U.S. patent application number 17/241989 was filed with the patent office on 2021-08-12 for heat-stable dry powder pharmaceutical compositions and methods.
The applicant listed for this patent is MannKind Corporation. Invention is credited to Kieran Curley, Karine Fabio, Marshall L. Grant, Joseph J. Guarneri, Andrea Leone-Bay.
Application Number | 20210244667 17/241989 |
Document ID | / |
Family ID | 1000005542644 |
Filed Date | 2021-08-12 |
United States Patent
Application |
20210244667 |
Kind Code |
A1 |
Fabio; Karine ; et
al. |
August 12, 2021 |
HEAT-STABLE DRY POWDER PHARMACEUTICAL COMPOSITIONS AND METHODS
Abstract
Disclosed herein are heat-stable dry powders which include
peptides or protein such as oxytocin for use as a pharmaceutical
composition. The composition is highly stable at increased
temperatures and relatively high humid environments, and are
intended for storage at room temperature with an improved
shelf-life. In particular, the dry powders are intended for
inhalation, however, other routes of administration can be used
when reconstituted in solution.
Inventors: |
Fabio; Karine; (Woodbury,
CT) ; Guarneri; Joseph J.; (Stamford, CT) ;
Curley; Kieran; (Cos Cob, CT) ; Grant; Marshall
L.; (Newtown, CT) ; Leone-Bay; Andrea;
(Ridgefield, CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MannKind Corporation |
Westlake Village |
CA |
US |
|
|
Family ID: |
1000005542644 |
Appl. No.: |
17/241989 |
Filed: |
April 27, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16529040 |
Aug 1, 2019 |
11013691 |
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17241989 |
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15885416 |
Jan 31, 2018 |
10413513 |
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16529040 |
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14905236 |
Jan 14, 2016 |
9925144 |
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PCT/US14/47304 |
Jul 18, 2014 |
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15885416 |
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61847981 |
Jul 18, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/198 20130101;
A61K 9/1611 20130101; A61K 45/06 20130101; A61K 38/23 20130101;
A61K 38/12 20130101; A61K 9/0073 20130101; A61K 38/28 20130101;
A61K 31/4172 20130101; A61K 38/26 20130101; A61K 9/1617 20130101;
A61K 9/1623 20130101; A61K 38/29 20130101; A61K 38/095
20190101 |
International
Class: |
A61K 9/16 20060101
A61K009/16; A61K 38/095 20060101 A61K038/095; A61K 9/00 20060101
A61K009/00; A61K 38/23 20060101 A61K038/23; A61K 38/26 20060101
A61K038/26; A61K 38/28 20060101 A61K038/28; A61K 38/29 20060101
A61K038/29; A61K 45/06 20060101 A61K045/06; A61K 31/198 20060101
A61K031/198; A61K 31/4172 20060101 A61K031/4172; A61K 38/12
20060101 A61K038/12 |
Claims
1. A dry powder pharmaceutical formulation comprising a peptide, a
protein, or a derivative or analog of said peptide or protein; a
citrate or tartrate, a cationic salt, an amino acid, and/or a
pharmaceutically acceptable carrier or excipient.
2. The dry powder pharmaceutical formulation of claim 1, wherein
the peptide, protein, derivative, or analog thereof is oxytocin, an
oxytocin derivative or an oxytocin analog.
3. The dry powder pharmaceutical formulation of claim 1, wherein
the citrate is sodium citrate, or zinc citrate.
4. The dry powder pharmaceutical formulation of claim 1, wherein
the cationic salt is a divalent cation salt which is zinc chloride,
zinc citrate, zinc acetate, magnesium chloride or calcium
chloride.
5. The dry powder pharmaceutical formulation of claim 1, wherein
the pharmaceutically acceptable carrier or excipient is a sugar
selected from mannose, mannitol, trehalose, or sorbitol.
6. The dry powder pharmaceutical formulation of claim 1, wherein
the pharmaceutically acceptable carrier or excipient is
polyvinylpyrrolidone, polyethylene glycol, or a
diketopiperazine.
7. The dry powder pharmaceutical formulation of claim 6, wherein
the diketopiperazine is fumaryl diketopiperazine or succinyl
diketopiperazine.
8. The dry powder pharmaceutical formulation of claim 1, wherein
the citrate salt is in an amount ranging from 100 to 20 equivalents
per mole of the oxytocin, the oxytocin analog or derivative
thereof.
9. The dry powder pharmaceutical formulation of claim 1, wherein
the cationic salt is in an amount ranging from 50 to 5 equivalents
per mole of oxytocin in the composition.
10. The dry powder pharmaceutical formulation of claim 1, wherein
said peptide is an endocrine hormone, a growth factor, or a
nociceptive peptide.
11. The dry powder pharmaceutical formulation of claim 10, wherein
said endocrine hormone is insulin, parathyroid hormone, calcitonin,
glucagon, glucagon-like peptide 1, oxyntomodulin, an analog or
active fragment of said endocrine hormone.
12. The dry powder pharmaceutical formulation of claim 10, wherein
said amino acid is leucine, isoleucine, trileucine, cysteine,
lysine, glycine, arginine, methionine, or histidine.
13. A method of making a dry powder formulation comprising
spray-drying a solution in a nitrogen gas chamber, comprising a
peptide, protein, fragments thereof and/or analogs thereof, wherein
the dry powder formulation comprises a mixture of the peptide,
protein, fragments thereof and/or analogs thereof; a citrate or
tartrate and a cationic salt at a pH ranging from pH 4.5 to pH 6.5,
and wherein the cationic salt is a divalent cationic salt.
14. The method of claim 13, wherein said dry powder formulation
comprises oxytocin.
15. The method of claim 13, wherein the citrate is sodium
citrate.
16. The method of claim 13, further comprising an amino acid or a
sugar.
17. The method of claim 16, wherein the amino acid is leucine,
isoleucine, trileucine, cysteine, lysine, glycine, arginine,
methionine, or histidine.
18. The method of claim 16, wherein the sugar is trehalose or
mannitol.
19. The method of claim 13, wherein the tartrate is sodium
tartrate.
20. A pharmaceutical dry powder formulation for inhalation
comprising oxytocin, an analog of oxytocin or a derivative of
oxytocin; sodium citrate, zinc chloride and a pharmaceutically
acceptable excipient.
21. The pharmaceutical dry powder formulation of claim 20, wherein
the sodium citrate is greater than 0.5% (w/w); greater than 5%
(w/w); greater than 10% (w/w), or greater than 20% (w/w) in the dry
powder formulation.
22. The method of claim 13, wherein the spray-drying comprises
atomizing the flow of solution into a drying nitrogen gas flow at
an inlet temperature of about 120.degree. C. to 150.degree. C.
23. The method of claim 17 wherein the amino acid is isoleucine.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. patent application
Ser. No. 15/885,416, filed Jan. 31, 2018, which is a continuation
of U.S. patent application Ser. No. 14/905,236, filed Jan. 14,
2106, now U.S. Pat. No. 9,925,144, which is a 371 of
PCT/US2014/047304, filed Jul. 18, 2014, which claims benefit under
35 U.S.C. .sctn. 119(e) from U.S. Provisional Patent Application
Ser. No. 61/847,981, filed Jul. 18, 2013, the content of each of
which is incorporated herein by reference in its entirety.
TECHNICAL FIELD
[0002] Disclosed herein are heat-stable dry powder compositions and
methods for delivering biodegradable substances, including peptides
and proteins, and systems and methods for delivering the dry
powders. In particular, the dry powders are preferably intended for
pulmonary delivery by inhalation to treat certain disorders and/or
diseases, including post-partum hemorrhaging.
BACKGROUND
[0003] Delivery of drugs has been a major problem for many years,
particularly when the compound to be delivered is unstable under
the conditions encountered in the gastro-intestinal tract when
administered orally to a subject, prior to reaching its targeted
location. For example, it is preferable in many cases to administer
drugs orally especially in terms of ease of administration, patient
compliance, and decreased cost. However, many compounds are
ineffective or exhibit low or variable potency when they are
administered orally. Presumably this is because the drugs are
unstable under conditions in the digestive tract or because they
are inefficiently absorbed. For biologic products, in particular
peptides and proteins, the acidic environment in the stomach is
detrimental to maintain function as most proteins are degraded
readily.
[0004] Isolated biological substances, including, certain proteins
and peptides can readily and completely lose functional activity,
for example, by taking them out of -20.degree. C. storage once.
Other isolated proteins and peptides undergo significant
degradation when stored at 4.degree. C., without the addition of
protease inhibitors. Most mammalian proteins and peptides degrade
at a temperature greater than 43.degree. C. It has been well
established that at 55.degree. C., most proteins undergo complete
denaturation in about 1-2 hours. In some cases, complete
denaturation and destabilization of an isolated protein also occurs
at room temperature.
[0005] Due to the problems associated with oral drug delivery of
drugs and in particular, biologically-derived products, drug
delivery to the lungs has been explored. For example, drugs
delivered to the lungs are designed to have an effect on the tissue
of the lungs, for example, vasodilators, surfactants,
chemotherapeutic agents or vaccines for flu or other respiratory
illnesses. Drug formulations for treating pulmonary diseases such
as asthma are available by several methods, including, using
nebulizers such as treatment with PULMOZYME.RTM., using
metered-dose inhalers such as SYMBICORT.RTM., and dry powder
inhalers such as ADVAIR DISKUS.RTM., PULMICORT FLEXAHER.RTM.. Other
drugs, including nucleotide drugs, have been delivered to the lungs
because they represent a tissue particularly appropriate for
treatment, for example, for genetic therapy in cystic fibrosis,
where retroviral vectors expressing an effective adenosine
deaminase are administered to the lungs.
[0006] Currently, formulations for treating systemic disease using
biologic products are available primarily through injectable
compositions. Dry powder compositions for pulmonary inhalation and
systemic delivery of insulin have been used including EXUBERA.RTM.,
and AFREZZA.RTM. in clinical trials. There is the desire, however,
to improve the shelf-life at room temperature for dry powder
compositions, especially those comprising a biologic molecule,
including peptides and nucleic acids, to further prolonged their
life, facilitate their storage and delivery prior to patient use,
particularly if refrigeration is not available.
[0007] For example, according to the World Health Organization, 800
women die every day from pregnancy or childbirth-related
complications. Among the major causes of death is severe bleeding
(post-partum hemorrhage) that can be prevented by the use of a
peptide hormone, oxytocin, a biologic molecule. Commercially
available oxytocin compositions are provided as liquid formulations
under the trade names PITOCIN.RTM. and SYNTOCINON.RTM. or as
generic oxytocin; the peptide in solution degrades readily at
ambient temperature, requires storage below 25.degree. C. prior to
use, and is administered only by injection. The preparations of
injectable formulations and special storage needed create
challenges, which prohibit their use in subtropical and tropical
climates where there is a great need, and refrigeration and
sterilization are not always readily available.
[0008] Accordingly, there is room for improvement in the
development of pharmaceutical formulations comprising biologic
molecules in particular for pulmonary delivery in the treatment of
disease.
SUMMARY
[0009] The present disclosure provides dry powder compositions for
inhalation which are stable at room temperature or higher
temperatures for prolonged periods of time without substantially
losing their biological activity. In one embodiment, a
pharmaceutical formulation is provided comprising a dry powder for
inhalation comprising a biologic molecule, wherein the biologic
molecule comprises a peptide or a protein for systemic delivery
using a dry powder inhalation system comprising an inhaler that can
be used with a unit dose cartridge or capsule for multiple use, a
single use inhaler with an integrally built-in container for single
use, or a multidose inhaler provided with a plurality of doses
integrally configured with the inhaler.
[0010] In one embodiment, a heat-stable pharmaceutical formulation
is provided comprising, a dry powder comprising a protein or a
peptide and one or more pharmaceutically acceptable carriers and/or
excipients, which formulations are stable at high temperatures and
high humidity. In one embodiment, the pharmaceutical formulation is
stable for a long period of time at temperatures, for example,
temperatures greater than 20.degree. C., greater than 25.degree.
C., greater than 30.degree. C., or greater than 35.degree. C.; and
relative humid environments such as environments having a relative
humidity greater than 5%, greater than 10%, greater than 30%,
greater than 50%, greater than 60%, or greater than 70%; wherein
the pharmaceutically acceptable carriers and/or excipients include,
for example, buffers, salts, polymers, diketopiperazines and/or
salts thereof, and the like. In one embodiment, the dry powder
compositions can optionally include surfactants such as
polysorbates, for example, polysorbate 80 and Tween.
[0011] In a certain embodiments, the formulation comprises a dry
powder comprising a peptide, including, for example, oxytocin, an
oxytocin derivative or an analog thereof such as carbotecin; a
buffer, and a monovalent or divalent cationic salt, and optionally
a sugar and/or an amino acid. In a particular embodiment, the
formulation comprises a dry powder comprising oxytocin, an oxytocin
derivative, or an oxytocin analog; a buffer and/or a divalent
cation or monovalent cation provided by a salt, including, zinc
citrate, zinc acetate, disodium tartrate, mono-sodium tartrate,
sodium citrate, disodium citrate, trisodium citrate, zinc chloride,
calcium chloride, magnesium chloride, sodium hydroxide, and the
like. In one embodiment, the formulation further comprises one or
more amino acids, including leucine, isoleucine, trileucine,
cystine, arginine, lysine, methionine, and/or histidine. In an
embodiment, the monovalent cation in the formulation can include
sodium, potassium and lithium. In an alternate embodiment, the
formulation may be provided with citric acid.
[0012] In a specific embodiment, a dry powder composition is
provided comprising oxytocin, sodium citrate, including,
monovalent, divalent or trivalent form, in an amount less than 40%
(w/w), less than 30% (w/w), less than 20% (w/w), or less than 10%
(w/w), and zinc chloride or zinc citrate in an amount less than 35%
(w/w), less than 20% (w/w), or less than 10% (w/w) in the
composition. In a particular embodiment, the zinc chloride is used
in an amount ranging from about 1% to about 7% (w/w) of the
composition. In an alternative embodiment, the zinc citrate is used
in an amount ranging from about 9% to about 35% (w/w) of the
composition.
[0013] In a specific embodiment, a dry powder composition is
provided comprising oxytocin, sodium tartrate, including,
monovalent, or divalent form, in an amount less than 40% (w/w),
less than 30% (w/w), less than 20% (w/w), or less than 10% (w/w),
and zinc chloride or zinc citrate in an amount less than 35% (w/w),
less than 20% (w/w), or less than 10% (w/w) in the composition. In
a particular embodiment, the zinc chloride is used in an amount
ranging from about 1% to about 7% (w/w) of the composition. In an
alternative embodiment, the zinc tartrate is used in an amount
ranging from about 9% to about 35% (w/w) of the composition.
[0014] In one embodiment, the dry powder composition comprises
citrate salts in an amount ranging from 100 to 20 equivalents per
mole of oxytocin, an oxytocin analog or derivative thereof; and the
amount of zinc salts can range from 50 to 5 equivalents per mole of
oxytocin in the composition. In some embodiments, concentrated
sodium citrate buffers were used as the source of citrate; wherein
the citrate buffers had a concentration up to 0.1 M or 0.75 M and
range in pH values of 4.0 to 6.5.
[0015] In one embodiment, the dry powder composition comprises
oxytocin or an analog or derivative thereof; zinc and citrate,
wherein the oxytocin, analog or derivative thereof is in an amount
up to 200 IU in a single inhalable dose. In some embodiments, the
dry powder composition comprises 150 IU, 100 IU, 50 IU, 40 IU, 20
IU, 10 IU, 5 IU, 1 IU, 0.05 IU, or 0.005 IU of oxytocin, an analog
or a derivative thereof in a single inhalable dose.
[0016] A method of making a dry powder formulation comprising
mixing or homogenizing a solution comprising a peptide or protein
or analog thereof, wherein the solution comprises citrate salts in
an amount ranging from 100 to 20 equivalents per mole of the
peptide or protein; and an amount of zinc salts can range from 50
to 5 equivalents per mole of the peptide or protein or analog
thereof in the composition. In some embodiments, concentrated
sodium citrate buffers were used as the source of citrate; and
spray-drying a solution in a nitrogen gas chamber, comprising a
peptide, protein, fragments thereof and/or analogs thereof, wherein
the dry powder formulation comprises a mixture of the peptide,
protein, fragments thereof and/or analogs thereof; a citrate or
tartrate and a cationic salt at a pH ranging from pH 4.5 to pH 6.5,
and wherein the cationic salt is a divalent cationic salt.
[0017] Embodiments include a method for treating post-partum
hemorrhaging comprising administering to a subject in need of
treatment a dry powder formulation by inhalation, the composition
comprising oxytocin, an analog thereof or derivative thereof; a
citrate or tartrate and a source of a cation, including, zinc
within 24 hours post-partum. In one embodiment, the treatment
comprises administrating one or more doses of the dry powder
formulation described herewith immediately upon childbirth.
[0018] In an alternate embodiment, a method of preventing
post-partum hemorrhage comprising administering to a subject
susceptible of post-partum hemorrhage a dry powder formulation
comprising oxytocin, an analog thereof or derivative thereof; a
citrate or tartrate, and a source of a cation, including, zinc
within 24 hours or immediately after childbirth.
[0019] In other embodiments described herewith, there are disclosed
methods for making heat-stable and humidity-stable formulations and
methods for using the formulations in the treatment of diseases
and/or disorders including, for example, post-partum hemorrhaging,
autism, social anxiety disorders; mood disorders, and other
hormone-related diseases, in embodiments using an inhalation
system. In an exemplary embodiment, the inhalation system is a high
resistance inhaler for single dose usage.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] The following drawings form part of the present
specification and are included to further demonstrate certain
aspects of the examples disclosed herein. The disclosure may be
better understood by reference to one or more of these drawings in
combination with the detailed description of specific embodiments
presented herein.
[0021] FIGS. 1A and 1B are scanning electron micrographs of an
amorphous dry powder formulation embodiment comprising 1% oxytocin,
87% trehalose; 10% isoleucine and 10% polyvinylpirrolidone (PVP) at
low (1A) and high magnification (1B).
[0022] FIG. 2A is a scanning electron micrograph of a control
powder similar to FIG. 1B at high magnification. FIGS. 2B, 2C and
2D are scanning electron micrographs of an amorphous dry powder
formulation embodiments comprising 1% oxytocin; citrate and a zinc
salt at high magnification and containing differing amounts of
divalent zinc salts and citrate salts.
[0023] FIG. 3 provides a graphic representation of data obtained
from stability studies data for dry powder composition embodiments
comprising 1% oxytocin incubated at 40.degree. C. and 75% relative
humidity for a period of approximately 11 months compared to a
control.
[0024] FIG. 4 provides a graphic representation of data obtained
from X-ray diffraction studies of dry powders showing amorphous
content of the powders by their characteristic scan patterns.
[0025] FIG. 5 is a graphic representation of dry powder samples
from the stability studies wherein the samples contained divalent
zinc salt and citrate salts at various concentrations.
DETAILED DESCRIPTION
[0026] Drug delivery to the lungs offers many advantages. It is
difficult to deliver drugs into the lungs due to problems in
transporting the drugs past natural physical barriers in a uniform
volume and weight of the drug and the drug physical and chemical
characteristics. Disclosed herein are heat-stable formulations
comprising, a buffer, including, citrate, and a monovalent, or
divalent cation, and one or more pharmaceutically acceptable
carriers and/or excipients. Embodiments disclosed herein show that
the dry powder formulations are stable at high heat and humidity
and thus they facilitate and overcome the storage and refrigeration
challenges posed by prior art formulations. A method of making the
dry powder composition for extended storage at temperatures greater
than 20.degree. C. and humid environments if also provided.
[0027] As used herein, the term "microparticle" refers to a
particle with a diameter of about 0.5 to about 1000 .mu.m,
irrespective of the precise exterior or interior structure.
Microparticles having a diameter of between about 0.5 and about 10
microns can reach the lungs, successfully passing most of the
natural barriers. A diameter of less than about 10 microns is
required to navigate the turn of the throat and a diameter of about
0.5 microns or greater is required to avoid being exhaled. To reach
the deep lung (or alveolar region) where most efficient absorption
is believed to occur, it is preferred to maximize the proportion of
particles contained in the "respirable fraction" (RF), generally
accepted to be those particles with an aerodynamic diameter of
about 0.5 to about 6 microns, though some references use somewhat
different ranges, as measured using standard techniques, for
example, with an Anderson Cascade Impactor. Other impactors can be
used to measure aerodynamic particle size such as the NEXT
GENERATION IMPACTOR.TM. (NGI.TM., MSP Corporation), for which the
respirable fraction is defined by similar aerodynamic size, for
example <6.4 .mu.m. In some embodiments, a laser diffraction
apparatus is used to determine particle size, for example, the
laser diffraction apparatus disclosed in U.S. patent application
Ser. No. 12/727,179, filed on Mar. 18, 2010, which is incorporated
herein in its entirety for its relevant teachings related to laser
diffraction, wherein the volumetric median geometric diameter
(VMGD) of the particles is measured to assess performance of the
inhalation system. For example, in various embodiments cartridge
emptying of .gtoreq.80%, 85%, or 90% and a VMGD of the emitted
particles of .ltoreq.12.5 .mu.m, .ltoreq.7.0 .mu.m, or .ltoreq.4.8
.mu.m can indicate progressively better aerodynamic
performance.
[0028] As used herein, the term "about" is used to indicate that a
value includes the standard deviation of the measurement for the
device or method being employed to determine the value.
[0029] Respirable fraction on fill (RF/fill) represents the % of
powder in a dose that is emitted from an inhaler upon discharge of
the powder content filled for use as the dose, and that is suitable
for respiration, i.e., the percent of particles from the filled
dose that are emitted with sizes suitable for pulmonary delivery,
which is a measure of microparticle aerodynamic performance. As
described herein, a RF/fill value of 40% or greater than 40%
reflects acceptable aerodynamic performance characteristics. In
certain embodiments disclosed herein, the respirable fraction on
fill can be greater than 50%. In an exemplary embodiment, a
respirable fraction on fill can be up to about 80%, wherein about
80% of the fill is emitted with particle sizes <5.8 .mu.m as
measured using standard techniques.
[0030] As used herein, the term "dry powder" refers to a fine
particulate composition that is not suspended or dissolved in a
propellant, or other liquid. It is not meant to necessarily imply a
complete absence of all water molecules.
[0031] As used herein, "amorphous powder" refers to dry powders
lacking a definite repeating form, shape, or structure, including
all non-crystalline powders.
[0032] In one embodiment, the dry powder is a relatively cohesive
powder which requires optimal deagglomeration condition. In one
embodiment, the inhalation system provides a re-useable, miniature
breath-powered inhaler in combination with single-use cartridges
containing pre-metered doses of a dry powder formulation.
[0033] As used herein the term "a unit dose inhaler" refers to an
inhaler that is adapted to receive or comprises a single container
comprising a dry powder formulation and delivers a single dose of a
dry powder formulation by inhalation from the container to a user.
In some instances multiple unit doses will be required to provide a
user with a specified dosage. In one embodiment, the inhaler is a
dry powder inhaler, which can be disposable for single use, or
reusable for multiple uses with a single unit dose container.
[0034] As used herein the term "a multiple dose inhaler" refers to
an inhaler having a plurality of containers, each container
comprising a pre-metered dose of a dry powder medicament and the
inhaler delivers a single dose of a medicament powder by inhalation
at any one time.
[0035] As used herein a "container" is an enclosure configured to
hold or contain a dry powder formulation, a powder containing
enclosure, and can be a structure with or without a lid. This
container can be provided separately from the inhaler or can be
structurally integrated within the inhaler (e.g. non-removable).
Further, the container can be filled with a dry powder. A cartridge
can also include a container.
[0036] As used herein a "powder mass" refers to an agglomeration of
powder particles or agglomerate having irregular geometries such as
width, diameter, and length.
[0037] As used herein, the term "microparticle" refers to a
particle with a diameter of about 0.5 to about 1000 .mu.m,
irrespective of the precise exterior or interior structure. However
four pulmonary delivery microparticles that are less than 10 .mu.m
are generally desired, especially those with mean particles sizes
of less than about 5.8 .mu.m in diameter.
[0038] In an exemplary embodiment, a dry powder formulation is
provided, comprising, a peptide or a protein, wherein the peptide
or protein is sensitive to degradation by heat. In a particular
embodiment, the dry powder formulation comprises a peptide
including, oxytocin, an oxytocin derivative, or an oxytocin analog;
a citrate, including; sodium citrate and zinc citrate; a divalent
salt; including zinc chloride; calcium chloride and magnesium
chloride; and one or more pharmaceutically acceptable carriers
selected from sugars, for example, saccharides, disaccharides;
oligosaccharides; an amino acid; wherein the sugar is, for example,
trehalose, mannose, mannitol or sorbitol, and the carrier is
polyethylene glycol, polyvinylpyrrolidone, or a diketopiperazine
capable of forming microparticles, including, fumaryl
diketopiperazine, succinyl diketopiperazine, maleyl
diketopiperazine, malonyl diketopiperazine and oxalyl
diketopiperazine, or the disodium or magnesium salt thereof, and
derivatives thereof.
[0039] In another embodiment, the formulation comprises a peptide,
including, growth hormone, calcitonin, glucagon, parathyroid
hormone, parathyroid hormone (1-34), glucagon-like peptide-1,
interferon, interleukin, erythropoietin, luteinizing
hormone-releasing hormone, somatostatin, vasopressin, enkephalin,
adrenocorticotropic hormone, growth hormone-releasing hormone,
growth factors, including, granulocyte colony formation-stimulating
factor; thyroid stimulating hormone, thyroid-stimulating
hormone-releasing hormone, antinociceptive peptides, angiotensin,
prolactin, luteinizing hormone, rennin, gastric inhibitory
polypeptide (GIP), and C-peptide.
[0040] In another embodiment, the formulation comprises a peptide,
wherein the peptide is oxytocin, insulin, growth hormone,
calcitonin, glucagon, parathyroid hormone, glucagon-like peptide-1,
glucagon like-peptide-2, parathyroid hormone (1-34), or parathyroid
hormone releasing hormone, oxyntomodulin, peptide YY, leptin,
deoxyribonuclease, ribonuclease, and follicle stimulating
hormone.
[0041] In one embodiment, the formulation comprises one or more
peptides, one or more amino acid, wherein the amino acid is
isoleucine, leucine, trileucine, cystine, cysteine, glycine,
lysine, arginine, histidine, or methionine; and one or more sugars,
including, lactose, mannitol, mannose, sorbitol, trehalose, and the
like. In this and other embodiments, the carrier can be
polyethylene glycol, polyvinylpyrrolidone, or a saccharide, an
oligosaccharide, or a polysaccharides, including lactose,
trehalose, mannose, mannitol, or sorbitol; zinc citrate and zinc
chloride; wherein the formulation is made by a spray-drying process
wherein the peptide is in a buffered solution having a pH ranging
from about pH 3.5 to about pH 7; or pH 4.5 to pH 6.5.
[0042] In a particular embodiment, the formulation comprises
oxytocin in concentration from about 0.005 IU to about 40 IU, from
1 IU to about 15 IU; or from about 5 IU to about 20 IU. In one
embodiment, oxytocin is administered to a patient to prevent
post-partum hemorrhaging a few minutes after giving birth in a
formulation comprising oxytocin in an amount ranging from 5 to
about 40 IU in a single inhalation. In this embodiment, the content
of oxytocin that can be provided in the formulation ranges from
about 0.1% (w/w) to about 50% (w/w), from about 0.5% (w/w) to about
40% (w/w), from about 0.5% (w/w) to about 20% (w/w), or from about
1% (w/w) to about 10% (w/w). In certain embodiments, the amount of
oxytocin can be greater than 40 IU depending in the need of the
subject to be treated.
[0043] In one embodiment, there is provided a method for the
effective delivery of a formulation to the blood stream of a
subject, comprising providing to a subject in need of treatment an
inhalation system comprising an inhaler including a cartridge
containing a formulation comprising a dry powder formulation
comprising a peptide including, oxytocin, a citrate buffer or
tartrate buffer and a divalent cation salt, wherein the divalent
cation is zinc. In this and other embodiments, the inhalation
system delivers a powder plume comprising particles having a
volumetric median geometric diameter (VMGD) less than 8 .mu.m. In
an example embodiment, the VMGD of the microparticles can range
from about 4 .mu.m to 6 .mu.m. In an example embodiment, the VMGD
of the powder particles can be from 3 .mu.m to about 6 .mu.m in a
single inhalation of the formulation of fill mass ranging between 1
mg and 10 mg of dry powder. In this and other embodiments, the
inhalation system delivers greater than 40%; or greater than 60% of
the dry powder formulation from the cartridge.
[0044] In a further embodiment, the formulation is an amorphous dry
powder comprising microparticles of disodium fumaryl
diketopiperazine comprising oxytocin, a citrate buffer; zinc
chloride, an amino acid, such as leucine, isoleucine, trileucine or
cystine and mannitol or trehalose, or a combination thereof.
[0045] In an embodiment, the formulation comprises an amorphous dry
powder comprising a peptide, including, a heat-sensitive peptide,
including oxytocin; wherein the dry powder is formed by mixing
oxytocin in a solution containing a citrate or acetate buffer at an
adjusted pH ranging from 4.5 to 6.5 and adding a divalent cationic
salt, including zinc chloride and optionally a sugar such as
trehalose or mannitol prior to drying.
[0046] In a particular embodiment, the formulation comprises an
amorphous dry powder comprising oxytocin; wherein the dry powder is
formed by mixing oxytocin in a solution containing citrate salts
and/or citric acid and adding a divalent cationic salt, including,
zinc chloride and optionally a sugar such as trehalose or mannitol
and optionally, one or more carriers.
[0047] Further embodiments concern drug delivery systems comprising
an inhaler, a unit dose dry powder medicament container, and a dry
powder comprising a heat-sensitive peptide as disclosed herein and
zinc citrate.
[0048] One embodiment discloses a formulation comprising oxytocin,
a derivative thereof, or an analog thereof, wherein the formulation
further comprises diketopiperazine microparticles, including,
microparticles of fumaryl diketopiperazine having a specific
surface area (SSA) of less than about 67 m.sup.2/g. Another
embodiment includes diketopiperazine microparticles in which the
specific surface area is from about 35 to about 67 m.sup.2/g,
within a 95% confidence limit. Another embodiment includes
diketopiperazine microparticles in which the specific surface area
is from about 35 to about 62 m.sup.2/g. Another embodiment includes
diketopiperazine microparticles in which the specific surface area
is from about 40 to about 62 m.sup.2/g.
[0049] In alternative embodiments, the FDKP microparticles comprise
a drug or active agent. In various embodiments of the FDKP
microparticles, the drug can be, for example, a peptide, including,
oxytocin, insulin, glucagon-like peptide-1 (GLP-1), glucagon,
exendin, parathyroid hormone, calcitonin, oxyntomodulin,
derivatives and/or analogs thereof, and the like. In another
embodiment of the FDKP microparticles, the peptide content can vary
depending on downstream processing conditions. In a particular
example, the FDKP microparticles can be prepared to have a
drug/peptide content that can vary depending on the dose to be
targeted or delivered. For example, wherein the drug is insulin,
the insulin component can be from about 3 U/mg to about 6 U/mg in
the powder formulation comprising the microparticles and the zinc
salt and citrate can be added to solution prior to forming the
particles. In certain embodiments, the drug is adsorbed to the
surfaces of pre-formed microparticles.
[0050] Further embodiments concern drug delivery systems comprising
a combination of an inhaler, a unit dose dry powder medicament
container, for example, a cartridge, and comprising the dry powder
formulations disclosed herein and an active agent. In one
embodiment, the delivery system for use with the dry powders
includes an inhalation system comprising a high resistance inhaler
having air conduits which impart a high resistance to airflow
through the conduits for deagglomerating and dispensing the powder.
In one embodiment, the inhalation system has a resistance value of,
for example, approximately 0.065 to about 0.200 (IkPa)/liter per
minute. In certain embodiments, the dry powders can be delivered
effectively by inhalation with an inhalation system wherein the
peak inhalation pressure differential can range from about 2 to
about 20 kPa, which can produce resultant peak flow rates of about
between 7 and 70 liters per minute. In certain embodiments, the
inhalation system are configured to provide a single dose by
discharging powder from the inhaler as a continuous flow, or as one
or more pulses of powder delivered to a patient. In some
embodiments disclosed herewith, the dry powder inhaler system
comprises a predetermined mass flow balance within the inhaler. For
example, a flow balance of approximately 10% to 70% of the total
flow exiting the inhaler and into the patient is delivered by one
or more dispensing ports, which airflow passes through the area
containing the powder formulation, and wherein approximately 30% to
90% air flow is generated from other conduits of the inhaler.
Moreover, bypass flow, or flow not entering and exiting the area of
powder containment such as through a cartridge, can recombine with
the flow exiting the powder dispensing port within the inhaler to
dilute, accelerate and ultimately deagglomerate the fluidized
powder prior to exiting the mouthpiece. In one embodiment, flow
rates ranging from about 7 to 70 liters per minute result in
greater than 75% of the container or the cartridge contents
dispensed in fill masses between 1 mg and 50 mg; or 1 mg to 30 mg.
In certain embodiments, an inhalation system as described above can
emit a respirable fraction/fill of a powder dose at percentages
greater than 40% in a single inhalation, greater than 50%, greater
than 60%, or greater than 70%.
[0051] In particular embodiments, an inhalation system is provided
comprising a dry powder inhaler, and a dry powder formulation. In
some aspects of this embodiment of the inhalation system, the dry
powder formulation is provided in a unit dose cartridge.
Alternatively, the dry powder formulation can be preloaded in the
inhaler. In this embodiment, the structural configuration of the
inhalation system allows the deagglomeration mechanism of the
inhaler to produce respirable fractions greater than 50%; that is,
more than half of the powder contained in the inhaler (cartridge)
is emitted as particles of less than 5.8 .mu.m. The inhalers can
discharge greater than 85% of a powder medicament contained within
a container during dosing. In certain embodiments, the inhalers can
discharge greater than 85% of a powder medicament contained in a
single inhalation. In one embodiment, the inhalers can discharge
greater that 90% of the cartridge contents or container contents in
less than 3 seconds at pressure differentials between 2 and 5 kPa
with fill masses ranging up to 30 mg.
[0052] Another embodiment disclosed herein includes a method of
making microparticles suitable for pulmonary administration as a
dry powder formulation comprising, a carrier particle, including,
diketopiperazine microparticles. In this and other embodiments, the
dry powder formulation is obtained by spray-drying a solution
containing a peptide, wherein the one or more excipients is
dissolved in an aqueous solution comprising the zinc salt and
citrate and mixed, followed by adding the amount of the peptide
with mixing to form a feed solution; atomizing the flow of solution
into a drying nitrogen gas flow at an inlet temperature of about
120.degree. C. to 150.degree. C. and an outlet temperature of about
60.degree. C. to 65.degree. C., or 50.degree. C. to 75.degree. C.,
or 40.degree. C. to 85.degree. C., or the like.
[0053] In some embodiments, the method of making diketopiperazine
microparticles having the specific surface area of less than about
67 m.sup.2/g, and/or a trans isomer ratio of about 45% to 65%,
which utilizes a diketopiperazine having the formula
3,6-bis(N--X-4-aminobutyl)-2,5-diketopiperazine disodium salt or
magnesium salt, wherein X is selected from the group consisting of
fumaryl, succinyl, maleyl, and glutaryl. In an exemplary
embodiment, the diketopiperazine has the formula
(bis-3,6-bis(N-fumaryl-4-aminobutyl)-2,5-diketopiperazine or
3,6-bis(N-fumaryl-4-amino-butyl)-2,5-diketopiperazine.
[0054] Another embodiment disclosed herein includes a method of
delivering a drug, for example, a peptide such as insulin to a
patient in need thereof comprising administering a dry powder to
the deep lung by inhalation of the dry powder by the patient;
wherein the dry powder comprises diketopiperazine microparticles
comprising insulin, a zinc salt and citrate; wherein the
microparticles are formed of a diketopiperazine and have a SSA
ranging from about 35 to about 67 m.sup.2/g or about 40 to about 67
m.sup.2/g and/or in microparticles having a trans isomer content
raging from about 45% to about 65%. In aspects of this embodiment,
particular features of an inhaler system are specified. Further
embodiments involve methods of treating an insulin-related disorder
comprising administering a dry powder described above to a person
in need thereof. In various embodiments an insulin-related disorder
can specifically include or exclude any or all of pre-diabetes,
type 1 diabetes mellitus (honeymoon phase, post-honeymoon phase, or
both), type 2 diabetes mellitus, gestational diabetes,
hypoglycemia, hyperglycemia, insulin resistance, secretory
dysfunction, impaired early-phase release of insulin, loss of
pancreatic .beta.-cell function, loss of pancreatic .beta.-cells,
and metabolic disorder.
[0055] One embodiment comprises a method of treating an
endocrine-related disease or disorder comprising administering to a
person in need thereof a dry powder formulation comprising a
peptide hormone, including, oxytocin, GLP-1 and the like, citrate
and a divalent cationic salt. In one embodiment, the dry powder
formulation can comprise disodium FDKP microparticles or FDKP
microparticles having a specific surface area of less than about 67
m.sup.2/g and a drug suitable to treat said disease or disorder.
Further embodiments include a method of treating an insulin-related
disorder comprising administering a dry powder comprising
microparticles of disodium FDKP or FDKP, a citrate, acetate or
tartrate buffer and a divalent cation, including zinc, magnesium
and calcium or a monovalent cation, including, sodium, potassium
and lithium described above to a person in need thereof. The method
can comprise administering to a subject a dry powder formulation.
In various embodiments, a hormone-related disorder such as
post-partum hemorrhage, or any other oxytocin-related disease can
be treated with the formulation comprising oxytocin. In embodiments
wherein an insulin-related disorder is to be treated, the
formulation comprising insulin the subject to be treated can
specifically include or exclude any or all of pre-diabetes, type 1
diabetes mellitus (honeymoon phase, post-honeymoon phase, or both),
type 2 diabetes mellitus, gestational diabetes, hypoglycemia,
hyperglycemia, insulin resistance, secretory dysfunction, impaired
early-phase release of insulin, loss of pancreatic .beta.-cell
function, loss of pancreatic .beta.-cells, and metabolic disorder.
In one embodiment, the dry powder comprises insulin. In other
embodiments, the dry powder comprises glucagon, an exendin, or
GLP-1, PTH, PTHrP, combinations thereof, and the like.
[0056] In embodiments herewith, specific RF/fill values can depend
on the inhaler used to deliver the powder. Powders generally tend
to agglomerate and crystalline DKP microparticles form particularly
cohesive powders. One of the functions of a dry powder inhaler is
to deagglomerate the powder so that the resultant particles
comprise a respirable fraction suitable for delivering a dose by
inhalation. However, deagglomeration of cohesive powders is
typically incomplete so that the particle size distribution seen
when measuring the respirable fraction as delivered by an inhaler
will not match the size distribution of the primary particles, that
is, the profile will be shifted toward larger particles. Inhaler
designs vary in their efficiency of deagglomeration and thus the
absolute value of RF/fill observed using different designs will
also vary. However, optimal RF/fill as a function of specific
surface area will be similar from inhaler to inhaler.
[0057] One class of drug delivery agents that has been used to
overcome problems in the pharmaceutical arts such as drug
instability and/or poor absorption are the 2,5-diketopiperazines.
2,5-Diketopiperazines can be formed into microparticles that
incorporate a drug or microparticles onto which a drug can be
adsorbed. The combination of a drug and a diketopiperazine can
impart improved drug stability and/or absorption characteristics.
These microparticles can be administered by various routes of
administration. As dry powders these microparticles can be
delivered by inhalation to specific areas of the respiratory
system, including the lungs.
[0058] Such microparticles are typically obtained by pH-based
precipitation of the free acid (or base) resulting in
self-assembled microparticles comprised of aggregated crystalline
plates. The stability of the particle can be enhanced by small
amounts of a surfactant, such as polysorbate-80, in the DKP
solution from which the particles are precipitated (see for example
US Patent Publication No. 2007/0059373 entitled "Method of drug
formulation based on increasing the affinity of crystalline
microparticle surfaces for active agents" which is incorporated
herein by reference in its entirety for all that it teaches
regarding the formation and loading of DKP microparticles and dry
powders thereof). Ultimately solvent can be removed to obtain a dry
powder. Appropriate methods of solvent removal include
lyophilization and spray drying (see for example US Patent
Publication No. 2007/0196503 entitled "A method for improving the
pharmaceutic properties of microparticles comprising
diketopiperazine and an active agent" and U.S. Pat. No. 6,444,226
entitled "Purification and stabilization of peptide and protein
pharmaceutical agents" each of which is incorporated herein by
reference in its entirety for all that it teaches regarding the
formation and loading of DKP microparticles and dry powders
thereof). The microparticles disclosed herein can be composed of
DKP free acid or bases or composed of DKP salts. Such particles are
typically formed (as opposed to dried) by spray drying, resulting
in spheres and/or collapsed spheres of an amorphous salt (as
opposed to a free acid or base) so that they are chemically,
physically, and morphologically distinct entities. In embodiments
the present disclosure refers to FDKP as the free acid or the
dissolved anion. In embodiments herewith, an exemplary embodiment
includes the disodium salt of FDKP or FDKP disodium salt as
disclosed and contemplated in U.S. Pat. Nos. 7,820,676 and
8,278,308, which are incorporated herein by reference in its
entirety.
[0059] Methods for synthesizing diketopiperazines are described in,
for example, Katchalski, et al., J. Amer. Chem. Soc. 68, 879-880
(1946) and Kopple, et al., J. Org. Chem. 33(2), 862-864 (1968), the
teachings of which are incorporated herein by reference for all
they disclose relating to diketopiperazine synthesis.
2,5-Diketo-3,6-di(aminobutyl)piperazine (Katchalski et al. refer to
this as lysine anhydride) can also be prepared via
cyclodimerization of N-.epsilon.-P-L-lysine in molten phenol,
similar to the Kopple method, followed by removal of the blocking
(P)-groups with an appropriate reagent and conditions. For example,
CBz-protecting groups can be removed using 4.3 M HBr in acetic
acid. This route can be preferred because it uses a commercially
available starting material, it involves reaction conditions that
are reported to preserve stereochemistry of the starting materials
in the product and all steps can be easily scaled up for
manufacture. Methods for synthesizing diketopiperazines are also
described in U.S. Pat. No. 7,709,639, entitled, "Catalysis of
Diketopiperazine Synthesis," which is also incorporated by
reference herein for its teachings regarding the same.
[0060] Fumaryl diketopiperazine
(3,6-bis(N-fumaryl-4-aminobutyl)-2,5-diketopiperazine; FDKP) is one
preferred diketopiperazine for pulmonary applications:
##STR00001##
[0061] FDKP provides a beneficial microparticle matrix because it
has low solubility in acid but is readily soluble at neutral or
basic pH. These properties allow FDKP to crystallize and the
crystals to self-assemble into form microparticles under acidic
conditions. The particles dissolve readily under physiological
conditions where the pH is neutral. As noted, microparticles having
a diameter of between about 0.5 and about 10 microns can reach the
lungs, successfully passing most of the natural barriers. Particles
in this size range can be readily prepared from FDKP.
[0062] As noted, microparticles having a diameter of between about
0.5 and about 10 microns can reach the lungs, successfully passing
most of the natural barriers. Particles in this size range can be
readily prepared from diketopiperazines with acidic groups, such as
the carboxylate groups in FDKP (as well as in related molecules
such as 2,5-diketo-3,6-di(4-X-aminobutyl)piperazine wherein X is
succinyl, glutaryl, or maleyl). Upon acid precipitation
self-assembled particles composed of aggregates of crystalline
plates are obtained. The size of these plates relates to the
specific surface area of the particles which in turn is implicated
in effects on the structure, loading capacity, and aerodynamic
performance of the particles.
[0063] The SSA of DKP microparticles is a measure of average
crystal size and can be used to gauge the relative contributions of
crystal nucleation and growth to microparticle characteristics. SSA
depends on the size of microparticle crystals and the density
(.rho.) of the microparticle matrix and is inversely proportional
to the characteristic size, L, of the crystals. Embodiments
disclosed herein show that microparticles with a specific surface
area less than about 67 m.sup.2/g exhibit characteristics
beneficial to delivery of drugs to the lungs such as improved
aerodynamic performance with moderately efficient inhalers such as
the MEDTONE.RTM. inhaler disclosed in U.S. Pat. No. 7,464,706
entitled, "Unit Dose Cartridge and Dry Powder Inhaler," which is
incorporated by reference herein for its teachings regarding the
same. An alternate embodiment with a specific surface area less
than about 62 m.sup.2/g provides a greater level of assurance that
a batch of particles will meet a minimum aerodynamic performance
standard. As SSA also affects drug loading/content capacity,
various embodiments require SSA .gtoreq.35, 40, or 45 m.sup.2/g for
improved drug adsorption capacity. Additionally, as SSA falls below
about 35 m.sup.2/g inconsistent cartridge emptying is observed even
with high efficiency inhalers such as those disclosed in U.S.
patent application Ser. No. 12/484,125 (now U.S. Pat. No.
8,499,757, entitled, "A Dry Powder Inhaler and System for Drug
Delivery," filed on Jun. 12, 2009, and U.S. patent application Ser.
No. 12/717,884, now U.S. Pat. No. 8,485,180, entitled, "Improved
Dry Powder Drug Delivery System," filed on Mar. 4, 2010, which
disclosures are herein incorporated by reference for its teachings
regarding the same.
[0064] FDKP Microparticle Formation.
[0065] The first step in the manufacture of FDKP microparticles is
the formation of the microparticles by pH-induced crystallization
of FDKP and the self-assembly of the FDKP crystals into
microparticles having an overall spherical morphology (FIG. 2).
Accordingly, the manufacture of microparticles is essentially a
crystallization process. Excess solvent can be removed by washing
the suspension by repeated centrifugation, decantation and
re-suspension, or by diafiltration.
[0066] In one embodiment, to form peptide-loaded FDKP
microparticles, for example, insulin can be adsorbed directly onto
the microparticles while in suspension (i.e. prior to freeze
drying) by adding an insulin stock solution to the FDKP
microparticle suspension comprising a citrate buffer. In
embodiments, a pH control step can also be performed after the
addition of the insulin stock solution. This step can promote
insulin adsorption onto the microparticles in suspension prior to
further processing. Increasing the pH of the suspension to about
4.5 promotes complete insulin adsorption onto the microparticles in
suspension without excessive dissolution of the FDKP from the
particle matrix and also improves the stability of insulin in the
bulk drug product. The suspension can be flash-frozen drop-wise
(i.e. cryo-pelletized) in liquid nitrogen and lyophilized to remove
the solvent and obtain a dry powder. In alternative embodiments the
suspension can be spray-dried to obtain the dry powder.
[0067] In one embodiment, a manufacturing process for making the
present FDKP microparticles containing insulin is provided. In
summary, using a high shear mixer such as a Dual-feed SONOLATOR.TM.
at 2000 psi through a 0.001-in.sup.2 orifice, or for example, the
high shear mixer as disclosed in U.S. Provisional Patent
Application Ser. No. 61/257,311, filed on Nov. 2, 2009, which
disclosure is incorporated herein by reference in its entirety for
all that it teaches regarding the production of DKP
microparticlesparticles, equal masses of about 10.5 wt % acetic
acid and about 2.5 wt % FDKP solutions at about 16.degree.
C..+-.about 2.degree. C. can be fed at 2000 psi through a
0.001-in.sup.2 orifice. The precipitate can be collected in a
deionized (DI) water reservoir of about equal mass and temperature.
The resultant suspension comprises about 0.8% solids. The
precipitate can be concentrated and washed by tangential flow
filtration. The precipitate can be first concentrated to about 4%
solids then washed with deionized water. The suspension can be
finally concentrated to about 10% solids based on the initial mass
of FDKP. The concentrated suspension can be assayed for solids
content by an oven drying method. In this embodiment, the FDKP
microparticles in suspension are homogenized with zinc and citrate
solution containing the insulin to form the powder particles then
sprayed dried or lyophilized.
[0068] The microparticles described herein can comprise one or more
active agents. As used herein "active agent", used interchangeably
with "drug", refers to pharmaceutical substances, including small
molecule pharmaceuticals, biologicals and bioactive agents. Active
agents can be naturally occurring, recombinant or of synthetic
origin, including proteins, polypeptides, peptides, nucleic acids,
organic macromolecules, synthetic organic compounds,
polysaccharides and other sugars, fatty acids, and lipids, and
antibodies and fragments thereof, including, but not limited to,
humanized or chimeric antibodies, F(ab), F(ab).sub.2, a
single-chain antibody alone or fused to other polypeptides or
therapeutic or diagnostic monoclonal antibodies to cancer antigens.
The active agents can fall under a variety of biological activity
and classes, such as vasoactive agents, neuroactive agents,
hormones, anticoagulants, immunomodulating agents, cytotoxic
agents, antibiotics, antiviral agents, antigens, infectious agents,
inflammatory mediators, hormones, and cell surface antigens. More
particularly, active agents can include, in a non-limiting manner,
cytokines, lipokines, enkephalins, alkynes, cyclosporins, anti-IL-8
antibodies, IL-8 antagonists including ABX-IL-8; prostaglandins
including PG-12, LTB receptor blockers including LY29311, BIIL 284
and CP105696, triptans such as sumatriptan and palmitoleate,
insulin and analogs thereof, growth hormone and analogs thereof,
parathyroid hormone (PTH) and analogs thereof, parathyroid hormone
related peptide (PTHrP), ghrelin, obestatin, enterostatin,
granulocyte macrophage colony stimulating factor (GM-CSF), amylin,
amylin analogs, glucagon-like peptide 1 (GLP-1), Texas Red,
clopidogrel, PPACK (D-phenylalanyl-L-prolyl-L-arginine chloromethyl
ketone), oxyntomodulin (OXM), peptide YY(3-36) (PYY), adiponectin,
cholecystokinin (CCK), secretin, gastrin, glucagon, motilin,
somatostatin, brain natriuretic peptide (BNP), atrial natriuretic
peptide (ANP), IGF-1, growth hormone releasing factor (GHRF),
integrin beta-4 precursor (ITB4) receptor antagonist, nociceptin,
nocistatin, orphanin FQ2, calcitonin, CGRP, angiotensin, substance
P, neurokinin A, pancreatic polypeptide, neuropeptide Y,
delta-sleep-inducing peptide and vasoactive intestinal peptide.
[0069] The drug content to be delivered depends on the need of the
subject and the potency of the drug. In certain embodiments,
microparticles formed from FDKP having a trans isomer content
between 45% and 65% is typically greater than 0.01% are used. In
one embodiment, the drug content to be delivered with the
microparticles having the aforementioned trans isomer content, can
range from about 0.01% to about 20%, which is typical for peptides
such as insulin. For example, if the drug is insulin, the present
microparticles typically comprise 3-6 U/mg (approximately 10 to
15%) insulin. In certain embodiments, the drug content of the
particles can vary depending on the form and size of the drug to be
delivered.
[0070] The range of loading of the drug to be delivered is
typically between about 0.01% and about 90%, depending on the form
and size of the drug to be delivered and the potency of the dose
required. For oxytocin, preferred loads are about 0.5% to about 50%
(w/w); or from about 0.5% (w/w) to about 20% (w/w).
[0071] As long as the DKP microparticles described herein retain
the required isomer content, they can adopt other additional
characteristics beneficial for delivery to the lung and/or drug
adsorption. U.S. Pat. No. 6,428,771 entitled "Method for Drug
Delivery to the Pulmonary System" describes DKP particle delivery
to the lung and is incorporated by reference herein for its
teachings regarding the same. U.S. Pat. No. 6,444,226, entitled,
"Purification and Stabilization of Peptide and Protein
Pharmaceutical Agents" describes beneficial methods for adsorbing
drugs onto microparticle surfaces and is also incorporated by
reference herein for its teachings regarding the same.
Microparticle surface properties can be manipulated to achieve
desired characteristics as described in U.S. patent application
Ser. No. 11/532,063, now U.S. Pat. No. 7,799,344, entitled "Method
of Drug Formulation based on Increasing the Affinity of Crystalline
Microparticle Surfaces for Active Agents" which is incorporated by
reference herein for its teachings regarding the same. U.S. patent
application Ser. No. 11/532,065, entitled "Method of Drug Formation
based on Increasing the Affinity of Active Agents for Crystalline
Microparticle Surfaces" describes methods for promoting adsorption
of active agents onto microparticles. U.S. patent application Ser.
No. 11/532,065, now U.S. Pat. No. 7,803,404 is also incorporated by
reference herein for its teachings regarding the same.
EXAMPLES
[0072] The following examples are included to demonstrate
embodiments of the disclosed microparticles. It should be
appreciated by those of skill in the art that the techniques
disclosed in the examples which follow represent techniques
discovered by the inventor to function well in the practice of the
present disclosure, and thus can be considered to constitute
preferred modes for its practice. However, those of ordinary skill
in the art should, in light of the present disclosure, appreciate
that many changes can be made in the specific embodiments which are
disclosed and still obtain a like or similar result.
Example 1
[0073] Preparation, Characterization and Stability of Oxytocin
Spray-Dried Powders
[0074] Fourteen powders containing 1% (w/w) oxytocin and varying
amounts of buffers, salts, carriers, excipients, including,
trehalose, PVP, isoleucine, cystine, trileucine, FDKP, sodium
citrate and zinc salt, obtained from various vendors as described
in Table 1 below, were prepared at the 7 g scale as shown on Table
2 below. The samples were prepared by weighing the amounts required
as stated in Table 2 and dissolved in deionized water to form a
solution or suspension, the oxytocin was added and mixed prior to
spray drying. In the samples using the citrate buffer and the
divalent cation, oxytocin was dissolved in the citrate buffer prior
to adding the rest of the ingredients in the mixture. The solution
or suspension was then spray-dried using the parameters as
described in Table 3 below. Suspensions were homogenized in a high
shear mixer prior to spray drying. The solutions were filtered
through a 0.2 .mu.m membrane prior to spray-drying.
[0075] The dry powders were collected and used in the experiments
described below. Experiments were conducted to characterize the
powders obtained using various techniques, including to measure the
oxytocin content of various samples before and after incubation to
determine yields, loss on drying (LOD), aerodynamic performance,
particle size and particle morphology were evaluated. Stability
studies were carried out using aliquots from each of the dry powder
samples prepared, which had been incubated at 40.degree. C. in a
relative humidity of 75% (40.degree. C./75% RH) in scintillation
vials sealed with a fluoropolymer resin lined screw cap which had
been placed in heat-sealed aluminum pouches for the time periods
studied. Samples of the incubated material were taken at various
times after onset of the experiments and up to approximately 7
months. The samples were evaluated by high performance liquid
chromatography (HPLC) assay (see preparation described below) to
determine the presence of the oxytocin in the samples and the
degradation products. Oxytocin stability studies were performed up
to 11 months for certain powders, including, Sample ID Nos. 4, 6
and 13 in Table 2.
TABLE-US-00001 TABLE 1 Formulation Components Chemicals Supplier
FDKP Na MannKind PVP K30 Spectrum Trehalose Alfa Aesar L-isoleucine
Alfa Aesar Trileucine Bachem Cystine Alfa Aesar Citric acid
anhydrous EMD Trisodium citrate dihydrate Alfa Aesar Zinc chloride
Sigma Aldrich Zinc citrate Sigma Aldrich Oxytocin American
peptide
Buchi mini spray dryer B 290 Filtration unit fast PES membrane (0.2
.mu.m) 150 mL system (Nalgene)
Homogenizer (Tekmar Tissumizer)
TABLE-US-00002 [0076] TABLE 2 Sample Formulations Contents Sample
Wt. % Target (dry basis) ID No. Citrate ZnCl.sub.2 Trehalose
Na.sub.2FDKP ILE* PVP 1 -- -- 87.00 -- 10.00 2.00 2 24.03 6.76
59.94 -- 6.89 1.38 3 12.01 3.38 73.47 -- 8.45 1.69 4 4.81 1.35
81.59 -- 9.38 1.88 5 24.03 -- 65.89 -- 7.57 1.51 6 27.73 6.76 56.69
-- 6.52 1.30 7 27.73 6.76 38.44 19.55 6.52 -- 8 -- -- 59.00 30.00
10.00 -- 9 -- -- 44.00 44.00 10.00 1.00 10 -- -- 89.00 -- 10.00 --
11 -- -- 87.00 -- TLE* 2.00 10.00 12 -- -- 87.00 -- CYS* 10.00 2.00
13 24.03 Zn citrate 39.26 -- ILE 0.90 30.30 4.51 *ILE: isoleucine,
TLE: trileucine, CYS: cystine.
[0077] Spray-dried powders were prepared with a target oxytocin
content of 1%. Formulation contents are detailed in Table 2. A
mixture having a ratio of 87:10:2 by weight of trehalose,
isoleucine and PVP served as a matrix for control formulations of
Samples ID Nos. 1 to 6. To this mixture were added sodium citrate
and zinc. The quantities of citrate salts were varied from 100 to
20 equivalents per mole of oxytocin (24 to 4.8% of total weight).
The quantities of the zinc salts were varied from 50 to 10
equivalents per mole of oxytocin (6.7 to 1.3% of total weight).
Concentrated sodium citrate buffers (75 mM pH 4.5 and 6.5) were
used as the source of citrate.
[0078] The solids content of the feed solutions was kept constant
at 5%. Feed solutions were filtered prior spray-drying. One
formulation containing FDKP appeared cloudy and was not filtered
(Sample ID No. 7); mixtures containing trileucine, cystine and zinc
citrate (Samples ID No. 11, 12 and 13) required homogenization and
the resulting suspensions were kept under constant stirring during
the spray-drying process. A suspension containing zinc citrate and
citrate buffer was prepared as feed solution and homogenized in a
high sheer mixing (Tissumizer homogenizer). A second solution
containing oxytocin and the remaining excipients in water were
added to the suspension and the final weight was adjusted to 140 g
with deionized water.
TABLE-US-00003 TABLE 3 Spray-drying conditions Inlet: 130.degree.
C. Outlet: 6.degree. C. Drying gas flow: 60 mbar (nitrogen)
Atomization flow: ~59.9 g/h Aspirator 80% Pump 5%
[0079] The oxytocin stability study results from are shown in Table
4 below. The data is shown as the percent (%) of oxytocin remaining
in the sample compared to the starting amount of material used. As
seen in Table 4 and FIG. 3, three of the powder formulations
(Sample ID Nos. 4, 6, and 13) tested maintained more than about 90%
of the oxytocin as assayed after 32 weeks of incubation. The data
also show that the combination of sodium citrate and zinc salt led
to the highest stability (less degradation of oxytocin) in solid or
dry powder form (about 100%, Sample ID No. 6). Moreover, the
addition of sodium citrate and zinc also led to higher respirable
fraction per fill content of powder (RF/fill) used, with a maximum
RF/fill of 60.2% for a powder containing 12% (w/w) sodium citrate,
3.4% (w/w) zinc chloride, 73.5% (w/w) trehalose, 8.4% (w/w)
isoleucine and 1.69% (w/w) PVP. The control powder formulated
without zinc and citrate, had a RF/fill of 40.9% (Sample ID No. 1),
but its oxytocin degradation rate was more rapid as there was only
51.6% oxytocin remaining in the sample after 32 weeks of
incubation.
[0080] Scanning electron micrographs (SEM) of sample control
powders were studied and shown in FIG. 1A (low magnification) and
FIG. 1B. (high magnification The SEMs show regularly-shaped,
substantially spherical particles, which appear substantially
homogeneous in size with small surface indentations and typical of
amorphous powders.
TABLE-US-00004 TABLE 4 Stability results for oxytocin powders at
40.degree. C./75% RH ID No. t = 0 t = 2 W t = 4 W t = 8 W t = 12 W
t = 16 W t = 20 W t = 24 W t = 28 W t = 32 W t = 36 W t = 40 W t =
44 W 1 100 87.8 68.3 67.6 69.1 69.5 59.8 61.3 57.0 51.6 2 100 90.3
87.1 86.4 83.2 81.9 79.9 77.8 12.7 75.7 3 100 91.9 89.8 88.2 86.8
84.6 79.3 81.1 78.1 79.1 4 100 94.1 95.8 94.0 94.1 89.5 90.0 87.3
90.2 89.2 82.8 82.3 81.6 5 100 98.1 94.0 93.5 88.8 87.0 84.9 80.5
82.5 81.6 6 100 99.1 97.8 97.8 93.2 95.6 94.9 91.5 92.9 101.4 101.4
89.4 90.0 7 100 91.4 90.1 88.9 86.5 78.2 73.2 70.1 73.0 67.6 8 100
95.9 91.1 86.4 55.3 71.7 60.7 71.7 63.4 60.3 9 100 94.9 94.1 92.5
88.0 82.2 81.3 78.3 74.2 66.1 10 100 89.6 85.6 67.3 59.8 51.1 50.0
49.1 46.4 46.6 11 100 99.3 89.5 88.2 83.5 77.5 73.8 76.2 73.6 72.4
12 100 94.4 93.1 87.0 84.5 78.9 83.7 82.0 79.1 76.4 13 100 98.8
94.2 93.3 91.0 91.7 89.5 90.3 92.5 92.0 93.6 87.6 83.2
Dry Powder Characterizations: LOD), RF/Fill, SEM, and Oxytocin
Assay
[0081] Loss on drying (LOD) was measured by thermogravimetric
analysis (TGA) with a heat and hold method (20.degree. C./min,
110.degree. C. isotherm for 30 minutes). The powders were obtained
with an average yield of 73.8% and a minimum LOD of 4.63%.
[0082] Aerodynamic performance of the spray-dried powders was
measured by Andersen Cascade Impaction with the Gen2C inhaler (30
Lpm, 8 s, MannKind Corp.) and the results are shown in Table 5.
Geometric particle size was determined by laser diffraction using a
Sympatec RODOS M powder disperser set at 0.5 bar and 3 bar
dispersing pressures. Particle morphology was assessed by field
emission scanning electron microscopy. Table 5 shows that particles
range in size from about 3.8 to 5.6 .mu.m at 0.5 bar and 3 bar
atmospheric pressures tested and had a % RF/fill of about 40 to
about 60%. As see in Table 5, the samples containing citrate and
zinc (Sample ID No. 2, 3 and 4) performed best as shown by the
cartridge emptying data (>70%) and emitted dose of 56% to 60%.
Density of the powders was evaluated with a tapped density analyzer
(Autotap) after 3000 taps. Density of the bulk powder didn't exceed
0.5 g/ml regardless of salt contents.
[0083] Oxytocin content was evaluated using an HPLC method.
Oxytocin standard solutions were prepared at approximately 250
.mu.g/mL in 0.1M sodium bicarbonate pH 9.5 (6.25 mg of oxytocin raw
material in 25.0 mL). Powders were prepared by dissolving 10.+-.1.0
mg in 0.1M sodium bicarbonate pH 9.5 to give a final oxytocin
concentration of 0.250 mg/mL. Initial drug content was assayed to
ascertain the starting material. Powders were prepared with a
target drug content of 1% and assays confirmed the oxytocin content
between 0.92 and 1.13%.
Oxytocin Stability in Powder Form
[0084] The powders were weighed into 20 mL glass vials that were
then closed, wrapped in foil, and heat sealed. The foil pouches
were placed on a stability chamber at 40.degree. C./75% RH. Samples
were pulled at 2 and 4 weeks; then pulled every 4 weeks and up to
32 weeks after incubation. Samples were stored frozen (-20.degree.
C.) until assayed by HPLC as discussed above.
TABLE-US-00005 TABLE 5 Aerodynamic performance, particle size and
density characterizations Aerodynamic performance Particle size
Density ID (30 LPM, 8 s) (.mu.m) (g/l) No. % RF % RF/fill % CE 0.5
bar 3 bar D.sub.Bulk D.sub.Tap 1 58.4 40.9 70.0 4.06 3.85 0.411
0.553 2 76.5 59.1 77.2 4.09 3.86 0.472 0.594 3 77.5 60.3 77.8 4.13
3.98 0.408 0.582 4 71.4 55.4 77.3 4.12 4.07 0.456 0.623 5 74.3 55.9
75.3 4.18 3.96 0.454 0.649 6 69.4 59.8 86.2 4.22 4.16 0.389 0.620 7
72.8 42.5 58.3 5.62 5.61 -- -- 8 -- -- -- 4.10 4.02 -- -- 9 -- --
-- 4.38 4.29 -- -- 10 68.8 59.5 86.4 4.05 3.86 -- -- 11 56.4 50.6
89.6 5.25 4.90 -- -- 12 -- -- -- 4.07 3.83 -- -- 13 70.8 53.2 75.1
4.17 4.00 0.397 0.580
Aerodynamic Performance, Particle Size and Morphology
[0085] Aerodynamic testing on selected powders highlighted the
beneficial effect of combining sodium citrate and zinc with
trehalose, isoleucine and PVP. The effect was observed with citrate
and zinc contents as low as 4.8% and 1.3% respectively (Table 5,
Sample ID No. 4). The maximum effect (60.3% RF/fill) was obtained
with 12% citrate content and 3.4% zinc. Powder Sample ID No. 2,
prepared with twice the amount of citrate and zinc, had an RF/fill
of 59.1%. Both Sample powders ID Nos. 2 and 4 when tested in the
inhaler were delivered out of the inhaler at about 77% of the
original content. The RF/fill of the control powder (Sample ID No.
1) formulated with trehalose, isoleucine and PVP (87/10/2) was
40.9% and at a rate less than the samples containing zinc and
citrate.
[0086] Particle morphology studied by scanning electron microscopy
shows that spray-drying of the control formulation (Sample ID No.
1) containing trehalose, PVP and isoleucine produced slightly
corrugated, spherical particles typical of leucine-containing
powders (FIG. 2A). The corrugated substantially spherical
morphology was maintained with the addition of salts (zinc and
citrate salts) to the mixture containing trehalose, PVP and
isoleucine (FIGS. 2B; 2C and 2D). However, the particles containing
zinc and citrate differ from the controls as they appear slightly
more corrugated and less spherical. As shown in the SEMs, the
particles formed with zinc and/or citrate appear substantially
spherical and have a slightly more indentations, corrugated surface
or wrinkle appearance, and less regular pattern. It was observed
that the particles containing oxytocin, zinc and citrate appeared
to be more fragile or more collapsible than the controls during
preparation in particular during vacuum drying step.
[0087] FIG. 4 provides a graphic representation of data obtained
from X-ray diffraction studies of dry powders showing amorphous
content of the powders by their characteristic scan patterns. The
data demonstrates that the X-ray diffraction analysis confirmed
that the spray-dried powders all appear as uniform amorphous in
content as demonstrated by data scans depicted in FIG. 4.
[0088] The data also demonstrates that the addition of citrate/zinc
to a powder containing FDKP (Sample ID No. 7) (19.5% w/w),
trehalose (38.4% w/w) and isoleucine (6.5% w/w) produced a powder
with improved properties (42% RF/fill) over the powder without
citrate and zinc. The powder containing citrate yielded a 17%
improvement in RF/fill over a powder formulated without
citrate/zinc (25.6% RF/fill, 78.4% CE).
[0089] The present powders were not excessively cohesive because
their median geometric particle sizes were similar at 0.5 bar and 3
bar RODOS dispersing pressures. The average values were 4.34 and
4.18 .mu.m at 0.5 and 3 bars.
[0090] The data in Table 5 show the aerodynamic performance of the
powders. Table 5 shows that the powders containing citrate and zinc
yielded high respirable fractions (>70%) and cartridge emptying
data in some instances were greater than >90% (data not shown).
Sample testing in an anatomically correct airway model showed that
about 73% of the dose in an inhaler containing the powders is
delivered to the lungs.
Oxytocin Stability Studies
[0091] The data indicate that out of the fourteen powders prepared,
three maintained more than 89% of the original total oxytocin
content obtained after 32 weeks of incubation at 40.degree. C./75%
RH (Table 4). The degradation rate appears to be the highest before
4 weeks for the most stable powders (FIG. 3) then plateaus; this
early onset of degradation is probably due to moderate to high
residual water content in the powders. The most stable powders was
prepared from a pH 6.5 citrate buffer and zinc chloride as a source
of zinc divalent cations. Overall, powders prepared in presence of
both citrate and zinc salts exhibited the highest stability. The
stabilizing effect of this combination was even observed at low
salts contents (Sample ID No. 1 vs. Sample ID Nos. 2, 3, 4). Among
the "non-buffered" formulations, the addition of disodium FDKP or
replacing isoleucine by trileucine or cystine enhanced the
stability of the powders.
Example 2
[0092] Preparation, Characterization and Stability of Alternative
Oxytocin Spray-Dried Powder Embodiments
[0093] Preparation of oxytocin spray-dried powders was performed as
in Example 1 above. In these experiments, fourteen powders
containing 1% (w/w) oxytocin and varying amounts of buffers, salts,
carries excipients, including, trehalose, PVP, isoleucine, sodium
citrate, citric acid, sodium tartrate, tartaric acid and zinc salt,
obtained from various vendors as described in Table 6 below, were
prepared at the 2.5 g scale as shown on Table 1 below. In these
experiments, L-(+)-tartaric acid and sodium L-(+)-tartrate
dihydrate were used and obtained from Alfa Aesar. Unlike in Example
1, bulk solid sodium citrate salt and citric acid were used as
source of sodium citrate. Samples containing 1% (w/w) oxytocin were
made as described in Example 1 and the solutions or suspensions
were then spray-dried using the parameters as described in Table 6
below.
[0094] The dry powders were collected and used in the experiments
described below. Powders identified with sample numbers ID Nos. 14
to 28 were obtained with an average yield of 76.7% and a minimum
loss on drying (LOD) of 5.73% (measured by Karl Fisher titration).
Powders ID Nos. 14 to 28 were submitted to an additional drying
step under vacuum pump that led to a minimum LOD of 2.90%.
[0095] Spray-dried powders containing a target oxytocin content of
1% were assayed and the data confirmed the oxytocin values ranged
between 0.87 to 1.01%. The components of the prepared formulations
are detailed in Table 7 showing the contents of each sample made
and tested. In certain embodiments, a mixture having a ratio of
87:10:2 by weight of trehalose, isoleucine and PVP served as a
matrix for all the formulations except samples ID No. 20 and 21. To
this mixture sodium citrate, citric acid and zinc were added. The
quantities of citrate salts were varied from 100 to 50 equivalents
per mole of oxytocin (29.2 to 14.6% of total weight). The
quantities of the zinc salts were varied from 50 to 5 equivalents
per mole of oxytocin (30.3 to 0.7% of total weight). In some
embodiments, the zinc cation appeared to be essential to the
composition characteristics as exemplified by zinc chloride use
alone (Sample ID No. 22) also provided improved stability of the
powders.
TABLE-US-00006 TABLE 6 Drying Conditions Inlet: 150.degree. C.
Outlet: 70.degree. C. Drying gas flow: 60 mbar (nitrogen)
Atomization flow: ~59.9 g/h Aspirator 80% Pump 5%
TABLE-US-00007 TABLE 7 Oxytocin Sample compositions Wt. % Target
(dry basis) Sample Sodium Citric ID No. citrate acid ZnCl.sub.2
Trehalose ILE PVP 14 29.2 -- 6.8 56.3 6.5 1.3 15 29.2 -- 2.7 59.8
6.9 1.4 16 29.2 -- 1.4 61.0 7.0 1.4 17 29.2 -- 0.7 61.6 7.1 1.4 18
14.6 -- 6.8 69.1 7.9 1.6 19 24.3 3.16 6.8 57.8 6.6 1.3 20 29.2 --
6.8 64.0 -- -- 21 -- -- 6.8 57.6 6.4 -- 22 -- -- 6.8 81.94 9.42
1.88 Zinc citrate 23 -- -- 30.3 61.2 7.0 1.4 24 -- -- 9.1 79.9 9.2
1.8 25 24.3 3.16 30.30 37.07 4.26 0.85 26 14.6 -- 30.30 48.42 5.57
1.11 Sodium Tartaric tartrate acid 27 22.8 -- 6.8 61.9 7.1 1.4 28
19.0 3.0 6.8 62.6 7.2 1.4
[0096] Aerodynamic Performance of Dry Powders
[0097] Aerodynamic performance of the spray-dried powders was
measured by Andersen Cascade Impaction with the Gen2C inhaler (21.6
Lpm, 4 s, MannKind Corp.) and the results are shown in Table 8.
High % Rf/fill (>50%) were obtained even under a low peak
inspiratory pressure. The data in Table 8 Shows the % RF/fill
ranging from about 20 to about 60% and cartridge emptying of total
contents was up to 77% (Sample ID No. 22). The highest % Rf/Fill
were obtained for powders containing zinc citrate, zinc chloride
with or without PVP. As seen in Table 8, % Rf/fill was improved by
the addition of isoleucine (Sample ID Nos. 14 and 20). The samples
containing zinc citrate or zinc chloride alone (Sample ID Nos. 22,
23 and 24) had high % Rf/fill of about 50 to 60% and cartridge
emptying greater than 70%.
[0098] Aerodynamic testing on selected powders highlighted the
beneficial effect of combining sodium citrate and zinc chloride
with trehalose, isoleucine with or without PVP as exemplified by
Sample ID No. 18, 19, 20 and 21. The improved performance of the
powders was observed with citrate and zinc contents as low as 14.6%
and 1.4% respectively (Sample ID Nos. 16 and 18). The maximum
effect (53.0% RF/fill) was obtained with 14.6% content in sodium
citrate and 6.8% in zinc (Sample ID No. 18). The beneficial effect
of combining zinc citrate with trehalose, isoleucine and PVP is
exemplified by the performance of powders Sample ID Nos. 23 and 24,
which yielded % Rf/fill greater than 50% and cartridge emptying of
about 73%.
TABLE-US-00008 TABLE 8 Aerodynamic performance by Andersen cascade
impactor. Sample ID No. % RF/fill % CE 14 43.3 57.2 15 47.3 62.1 16
49.8 61.2 17 22.4 35.3 18 53.0 71.9 19 44.8 61.6 20 36.0 52.4 21
50.3 66.9 22 51.2 77.0 23 51.5 73.0 24 60.3 72.7 25 37.5 55.0 26
37.7 59.3 27 57.3 74.7 28 45.1 59.6
[0099] Oxytocin Formulation Stability Studies
[0100] Stability of oxytocin spray-dried powders was performed as
in Example 1 above. Stability testing was performed up to 40 weeks.
The oxytocin stability study results from the assays are shown in
Table 9 below and FIG. 5. The data is shown as the percent (%)
remaining of sample compared to the starting material used. As seen
in Table 9, only 3 of the powder formulations (Sample ID Nos. 16,
27, and 28) tested maintained less than about 90% of the oxytocin
when aliquots of the sample were assayed after 40 weeks incubation.
The combination of citrate and zinc salts led to the highest
stability in solid state (greater than about 90%). The highest
stability was achieved with the combination containing 14.6% sodium
citrate and 6.8% zinc chloride (Sample ID No. 18). Powders
containing a minimum 9.1% content in zinc citrate with or without
sodium citrate maintained more than 97% of the oxytocin after 40
weeks incubation. FIG. 5 is a graphic representation of dry powder
samples from the stability studies wherein the samples containing
divalent zinc salt and citrate salts at various concentrations
showed a slow degradation of the oxytocin over a period of 40
weeks; wherein the samples tested retained greater than 90% of the
oxytocin content.
[0101] In the alternative embodiment using tartrate, powders
containing zinc and tartrate salts maintained also about 90% of the
oxytocin content after 24 weeks of incubation and greater than 85%
of the oxytocin content after 32 weeks of sample incubation.
TABLE-US-00009 TABLE 8 Stability results for oxytocin powders at
40.degree. C./75% RH ID No. t = 0 t = 2 W t = 4 W t = 8 W t = 14 W
t = 16 W t = 20 W t = 24 W t = 32 W t = 40 W 14 100 99.9 101.9
100.0 99.1 94.6 100.9 83.5 85.8 91.4 15 100 98.5 104.2 97.8 97.5
96.0 93.5 78.0 74.3 95.7 16 100 100.2 102.2 94.9 96.0 100.8 55.1
79.7 41.7 33.9 17 100 97.0 99.7 96.7 96.5 93.7 94.3 91.1 85.3 90.1
18 100 105.2 109.5 103.3 107.0 104.2 103.2 101.4 103.1 103.8 19 100
103.5 102.2 102.4 102.1 101.5 101.6 92.8 94.6 96.9 20 100 103.5
104.5 103.1 103.2 103.3 105.1 93.3 101.4 96.0 21 100 100.2 100.9
101.2 101.2 101.2 99.6 99.3 93.3 92.9 22 100 101.4 100.5 98.0 96.3
100.1 95.7 95.6 92.4 94.2 23 100 99.6 97.6 100.1 99.0 101.0 100.8
97.7 88.7 97.3 24 100 101.4 104.9 98.4 103.8 106.0 95.6 84.2 97.8
103.1 25 100 103.8 108.4 103.8 103.6 104.4 103.7 89.7 96.2 100.0 26
100 100.9 100.7 96.0 100.0 96.1 96.7 96.6 96.9 99.8 27 100 97.3
100.1 98.4 97.6 98.0 97.5 91.8 88.4 66.0 28 100 102.6 102.6 100.0
99.8 98.7 94.3 93.4 86.8 88.7
[0102] The addition of citrate or tartrate and zinc salts to the
formulations of oxytocin appeared beneficial for both the
aerodynamic performance and oxytocin stability.
Example 3
[0103] A pregnant woman, 35 year old and in her second pregnancy
has a history of mild post-partum hemorrhaging in her first
pregnancy is admitted to the hospital at 40 weeks of pregnancy and
in labor. Contractions are monitor to occur 5 minutes apart by the
attending obstetrician. The pregnant woman is noted to be bleeding
and delivers a healthy baby. In the operating room and immediately
upon childbirth, the woman is administered by oral inhalation a dry
powder formulation containing a single dose of 100 IU of oxytocin,
28% (w/w) citrate and 7% (w/w) zinc chloride, in a single
inhalation, using an inhalation system comprising a high resistance
inhaler as described in U.S. Pat. No. 8,484,129, which disclosure
is incorporated herein by reference in its entirety for its
teaching of the relevant subject matter. The woman was kept in the
hospital for 3 days and did not suffer any severe bleeding and was
released with her newborn baby.
[0104] Unless otherwise indicated, all numbers expressing
quantities of ingredients, properties such as molecular weight,
reaction conditions, and so forth used in the specification, and
claims are to be understood as being modified in all instances by
the term "about." Accordingly, unless indicated to the contrary,
the numerical parameters set forth in the following specification
and attached claims are approximations that may vary depending upon
the desired properties sought to be obtained by the present
invention. At the very least, and not as an attempt to limit the
application of the doctrine of equivalents to the scope of the
claims, each numerical parameter should at least be construed in
light of the number of reported significant digits and by applying
ordinary rounding techniques. Notwithstanding that the numerical
ranges and parameters setting forth the broad scope of the
invention are approximations, the numerical values set forth in the
specific examples are reported as precisely as possible. Any
numerical value, however, inherently contains certain errors
necessarily resulting from the standard deviation found in their
respective testing measurements.
[0105] The terms "a" and "an" and "the" and similar referents used
in the context of describing the invention (especially in the
context of the following claims) are to be construed to cover both
the singular and the plural, unless otherwise indicated herein or
clearly contradicted by context. Recitation of ranges of values
herein is merely intended to serve as a shorthand method of
referring individually to each separate value falling within the
range. Unless otherwise indicated herein, each individual value is
incorporated into the specification as if it were individually
recited herein. All methods described herein can be performed in
any suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g. "such as") provided herein is intended
merely to better illuminate the invention and does not pose a
limitation on the scope of the invention otherwise claimed. No
language in the specification should be construed as indicating any
non-claimed element essential to the practice of the invention.
[0106] The use of the term "or" in the claims is used to mean
"and/or" unless explicitly indicated to refer to alternatives only
or the alternatives are mutually exclusive, although the disclosure
supports a definition that refers to only alternatives and
"and/or."
[0107] Groupings of alternative elements or embodiments of the
invention disclosed herein are not to be construed as limitations.
Each group member may be referred to and claimed individually or in
any combination with other members of the group or other elements
found herein. It is anticipated that one or more members of a group
may be included in, or deleted from, a group for reasons of
convenience and/or patentability. When any such inclusion or
deletion occurs, the specification is herein deemed to contain the
group as modified thus fulfilling the written description of all
Markush groups used in the appended claims.
[0108] Preferred embodiments of this invention are described
herein, including the best mode known to the inventors for carrying
out the invention. Of course, variations on those preferred
embodiments will become apparent to those of ordinary skill in the
art upon reading the foregoing description. The inventor expects
those of ordinary skill in the art to employ such variations as
appropriate, and the inventors intend for the invention to be
practiced otherwise than specifically described herein.
Accordingly, this invention includes all modifications and
equivalents of the subject matter recited in the claims appended
hereto as permitted by applicable law. Moreover, any combination of
the above-described elements in all possible variations thereof is
encompassed by the invention unless otherwise indicated herein or
otherwise clearly contradicted by context.
[0109] Specific embodiments disclosed herein may be further limited
in the claims using consisting of or consisting essentially of
language. When used in the claims, whether as filed or added per
amendment, the transition term "consisting of" excludes any
element, step, or ingredient not specified in the claims. The
transition term "consisting essentially of" limits the scope of a
claim to the specified materials or steps and those that do not
materially affect the basic and novel characteristic(s).
Embodiments of the invention so claimed are inherently or expressly
described and enabled herein.
[0110] Furthermore, numerous references have been made to patents
and printed publications throughout this specification. Each of the
above cited references and printed publications are herein
individually incorporated by reference in their entirety.
[0111] Further, it is to be understood that the embodiments of the
invention disclosed herein are illustrative of the principles of
the present invention. Other modifications that may be employed are
within the scope of the invention. Thus, by way of example, but not
of limitation, alternative configurations of the present invention
may be utilized in accordance with the teachings herein.
Accordingly, the present invention is not limited to that precisely
as shown and described.
* * * * *