about 24.4.+-.0.5 degrees 2.theta..
[0020] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2, at about
18.5, and at about 20.2.+-.0.5 degrees 2.theta..
[0021] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 18.5, at about
10.1, at about 8.2, at about 20.2, and at about 24.4.+-.0.5 degrees
2.theta..
[0022] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising one or more peaks as shown in
FIG. 4.
[0023] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising one or more d-spacing values
at about 10.7, at about 8.7, at about 4.7, at about 4.1, and at
about 3.6.+-.0.5 degrees angstroms.
[0024] In some embodiments, a pharmaceutical composition comprising
a crystalline form of SEQ. ID. NO. 1 is provided.
[0025] In some embodiments, a pharmaceutical composition comprising
a crystalline Form I of SEQ. ID. NO. 1 is provided.
[0026] In some embodiments, the pharmaceutical composition
comprises Form I, wherein Form I is a peptide or a peptide mimetic
of SEQ. ID. NO. 1.
[0027] In some embodiments, the pharmaceutical composition
comprises Form I, wherein the peptide or a peptide mimetic is
cyclic.
[0028] In some embodiments, the pharmaceutical composition
comprises Form I, wherein the peptide or a peptide mimetic is
dimerized.
[0029] In some embodiments, the pharmaceutical composition
comprises Form I, wherein the peptide or a peptide mimetic is
trimerized.
[0030] In some embodiments, the pharmaceutical composition
comprises Form I, further comprising an additional drug for the
treatment of a cardiovascular or a cardio renal disorder.
[0031] In some embodiments, a process for preparing a crystalline
form of SEQ. ID. NO. 1, comprising crystallizing SEQ. ID. NO. 1 to
form Form I and optionally isolating the Form I of SEQ. ID. NO. 1
is provided.
[0032] In some embodiments, a process for preparing SEQ. ID. NO. 1,
comprising precipitating SEQ. ID. NO. 1 and optionally isolating
SEQ. ID. NO. 1 is provided.
[0033] In some embodiments, a pharmaceutical composition comprising
SEQ. ID. NO.1 prepared by precipitating SEQ. ID. NO.1 is
provided.
[0034] In some embodiments, a method of treating a cardiovascular
or a cardiorenal disorder comprising administering to a patient in
need thereof, a crystalline or an amorphous form of SEQ. ID. NO. 1
is provided.
[0035] In some embodiments, a method of treating a cardiovascular
or a cardiorenal disorder comprising administering to a patient in
need thereof, a crystalline Form I of SEQ. ID. NO. 1 is
provided.
[0036] The details of one or more embodiments are set forth in the
description below. Other features, objects, and advantages of the
present teachings will be apparent from the description of examples
and also from the appending claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0037] FIG. 1 shows X-ray powder diffraction pattern of amorphous
SEQ ID. NO. 1.
[0038] FIG. 2 shows Differential Scanning Calorimetry (DSC)
thermogram of amorphous SEQ. ID. NO. 1.
[0039] FIG. 3 shows Sorption/Desorption profile of amorphous SEQ
ID. NO. 1.
[0040] FIG. 4 shows X-ray powder diffraction pattern of crystalline
Form I of SEQ ID. NO. 1.
[0041] FIG. 5 shows Differential Scanning Calorimetry (DSC)
thermogram of crystalline Form I of SEQ. ID. NO. 1.
[0042] FIG. 6 shows HPLC chromatogram of Form I of SEQ ID. NO.
1.
[0043] FIG. 7 shows overlaid X-ray powder diffraction patterns of
amorphous SEQ ID. NO. 1 and crystalline Form I of SEQ ID. NO.
1.
[0044] FIG. 8 shows HPLC chromatograms of amorphous and crystalline
Form I of SEQ ID. NO. 1.
[0045] FIG. 9 shows microscopic image of crystalline Form I of SEQ
ID. NO. 1 under cross polarized light.
[0046] FIG. 10 shows microscopic image and particle size of
crystalline Form I of SEQ ID. NO. 1 under plane polarized
light.
[0047] FIG. 11 shows the overlaid X-ray powder diffraction patterns
for the precipitate of SEQ. ID. NO. 1 from ethanol and amorphous
SEQ. ID. NO. 1.
[0048] FIG. 12 shows the X-ray powder diffraction pattern for the
precipitate of SEQ. ID. NO. 1 from water and acetone.
[0049] FIG. 13 shows the X-ray powder diffraction pattern for the
precipitate of SEQ. ID. NO. 1 from water and isopropyl alcohol.
[0050] FIG. 14 shows overlaid X-ray powder diffraction patterns for
Form I of SEQ. ID. NO. 1 and amorphous SEQ. ID. NO. 1.
Corresponding DSC thermogram is shown in FIG. 16.
[0051] FIG. 15 shows overlaid X-ray powder diffraction pattern for
Form I of SEQ. ID. NO. 1 and amorphous SEQ. ID. NO. 1.
Corresponding DSC thermogram is shown in FIG. 16.
[0052] FIG. 16 shows a DSC thermogram for compounds produced
according to Examples 1E, 1F, 1G, and 1H.
[0053] FIG. 17 shows overlaid X-ray powder diffraction pattern for
Form I of SEQ. ID. NO. 1 and amorphous SEQ. ID. NO. 1.
Corresponding DSC thermogram is shown in FIG. 16.
[0054] FIG. 18 shows overlaid X-ray powder diffraction pattern for
Form I of SEQ. ID. NO. 1 and amorphous SEQ. ID. NO. 1.
Corresponding DSC thermogram is shown in FIG. 16.
[0055] FIG. 19 shows a TGA profile of crystalline SEQ. ID.
NO.1.
[0056] FIG. 20 shows FIG. 20 a DVS profile of crystalline SEQ. ID.
NO. 1.
[0057] FIG. 21 shows the PXRD of crystalline form SEQ. ID. NO.1
after completion of DVS analysis.
[0058] FIG. 22 shows PXRD patterns of amorphous SEQ. ID. NO. 1 at
various temperature and humidity conditions.
[0059] FIG. 23 shows PXRD patterns of crystalline SEQ. ID. NO.1 at
various temperature and humidity conditions.
DEFINITIONS
[0060] The terms "peptidyl" and "peptidic" include active
derivatives, variants, and/or mimetics of the peptides according to
the present embodiments. Peptidic compounds are structurally
similar bioactive equivalents of the peptides according to the
present embodiments.
[0061] The term "structurally similar bioactive equivalent" means a
peptidyl compound with structure sufficiently similar to that of an
identified bioactive peptide to produce substantially equivalent
therapeutic effects. For example, peptidic compounds derived from
the amino acid sequence of the peptide, or having an amino acid
sequence backbone of the peptide, are considered structurally
similar bioactive equivalents of the peptide.
[0062] The term "variant" refers to a protein or polypeptide in
which one or more amino acid substitutions, deletions, and/or
insertions are present as compared to the amino acid sequence of a
protein or peptide and include naturally occurring allelic variants
or alternative splice variants of a protein or peptide.
[0063] The term "variant" includes the replacement of one or more
amino acids in a peptide sequence with a similar or homologous
amino acid(s) or a dissimilar amino acid(s). In some embodiments,
variants include alanine substitutions at one or more of amino acid
positions. Other preferred substitutions include conservative
substitutions that have little or no effect on the overall net
charge, polarity, or hydrophobicity of the protein.
[0064] The term "variant" also encompasses polypeptides that have
the amino acid sequence of the proteins/peptides of the present
compounds with at least one and up to 25 (e.g., 1, 2, 3, 4, 5, 6,
7, 8, 9, 10, 15, 20) additional amino acids flanking either the 3'
or 5' end of the amino acid sequence or both. The term "variant"
also refers to a protein that is at least 60 to 99 percent
identical (e.g., 60, 65, 70, 75, 80, 85, 90, 95, 98, 99, or 100%,
inclusive) in its amino acid sequence of the proteins of the
present compounds as determined by standard methods that are
commonly used to compare the similarity in position of the amino
acids of two polypeptides. The degree of similarity or identity
between two proteins can be readily calculated by known
methods.
[0065] The term "derivative" refers to a chemically modified
protein or polypeptide that has been chemically modified either by
natural processes, such as processing and other post-translational
modifications, but also by chemical modification techniques, as for
example, by addition of one or more polyethylene glycol molecules,
sugars, phosphates, and/or other such molecules, where the molecule
or molecules are not naturally attached to wild-type proteins.
Derivatives include salts. Such chemical modifications are well
described in basic texts and in more detailed monographs, as well
as in research literature and they are well known to those of skill
in the art. It will be appreciated that the same type of
modification may be present in the same or varying degree at
several sites in a given protein or polypeptide. Also, a given
protein or polypeptide may contain many types of modifications.
Modifications can occur anywhere in a protein or polypeptide,
including the peptide backbone, the amino acid side-chains, and the
amino or carboxyl termini. Modifications include, for example,
acetylation, acylation, ADP-ribosylation, amidation, covalent
attachment of flavin, covalent attachment of a heme moiety,
covalent attachment of a nucleotide or nucleotide derivative,
covalent attachment of a lipid or lipid derivative, covalent
attachment of phosphotidylinositol, cross-linking, cyclization,
disulfide bond formation, demethylation, formation of covalent
cross-links, formation of cysteine, formation of pyroglutamate,
formylation, gamma-carboxylation, glycosylation, GPI anchor
formation, hydroxylation, iodination, methylation, myristoylation,
oxidation, proteolytic processing, phosphorylation, prenylation,
racemization, glycosylation, lipid attachment, sulfation,
gamma-carboxylation of glutamic acid residues, hydroxylation and
ADP-ribosylation, selenoylation, sulfation, transfer-RNA mediated
addition of amino acids to proteins, such as arginylation, and
ubiquitination. See, for instance, Proteins-Structure and Molecular
Properties, 2.sup.nd Ed., T. E. Creighton, W. H. Freeman and
Company, New York (1993) and Wold, F., "Posttranslational Protein
Modifications: Perspectives and Prospects," pages. 1-12 in
Posttranslational Covalent Modification Of Proteins, B. C. Johnson,
Ed., Academic Press, New York (1983); Seifter et al., Meth.
Enzvmol. 182:626-646 (1990) and Rattan et al., "Protein Synthesis:
Posttranslational Modifications and Aging," Ann N.Y. Acad. Sci.
663: 48-62 (1992).
[0066] The term "derivatives" include chemical modifications
resulting in the protein or polypeptide becoming branched or
cyclic, with or without branching. Cyclic, branched and branched
circular proteins or polypeptides may result from
post-translational natural processes and may be made by entirely
synthetic methods, as well.
[0067] The term "peptide mimetic" or "mimetic" refers to
biologically active compounds that mimic the biological activity of
a peptide or a protein but are no longer peptidic in chemical
nature, that is, they no longer contain any peptide bonds (that is,
amide bonds between amino acids). The term peptide mimetic is used
in a broader sense to include molecules that are no longer
completely peptidic in nature, such as pseudo-peptides,
semi-peptides and peptoids. Whether completely or partially
non-peptide, peptide mimetics according to the embodiments provide
a spatial arrangement of reactive chemical moieties that closely
resemble the three-dimensional arrangement of active groups in the
peptide on which the peptide mimetic is based. As a result of this
similar active-site geometry, the peptide mimetic has effects on
biological systems that are similar to the biological activity of
the peptide.
[0068] The peptide mimetics of the embodiments are preferably
substantially similar in both three-dimensional shape and
biological activity to the peptide described herein. According to
some embodiments, peptide mimetics of the present compounds have
protective groups at one or both ends of the compounds, and/or
replacement of one or more peptide bonds with non-peptide bonds.
Such modifications may render the compounds less susceptible to
proteolytic cleavage than the compound itself. For instance, one or
more peptide bonds can be replaced with an alternative type of
covalent bond (e.g., a carbon-carbon bond or an acyl bond). Peptide
mimetics can also incorporate amino-terminal or carboxyl terminal
blocking groups such as t-butyloxycarbonyl, acetyl, alkyl,
succinyl, methoxysuccinyl, suberyl, adipyl, azelayl, dansyl,
benzyloxycarbonyl, fluorenylmethoxycarbonyl, methoxyazelayl,
methoxyadipyl, methoxysuberyl, and 2,4,-dinitrophenyl, thereby
rendering the mimetic less susceptible to proteolysis. Non-peptide
bonds and carboxyl- or amino-terminal blocking groups can be used
singly or in combination to render the mimetic less susceptible to
proteolysis than the corresponding peptide/compound. Additionally,
substitution of D-amino acids for the normal L-stereoisomer can be
effected, e.g., to increase the half-life of the molecule.
[0069] The term "salt" or "salts" may refer to any acid addition
salts, including addition salts of free acids or addition salts of
free bases. All of these salts (or other similar salts) may be
prepared by conventional means. All such salts are acceptable
provided that they are non-toxic and do not substantially interfere
with the desired pharmacological activity.
[0070] The term "therapeutically effective amount" means the amount
of a compound that, when administered to a mammal for treating a
state, disorder or condition is sufficient to effect a treatment
(as defined below). The "therapeutically effective amount" will
vary depending on the compound, the disease and its severity, the
age, weight, physical condition and responsiveness of the mammal to
be treated.
[0071] The term "pharmaceutically acceptable" means biologically or
pharmacologically compatible for in vivo use in animals or humans,
and preferably means approved by a regulatory agency of the Federal
or a State government or listed in the U.S. Pharmacopeia or other
generally recognized pharmacopeia for use in animals, and more
particularly in humans.
[0072] The term "treat", in all its verb forms, means to relieve or
alleviate at least one symptom of a cardiovascular disorder or a
cardiorenal disorder in a subject, including chronic hypertension,
hypertensive crisis, acute congestive heart failure, angina, acute
myocardial infarction, left ventricular failure, cerebrovascular
insufficiency, intracranial haemorrhage, heart failure, acute
decompensated heart failure, essential hypertension, post-operative
hypertension, hypertensive heart disease, hypertensive renal
disease, renovascular hypertension, malignant hypertension,
post-renal transplant patient stabilization, dilated
cardiomyopathy, myocarditis, post-cardiac transplant patient
stabilization, disorders associated with post-stent management,
neurogenic hypertension, pre-eclampsia, abdominal aortic aneurysm,
and any cardiovascular disorder with a hemodynamic component.
Specifically in some aspects, the cardiovascular disorder is an
acute cardiovascular disorder. In other specific aspects, the acute
cardiovascular disorder is acute hypertensive crisis, toxemia of
pregnancy, acute myocardial infarction, acute congestive heart
failure, acute ischaemic heart disease, pulmonary hypertension,
post-operative hypertension, migraine, retinopathy and
post-operative cardiac/valve surgery.
[0073] The term "synergy" is defined as the interaction of two or
more agents so that their combined effect is greater than the sum
of their individual effects. For example, if the effect of drug A
alone in treating a disease is 25%, and the effect of drug B alone
in treating a disease is 25%, but when the two drugs are combined
the effect in treating the disease is 75%, the effect of A and B is
synergistic.
[0074] The term "additive" is defined as the interaction of two or
more agents so that their combined effect is the same as the sum of
their individual effects. For example, if the effect of drug A
alone in treating a disease is 25%, and the effect of drug B alone
in treating a disease is 25%, but when the two drugs are combined
the effect in treating the disease is 50%, the effect of A and B is
additive.
[0075] The term "pharmaceutically acceptable" or "therapeutically
acceptable" refers to molecular entities and compositions that are
physiologically tolerable and preferably do not typically produce
an allergic or similar untoward reaction, such as gastric upset,
dizziness and the like, when administered to a human. Preferably,
as used herein, the term "pharmaceutically acceptable" means
approved by a regulatory agency of the Federal or a State
government or listed in the U.S. Pharmacopeia or other generally
recognized pharmacopeia (e.g., Remington's Pharmaceutical Sciences,
Mack Publishing Co. (A. R. Gennaro edit. 1985)) for use in animals,
and more particularly in humans.
[0076] The term "approximately" means plus or minus 5%.
[0077] Approximately 80% and 20% v/v of ethanol and the remainder
of the solution approximately is water means a solution comprising
between approximately 80% v/v ethanol and 20% v/v water to
approximately 20% v/v ethanol and 80% v/v water.
[0078] Approximately 80% and 20% v/v isopropyl alcohol and the
remainder of the solution approximately is water means a solution
comprising between approximately 80% v/v isopropyl alcohol and 20%
v/v water to approximately 20% v/v isopropyl alcohol and 80% v/v
water.
[0079] Approximately 80% and 20% v/v of acetone and the remainder
of the solution approximately is water means a solution comprising
between approximately 80% v/v acetone and 20% v/v water to
approximately 20% v/v acetone and 80% v/v water.
DETAILED DESCRIPTION
[0080] The present embodiments relate to a synthetic octapeptide,
namely SEQ. ID. NO. 1, having the amino sequence structure as
follows: NH.sub.2-Sar Arg Val Tyr Ile His Pro D-Ala-OH. SEQ. ID NO.
1 is an agonist of .beta.-arrestin/GRK-mediated signal transduction
via the AT1 angiotensin receptor.
[0081] An amorphous form of SEQ ID. NO. 1 can be prepared using
solid-phase peptide synthesis using FMOC based solid state
synthesis on a chlortriyl resin. The crude peptide can be purified
by reverse phase chromatography and ion-exchange can be performed
to remove trifluroacetic acid and to replace it with acetic acid.
The amorphous form of SEQ ID. NO. 1 can then be isolated using
lyophilization. Lyophilization may not be feasible for a large
scale manufacturing of the peptide for commercial production.
[0082] The present application relates to novel methods of
precipitating SEQ. ID. NO. 1 and novel crystalline forms of SEQ.
ID. NO. 1.
[0083] An example of an amorphous form of SEQ. ID. NO. 1, is
illustrated in FIG. 1, which shows a X-ray powder diffraction
pattern of amorphous SEQ ID. NO. 1. FIG. 2 shows an example of a
Differential Scanning Calorimetry (DSC) thermogram of amorphous
SEQ. ID. NO. 1. FIG. 3 shows an example of Sorption/Desorption
profile of amorphous SEQ ID. NO. 1.
[0084] DSC thermogram shown in FIG. 2 exhibits two broad endotherm
at around 76.degree. C. and at around 159.degree. C., as well as
sharp endotherm at around 250.degree. C. SEQ. ID. NO. 1 is highly
hygroscopic as it adsorbed moisture with increase in % RH. More
than about 25% (by weight) moisture was adsorbed at 95% RH at
25.degree. C. as shown in FIG. 3. FIGS. 1-3 show that the peptide
of SEQ. ID. NO. 1 is amorphous.
[0085] In some embodiments, crystalline forms of SEQ ID NO. 1 are
provided. In some embodiments crystalline Form I of SEQ ID NO. 1 is
provided.
[0086] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern substantially as shown in FIG. 4. In
some embodiments, Form I is characterized by an X-ray powder
diffraction pattern comprising one or more peaks as provided in
Table 1. In some embodiments, Form I is characterized by an X-ray
powder diffraction pattern comprising substantially all of, or all
of, the peaks as provided in Table 1.
[0087] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 8.2.+-.0.5
degrees 28.
[0088] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 10.1.+-.0.5
degrees 2.theta..
[0089] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 15.4.+-.0.5
degrees 2.theta..
[0090] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 16.5.+-.0.5
degrees 2.theta..
[0091] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 18.5.+-.0.5
degrees 2.theta..
[0092] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 20.2.+-.0.5
degrees 2.theta..
[0093] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 23.1.+-.0.5
degrees 2.theta..
[0094] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 24.4.+-.0.5
degrees 2.theta..
[0095] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a peak at about 30.8.+-.0.5
degrees 2.theta..
[0096] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2.+-.0.5
degrees 2.theta., about 10.1.+-.0.5 degrees 2.theta., about
18.5.+-.0.5 degrees 2.theta., about 20.2.+-.0.5 degrees 2.theta.,
about 24.4.+-.0.5 degrees 2.theta..
[0097] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 6.1.+-.0.5
degrees 2.theta., about 8.2.+-.0.5 degrees 2.theta., about
10.1.+-.0.5 degrees 2.theta., about 11.4.+-.0.5 degrees 2.theta.,
about 13.2.+-.0.5 degrees 2.theta., about 16.1.+-.0.5 degrees
2.theta., about 18.5.+-.0.5 degrees 2.theta., about 20.2.+-.0.5
degrees 2.theta., about 23.1.+-.0.5 degrees 2.theta., about
24.4.+-.0.5 degrees 2.theta., and about 30.8.+-.0.5 degrees
2.theta..
[0098] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 6.1.+-.0.5
degrees 2.theta., about 8.2.+-.0.5 degrees 2.theta., about
10.1.+-.0.5 degrees 2.theta., about 10.7.+-.0.5 degrees 2.theta.,
about 12.3.+-.0.5 degrees 2.theta., about 14.0.+-.0.5 degrees
2.theta., about 15.4.+-.0.5 degrees 2.theta., about 16.1.+-.0.5
degrees 2.theta., about 17.3.+-.0.5 degrees 2.theta., about
18.5.+-.0.5 degrees 2.theta., about 19.1.+-.0.5 degrees 2.theta.,
about 20.2.+-.0.5 degrees 2.theta., about 20.9.+-.0.5 degrees
2.theta., about 21.5.+-.0.5 degrees 2.theta., about 24.4.+-.0.5
degrees 2.theta., and about 30.8.+-.0.5 degrees 2.theta..
[0099] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2.+-.0.5
degrees 2.theta., and at about 10.1.+-.0.5 degrees 2.theta.. In
some embodiments, the Form I is characterized by an X-ray powder
diffraction pattern comprising peaks at about 10.1.+-.0.5 degrees
2.theta. and at about 18.5.+-.0.5 degrees 2.theta.. In some
embodiments, the Form I is characterized by an X-ray powder
diffraction pattern comprising peaks at about 18.5.+-.0.5 degrees
2.theta. and at about 20.2-0.5 degrees 2.theta.. In some
embodiments, the Form I is characterized by an X-ray powder
diffraction pattern comprising peaks at about 20.2.+-.0.5 degrees
2.theta. and at about 24.4.+-.0.5 degrees 2.theta..
[0100] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2.+-.0.5
degrees 2.theta., at about 10.1.+-.0.5 degrees 2.theta. and at
about 18.5.+-.0.5 degrees 2.theta..
[0101] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 18.5.+-.0.5
degrees 2.theta., at about 20.2.+-.0.5 degrees 2.theta. and at
about 24.4.+-.0.5 degrees 2.theta..
[0102] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2.+-.0.5
degrees 2.theta., at about 10.1.+-.0.5 degrees 2.theta., at about
18.5.+-.0.5 degrees 2.theta. and at about 20.2.+-.0.5 degrees
28.
[0103] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2.+-.0.5
degrees 2.theta., at about 10.1.+-.0.5 degrees 2.theta., at about
18.5.+-.0.5 degrees 2.theta., and optionally one or more peaks at
about 16.5-0.5 degrees 2.theta. and/or about 23.1.+-.0.5 degrees
2.theta..
[0104] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2.+-.0.5
degrees 2.theta., about 10.1.+-.0.5 degrees 2.theta., about
18.5.+-.0.5 degrees 2.theta., about 20.2.+-.0.5 degrees 2.theta.,
about 24.4.+-.0.5 degrees 2.theta. and optionally having one or
more peaks at about at about 6.1.+-.0.5 degrees 2.theta., about
11.4.+-.0.5 degrees 2.theta., about 13.2.+-.0.5 degrees 2.theta.,
about 16.1.+-.0.5 degrees 2.theta., about 23.1.+-.0.5 degrees
2.theta., about 30.8.+-.0.5 degrees 2.theta..
[0105] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks at about 8.2.+-.0.5
degrees 2.theta., about 10.1.+-.0.5 degrees 2.theta., about
18.5.+-.0.5 degrees 2.theta., about 20.2.+-.0.5 degrees 2.theta.,
about 24.4.+-.0.5 degrees 2.theta. and optionally having one or
more peaks at about 6.1.+-.0.5 degrees 2.theta., about 10.7.+-.0.5
degrees 2.theta., about 12.3.+-.0.5 degrees 2.theta., about
14.0.+-.0.5 degrees 2.theta., about 15.4.+-.0.5 degrees 2.theta.,
about 16.1.+-.0.5 degrees 2.theta., about 17.3.+-.0.5 degrees
2.theta., about 19.1.+-.0.5 degrees 2.theta., about 20.2.+-.0.5
degrees 2.theta., about 20.9.+-.0.5 degrees 2.theta., about
21.5.+-.0.5 degrees 2.theta., about 30.8.+-.0.5 degrees
2.theta..
[0106] As used herein, unless otherwise indicated, the phrase "one
or more peaks" should be understood to be inclusive of (i)
crystalline forms that have XRD peaks at every peak value recited
after this phrase, (ii) crystalline forms that have an XRD peak at
only one of the peak values recited after this phrase, as well as
(iii) crystalline forms that have XRD peaks at two or more (e.g.,
three or more, four or more, five or more, six or more, or even
seven or more) of the peak values recited after this phrase.
[0107] In some embodiments, the Form I is characterized by a DSC
thermogram as shown in FIG. 5.
[0108] In some embodiments, the Form I is characterized by any
combination of the above data.
[0109] In some embodiments, the X-ray powder diffraction peaks
recited herein for particular embodiments can vary by .+-.0.4
degrees 2.theta., by .+-.0.3 degrees 2.theta., by .+-.0.2 degrees
2.theta., or by .+-.0.1 degrees 2.theta..
[0110] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising peaks having d-spacing values
at about 10.7, at about 8.7, at about 4.7, at about 4.3, and at
about 3.6.+-.0.5 angstroms.
[0111] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
10.7.+-.0.5 angstroms.
[0112] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
8.7.+-.0.5 angstroms.
[0113] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
5.7.+-.0.5 angstroms.
[0114] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
5.3.+-.0.5 angstroms.
[0115] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
4.7.+-.0.5 angstroms.
[0116] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
4.3.+-.0.5 angstroms.
[0117] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
3.8.+-.0.5 angstroms.
[0118] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value at about
3.6.+-.0.5 angstroms.
[0119] In some embodiments, the Form I is characterized by an X-ray
powder diffraction pattern comprising a d-spacing value
substantially as shown in Table 1.
[0120] In some embodiments, the X-ray powder diffraction peaks
recited herein for particular embodiments having d-spacing values
can vary by .+-.4% nm, by .+-.3% nm, by .+-.2% nm, or by .+-.1% nm
or by .+-.4% angstroms, by .+-.3% angstroms, by .+-.2% angstroms,
or by +1% angstroms.
[0121] One skilled in the art will understand that the relative
intensities and positions of the peaks obtained by X-ray powder
diffraction may vary depending upon, inter alia, the sample
preparation technique, the sample mounting procedure, and the
particular instrument employed. For example, in some embodiments,
the listed X-ray powder diffraction pattern peaks for the
crystalline Form I of SEQ ID. NO. 1 is about .+-.0.2 degrees
2.theta..
[0122] In some embodiments, the crystalline Form I of SEQ. ID. NO.
1 is characterized using High Performance Liquid Chromatography and
using microscopy. FIG. 6 shows an example of a HPLC chromatogram of
Form I. Other methods for characterizing Form I could also be
used.
[0123] Form I can have any desired degree of purity, relative to
other substances or components in the preparation. In some
embodiments, Form I is provided such that it is substantially pure,
such as, for example, having greater than 30%, greater than
40.degree. 0, greater than 500, greater than 60%, greater than 70%,
greater than 80%, greater than 85%, greater than 90%, greater than
95%, greater than 96%, greater than 97%, greater than 98%, greater
than 99%, greater than 99.2%, greater than 99.4%, greater than
99.5%, greater than 99.6%, greater than 99.7%, or greater than
99.9% purity, relative to other substances or components in the
preparation.
[0124] In exemplary embodiments, the Form I is about 45% to 95%
pure, such as, for example, about 50% to 95% pure, about 55% to 90%
pure, about 60% to 95% pure, or about 70% to 99% pure, relative to
other substances or components in the preparation. In some
embodiments, the Form I is about 95% to 99% pure. In some
embodiments, Form I is about 90% to 95% pure. In some embodiments,
the Form I is about 85% to 90% pure. In some embodiments, the Form
I is about 80% to 85% pure. In some embodiments, the Form I is
about 75% to 80% pure. In some embodiments, the Form I is about 70%
to 75% pure. In certain embodiments, the Form I is about 65% to 70%
pure. In some embodiments, the Form I is about 60% to 65% pure. In
other embodiments, the Form I is about 55% to 60% pure. In yet
other embodiments, the Form I is about 50% to 55% pure. In some
embodiments, Form I is about 45% to 50% pure.
[0125] In some embodiments, the Form I may comprise one or more
impurities and/or a degradation product, such as a hydrolysis
product, acetylation product, a formylation product, an oxidation
product, a water-mediated degradation product, and/or a deamidation
product. In some embodiments, a composition comprising Form I may
comprise one or more impurities and/or a degradation product, such
as a hydrolysis product, acetylation product, a formylation
product, an oxidation product, a water-mediated degradation
product, and/or a deamidation product. In some embodiments, the one
or more impurities may be biologically active.
[0126] In some embodiments, Form I and/or the composition
comprising Form I can contain any desired purity relative to
hydrolysis product(s). In some embodiments, the composition
comprises less than about 10% by weight of hydrolysis product(s),
relative to the total weight of Form I and/or the composition, such
as, for example, less than about 7.5 wt. %, less than about 5 wt.
%, or less than about 2 wt. % of hydrolysis product(s). In some
embodiments, Form I and/or the composition comprises from about
0.05% to about 5% by weight of hydrolysis product(s). In some
embodiments, Form I and/or the composition comprises from about
0.05% to about 2% by weight of the hydrolysis product(s). In some
embodiments, Form I and/or the composition comprises from about
0.1% to about 2% by weight of the hydrolysis product(s). In some
embodiments, Form I and/or the composition comprises from about
0.01% to about 2% by weight of the hydrolysis product(s).
[0127] Alternatively, or in addition, Form I and/or the composition
comprising Form I can contain any desired purity relative to
acetylation product(s). In some embodiments, the acetylation
product may comprise less than 10% by weight of the Form I and/or
the composition. In some embodiments, the acetylation product may
comprise less than 7.5% by weight of the Form I and/or the
composition. In some embodiments, the acetylation product may
comprise less than 5% by weight of the Form I and/or the
composition. In some embodiments, the acetylation product may
comprise less than 2% by weight of the Form I and/or the
composition. In some embodiments, the acetylation product may
comprise less than 1% by weight of the Form I and/or the
composition. In some embodiments, the acetylation product may
comprise less than 0.5% by weight of Form I and/or the composition.
In some embodiments, the acetylation product may comprise from
about 0.05% to about 5% by weight of Form I and/or the composition.
In some embodiments, the acetylation product may comprise from
about 0.05% to about 2% by weight of Form I and/or the composition.
In some embodiments, the acetylation product may comprise from
about 0.1% to about 2% by weight of Form I and/or the composition.
In some embodiments, the acetylation product may comprise from
about 0.01% to about 2% by weight of the composition.
[0128] Alternatively, or in addition, Form I and/or the composition
comprising Form I can contain any desired purity relative to
formylation product(s). In some embodiments, the formylation
product may comprise less than 10% by weight of Form I and/or the
composition. In some embodiments, the formylation product may
comprise less than 7.5% by weight of Form I and/or the composition.
In some embodiments, the formylation product may comprise less than
5% by weight of Form I and/or the composition. In some embodiments,
the formylation product may comprise less than 2% by weight of Form
I and/or the composition. In some embodiments, the formylation
product may comprise from about 0.05% to about 5% by weight of Form
I and/or the composition. In some embodiments, the formylation
product may comprise from about 0.05% to about 2% by weight of Form
I and/or the composition. In some embodiments, the formylation
product may comprise from about 0.1% to about 2% by weight of Form
I and/or the composition.
[0129] Alternatively, or in addition, Form I and/or the composition
comprising Form I can contain any desired purity relative to
oxidation product(s). In some embodiments, the oxidation product
may comprise less than 10% by weight of Form I and/or the
composition. In some embodiments, the oxidation product may
comprise less than 7.5% by weight of Form I and/or the composition.
In some embodiments, the oxidation product may comprise less than
5% by weight of Form I and/or the composition. In some embodiments,
the oxidation product may comprise less than 2% by weight of Form I
and/or the composition. In some embodiments, the oxidation product
may comprise from about 0.05% to about 5% by weight of Form I
and/or the composition. In some embodiments, the oxidation product
may comprise from about 0.05% to about 2% by weight of Form I
and/or the composition. In some embodiments, the oxidation product
may comprise from about 0.1% to about 2% by weight of Form I and/or
the composition. In some embodiments, the oxidation product may
comprise from about 0.01% to about 2% by weight of Form I and/or
the composition.
[0130] Alternatively, or in addition, Form I and/or the composition
comprising Form I can contain any desired purity relative to
water-mediated degradation product(s). In some embodiments, the
water-mediated degradation product(s) may comprise less than 10% by
weight of Form I and/or the composition. In some embodiments, the
water-mediated degradation product(s) may comprise less than 7.5%
by weight of Form I and/or the composition. In some embodiments,
the water-mediated degradation product(s) may comprise less than 5%
by weight of Form I and/or the composition. In other embodiments,
the water-mediated degradation product(s) may comprise less than 2%
by weight of Form I and/or the composition. In some embodiments,
the water-mediated degradation product(s) may comprise from about
0.05% to about 5% by weight of Form I and/or the composition. In
exemplary embodiments, the water-mediated degradation product(s)
may comprise from about 0.05% to about 2% by weight of Form I
and/or the composition. In some embodiments, the water-mediated
degradation product(s) may comprise from about 0.1% to about 2% by
weight of Form I and/or the composition. In some embodiments, the
water-mediated degradation product(s) may comprise from about 0.01%
to about 2% by weight of Form I and/or the composition
[0131] Alternatively, or in addition, Form I and/or the composition
comprising Form I can contain any desired purity relative to
deamidation product(s). In some embodiments, the deamidation
product may comprise less than 10% by weight of Form I and/or the
composition. In some embodiments, the deamidation product may
comprise less than 7.5% by weight of Form I and/or the composition.
In some embodiments, the deamidation product may comprise less than
5% by weight of Form I and/or the composition. In other
embodiments, the deamidation product may comprise less than 2% by
weight of Form I and/or the composition. In some embodiments, the
deamidation product may comprise from about 0.05% to about 5% by
weight of Form I and/or the composition. In some embodiments, the
deamidation product may comprise from about 0.05% to about 2% by
weight of Form I and/or the composition. In some embodiments, the
deamidation product may comprise from about 0.1% to about 2% by
weight of Form I and/or the composition. In some embodiments, the
deamidation product may comprise from about 0.01% to about 2% by
weight of Form I and/or the composition.
[0132] In some embodiments, a composition is provided comprising
Form I and less than 10 wt. % such as less than 8 wt. 0%, less than
6 wt. %, less than 5 wt. %, less than 4 wt. %, less than 3 wt. %,
less than 2 wt. %, less than 1 wt. %, less than 0.5 wt. %, or less
than 0.25 wt. % of a combined total of a degradation product, such
as a hydrolysis product, a formylation product, an oxidation
product, a water-mediated degradation product, and/or a deamidation
product.
[0133] In some embodiments, a composition is provided comprising
Form I and less than 20 wt. % such as less than 18 wt. %, less than
16 wt. %, less than 14 wt. %, less than 12 wt. %, less than 10 wt.
%, less than 8 wt. %, less than 6 wt. %, less than 5 wt. %, less
than 4 wt. %, less than 3 wt. %, less than 2 wt. %, less than 1 wt.
%, less than 0.5 wt. %, or less than 0.25 wt. % of a combined total
of a degradation product, such as a hydrolysis product, an
acetylation product, a formylation product, an oxidation product, a
water-mediated degradation product, and/or a deamidation
product.
[0134] In some embodiments, a composition is provided comprising
Form I and less than 10 wt. % such as less than 8 wt. %, less than
6 wt. 0%, less than 5 wt. %, less than 4 wt. %, less than 3 wt. %,
less than 2 wt. %, less than 1 wt. %, less than 0.5 wt. %, or less
than 0.25 wt. % of a combined total of one or more impurities
and/or a degradation product, such as a hydrolysis product, a
formylation product, an oxidation product, a water-mediated
degradation product, and/or a deamidation product.
[0135] In some embodiments, a composition is provided comprising
Form I and less than 20 wt. % such as less than 18 wt. %, less than
16 wt. %, less than 14 wt. %, less than 12 wt. %, less than 10 wt.
%, less than 8 wt. %, less than 6 wt. %, less than 5 wt. %, less
than 4 wt. %, less than 3 wt. %, less than 2 wt. %, less than 1 wt.
%, less than 0.5 wt. %, or less than 0.25 wt. % of a combined total
of one or more impurities and/or a degradation product, such as a
hydrolysis product, an acetylation product, a formylation product,
an oxidation product, a water-mediated degradation product, and/or
a deamidation product.
[0136] In some embodiments, a composition is provided comprising
Form I and multimers. In some embodiments, the multimers may be
formed due to disulfide linkages. In some embodiments, the
multimers may be formed due to non-disulfide linkages. In some
embodiments, the composition may contain any desired purity
relative to multimers. In some embodiments, the composition
comprises less than about 20 wt. % of multimers, such as, for
example, less than about 18 wt. %, less than about 16 wt. %, less
than about 14 wt. %, less than about 12 wt. %, less than about 10
wt. %, less than about 8 wt. %, less than about 6 wt. %, less than
about 5 wt. %, less than about 4 wt. %, less than about 3 wt. %,
less than about 2 wt. %, less than about 1 wt. %, less than about
0.5 wt. %, or less than about 0.1 wt. % of multimers.
[0137] In some embodiments, a composition is provided comprising
Form I and/or one more peptide mimetic of SEQ ID. NO. 1. In some
embodiments, the composition may contain any desired purity
relative to peptide mimetic of SEQ ID. NO. 1. For example, the
composition may comprise less than about 20 wt. % of peptide
mimetic, such as, for example, less than about 18 wt. %, less than
about 16 wt. %, less than about 14 wt. %, less than about 12 wt. %,
less than about 10 wt. %, less than about 8 wt. %, less than about
6 wt. %, less than about 5 wt. %, less than about 4 wt. %, less
than about 3 wt. %, less than about 2 wt. %, less than about 1 wt.
%, less than about 0.5 wt. %, or less than about 0.1 wt. % of
peptide mimetic. In some embodiments, the peptide mimetic is
cyclic. In some embodiments, the peptide mimetic is a dimer. In
some embodiments, the peptide mimetic is a trimer.
[0138] In some embodiments, the compositions may comprise Form I
and a dimer. In some embodiments, the composition comprises less
than about 20 wt. % of dimers, such as, for example, less than
about 18 wt. %, less than about 16 wt. %, less than about 14 wt. %,
less than about 12 wt. %, less than about 10 wt. %, less than about
8 wt. %, less than about 6 wt. %, less than about 5 wt. %, less
than about 4 wt. %, less than about 3 wt. %, less than about 2 wt.
%, less than about 1 wt. %, less than about 0.5 wt. %, or less than
about 0.1 wt. % of dimers.
[0139] In some embodiments, the compositions may comprise Form I
and a trimer. In some embodiments, the composition comprises less
than about 20 wt. % of trimers, such as, for example, less than
about 18 wt. %, less than about 16 wt. %, less than about 14 wt. %,
less than about 12 wt. %, less than about 10 wt. %, less than about
8 wt. %, less than about 6 wt. %, less than about 5 wt. %, less
than about 4 wt. %, less than about 3 wt. %, less than about 2 wt.
%, less than about 1 wt. %, less than about 0.5 wt. %, or less than
about 0.1 wt. % of trimers.
[0140] In some embodiments, a composition is provided comprising
Form I and an isomer. In some embodiments, the composition
comprises less than about 20 wt. % of isomers, such as, for
example, less than about 18 wt. %, less than about 16 wt. %, less
than about 14 wt. %, less than about 12 wt. %, less than about 10
wt. %, less than about 8 wt. %, less than about 6 wt. %, less than
about 5 wt. %, less than about 4 wt. %, less than about 3 wt. %,
less than about 2 wt. %, less than about 1 wt. %, less than about
0.5 wt. %, or less than about 0.1 wt. % of isomers.
[0141] In some embodiments, a composition is provided comprising
Form I and less than about 40 wt %, such as less than about 30 wt.
%, less than about 20 wt. %, less than about 15 wt. %, less than
about 10 wt. %, less than about 8 wt. %, less than about 6 wt. %,
less than about 5 wt. %, less than about 4 wt. %, less than about 3
wt. %, less than about 2 wt. %, less than about 1 wt. %, less than
about 0.5 wt. %, less than about 0.1 wt. %, or less than about 0.01
wt. % of amorphous SEQ. ID. NO. 1.
[0142] In some embodiments, a composition is provided comprising
from about 50:50 and 99:1 Form I to amorphous SEQ. ID. NO. 1, such
as, for example, from about 55:45 and 95:5 Form I to amorphous SEQ.
ID. NO. 1, from about 60:40 and 90:10 Form I to amorphous SEQ. ID.
NO. 1, from about 70:30 and 85:15 Form I to amorphous SEQ. ID. NO.
1, or from about 75:25 and 99:1 Form I to amorphous SEQ. ID. NO.
1.
[0143] In some embodiments, processes for preparing crystalline
forms of SEQ ID. NO. 1 are provided. In some embodiments, the
crystalline Form I is produced by precipitating and crystallizing
SEQ ID. NO. 1 and optionally isolating the Form I. In some
embodiments, the Form I is prepared by precipitating and
crystallizing SEQ ID. NO. 1 in an aqueous solution and optionally
isolating the Form I. In some embodiments, the Form I is prepared
by precipitating and crystallizing SEQ ID. NO. 1 in a super
saturated aqueous solution and optionally isolating the Form I.
[0144] Any suitable aqueous solution can be used in this regard,
such as, for example, water, DMSO, acids, and polar solvents, at
various strengths or concentrations. Such solutions may include,
but are not limited to, DMSO, water, ethanol, butanol, methanol,
dicholoromethane, tetrahydrofuran, ethyl acetate, acetone,
dimethylformamide, acetonitrile, dimethyl sulfoxide, propylene
carbonate, formic acid, isopropanol, propanol, acetic acid, and
nitromethane. In some embodiments, the aqueous solution is selected
from water, ethanol, propanol, dimethyl sulfoxide, acetone, and
isopropanol. In some embodiments, the water does not have any
additional components or solvents added to it. In some embodiments,
the aqueous solution comprises water and acetone. In some
embodiments, the aqueous solution comprises water and isopropyl
alcohol. In some embodiments, the aqueous solution does not contain
ethanol. In some embodiments, the aqueous solution does not contain
acetone. In some embodiments, the aqueous solution does not contain
organic solvents.
[0145] The crystalline Form I of SEQ ID NO. 1 may be identified,
characterized, and distinguished from amorphous form using any
suitable manner. One skilled in the art will know many different
methods of identification and characterization of Form I of SEQ ID
NO. 1. For example, the crystalline Form I of SEQ ID NO. 1 may be
identified and characterized based on differences in diffraction,
thermal, intensity, and/or spectroscopic properties of the
amorphous and crystalline form. Suitable methods include, but are
not limited to, X-ray diffractometry, capillary melting point
determination, thermogravimetric analysis (TGA), differential
scanning calorimetry (DSC), and/or spectroscopic methods.
[0146] In some embodiments, Form I is precipitated from water.
FIGS. 4 and 5 shows an example characterization of Form I using
X-ray powder diffraction and DSC. In some embodiments, Form I was
precipitated from water and was characterized by HPLC and
microscopy. FIG. 6 shows an example HPLC chromatogram of Form
I.
[0147] FIG. 7 shows overlaid X-ray powder diffraction patterns of
amorphous SEQ ID. NO. 1 and crystalline Form I of SEQ ID. NO. 1
obtained using aqueous solution. In some embodiments, Form I is
precipitated from supersaturated aqueous solution. XRD
characterization from supersaturated aqueous solution indicates
that Form I is crystalline material as shown in FIG. 7. DSC
thermogram of Form I in FIG. 5 also showed different endothermic
transitions from the amorphous SEQ. ID. NO. 1, with the last
endothermic peak appeared at around 255.degree. C.-259.degree.
C.
[0148] FIG. 8 shows exemplary HPLC chromatograms of amorphous and
crystalline Form I of SEQ ID. NO. 1. The HPLC chromatograms shown
in FIG. 8 confirmed that the crystalline form prepared from aqueous
solution is chemically the same as SEQ. ID. NO. 1 amorphous form.
These results demonstrate that Form I is a crystalline form of SEQ.
ID. NO. 1.
[0149] Form I was further examined using polarized microscopy. FIG.
9 shows an example microscopic image of crystalline Form I under
cross polarized light. FIG. 10 shows an example microscopic image
and particle size of crystalline Form I under plane polarized
light. These results confirmed that Form I was crystalline material
as birefringence was observed in the sample under cross polarized
light after turning the sample stage from 0.degree. to 90.degree.
angle as shown in FIG. 9. The particle size was also determined as
shown in FIG. 10 and Table 2.
[0150] In some embodiments, process for preparing the SEQ ID. NO. 1
are provided. In some embodiments, amorphous SEQ. ID. NO. 1 is
prepared by precipitating SEQ ID. NO. 1 and optionally isolating
SEQ. ID. NO.1. In some embodiments, amorphous SEQ. ID. NO. 1 was
precipitated from a solution comprising ethanol. In some
embodiments, amorphous SEQ. ID. NO. 1 was precipitated from a
solution comprising acetone. In some embodiments, amorphous SEQ.
ID. NO. 1 was precipitated from a solution comprising isopropyl
alcohol.
[0151] In some embodiments. SEQ. ID. NO. 1 can be dissolved in an
aqueous solution of, for example, without limiting, ethanol,
wherein the percentage of ethanol in the solution may be from
approximately 80% and 20% v/v and the remainder of the solution
approximately is water. In some embodiments, the percentage of
ethanol in the solution may be approximately 60% v/v and the
remainder of the solution approximately is water. In some
embodiments, the percentage of ethanol in the solution may be
approximately 50% v/v and the remainder of the solution
approximately is water. In some embodiments, the percentage of
ethanol in the solution may be approximately 55% v/v and the
remainder of the solution approximately is water. In some
embodiments, the percentage of ethanol in the solution may be
approximately 40% v/v and the remainder of the solution
approximately is water.
[0152] In some embodiments, SEQ. ID. NO.1 can be dissolved in an
aqueous solution of, for example, without limiting, isopropyl
alcohol, wherein the percentage of isopropyl alcohol in the
solution may be from approximately 80% and 30% v/v and the
remainder of the solution approximately is water. In some
embodiments, the percentage of isopropyl alcohol in the solution
may be approximately 60% v/v and the remainder of the solution
approximately is water. In some embodiments, the percentage of
isopropyl alcohol in the solution may be approximately 50% v/v and
the remainder of the solution approximately is water. In some
embodiments, the percentage of isopropyl alcohol in the solution
may be approximately 55% v/v and the remainder of the solution
approximately is water. In some embodiments, the percentage of
isopropyl alcohol in the solution may be approximately 40% v/v and
the remainder of the solution approximately is water.
[0153] In some embodiments, SEQ. ID. NO.1 can be dissolved in an
aqueous solution of, for example, without limiting, acetone,
wherein the percentage of acetone in the solution may be from
approximately 80% and 20% v/v and the remainder of the solution
approximately is water. In some embodiments, the percentage of
acetone in the solution may be approximately 60% v/v and the
remainder of the solution approximately is water. In some
embodiments, the percentage of acetone in the solution may be
approximately 50% v/v and the remainder of the solution
approximately is water. In some embodiments, the percentage of
acetone in the solution may be approximately 55% v/v and the
remainder of the solution approximately is water. In some
embodiments, the percentage of acetone in the solution may be
approximately 40% v/v and the remainder of the solution
approximately is water.
[0154] Additionally, any suitable aqueous solution can be used in
this regard, such as, for example, water, DMSO, acids, and polar
solvents, at various strengths or concentrations. Such solutions
may include, but are not limited to, DMSO, water, ethanol, butanol,
methanol, dicholoromethane, tetrahydrofuran, ethyl acetate,
acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide,
propylene carbonate, formic acid, isopropanol, propanol, acetic
acid, and nitromethane. In some embodiments, the solvent is
selected from water, ethanol, propanol, dimethyl sulfoxide,
acetone, and isopropanol.
[0155] Precipitates from, for example, but not limited to, the
solutions described herein, including, but not limited to, ethanol,
acetone/water and isopropyl alcohol/water produce precipitate
showing the same XRD pattern as the amorphous SEQ. ID. NO. 1. The
results described herein demonstrate that the precipitates contain
amorphous, mostly amorphous, a mixture of amorphous and crystalline
forms, one or more crystalline forms, or a mixture of amorphous and
one or more crystalline forms. Each of the preceding are considered
as separate embodiments.
[0156] In some embodiments, the precipitate of SEQ. ID. NO.1 from,
for example, but not limited to, ethanol, acetone or isopropyl
alcohol may comprise amorphous, mostly amorphous, a mixture of
amorphous and crystalline forms, one or more crystalline forms, or
a mixture of amorphous and one or more crystalline forms. Each of
which are considered as a separate embodiments.
[0157] In some embodiments, a precipitate comprising between about
50:50 and 99:1 Form I to amorphous SEQ. ID. NO. 1 is provided. In
some embodiments, the precipitate may comprise amorphous SEQ. ID.
NO. 1. In some embodiments, for example, the precipitate may
comprise mostly amorphous SEQ. ID. NO.1 at about 99% to about 1% by
weight, ranging from about less than 90%, less than 80%, less than
70%, less than 60%, less than 40%, less than 30%, less than 20%,
less than 10%, less than 5%, less than 1%, less than 0.50%, less
than 0.25% by weight.
[0158] In some embodiments, the precipitate may comprise mostly
crystalline SEQ. ID. NO. 1 at about 99% to about 1% by weight,
ranging from about less than 90%, less than 80%, less than 70%,
less than 60%, less than 40%, less than 30%, less than 20%, less
than 100, less than 5%, less than 1%, less than 0.50%, less than
0.25% by weight. In some other embodiments, for example, the
precipitate may comprise one or more crystalline forms of SEQ. ID.
NO. 1.
[0159] In some embodiments, the precipitate may comprise a mixture
of one or more crystalline forms of SEQ. ID. NO. 1 and amorphous
SEQ. ID. NO. 1. For example, the precipitate may comprise
crystalline SEQ. ID. NO.1 at about 99% to about 1% by weight,
ranging from about less than 90%, less than 80%, less than 70%,
less than 60%, less than 40%, less than 30%, less than 20%, less
than 10%, less than 5%, less than 100%, less than 0.50%, or less
than 0.25% by weight of the mixture. Alternatively, the precipitate
may comprise amorphous SEQ. ID. NO.1 at about 99% to about 1% by
weight, ranging from about less than 90%, less than 80%, less than
70%, less than 60%, less than 40%, less than 30%, less than 20%,
less than 10%, less than 5%, less than 1%, less than 0.50%, or less
than 0.25% by weight of the mixture.
[0160] The methods of precipitating can also comprise heating the
solution comprising SEQ. ID. NO. 1 and then allowing the solution
to cool to ambient temperature. In some embodiments, ambient
temperature is 20-25.degree. C. In some embodiments, the solution
is cooled to 10-20.degree. C. In some embodiments, the solution is
heated to at least, or about, 10, 20, 30, 40, 50, or 60.degree. C.
before being cooled or allowed to cool to ambient temperature or a
specific temperature. In some embodiments, the solution is heated
to about 10-20, 10-30, 10-40, 10-50, 20-30, 20-40, 20-50, 20-60,
30-40, 30-50, 30-60, 40-50, 40-60, 50-60, 25-45, 35-45, or 35-50,
45-55, or 45-60.degree. C. In some embodiments, the solution is
heated at given temperature for about 0.5 to about 2 hours, about 1
to about 2 hours, about 0.5 to about 1.5 hours, or about 1 to about
1.5 hours.
[0161] In some embodiments, the crystal form is precipitated at a
temperature of about 15 to about 25.degree. C., about 15 to about
23.degree. C., about 15 to about 20.degree. C., about 15 to about
18.degree. C., about 17 to about 25.degree. C., about 17 to about
23.degree. C., about 17 to about 21.degree. C., about 17 to about
20.degree. C., about 18 to about 25.degree. C., about 18 to about
23.degree. C., about 18 to about 21.degree. C., about 18 to about
20.degree. C., about 19 to about 25.degree. C., about 19 to about
23.degree. C., or about 19 to about 21.degree. C., or any
temperature between the respective ranges. In some embodiments, the
precipitates are allowed to form for about, or at least, 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 12, 18, or 24 hours. In some embodiments, the
precipitates are allowed to form for about 2 to about 24, about 2
to about 18, about 2 to about 12, about 2 to about 10, about 2 to
about 8, about 2 to about 6, about 2 to about 4, about 4 to about
24, about 4 to about 18, about 2 to about 12, about 4 to about 10,
about 4 to about 8, about 4 to about 6, about 5 to about 24, about
5 to about 18, about 5 to about 12, about 5 to about 10, about 5 to
about 8, about 5 to about 6, about 6 to about 24, about 6 to about
18, about 6 to about 12, about 6 to about 10, about 6 to about 8,
about 8 to about 24, about 8 to about 18, about 8 to about 12, or
about 8 to about 10 hours.
[0162] In some embodiments, an additional volume of water is added
once the solution is heated and before it is cooled to the ambient
or near ambient temperatures. In some embodiments, the solution is
further cooled to about, or less than, 15, 10, 5, 0, -5, or
-10.degree. C. In some embodiments, the solution is cooled to about
-5.degree. C. to about 15.degree. C., about -5.degree. C. to about
10.degree. C., about -5.degree. C. to about 5.degree. C., about
-10.degree. C. to about 15.degree. C., about -10.degree. C. to
about 10.degree. C., about -10.degree. C. to about 5.degree. C.,
about -10.degree. C. to about 0.degree. C., about -10.degree. C. to
about -5.degree. C., about -5.degree. C. to about 15.degree. C.,
about -5.degree. C. to about 10.degree. C., about -5.degree. C. to
about 5.degree. C., about -5.degree. C. to about 0.degree. C.,
about 0.degree. C. to about 15.degree. C., about 0.degree. C. to
about 10.degree. C., about 0.degree. C. to about 5.degree. C.,
about 5.degree. C. to about 15.degree. C., or about 5.degree. C. to
about 10.degree. C. In some embodiments, the solution is cooled for
about, or at least, 1, 2, 3, 4, 5, 6, 12, 18, or 24 hours.
[0163] In some embodiments, the process comprises drying the
precipitate. In some embodiments, the precipitate is dried under
vacuum. In some embodiments, the precipitate is dried at a
temperature of about 30.degree. C. to about 50.degree. C., about
35.degree. C. to about 50.degree. C., about 30.degree. C. to about
45.degree. C., about 35.degree. C. to about 45.degree. C., or about
40.degree. C. to about 45.degree. C. In some embodiments, the
precipitate is dried at a temperature of about 30.degree. C., about
35.degree. C., about 40.degree. C., or about 45.degree. C. to about
50.degree. C. For the avoidance of doubt, the drying can at a
specific temperature can be performed under vacuum. In some
embodiments, the dried material is also lyophilized.
[0164] In some embodiments, the precipitations steps described
above can be repeated. In some embodiments, the process is repeated
one, two, or three times.
Pharmaceutical Compositions/Formulations
[0165] Embodiments described herein can be used in pharmaceutical
compositions and can be formulated by standard techniques using one
or more physiologically acceptable carriers or excipients. In some
embodiments, the formulations may contain a buffer and/or a
preservative. Form I and their physiologically acceptable salts,
anhydrates, hydrates and/or solvates, can be formulated for
administration by any suitable route, including via inhalation,
topically, nasally, orally, parenterally (for example,
intravenously, intraperitoneally, intravesically or intrathecally)
or rectally in a vehicle comprising one or more pharmaceutically
acceptable carriers, the proportion of which is determined by the
route of administration and standard biological practice. Other
routes of administration are also described herein and can be used
as well.
[0166] In some embodiments, pharmaceutical compositions are
provided comprising effective amounts of Form I with, for example,
pharmaceutically acceptable diluents, preservatives, solubilizers,
emulsifiers, adjuvants and/or other carriers. Such compositions are
known to one skilled in the art and the compositions can be
formulated using standard techniques. For example, diluents of
various buffer content such as, but not limited to, TRIS or other
amines, carbonates, phosphates, amino acids, for example,
glycinamide hydrochloride (especially in the physiological pH
range), N-glycylglycine, sodium or potassium phosphate (dibasic,
tribasic), etc. or TRIS-HCl or acetate), pH and ionic strength;
additives such as detergents and solubilizing agents (e.g.,
surfactants such as Pluronics, Tween 20, Tween 80 (Polysorbate 80),
Cremophor, polyols such as polyethylene glycol, propylene glycol,
etc.), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite),
preservatives (e.g., Thimersol, benzyl alcohol, parabens, etc.) and
bulking substances (e.g., sugars such as sucrose, lactose,
mannitol, polymers such as polyvinylpyrrolidones or dextran, etc.);
and/or incorporation of the material into particulate preparations
of polymeric compounds such as polylactic acid, polyglycolic acid,
etc. or into liposomes may be used. Hyaluronic acid may also be
used. Such compositions can be employed to influence the physical
state, stability, rate of in vivo release, and rate of in vivo
clearance of a composition comprising Form I as described herein.
See, e.g., Remington's Pharmaceutical Sciences, 18th Ed. (1990,
Mack Publishing Co., Easton, Pa. 18042) pages 1435-1712 which are
herein incorporated by reference. Where a buffer is to be included
in the formulations, the buffer can be, for example, but not
limited to, sodium acetate, sodium carbonate, citrate,
glycylglycine, histidine, glycine, lysine, arginine, sodium
dihydrogen phosphate, disodium hydrogen phosphate, sodium
phosphate, and tris(hydroxymethyl)-aminomethan, or mixtures
thereof. Each buffer can be used independently or in combination
with another buffer. In some embodiments, the buffer is
glycylglycine, sodium dihydrogen phosphate, disodium hydrogen
phosphate, sodium phosphate or mixtures thereof.
[0167] Where a pharmaceutically acceptable preservative is to be
included in the formulations, the preservative can be, but is not
limited to, phenol, m-cresol, methyl p-hydroxybenzoate, propyl
p-hydroxybenzoate, 2-phenoxyethanol, butyl p-hydroxybenzoate,
2-phenylethanol, benzyl alcohol, chlorobutanol, and thiomerosal, or
mixtures thereof. In some embodiments the preservative is phenol
and/or m-cresol.
[0168] In some embodiments the preservative is present in a
concentration from about 0.1 mg/ml to about 100 mg/ml, more
preferably in a concentration from about 0.1 mg/ml to about 50
mg/ml, about 0.1 mg/ml to about 25 mg/ml. In some embodiments, the
preservative is present in a concentration from about 0.1 mg/ml to
about 10 mg/ml.
[0169] The use of a preservative in pharmaceutical compositions is
well-known to the skilled person. For convenience reference is made
to Remington: The Science and Practice of Pharmacy, 19th edition,
1995.
[0170] In some embodiments, the formulation may further comprise a
chelating agent where the chelating agent may be salts of
ethlenediaminetetraacetic acid (EDTA), citric acid, and aspartic
acid, and mixtures thereof.
[0171] In some embodiments, the chelating agent is present in a
concentration from 0.1 mg/ml to 10 mg/ml, particularly in a
concentration from 0.1 mg/ml to 5 mg/ml. In some embodiments, the
chelating agent is present in a concentration from 0.1 mg/ml to 2
mg/ml. In some embodiments, the chelating agent is present in a
concentration from 2 mg/ml to 5 mg/ml.
[0172] The use of a chelating agent in pharmaceutical compositions
is well-known to the skilled person. For convenience reference is
made to Remington: The Science and Practice of Pharmacy, 19th
edition, 1995.
[0173] In some embodiments, the formulation may further comprise a
stabilizer selected from the group of high molecular weight
polymers or low molecular compounds where such stabilizers include,
but are not limited to, polyethylene glycol (e.g., PEG 3350),
polyvinylalcohol (PVA), polyvinylpyrrolidone,
carboxymethylcellulose, different salts (e.g. sodium chloride),
L-glycine, L-histidine, imidazole, arginine, lysine, isoleucine,
aspartic acid, tryptophan, threonine and mixtures thereof. In some
embodiments, the stabilizer is L-histidine, imidazole, arginine, or
any combination thereof.
[0174] In some embodiments, the high molecular weight polymer is
present in a concentration from 0.1 mg/ml to 100 mg/ml, in a
concentration from 0.1 mg/ml to 50 mg/ml. In some embodiments, the
high molecular weight polymer is present in a concentration from
0.1 mg/ml to 5 mg/ml. In some embodiments, the high molecular
weight polymer is present in a concentration from 5 mg/ml to 10
mg/ml. In some embodiments, the high molecular weight polymer is
present in a concentration from 10 mg/ml to 20 mg/ml. In some
embodiments, the high molecular weight polymer is present in a
concentration from 20 mg/ml to 30 mg/ml. In some embodiments, the
high molecular weight polymer is present in a concentration from 30
mg/ml to 50 mg/ml.
[0175] In some embodiments, the low molecular weight polymer is
present in a concentration from 0.1 mg/ml to 100 mg/ml. In some
embodiments, the low molecular weight polymer is present in a
concentration from 0.1 mg/ml to 50 mg/ml. In some embodiments, the
low molecular weight polymer is present in a concentration from 0.1
mg/ml to 5 mg/ml. In some embodiments, the low molecular weight
polymer compound is present in a concentration from 5 mg/ml to 10
mg/ml. In some embodiments, the low molecular weight polymer is
present in a concentration from 10 mg/ml to 20 mg/ml. In some
embodiments, the low molecular weight polymer is present in a
concentration from 20 mg/ml to 30 mg/ml. In some embodiments, the
low molecular weight polymer is present in a concentration from 30
mg/ml to 50 mg/ml. In some embodiments, the low molecular weight
polymer is present in a concentration from 50 mg/ml to 60 mg/ml. In
some embodiments, the low molecular weight polymer is present in a
concentration from 60 mg/ml to 80 mg/ml. In some embodiments, the
low molecular weight polymer is present in a concentration from 80
mg/ml to 100 mg/ml.
[0176] The use of a stabilizer in pharmaceutical compositions is
well-known to the skilled person. For convenience reference is made
to Remington: The Science and Practice of Pharmacy, 19th edition,
1995.
[0177] In some embodiments, the formulation may comprise a
surfactant where a surfactant can be a detergent, ethoxylated
castor oil, polyglycolyzed glycerides, acetylated monoglycerides,
sorbitan fatty acid esters, poloxamers, such as 188 and 407,
polyoxyethylene sorbitan fatty acid esters, polyoxyethylene
derivatives such as alkylated and alkoxylated derivatives (tweens,
e.g., Tween-20, or Tween-80), monoglycerides or ethoxylated
derivatives thereof, diglycerides or polyoxyethylene derivatives
thereof, glycerol, cholic acid or derivatives thereof, lecithins,
alcohols and phospholipids, glycerophospholipids (lecithins,
kephalins, phosphatidyl serine), glyceroglycolipids
(galactopyransoide), sphingophospholipids (sphingomyelin), and
sphingoglycolipids (ceramides, gangliosides), DSS (docusate sodium,
docusate calcium, docusate potassium, SDS (sodium dodecyl sulfate
or sodium lauryl sulfate), dipalmitoyl phosphatidic acid, sodium
caprylate, bile acids and salts thereof and glycine or taurine
conjugates, ursodeoxycholic acid, sodium cholate, sodium
deoxycholate, sodium taurocholate, sodium glycocholate,
N-Hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate, anionic
(alkyl-aryl-sulphonates) monovalent surfactants, palmitoyl
lysophosphatidyl-L-serine, lysophospholipids (e.g.,
1-acyl-sn-glycero-3-phosphate esters of ethanolamine, choline,
serine or threonine), alkyl, alkoxyl (alkyl ester), alkoxy (alkyl
ether)-derivatives of lysophosphatidyl and phosphatidylcholines,
e.g., lauroyl and myristoyl derivatives of lysophosphatidylcholine,
dipalmitoylphosphatidylcholine, and modifications of the polar head
group, that is cholines, ethanolamines, phosphatidic acid, serines,
threonines, glycerol, inositol, and the positively charged DODAC,
DOTMA, DCP, BISHOP, lysophosphatidylserine and
lysophosphatidylthreonine, zwitterionic surfactants (e.g.,
N-alkyl-N,N-dimethylammonio-1-propanesulfonates,
3-cholamido-1-propyldimethylammonio-1-propanesulfonate,
dodecylphosphocholine, myristoyl lysophosphatidylcholine, hen egg
lysolecithin), cationic surfactants (quarternary ammonium bases)
(e.g., cetyl-trimethylammonium bromide, cetylpyridinium chloride),
non-ionic surfactants, polyethyleneoxide/polypropyleneoxide block
copolymers (Pluronics/Tetronics, Triton X-100, Dodecyl
.beta.-D-glucopyranoside) or polymeric surfactants (Tween-40,
Tween-80, Brij-35), fusidic acid derivatives--(e.g., sodium
tauro-dihydrofusidate etc.), long-chain fatty acids and salts
thereof C6-C12 (e.g., oleic acid and caprylic acid), acylcarnitines
and derivatives, N.sub..alpha.-acylated derivatives of lysine,
arginine or histidine, or side-chain acylated derivatives of lysine
or arginine, N.sub..alpha.-acylated derivatives of dipeptides
comprising any combination of lysine, arginine or histidine and a
neutral or acidic amino acid, N.sub..alpha.-acylated derivative of
a tripeptide comprising any combination of a neutral amino acid and
two charged amino acids, imidazoline derivatives, or any mixture
thereof.
[0178] The use of a surfactant in pharmaceutical compositions is
well-known to the skilled person. For convenience reference is made
to Remington: The Science and Practice of Pharmacy, 19th edition,
1995.
[0179] The formulations may also comprise a pharmaceutically
acceptable sweetener. In some embodiments, the sweetener comprises
at least one intense sweetener such as, but not limited to,
saccharin, sodium or calcium saccharin, aspartame, acesulfame
potassium, sodium cyclamate, alitame, a dihydrochalcone sweetener,
monellin, stevioside or sucralose
(4,1',6'-trichloro-4,1',6'-trideoxygalactosucrose), preferably
saccharin, sodium or calcium saccharin, and optionally a bulk
sweetener such as sorbitol, mannitol, fructose, sucrose, maltose,
isomalt, glucose, hydrogenated glucose syrup, xylitol, caramel or
honey.
[0180] Intense sweeteners are conveniently employed in low
concentrations. For example, in the case of sodium saccharin, the
concentration may range from 0.04% to 0.1% (w/v) based on the total
volume of the final formulation, or from about 0.06% in the
low-dosage formulations and about 0.08% in the high-dosage ones.
The bulk sweetener can effectively be used in larger quantities
ranging from about 10% to about 35% or from about 10% to 15%
(w/v).
[0181] The formulations may be prepared by conventional techniques,
for example, as described in Remington's Pharmaceutical Sciences,
1985 or in Remington: The Science and Practice of Pharmacy, 19th
edition, 1995, where such conventional techniques of the
pharmaceutical industry involve dissolving and mixing the
ingredients as appropriate to give the desired end product.
[0182] Administration of the compound or the formulations described
herein may be carried out using any method known in the art. For
example, administration may be transdermal, parenteral,
intravenous, intra-arterial, subcutaneous, intramuscular,
intracranial, intraorbital, ophthalmic, intraventricular,
intracapsular, intraspinal, intracisternal, intraperitoneal,
intracerebroventricular, intrathecal, intranasal, aerosol, by
suppositories, inhalation, or by oral administration. In some
embodiments, the compound or formulation is administered
intravenously or by injection.
[0183] For oral administration, Form I or a therapeutically
acceptable salt thereof can be formulated in unit dosage forms such
as gelcaps, caplets, granules, lozenges, bulk powders, capsules or
tablets. The tablets or capsules may be prepared by conventional
means with pharmaceutically acceptable excipients, including
binding agents, for example, pregelatinised maize starch,
polyvinylpyrrolidone, or hydroxypropyl methylcellulose; fillers,
for example, lactose, microcrystalline cellulose, or calcium
hydrogen phosphate, lubricants, for example, magnesium stearate,
talc, or silica; disintegrants, for example, potato starch or
sodium starch glycolate; or wetting agents, for example, sodium
lauryl sulphate. Tablets can be coated by methods well known in the
art.
[0184] Liquid preparations for oral administration can take the
form of, for example, solutions, syrups, or suspensions, or they
can be presented as a dry product for constitution with water or
other suitable vehicle before use. Such liquid preparations can be
prepared by conventional means with pharmaceutically acceptable
additives, for example, suspending agents, for example, sorbitol
syrup, cellulose derivatives, or hydrogenated edible fats;
emulsifying agents, for example, lecithin or acacia; non-aqueous
vehicles, for example, almond oil, oily esters, ethyl alcohol, or
fractionated vegetable oils; and preservatives, for example, methyl
or propyl-p-hydroxybenzoates or sorbic acid. The preparations can
also contain buffer salts, flavoring, coloring, and/or sweetening
agents as appropriate. If desired, preparations for oral
administration can be suitably formulated to give controlled
release of the active compound.
[0185] For topical administration, Form I can be formulated in a
pharmaceutically acceptable vehicle containing 0.1 to 10 percent,
preferably 0.5 to 5 percent, of the active compound(s). Such
formulations can be in the form of a cream, lotion, sublingual
tablet, aerosols and/or emulsions and can be included in a
transdermal or buccal patch of the matrix or reservoir type as are
conventional in the art for this purpose.
[0186] For parenteral administration, Form I or an amorphous form
of the compound can be administered by either intravenous,
subcutaneous, or intramuscular injection, in compositions with
pharmaceutically acceptable vehicles or carriers. Form I can be
formulated for parenteral administration by injection, for example,
by bolus injection or continuous infusion. Formulations for
injection can be presented in unit dosage form, for example, in
ampoules or in multi-dose containers, with an added preservative.
The compositions can take such forms as suspensions, solutions, or
emulsions in oily or aqueous vehicles, and can contain formulatory
agents, for example, suspending, stabilizing, and/or dispersing
agents. Additionally, the compound can be precipitated and stored
in an ampule or other container and then dissolved in a solution
prior to being administered to a subject.
[0187] For administration by injection, the compound can be used in
solution, and, for example, in a sterile aqueous vehicle which may
also contain other solutes such as buffers or preservatives as well
as sufficient quantities of pharmaceutically acceptable salts or of
glucose to make the solution isotonic. In some embodiments, the
pharmaceutical compositions may be formulated with a
pharmaceutically acceptable carrier to provide sterile solutions or
suspensions for injectable administration. In particular,
injectables can be prepared in conventional forms, either as liquid
solutions or suspensions, solid forms suitable for solution or
suspensions in liquid prior to injection or as emulsions. Suitable
excipients are, for example, water, saline, dextrose, mannitol,
lactose, lecithin, albumin, sodium glutamate, cysteine
hydrochloride, or the like. In addition, if desired, the injectable
pharmaceutical compositions may contain minor amounts of nontoxic
auxiliary substances, such as wetting agents, pH buffering agents,
and the like. If desired, absorption enhancing preparations (e.g.,
liposomes) may be utilized. Suitable pharmaceutical carriers are
described in "Remington's pharmaceutical Sciences" by E. W.
Martin.
[0188] For administration by inhalation, the compound may be
conveniently delivered in the form of an aerosol spray presentation
from pressurized packs or a nebulizer, with the use of a suitable
propellant, for example, dichlorodifluoromethane,
trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide,
or other suitable gas. In the case of a pressurized aerosol, the
dosage unit can be determined by providing a valve to deliver a
metered amount. Capsules and cartridges of, for example, gelatin
for use in an inhaler or insufflator can be formulated containing a
powder mix of the compound and a suitable powder base, for example,
lactose or starch. For intranasal administration the compound may
be used, for example, as a liquid spray, as a powder or in the form
of drops.
[0189] The compound can also be formulated in rectal compositions,
for example, suppositories or retention enemas, for example,
containing conventional suppository bases, for example, cocoa
butter or other glycerides.
[0190] Furthermore, the compound can be formulated as a depot
preparation. Such long-acting formulations can be administered by
implantation (for example, subcutaneously or intramuscularly) or by
intramuscular injection. Thus, for example, the compound can be
formulated with suitable polymeric or hydrophobic materials (for
example as an emulsion in an acceptable oil) or ion exchange
resins, or as sparingly soluble derivatives, for example, as a
sparingly soluble salt.
[0191] The compositions can, if desired, be presented in a pack or
dispenser device that can contain one or more unit dosage forms
containing the active ingredient. The pack can, for example,
comprise metal or plastic foil, for example, a blister pack. The
pack can also contain individual vials or other containers. The
pack or dispenser device can be accompanied by instructions for
administration.
Dosages
[0192] Crystalline Form I may be administered to a patient at
therapeutically effective doses to prevent, treat, or control
diseases and disorders mediated, in whole or in part, by a
GPCR-ligand interaction described herein. Pharmaceutical
compositions comprising crystalline Form I may be administered to a
patient in an amount sufficient to elicit an effective protective
or therapeutic response in the patient. The dose will be determined
by the efficacy of the particular compound employed and the
condition of the subject, as well as the body weight or surface
area of the area to be treated. The size of the dose also will be
determined by the existence, nature, and extent of any adverse
effects that accompany the administration of a particular compound
or vector in a particular subject.
[0193] The amount and frequency of administration of the compound
comprising Form I or another amorphous form prepared according to a
method described herein and/or the pharmaceutically acceptable
salts thereof can be regulated according to the judgment of the
attending clinician considering such factors as age, condition and
size of the patient as well as severity of the symptoms being
treated. An ordinarily skilled physician or veterinarian can
readily determine and prescribe the effective amount of the drug
required to prevent, counter or arrest the progress of the
condition. In general it is contemplated that an effective amount
would be from 0.001 mg/kg to 10 mg/kg body weight, and in
particular from 0.01 mg/kg to 1 mg/kg body weight. More
specifically it is contemplated that an effective amount would be
to continuously infuse by intravenous administration from 0.01
micrograms/kg body weight/min to 100 micrograms/kg body weight/min
for a period of 12 hours to 14 days. It may be appropriate to
administer the required dose as two, three, four or more sub-doses
at appropriate intervals throughout the day. Sub-doses may be
formulated as unit dosage forms, for example, containing 0.01 to
500 mg, and in particular 0.1 mg to 200 mg of active ingredient per
unit dosage form.
[0194] In some embodiments, the pharmaceutical preparation is in a
unit dosage form. In such form, the preparation is subdivided into
suitably sized unit doses containing appropriate quantities of the
active component, e.g., an effective amount to achieve the desired
purpose. The quantity of active compound in a unit dose of
preparation may be varied or adjusted from about 0.01 mg to about
1000 mg, from about 0.01 mg to about 750 mg, from about 0.01 mg to
about 500 mg, or from about 0.01 mg to about 250 mg, according to
the particular application. The actual dosage employed may be
varied depending upon the requirements of the patient and the
severity of the condition being treated. Determination of the
proper dosage regimen for a particular situation is within the
skill of the art. For convenience, the total dosage may be divided
and administered in portions during the day as required.
Medical Use
[0195] A composition comprising crystalline Form I of SEQ ID. NO. 1
or an amorphous form prepared according to a method described
herein can be used for treating a cardiovascular or cardiorenal
disorder that, for example, would respond favorably to a decrease
in blood pressure.
[0196] In some embodiments, methods of treating cardiovascular
disorders are provided. In some embodiments, the method comprises
administering to a subject, or a subject in need thereof, a
therapeutically effective amount of crystalline Form I and/or
pharmaceutically acceptable salt thereof. In some embodiments, the
method comprises administering to a subject, or a subject in need
thereof, a therapeutically effective amount of an amorphous form of
SEQ ID. NO. 1 prepared according to a method described herein
and/or pharmaceutically acceptable salt thereof. These
cardiovascular disorders include, but are not limited to, chronic
hypertension, hypertensive crisis, acute congestive heart failure,
angina, acute myocardial infarction, left ventricular failure,
cerebrovascular insufficiency, intracranial haemorrhage, heart
failure, acute decompensated heart failure, which can also be
referred to as acute heart failure, essential hypertension,
post-operative hypertension, hypertensive heart disease,
hypertensive renal disease, renovascular hypertension, malignant
hypertension, post-renal transplant patient stabilization, dilated
cardiomyopathy, myocarditis, post-cardiac transplant patient
stabilization, disorders associated with post-stent management,
neurogenic hypertension, pre-eclampsia, abdominal aortic aneurysm,
and any cardiovascular disorder with a hemodynamic component.
[0197] In some embodiments, the cardiovascular disorder is an acute
cardiovascular disorder. In some embodiments, the acute
cardiovascular disorder is acute hypertensive crisis, toxemia of
pregnancy, acute myocardial infarction, acute congestive heart
failure, acute heart failure, acute ischaemic heart disease,
pulmonary hypertension, post-operative hypertension, migraine,
retinopathy and post-operative cardiac/valve surgery.
[0198] In some embodiments, methods of treating viral infectious
disease linked to AT1R are provided. In some embodiments, the
methods comprise administering to a subject in need thereof a
therapeutically effective amount of crystalline Form I and/or
pharmaceutically acceptable salt thereof. In specific embodiments,
the composition is administered by intravenous injection.
Combination Therapies
[0199] Methods are also provided for treating any cardiovascular or
cardiorenal disorder by administering crystalline Form I and/or an
amorphous form prepared according to a method described herein,
and/or pharmaceutically acceptable salts thereof, in combination
with other drugs for the treatment of cardiovascular and/or
cardiorenal disorders. These other drugs include diuretics such as
furosemide; vasodilators such as nitroglycerin, nitroprusside,
brain natriuretic peptide (BNP), or analogues thereof; inotropes
such as dobutamine; angiotensin converting enzyme (ACE) inhibitors
such as captopril and enalapril; 3 blockers such as carvedilol and
propranolol; angiotensin receptor blockers (ARBs) such as valsartan
and candesartan; and/or aldosterone antagonists such as
spironolactone.
[0200] In the combination therapies, crystalline Form I or the
amorphous form is co-administered with one or more drugs for the
treatment of cardiovascular and/or cardiorenal disorders to
increase efficacy of treatment of cardiovascular and/or cardiorenal
disorders and to reduce side effects associated with high doses of
these therapeutics.
[0201] The combination therapies described above have synergistic
and additive therapeutic effects. An improvement in the drug
therapeutic regimen can be described as the interaction of two or
more agents so that their combined effect reduces the incidence of
adverse event (AE) of either or both agents used in co-therapy.
This reduction in the incidence of adverse effects can be a result
of, e.g., administration of lower dosages of either or both agent
used in the co-therapy. For example, if the effect of Drug A alone
is 25% and has an adverse event incidence of 45% at labeled dose;
and the effect of Drug B alone is 25% and has an adverse event
incidence of 30% at labeled dose, but when the two drugs are
combined at lower than labeled doses of each, if the overall effect
is 35% (an improvement, but not synergistic or additive) and the
adverse incidence rate is 20%, there is an improvement in the drug
therapeutic regimen.
[0202] In some embodiments, the compounds described herein are
administered as a mono-therapy. In some embodiments, the compounds
described herein are administered as part of a combination therapy.
For example, a compound may be used in combination with other drugs
or therapies that are used in the treatment/prevention/suppression
and/or amelioration of the diseases or conditions for which
compounds are useful.
[0203] Such other drug(s) may be administered, by a route and in an
amount commonly used therefore, contemporaneously or sequentially
with the compounds described herein. When a compound described
herein is used contemporaneously with one or more other drugs, a
pharmaceutical unit dosage form containing such other drugs in
addition to the compound described herein may be employed.
Accordingly, the pharmaceutical compositions include those that
also contain one or more other active ingredients, in addition to
the compounds described herein.
[0204] A subject or patient in whom administration of the
therapeutic compound is an effective therapeutic regimen for a
disease or disorder is often a human, but can be any animal,
including a laboratory animal in the context of a clinical trial or
screening or activity experiment. Thus, as can be readily
appreciated by one of ordinary skill in the art, the methods,
compound and compositions are particularly suited to administration
to any animal, such as a mammal, and including, but by no means
limited to, humans, domestic animals, such as feline or canine
subjects, farm animals, such as but not limited to bovine, equine,
caprine, ovine, and porcine subjects, wild animals (whether in the
wild or in a zoological garden), research animals, such as mice,
rats, rabbits, goats, sheep, pigs, dogs, cats, etc., avian species,
such as chickens, turkeys, songbirds, etc., i.e., for veterinary
medical use.
[0205] The following examples are merely illustrative and should
not be construed as limiting the scope of the embodiments in any
way as many variations and equivalents that are encompassed by
these embodiments will become apparent to those skilled in the art
upon reading the present disclosure.
EXAMPLES
Example 1A
[0206] 100 mg SEQ. ID. NO. 1 was gradually dissolved in 0.5 ml of
aqueous solution (deionized water). The mixture was vortexed until
the sample completely dissolved. The mixture was allowed to sit at
ambient condition for precipitation. After a suitable formation
period, crystalline Form I was carefully isolated and dried. Form I
was then characterized using X-ray powder diffractometry. FIG. 4
shows the X-ray powder diffraction pattern of Form I.
Example 1B
[0207] 50 mg SEQ. ID. NO. 1 was gradually dissolved in 0.25 ml of
ethanol. The mixture was vortexed until the sample completely
dissolved. The mixture was allowed to sit at ambient condition for
precipitation. After a suitable formation period of about 12 hours
to 48 hours, the precipitate was carefully isolated and dried on an
evaporator under continuous flow of nitrogen for about 12 hours to
48 hours. The precipitate was then characterized using X-ray powder
diffractometry. FIG. 11 shows the X-ray powder diffraction pattern
of the precipitate from ethanol.
Example 1C
[0208] 500 mg SEQ. ID. NO. 1 was gradually dissolved in 2 ml of
water and the mixture was warmed to an appropriate temperature in
the range of 10.degree. C. to 60.degree. C. To this mixture,
acetone was added. The mixture was cooled to ambient temperature
and was stirred for about 2-5 hours. The suspension was carefully
filtered and isolated, and rinsed with acetone and dried under
vacuum. The precipitate was then characterized using X-ray powder
diffractometry. FIG. 12 shows the X-ray powder diffraction pattern
of the precipitate from water and acetone.
Example 1D
[0209] 500 mg SEQ. ID. NO. 1 was gradually dissolved in 2 ml of
water and the mixture was warmed to an appropriate temperature in
the range of 10.degree. C. to 60.degree. C. To this mixture, 5 ml
isopropyl alcohol was added. The mixture was cooled to ambient
temperature and was stirred for about 2-5 hours. The suspension was
carefully filtered and isolated, and rinsed with isopropyl alcohol
and dried under vacuum. The precipitate was then characterized
using X-ray powder diffractometry. FIG. 13 shows the X-ray powder
diffraction pattern of the precipitate from water and isopropyl
alcohol.
Example 1E
[0210] 500 mg SEQ. ID. NO. 1 was gradually dissolved in 1 ml of
water at an appropriate temperature in the range of 10.degree. C.
to 60.degree. C. The mixture was cooled to ambient temperature and
was stirred for about 2-5 hours. The suspension was carefully
isolated, and dried. Form I was then characterized using X-ray
powder diffractometry. FIG. 14 shows the X-ray powder diffraction
pattern of Form I of SEQ. ID. NO. 1. Corresponding DSC thermogram
is shown in FIG. 16.
Example 1F
[0211] 500 mg SEQ. ID. NO. 1 was gradually dissolved in 1.5 ml of
water at an appropriate temperature in the range of 10.degree. C.
to 60.degree. C. The mixture was cooled to ambient temperature and
was stirred for about 2-5 hours and acetone was added. The
suspension was carefully filtered and isolated, and dried. Form I
was then characterized using X-ray powder diffractometry. FIG. 15
shows the X-ray powder diffraction pattern of Form I. Corresponding
DSC thermogram is shown in FIG. 16.
Example 1G
[0212] 500 mg SEQ. ID. NO. 1 was gradually dissolved in 2.5 ml of
water at an appropriate temperature in the range of 10.degree. C.
to 60.degree. C. The mixture was cooled to ambient temperature. The
suspension was carefully isolated, and dried under vacuum. Form I
was then characterized using X-ray powder diffractometry. FIG. 17
shows the X-ray powder diffraction pattern of Form I. Corresponding
DSC thermogram is shown in FIG. 16.
Example 1H
[0213] 500 mg SEQ. ID. NO. 1 was gradually dissolved in 3.5 ml of
water at an appropriate temperature in the range of 10.degree. C.
to 60.degree. C. The mixture was cooled to ambient temperature. The
suspension was stirred at room temperature and was carefully
isolated by filtration, and dried under vacuum. Form I was then
characterized using X-ray powder diffractometry. FIG. 18 shows the
X-ray powder diffraction pattern of Form I. Corresponding DSC
thermogram is shown in FIG. 16.
[0214] The chemical identity of crystalline Form I was determined
using High Pressure Liquid Chromatography (HPLC) and Waters Sunfire
C18 column (5 .mu.m, 4.6.times.250 mm, Part #186002560) with UV
detection at 215 nm. The column temperature was set at about
30-65.degree. C. A gradient method consisting of 0.1%
trifluoroacetic acid in deionized water as mobile phase A and 0.1%
trifluoroacetic in methanol/DI water mixture (2:1 v/v) as mobile
phase B was used at a flow rate of 1.0 mL/min with a run time of 39
min for each sample. Samples of amorphous SEQ. ID. NO. 1 and
crystalline Form I were prepared in DI water at the same
concentration of around 1.2 mg/mL and injected at a volume of 20
.mu.L. Data was acquired and analyzed by TotalChrom.RTM.
Chromatography Data System software (Perkin Elmer, Inc., Waltham,
Mass.). Solubility of SEQ. ID. NO.1 in solvents as described above
was unexpected and surprising and it allowed for successful
precipitation and isolation of SEQ. ID. NO. 1, which is necessary
for large scale manufacturing for commercial production.
[0215] XRD characterization of Form I from supersaturated aqueous
solution indicated that it is crystalline material. DSC thermogram
of Form I also showed different endothermic transitions from
amorphous SEQ. ID. NO. 1, with the last endothermic peak appeared
to be around 255.degree. C.-259.degree. C. The HPLC chromatograms
confirmed that the crystalline form prepared from supersaturated
aqueous solution is chemical the same as SEQ. ID. NO. 1 amorphous
form.
[0216] These results suggest that Form I from water is a
crystalline form of SEQ. ID. NO. 1. Results also suggest that the
precipitates from ethanol, acetone/water and isopropyl
alcohol/water showed same XRD pattern (FIGS. 11-13) as the
amorphous SEQ. ID. NO. 1 and that they may comprise amorphous,
mostly amorphous, a mixture of amorphous and crystalline forms, one
or more crystalline forms, or a mixture of amorphous and one or
more crystalline forms.
Example 2
XRPD Analysis
[0217] The X-ray powder diffraction patterns of Example 1 were
determined using a bench-top X-ray diffractometer D8 Advance,
Bruker AXS Inc., Madison, Wis.). A small amount of sample obtained
from Example 1 was loaded onto Si-low background sample holder, and
exposed to CuK.alpha. radiation (40 kV.times.40 mA). The sample was
scanned in a locked coupled mode with spinner rotating at a speed
of 25 rpm. The angular range was 2.degree. to 40.degree. 20 in a
step size of 0.0069, number of steps of 5470 and time/step of 0.39
second. Data collection and analyses were performed with
commercially available software (Eva, version 2.0, Bruker AXS Inc.,
Madison, Wis.).
[0218] FIG. 4 shows the X-ray powder diffraction pattern for
crystalline Form I. Peak positions are provided in Table 1.
TABLE-US-00001 TABLE 1 Angle d value (2.theta.) (Angstrom) 6.182
14.28439 8.242 10.71841 8.633 10.2345 10.103 8.74851 10.71 8.24109
11.451 7.72162 12.364 7.15297 13.248 6.67774 14.081 6.28461 15.452
5.72976 16.103 5.49955 16.58 5.34234 17.323 5.11512 17.825 4.97193
18.516 4.78809 19.197 4.6198 20.14 4.37666 20.902 4.24645 21.517
4.12648 23.149 3.83918 24.423 3.64173 26.247 3.39266 1.812 3.20517
30.801 2.9006 32.472 2.75505 33.577 2.66691 38.428 2.34063
Example 3
Differential Scanning Calorimetry Analysis
[0219] Crystalline Form I was analyzed using Differential Scanning
Calorimetry. A differential scanning calorimeter (DSC Q2000, TA
Instruments, New Castle, Del.) with a refrigerated cooling
accessory was used for the analysis. Approximately 2 to 5 mg of
sample obtained from Example 1 was weighed and heated under dry
nitrogen purge (flow rate of 50 mL/min) from 25.degree. C. to
300.degree. C. at 10.degree. C./min. Data was analyzed using
Universal Analysis (TA Instruments, New Castle, Del.) 1.1.3 Dynamic
Vapor Sorption (DVS)
[0220] The moisture sorption-desorption profiles of amorphous SEQ.
ID. NO. 1 were obtained using a DVS Intrinsic Vapor Sorption
Analyzer (Surface Measurement Systems Ltd, Allentown, Pa.) and are
shown in FIG. 3. A small quantity of sample from Example 1 was
placed in a DVS sample holder. Two cycles of sorption/desorption
profile were recorded at 25.degree. C. in the range of 0% to 95% RH
(the first sorption cycle was started at 45% RH) with maximum
equilibration time of 120 min at each step (0%, 5%, 15%, 25%, 35%,
45%, 55%, 65%, 75%, 85%, 95%).
Example 4
Microscopic Analysis
[0221] A polarized light microscope (Nikon Eclipse E600 POL,
Morrell Instrument Company, Melville, N.Y.) with Plan Fluor
10.times. objective was used. A tiny amount of sample was placed
onto glass slide by spatula. The sample was then placed on circular
rotating stage of the microscope. Sample was first observed under
plane polarized light and then observed cross polarized light for
birefringence phenomena. Images were captured and particle size was
analyzed by Image-Pro.RTM. Plus version 5.0 (Media Cyberneics,
Inc., Rockville, Md.). Form I from water was further examined using
polarized microscopy. Results confirmed that the sample was
crystalline material as birefringence was observed in the sample
under cross polarized light after turning the sample stage from
0.degree. to 90.degree. angle as shown in FIG. 9. Particle size was
also determined as shown in FIG. 10 and Table 2.
TABLE-US-00002 TABLE 2 Size (length) Size (width) Obj..sup.# Area
.mu.m .mu.m 1 297 24.4 17.6 2 2383 95.4 47.2 4 638 35.2 25.2 5 992
50.3 34.2 6 2635 96.7 39.4 7 3467 93.5 61.9 9 475 44.8 14.6 10 2123
87.7 40.8
[0222] XRD characterization of Form I from supersaturated aqueous
solution indicated that it is crystalline material. DSC thermogram
of Form I also showed different endothermic transitions from
amorphous SEQ. ID. NO. 1, with the last endothermic peak appeared
to be around 255.degree. C.-259.degree. C. The HPLC chromatograms
confirmed that the crystalline form prepared from supersaturated
aqueous solution is chemical the same as SEQ. ID. NO. 1 amorphous
form. These results suggest that Form I from water is a crystalline
form of SEQ. ID. NO. 1. By contrast, the precipitate from ethanol
showed same XRD pattern as the amorphous SEQ. ID. NO. 1 indicating
that at least some amorphous material may be present in the
precipitate.
Example 5
Form I has Surprising and Unexpected Properties
[0223] Methods
[0224] Appearance: The appearance and flow property of crystalline
powder was examined visually and photographic image was taken using
a Nikon D3100 Digital SLR (14.2 MP with 18-55 mm f/3.5-5.6 AF-S DX
VR Nikkor Zoom Lens) digital camera.
[0225] Bulk Density: The bulk density of amorphous and crystalline
form is roughly determined by dividing the weight of powder in gram
by the volume of the weighted amount in mL. The powder is
accurately weighed on a calibrated balance; the volume is measured
by transferring the weighed amount in a graduated cylinder.
[0226] Thermogravimetry Analysis (TGA): A thermogravimetric
analyzer (TGA Q5000IR, TA Instruments, New Castle, Del.) with air
cooling was used. About 2 mg of sample was weighed in platinum TGA
pan and heated under dry nitrogen purge (flow rate 25 ml/min) at
10.degree. C./min. The data was analyzed using Universal Analysis
(TA instruments, New Castle, Del.).
[0227] Dynamic Vapor Sorption (DVS): The moisture
sorption-desorption profile of Form I was obtained using a DVS
Intrinsic Vapor Sorption Analyzer (Surface Measurement Systems Ltd,
Allentown, Pa.). A small quantity of sample was placed in a DVS
sample holder. 2 cycles of sorption/desorption profile were
recorded at 25.degree. C. in the range of 0% to 95% RH (the first
sorption cycle was started at 45% RH) with maximum equilibration
time of 120 min at each step (0%, 5%, 15%, 25%, 35%, 45%, 55%, 65%,
75%, 85%, 95%).
[0228] Stability Study: The effects of temperature and humidity on
amorphous and crystalline (Form I) SEQ. ID. NO. 1 was evaluated,
this is to provide assessment on the solid state stability of the
crystalline SEQ. ID. NO.1. Approximately 1.6 mg of SEQ. ID. NO.1
was kept in an open scintillation vial in various temperatures and
relative humidity conditions. The study design is provided in Table
3
TABLE-US-00003 TABLE 3 Stability Study Design Storage Temperature
Relative Humidity Time points 25.degree. C. 60% 2, 4, 8, and 12
weeks 40.degree. C. 11%, 32%, 75%, 90% 2, 4, 8, and 12 weeks
50.degree. C. 75% 2, 4, 8, and 12 weeks
[0229] Powder X-Ray Diffractometry (PXRD): A small amount of sample
was loaded onto Si-low background sample holder, and exposed to
CuK.alpha. radiation (40 kV.times.40 mA) in an X-ray diffractometer
(D8 Advance, Bruker AXS Inc., Madison, Wis.). The sample was
scanned in a locked coupled mode with spinner rotating at a speed
of 25 rpm. The angular range was 2.degree. to 40.degree. 20 in a
step size of 0.0069, number of steps of 5470 and time/step of 0.39
second. The data collection and analyses were performed with
commercially available software (Eva, version 2.0, Bruker AXS Inc.,
Madison, Wis.).
[0230] Determination of Degradation Product: The amount of
degradation products in the composition comprising SEQ. ID. NO. 1
was quantified by UPLC with UV detection at 205 nm using procedure
described in method PRD-TM-ANL-01105 version 1.0.
[0231] Results
[0232] Appearance: The color of crystalline form SEQ. ID. NO.1
(Form I) remained the same as the amorphous form. In addition, it
was found that the crystalline form had better flow property as
compared to the amorphous form.
[0233] Bulk Density: The bulk density of amorphous and crystalline
SEQ. ID. NO.1 was determined to be 0.07 g/ml and 0.4 g/ml,
respectively. Thus, the bulk density of crystalline SEQ. ID. NO. 1
was improved significantly and is approximately 6-fold higher than
the amorphous material. This significant increase in bulk density
of the crystalline form could not have been predicted.
[0234] TGA: The TGA weight loss curve of crystalline SEQ. ID. NO. 1
presented in FIG. 19 exhibits two weight-loss steps. The first
weight loss event of 7.1% w/w occurs between 25.degree. C. to
100.degree. C.; the second one of 7.4% w/w occurs between
100.degree. C. to 225.degree. C. The two weight loss events matches
the broad endotherms observed from the DSC thermogram and are
associated with the loss of water and residual acetic acid present
in the material. The data is shown in FIG. 19.
[0235] Dynamic Vapor Sorption (DVS): The DVS data (FIG. 20) shows
that crystalline SEQ. ID. NO.1 is not as hygroscopic as amorphous
SEQ. ID. NO.1. The crystalline form only adsorbed roughly 7.6% (by
weight) at 60% RH and 11% (by weight) at 95% RH. However, amorphous
form adsorbed more than 10% (by weight) at 60% RH and 30% (by
weight) at 95% RH. After completion of DVS analysis, the sample was
characterized by X-ray powder diffractometry. FIG. 21 shows that
the crystalline form remained unchanged after exposing to two
sorption/desorption cycles.
[0236] Stability Study: The PXRD patterns of amorphous and
crystalline SEQ. ID. NO.1 after 3 months of storage at various
conditions are displayed in FIG. 22 and FIG. 23, respectively. The
solid state form of both materials remains stable at all
time-points and various temperature/humidity conditions. The
results of total degradation of amorphous and crystalline SEQ. ID.
NO. 1 are provided in Table 4. The total degradation of amorphous
SEQ. ID. NO. 1 increase significantly as a function of time at high
temperature and humidity conditions. At elevated conditions of
40.degree. C./75% RH, 40.degree. C./90% RH and 50.degree. C./75%
RH, a high level of hydrolytic degradation product is detected,
which is the main contributor to the total degradation products of
amorphous SEQ. ID. NO. 1. The degradation products of crystalline
SEQ. ID. NO. 1 did not increase as significantly as compared to the
amorphous form. There was only approximately 0.4% total degradation
increased for crystalline SEQ. ID. NO.1 at 25.degree. C./60% RH,
40.degree. C./11% RH and 40.degree. C./32% RH. At elevated
conditions of 40.degree. C./75% RH, 40.degree. C./90% RH and
50.degree. C./75% RH, hydrolytic degradation product is also the
main contributor to the total degradation products, however, the
level observed in crystalline SEQ. ID. NO. 1 is significantly lower
than the one determined in amorphous SEQ. ID. NO. 1. Accordingly,
crystalline SEQ. ID. NO. 1 is significantly, and unexpectedly,
chemically more stable than amorphous SEQ. ID. NO. 1.
TABLE-US-00004 TABLE 4 Total degradation results of amorphous and
crystalline SEQ. ID. NO. 1 at various stability conditions Initial
2 wk 4 wk 8 wk 12 wk Amorphous SEQ. ID. NO. 1 25.degree. C./60% RH
0.75% 1.08% 1.60% 1.40% 1.86% 40.degree. C./11% RH 1.17% 1.50%
1.44% 2.07% 40.degree. C./32% RH 0.91% 1.73% 1.45% 1.65% 40.degree.
C./75% RH 1.39% 2.30% 3.79% 4.06% 40.degree. C./90% RH 3.36% 6.04%
9.93% 13.18% 50.degree. C./75% RH 2.10% 3.22% 34.75% 38.15%
Crystalline SEQ. ID. NO. 1 25.degree. C./60% RH 0.65% 0.73% 0.90%
1.05% 1.00% 40.degree. C./11% RH 0.72% 1.21% 0.89% 1.04% 40.degree.
C./32% RH 0.74% 1.01% 0.95% 1.08% 40.degree. C./75% RH 0.85% 1.16%
1.09% 1.44% 40.degree. C./90% RH 1.03% 1.34% 1.68% 1.50% 50.degree.
C./75% RH 0.80% 1.47% 4.52% 4.64%
[0237] As is demonstrated in the Examples, and as is discussed in
this application, the crystalline Form I has physical properties
that are surprising and unexpected as compared to amorphous form.
For example, the data demonstrates that crystalline Form I has
enhanced bulk density properties as compared to the amorphous form,
which enables the product to have better handling, easier storage
(does not take up as much space), and better flow in manufacturing.
The crystalline form also has better chemical stability as compared
to the amorphous form, and it is less hygroscopic. Thus, because of
these two improved stability factors, Form I is unexpectedly and
surprisingly more stable and, unexpectedly, can be stored at either
controlled refrigerated temperature (4 C) or at room temperature
(20-25 C), which is a significant and unexpected advantage over the
amorphous form, which is stored at -20 C. The ability to store Form
I at a higher temperature is unexpected, in part, because it is
unusual for a crystalline form of a peptide to have this increase
in stability at the higher temperatures (>-20 C) especially in
view of the hygroscopic properties of the amorphous form of SEQ.
ID. NO.1. The crystalline Form I of SEQ. ID. NO. 1 is also expected
to have synergy with other active or inactive components resulting
in enhanced performance characteristics or properties of
pharmaceutical compositions comprising one or more crystalline
forms of the compounds described herein.
EQUIVALENTS
[0238] While the embodiments have been depicted and described by
reference to exemplary embodiments, such a reference does not imply
a limitation on the scope, and no such limitation is to be
inferred. The embodiments are capable of considerable modification,
alteration, and equivalents in form and function, as will occur to
those ordinarily skilled in the pertinent arts having the benefit
of this disclosure.
[0239] The depicted and described embodiments are exemplary only,
and are not exhaustive of the scope.
[0240] All references cited herein are hereby incorporated by
reference in their entirety.
Sequence CWU 1
1
118PRTArtificial Sequencesynthetic
peptideMOD_RES(1)..(1)MeGlyMOD_RES(8)..(8)D-Ala 1Xaa Arg Val Tyr
Ile His Pro Xaa 1 5
* * * * *