U.S. patent application number 12/981036 was filed with the patent office on 2011-10-20 for pharmaceutical compositions and related methods of delivery.
Invention is credited to Yaron Ilan, Irina Karmeli, Roni Mamluk, Karen Marom, Paul Salama, Moshe Tzabari.
Application Number | 20110257095 12/981036 |
Document ID | / |
Family ID | 41277888 |
Filed Date | 2011-10-20 |
United States Patent
Application |
20110257095 |
Kind Code |
A1 |
Salama; Paul ; et
al. |
October 20, 2011 |
PHARMACEUTICAL COMPOSITIONS AND RELATED METHODS OF DELIVERY
Abstract
The pharmaceutical compositions described herein include a
suspension which comprises an admixture in solid form of a
therapeutically effective amount of a therapeutic agent and at
least one salt of a medium chain fatty acid and a hydrophobic
medium, e.g. castor oil or glyceryl tricaprylate or a mixture
thereof. The pharmaceutical compositions described herein contain
medium chain fatty acid salts and are substantially free of
alcohols. The pharmaceutical compositions may be encapsulated in a
capsule. Methods of treating or preventing diseases by
administering such compositions to affected subjects are also
disclosed.
Inventors: |
Salama; Paul; (Ashdod,
IL) ; Mamluk; Roni; (Modin, IL) ; Marom;
Karen; (Mevaseret Zion, IL) ; Karmeli; Irina;
(Maale Adummim, IL) ; Ilan; Yaron; (Jerusalem,
IL) ; Tzabari; Moshe; (Jerusalem, IL) |
Family ID: |
41277888 |
Appl. No.: |
12/981036 |
Filed: |
December 29, 2010 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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12561738 |
Sep 17, 2009 |
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12981036 |
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61097716 |
Sep 17, 2008 |
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61141686 |
Dec 31, 2008 |
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61161387 |
Mar 18, 2009 |
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Current U.S.
Class: |
514/10.9 ;
514/11.1 |
Current CPC
Class: |
A61K 9/0031 20130101;
A61P 1/18 20180101; A61P 1/14 20180101; A61K 47/32 20130101; A61P
31/14 20180101; A61K 38/212 20130101; A61P 9/12 20180101; A61P
31/04 20180101; A61K 38/14 20130101; A61K 47/14 20130101; A61P 9/08
20180101; A61K 31/7036 20130101; A61K 9/4866 20130101; A61K 38/28
20130101; A61P 9/00 20180101; A61K 9/4891 20130101; A61K 38/09
20130101; A61P 1/00 20180101; A61P 5/00 20180101; A61P 31/00
20180101; A61K 38/08 20130101; A61P 3/10 20180101; A61P 27/02
20180101; A61K 38/12 20130101; A61P 7/04 20180101; A61K 38/22
20130101; A61K 31/713 20130101; A61K 47/26 20130101; A61K 38/26
20130101; A61P 5/08 20180101; A61K 38/095 20190101; A61K 51/1021
20130101; A61P 35/00 20180101; A61K 9/0053 20130101; A61P 9/14
20180101; A61K 47/24 20130101; A61P 37/00 20180101; A61K 31/721
20130101; A61P 1/04 20180101; A61P 3/04 20180101; A61K 31/70
20130101; A61K 38/29 20130101; A61K 51/1045 20130101; A61K 47/12
20130101; A61K 51/083 20130101; A61P 31/20 20180101; A61P 13/12
20180101; A61K 51/1024 20130101; A61P 1/12 20180101; A61K 38/27
20130101; A61P 1/10 20180101; A61P 5/06 20180101; A61K 9/10
20130101; A61K 9/1623 20130101; A61P 1/16 20180101; A61P 3/00
20180101; A61P 5/02 20180101; A61K 9/4858 20130101; A61P 25/00
20180101; A61K 47/44 20130101 |
Class at
Publication: |
514/10.9 ;
514/11.1 |
International
Class: |
A61K 38/11 20060101
A61K038/11; A61K 38/12 20060101 A61K038/12; A61P 9/14 20060101
A61P009/14; A61P 9/12 20060101 A61P009/12; A61P 1/04 20060101
A61P001/04; A61K 38/31 20060101 A61K038/31; A61P 9/08 20060101
A61P009/08 |
Claims
1-195. (canceled)
196. A method of prophylactically treating a subject for variceal
bleeding, which comprises orally administering to the subject a
somatostatin analog or a composition comprising a somatostatin
analog.
197. The method of claim 196, wherein the somatostatin analog is
octeotride.
198. The method of claim 196, wherein the prophylaxis is
primary.
199. The method of claim 196, wherein the prophylaxis is
secondary.
200. The method of claim 196, wherein the variceal bleeding is
associated with portal hypertension.
201. The method of claim 196, wherein the varices are gastric or
esophageal.
202. The method of claim 196, wherein the somatostatin analog is
administered once daily.
203. The method of claim 196, wherein the somatostatin analog is
administered twice or more daily.
204. A method of treating a subject suffering from portal
hypertension or variceal bleeding, comprising orally administering
to the subject a somatostatin analog or a composition comprising a
somatostatin analog.
205. The method of claim 204, wherein the somatostatin analog is
octeotride.
206. The method of claim 204, wherein the subject suffers from
variceal bleeding.
207. The method of claim 204, wherein the varices are gastric or
esophageal.
208. The method of claim 204, wherein the subject suffers from
portal hypertension.
209. The method of claim 204, wherein the therapeutic agent is
administered once daily.
210. The method of claim 204, wherein the therapeutic agent is
administered twice or more daily.
211. A method of treating a subject suffering from abnormal GI
motility, flushing episodes associated with carcinoid syndrome, an
endocrine tumor (such as carcinoids, VIPoma), gastroparesis,
diarrhea, pancreatic leak or a pancreatic pseudo-cyst, or
hepato-renal syndrome, shock of hypovolemic (e.g. hemorrhagic) or
vasodilatory (e.g. septic) origin, cardiopulmonary resuscitation or
anesthesia-induced hypotension, comprising orally administering to
the subject a somatostatin analog or a composition comprising a
somatostatin analog.
212. The method of claim 211, wherein the somatostatin analog is
octeotride.
213. A method of prophylactically treating a subject for variceal
bleeding, which comprises orally administering to the subject a
vasopressin analog or a composition comprising a vasopressin
analog.
214. The method of claim 213, wherein the vasopressin analog is
terlipressin.
215. The method of claim 213, wherein the prophylaxis is
primary.
216. The method of claim 213, wherein the prophylaxis is
secondary.
217. The method of claim 213, wherein the variceal bleeding is
associated with portal hypertension.
218. The method of claim 213, wherein the therapeutic agent is
administered once daily.
219. The method of claim 213, wherein the therapeutic agent is
administered twice or more daily.
220. A method of treating a subject suffering from hepato-renal
syndrome (HRS), bleeding esophageal varices or portal hypertension,
which comprises orally administering to the subject a vasopressin
analog or a composition comprising a vasopressin analog.
221. The method of claim 220, wherein the subject suffers from
hepato-renal syndrome (HRS).
222. The method of claim 220, wherein the hepato-renal syndrome
(HRS) is HRS I.
223. The method of claim 220, wherein the hepato-renal syndrome
(HRS) is HRS II.
224. The method of claim 220, wherein the subject suffers from
bleeding esophageal varices.
225. The method of claim 220, wherein the subject suffers from
portal hypertension.
226. The method of claim 220, wherein the vasopres sin analog is
terlipressin.
227. The method of claim 220, wherein the variceal bleeding is
associated with portal hypertension.
228. The method of claim 220, wherein the varices are gastric or
esophageal.
229. The method of claim 220, wherein the therapeutic agent is
administered once daily.
230. The method of claim 220, wherein the therapeutic agent is
administered twice or more daily.
Description
CLAIM OF PRIORITY
[0001] This application claims priority from U.S. Ser. No.
61/097,716, filed Sep. 17, 2008, U.S. Ser. No. 61/141,686, filed
Dec. 31, 2008, and U.S. Ser. No. 61/161,387, filed Mar. 18, 2009,
each of which is incorporated herein by reference in its
entirety.
FIELD OF THE TECHNOLOGY
[0002] The present invention relates generally to pharmaceutical
compositions enabling improved delivery e.g. oral delivery and
methods of using such compositions.
BACKGROUND
[0003] Techniques enabling efficient transfer of a substance of
interest across a biological barrier are of considerable interest
in the fields of biotechnology and medicine. For example, such
techniques may be used for the transport of a variety of different
substances across a biological barrier regulated by tight junctions
(i.e., the mucosal epithelia, which include the intestinal and
respiratory epithelia, and the vascular endothelia, which include
the blood-brain barrier, nasal membrane, cornea and other eye
membranes, and genito-urinary membranes). In particular there is
great interest in oral delivery of therapeutic agents to avoid the
use of more invasive means of administration and hence improve
patient convenience and compliance.
[0004] Diverse drug delivery vehicles have been employed, among
them liposomes, lipidic or polymeric nanoparticles, and
microemulsions. These have improved the oral bioavailability of
certain drugs, mostly by the protective effect they offer. However,
for most relevant drugs, bioavailability remains very low and fails
to achieve the minimal therapeutic goals.
[0005] Hence, a need exists for an efficient, specific,
non-invasive, low-risk means to target various biological barriers
for the non invasive delivery of various therapeutic agents such as
peptides and polypeptides, macromolecule drugs and other
therapeutic agents which include small molecules with low
bioavailability.
SUMMARY
[0006] The inventors of the present invention have discovered that
the absorption of certain therapeutic agents in a subject can be
improved when administered in a composition described herein. For
example, a therapeutic agent administered in a formulation in
accordance with one or more embodiments exhibits an improved
bioavailability (BA) relative to the same therapeutic agent
administered via a similar route but in a composition substantially
free of the medium chain fatty acid salt component described herein
or having a lower amount of the medium chain fatty acid salt
component described herein. Such improvement in relative BA may be
on the order of at least about 1.5-, 2-, 3-, 5-, 10-, 50- or
100-fold. In some aspects, a composition described herein improves
the absorption in the gastrointestinal (GI) tract of a therapeutic
agent that is generally characterized by low or zero oral
bioavailability and/or absorption. These therapeutic agents may
have low or zero bioavailability, e.g., in aqueous solution, and in
other oral formulations known in the art. In at least one aspect, a
composition described herein improves bioavailability by enhancing
the GI wall/barrier permeability to the drug molecules. For
example, a composition described herein may facilitate absorption
by permeating the GI wall/barrier primarily via unsealing of the
tight junctions between GI epithelial cells, although it may also
work by transcellular absorption.
[0007] The present inventors have devised a process for producing a
pharmaceutical composition (bulk drug product) which involves
preparing a water soluble composition comprising a therapeutically
effective amount of at least one therapeutic agent and a medium
chain fatty acid salt (and other ingredients--see below), drying
(e.g. by lyophilization) the water soluble composition to obtain a
solid powder, and suspending the lyophilized material (the solid
powder) in a hydrophobic (oily) medium, preferably castor oil or
glyceryl tricaprylate (including other ingredients e.g. PVP and
surfactants and viscosity modifiers--see below), to produce a
suspension containing in solid form the therapeutic agent and the
medium chain fatty acid salt, thereby producing the bulk drug
product, which must contain at least 10% by weight of medium chain
fatty acid salt. The solid form may comprise a particle (e.g.,
consists essentially of particles, or consists of particles. The
particle may be produced by lyophilization or by granulation. The
bulk drug product may then be encapsulated in capsules which will
be coated by a pH sensitive coating and may be used for oral
delivery. A typical process for producing the claimed formulation
is shown in FIG. 1, where insulin is exemplified as the active
pharmaceutical ingredient (API) and the medium chain fatty acid
salt is sodium octanoate (Na--C8), also termed sodium
caprylate.
[0008] The present invention demonstrates delivery of the product
to the intestine, which is a model for oral delivery, and from
there to the bloodstream with high bioavailability.
[0009] Thus in one aspect the invention features a composition. The
composition includes a therapeutic agent and a medium chain fatty
acid salt associated with a substantially hydrophobic medium,
preferably castor oil, wherein the therapeutic agent and the medium
chain fatty acid salt thereof are in solid form, e.g. in the same
solid form such as a particle, obtained by drying from an aqueous
medium, e.g. by lyophilizing the aqueous medium, and wherein the
medium chain fatty acid salt is present at 10% by weight or more,
preferably 12-15%, e.g., about 12%, about 13%, about 14%, or about
15% or about 16%, or about 17%, and wherein the composition
contains other ingredients (as described herein) but is
substantially free of a "membrane fluidizing agent". "Membrane
fluidizing agents" are defined as various linear, branched,
aromatic and cyclic medium chain alcohols, in particular geraniol
and octanol.
[0010] The present compositions of the invention are not emulsions.
Almost all of the present compositions are oily suspensions and the
amount of water in the compositions is very low; a few of the
present compositions which are not suspensions incorporate a high
amount (about 78% octanoic acid) and are solutions.
[0011] In the compositions of the invention, the therapeutic agent
and medium chain fatty acid salt are in intimate contact with the
substantially hydrophobic medium. For example, a powder comprising
the therapeutic agent and medium chain fatty acid salt is coated,
immersed or suspended in the substantially hydrophobic medium.
[0012] During the production process the aqueous medium which
contains the therapeutic agent and the medium chain fatty acid salt
and the other ingredients is dried (e.g. by lyophilization) to
obtain the hydrophilic fraction which is a powder (e.g., a solid
form comprising a plurality of particles), and a particle in that
powder contains all the ingredients i.e. the therapeutic agent and
medium chain fatty acid salt are together in a single particle. The
solid form may be, for example, a granulated particle or a
lyophilized particle.
[0013] In some embodiments, the therapeutic agent is selected from
the group consisting of peptides, polysaccharides, polynucleotides,
and small molecules. The therapeutic agent may be a protein. For
example, the therapeutic agent may be insulin. In other
embodiments, the therapeutic agent is a polynucleotide e.g. DNA or
RNA compound. In some embodiments, the therapeutic agent is a small
molecule, a poorly soluble drug, or a highly crystalline drug. The
therapeutic agent may be a growth hormone. In at least one
embodiment, the therapeutic agent is teriparatide. In some
embodiments, the therapeutic agent may be leuprolide or alendronate
or octreotide.
[0014] In some embodiments, the composition includes a plurality of
medium chain fatty acid salts and derivatives thereof. For example,
the solid particle may further include a plurality of medium chain
fatty acid salts and derivatives thereof.
[0015] In some embodiments, the medium chain fatty acid salt is
selected from the group consisting of sodium hexanoate, sodium
heptanoate, sodium octanoate, sodium nonanoate, sodium decanoate,
sodium undecanoate, sodium dodecanoate, sodium tridecanoate, and
sodium tetradecanoate or a combination thereof. In accordance with
one or more embodiments, the composition is substantially free of
sodium dodecanoate, sodium tridecanoate, and sodium tetradecanoate.
In some embodiments, the medium chain fatty acid is sodium
octanoate and the sodium octanoate is present at a concentration of
above 10% e.g. about 11% to about 50% weight/weight (wt/wt).
[0016] In some embodiments, the substantially hydrophobic medium
comprises a triglyceride. For example, the triglyceride may be
selected from the group consisting of glyceryl tributyrate,
glyceryl monooleate, glyceryl monocaprylate and glyceryl
tricaprylate.
[0017] In some embodiments, the substantially hydrophobic medium
comprises mineral oil, castor oil, olive oil, corn oil, coconut
oil, peanut oil, soybean oil, cotton seed oil, sesame oil or canola
oil, or combinations thereof.
[0018] In some embodiments the water-soluble composition contains a
medium chain fatty acid salt and the hydrophobic medium contains
the corresponding medium chain fatty acid; in some particular
embodiments the medium chain fatty acid salt is a salt of octanoic
acid such as sodium octanoate and the medium chain fatty acid is
octanoic acid.
[0019] In some embodiments the water-soluble composition contains a
medium chain fatty acid salt and the hydrophobic medium contains
the corresponding medium chain monoglyceride or the corresponding
medium chain triglyceride or a combination thereof; in some
particular embodiments the medium chain fatty acid salt is sodium
octanoate and the monoglyceride is glyceryl monocaprylate and the
triglyceride is glyceryl tricaprylate.
[0020] In some embodiments, the composition further includes one or
more excipients. The excipients may be a salt e.g MgCl.sub.2 or an
amine containing compound or mannitol. In some embodiments, the
excipient is in the same solid form as the therapeutic agent.
[0021] In some embodiments the excipient is a stabilizer. The
inventors unexpectedly found that although polyvinylpyrolidine
(PVP) in particular PVP-12 is known in the art as a stabilizer, in
formulations of the invention it serves to increase the effect of
the permeability enhancer on absorbance of the therapeutic
agent.
[0022] In some embodiments, the composition further includes one or
more surfactants. For example, the surfactant may be selected from
the group consisting of sorbitan monopalmitate (Span-40.RTM.),
polyoxyethylenesorbitan monooleate (Tween80), lecithin, and
glyceryl monooleate (GMO). In one or more embodiments, the
surfactant comprises from about 0.1% to about 6% by weight of the
composition.
[0023] In preferred embodiments, the composition is an oral dosage
form. For example, the composition may be filled in a hard or soft
capsule. In some embodiments, the composition is in the form of a
suppository. In accordance with one or more embodiments, the
composition may be in the form of an enema fleet.
[0024] In some embodiments, the bioavailability of the therapeutic
agent, when administered to a subject, is at least 1.5-2% relative
to parenteral (subcutaneous or intravenous) administration. In some
embodiments, the composition, when administered to a subject,
provides above 2%, above 3%, above 5%, above 10%, or above 20% or
above 30% absorption of the therapeutic agent across a biological
barrier. The levels of absorption achieved produce the therapeutic
levels needed for the indication concerned.
[0025] In one aspect, the invention features a method of treating a
disorder in a subject. The method includes administering to the
subject any one of the compositions described herein.
[0026] In some embodiments, the composition is administered orally.
In other embodiments, the composition is administered rectally,
sublingually or via buccal administration.
[0027] In some embodiments, the disorder may be anemia. In
accordance with one or more embodiments, the disorder is
osteoporosis. The disorder may be female infertility. In other
embodiments, the disorder is growth failure or growth hormone
deficiency. In at least one embodiment, the disorder is HIV-related
weight loss or wasting, acromegaly or diabetes.
[0028] In some embodiments the therapeutic agent is octreotide and
the disorder is acromegaly, abnormal GI motility, gastroparesis,
diarrhea or portal hypertension.
[0029] In some embodiments, the method may include encapsulating
the suspension to form a capsule. The method may further include
coating the capsule.
[0030] In some embodiments, the method may include providing
instructions to administer the capsule to a subject. The
instructions may relate to administering the capsule to a subject
for any indication described herein. In one aspect, the invention
features capsules provided with instructions relating to
administering the capsule to a subject for any indication described
herein.
[0031] Still other aspects, embodiments, and advantages of these
exemplary aspects and embodiments, are discussed in detail below.
Moreover, it is to be understood that both the foregoing
information and the following detailed description are merely
illustrative examples of various aspects and embodiments, and are
intended to provide an overview or framework for understanding the
nature and character of the claimed aspects and embodiments. The
accompanying drawings are included to provide illustration and a
further understanding of the various aspects and embodiments, and
are incorporated in and constitute a part of this specification.
The drawings, together with the remainder of the specification,
serve to explain principles and operations of the described and
claimed aspects and embodiments.
[0032] Throughout this application, various publications, including
United States patents, are referenced by author and year and
patents and applications by number. The disclosures of these
publications and patents and patent applications in their
entireties are hereby incorporated by reference into this
application in order to more fully describe the state of the art to
which this invention pertains.
BRIEF DESCRIPTION OF THE DRAWINGS
[0033] Various aspects of at least one embodiment are discussed
below with reference to the accompanying Figures. In the Figures,
which are not intended to be drawn to scale, each identical or
nearly identical component that is illustrated in various figures
is represented by a like numeral. For purposes of clarity, not
every component may be labeled in every drawing. The Figures are
provided for the purposes of illustration and explanation and are
not intended as a definition of the limits of the invention. In the
Figures:
[0034] FIG. 1 presents a process for production of an insulin
formulation of a composition in accordance with one or more
embodiments as referenced in the accompanying Examples;
[0035] FIGS. 2-5 present data referenced in accompanying Examples 3
through 6;
[0036] FIG. 6 presents data referenced in accompanying Example
8;
[0037] FIG. 7 presents molecular weight marker permeability data
referenced in accompanying Example 33;
[0038] FIG. 8 presents time-course permeability data referenced in
accompanying Example 34; and
[0039] FIGS. 9 and 10 present data relating to administration of
octreotide to monkeys referenced in accompanying Example 35.
DETAILED DESCRIPTION
[0040] The compositions described herein can be administered to a
subject to provide for improved bioavailability of a therapeutic
agent.
[0041] Pharmaceutical compositions: The pharmaceutical compositions
described herein include a therapeutic agent and a medium chain
fatty acid salt in intimate contact or association with a
substantially hydrophobic medium. For example, the therapeutic
agent and the medium chain fatty acid or derivative thereof may be
coated, suspended, sprayed by or immersed in a substantially
hydrophobic medium forming a suspension. The compositions of the
invention are not emulsions. Almost all of the compositions are
oily suspensions and the amount of water in the compositions is
very low; a few of the present compositions which are not
suspensions incorporate a high amount (about 78% octanoic acid) and
are solutions by visual analysis. The suspension may be a liquid
suspension incorporating solid material, or a semi-solid suspension
incorporating solid material (an ointment).
[0042] Many of the compositions described herein comprise a
suspension which comprises an admixture of a hydrophobic medium and
a solid form wherein the solid form comprises a therapeutically
effective amount of a therapeutic agent and at least one salt of a
medium chain fatty acid, and wherein the medium chain fatty acid
salt is present in the composition at an amount of 10% or more by
weight. The solid form may comprise a particle (e.g., consist
essentially of particles, or consist of particles). The particle
may be produced by lyophilization or by granulation. In some
embodiments, preferably after milling, 90% (v/v) of the particles
are below 130 microns, and 50% (v/v) of the particles are below 45
microns.
[0043] A cargo compound is a therapeutic agent (e.g. insulin) or a
test compound (e.g. high molecular weight dextran) which is
formulated as described herein within the compositions of the
invention.
[0044] The inventors were particular to include in many of the
compositions of the invention only excipients which are generally
recognized as safe, based on available data on human use, animal
safety and regulatory guidelines (e.g. GRAS excipients). Some
compositions of the invention may have other types of excipients
(e.g. non-GRAS). In some embodiments the compositions of the
invention have amounts of excipients that are within the maximum
daily doses as noted in such available data for each specific
excipient.
[0045] The medium chain fatty acid salt may generally facilitate or
enhance permeability and/or absorption of the therapeutic agent. In
some embodiments the medium chain fatty acid salts include
derivatives of medium chain fatty acid salts. The therapeutic agent
and the medium chain fatty acid salt are in solid form, for
example, a solid particle such as a lyophilized particle,
granulated particle, pellet or micro-sphere. In preferred
embodiments, the therapeutic agent and the medium chain fatty acid
salt are both in the same solid form, e.g., both in the same
particle. In other embodiments, the therapeutic agent and the
medium chain fatty acid salt may each be in a different solid form,
e.g. each in a distinct particle. The compositions described herein
are substantially free of any "membrane fluidizing agents" defined
as linear, branched, aromatic and cyclic medium chain alcohols, in
particular geraniol and octanol. For example the compositions
preferably include no membrane fluidizing agents but certain
embodiments may include for example less than 1% or less than 0.5%
or less than 0.1% by weight of membrane fluidizing agents.
[0046] Unlike emulsions, where water is an essential constituent of
the formulation, the compositions described herein provide a solid
form such as a particle containing the therapeutic agent, which is
then associated with the hydrophobic (oily) medium. The amount of
water in the compositions is generally less than 3% by weight,
usually less than about 2% or about 1% or less by weight.
[0047] The compositions described herein are suspensions which
comprise an admixture of a hydrophobic medium and a solid form
wherein the solid form comprises a therapeutically effective amount
of a therapeutic agent and at least one salt of a medium chain
fatty acid. The solid form may be a particle (e.g., consist
essentially of particles, or consist of particles). The particle
may be produced by lyophilization or by granulation. The medium
chain fatty acid salt is generally present in the compositions
described herein at an amount of 10% or more by weight. In certain
embodiments the medium chain fatty acid salt is present in the
composition at an amount of 10%-50%, preferably 11%-18% or about
11%-17% or 12%-16% or 12%-15% or 13%-16% or 13%-15% or 14%-16% or
14%-15% or 15%-16% or most preferably 15% or 16% by weight, and the
medium chain fatty acid has a chain length from about 6 to about 14
carbon atoms preferably 8, 9 or 10 carbon atoms.
[0048] In some embodiments in the compositions described above, the
solid form including the therapeutic agent also includes a
stabilizer (e.g., a stabilizer of protein structure). Stabilizers
of protein structure are compounds that stabilize protein structure
under aqueous or non-aqueous conditions or can reduce or prevent
aggregation of the therapeutic agent, for example during a drying
process such as lyophilization or other processing step.
Stabilizers of structure can be polyanionic molecules, such as
phytic acid, polyvalent ions such as Ca, Zn or Mg, saccharides such
as a disaccharide (e.g., trehalose, maltose) or an oligo or
polysaccharide such as dextrin or dextran, or a sugar alcohol such
as mannitol, or an amino acid such as glycine, or polycationic
molecules, such as spermine, or surfactants such as polyoxyethylene
sorbitan monooleate (Tween 80) or pluronic acid. Uncharged
polymers, such as mannitol, methyl cellulose and polyvinyl alcohol,
are also suitable stabilizers.
[0049] Although polyvinylpyrrolidone (PVP) is known in the art as a
stabilizer, the inventors unexpectedly found that, in the
compositions of the invention described herein, PVP, in particular
PVP-12, serves to increase the effect of the permeability enhancer
in a synergistic manner; furthermore, increasing the level of
PVP-12 to 10% increased the absorption of the therapeutic agent
into the blood due to the improved activity of the formulations.
The inventors demonstrated that dextran had a similar (but lower)
effect as PVP did. Other matrix forming polymers have a similar
effect.
[0050] In some embodiments, such as when the therapeutic agent is a
small molecule, a bulking agent may be added, for example, mannitol
or glycin.
[0051] In certain embodiments of the compositions described herein
the therapeutic agent is a protein, a polypeptide, a peptide, a
glycosaminoglycan, a small molecule, a polysaccharide or a
polynucleotide inter alia, such as octreotide, growth hormone,
parathyroid hormone, parathyroid hormone amino acids 1-34
[PTH(1-34) termed teriparatide], a low molecular weight heparin or
fondaparinux inter alia. Low molecular weight heparins are defined
as heparin salts having an average molecular weight of less than
8000 Da and for which at least 60% of all chains have a molecular
weight less than 8000 Da.
[0052] In a particular embodiment of the compositions described
herein the salt of the fatty acid is sodium octanoate and the
hydrophobic medium is castor oil; in another particular embodiment
the composition further comprises glyceryl monooleate and sorbitan
monopalmitate or glyceryl monocaprylate and glyceryl tricaprylate
and polyoxyethylenesorbitan monooleate; in another particular
embodiment the composition further comprises glyceryl tributyrate,
lecithin, ethylisovalerate and at least one stabilizer. In
particular embodiments the therapeutic agent is octreotide, growth
hormone, parathyroid hormone, teriparatide, interferon-alfa
(IFN-.alpha.), a low molecular weight heparin, fondaparinux, siRNA,
somatostatin and analogs (agonists) thereof including
peptidomimetics, exenatide, vancomycin or gentamicin inter
alia.
Therapeutic Agents:
[0053] The pharmaceutical compositions described herein can be used
with a variety of therapeutic agents (also termed active
pharmaceutical ingredient=API). In some embodiments, the
pharmaceutical composition includes a plurality of therapeutic
agents (effectors). The therapeutic agents can either be in the
same solid form (e.g., in the same particle), or the therapeutic
agents can each be in an independent solid form (e.g., each in
different particles. In some embodiments, the therapeutic agent is
in the form of a particle, for example, a granulated or solid
particle. The particle is associated with or is in intimate contact
with a substantially hydrophobic medium, for example, a hydrophobic
medium described herein.
[0054] Therapeutic agents that can be used in the compositions
described herein include any molecule or compound serving as, for
example, a biological, therapeutic, pharmaceutical, or diagnostic
agent including an imaging agent. The therapeutic agents include
drugs and other agents including, but not limited to, those listed
in the United States Pharmacopeia and in other known pharmacopeias.
Therapeutic agents are incorporated into the formulations of the
invention without any chemical modification. Therapeutic agents
include proteins, polypeptides, peptides, polynucleotides,
polysaccharides and small molecules.
[0055] The term "small molecule" is understood to refer to a low
molecular weight organic compound which may be synthetically
produced or obtained from natural sources and typically has a
molecular weight of less than 2000 Da, or less than 1000 Da or even
less than 600 Da e.g. less than or about 550 Da or less than or
about 500 Da or less than or about 400 Da; or about 400 Da to about
2000 Da; or about 400 Da to about 1700 Da. Examples of small
molecules are ergotamine (molecular weight=582 Da), fondaparinux
(molecular weight=1727 Da), leuprolide (molecular weight=1209 Da),
vancomycin (molecular weight=1449 Da), gentamicin (molecular
weight=478 Da) and doxorubicin (molecular weight=544).
[0056] The term "polynucleotide" refers to any molecule composed of
DNA nucleotides, RNA nucleotides or a combination of both types
which comprises two or more of the bases guanidine, citosine,
timidine, adenine, uracil or inosine, inter alia. A polynucleotide
may include natural nucleotides, chemically modified nucleotides
and synthetic nucleotides, or chemical analogs thereof and may be
single-stranded or double-stranded. The term includes
"oligonucleotides" and encompasses "nucleic acids".
[0057] By "small interfering RNA" (siRNA) is meant an RNA molecule
(ribonucleotide) which decreases or silences (prevents) the
expression of a gene/mRNA of its endogenous or cellular
counterpart. The term is understood to encompass "RNA interference"
(RNAi), and "double-stranded RNA" (dsRNA).
[0058] By "polypeptide" is meant a molecule composed of covalently
linked amino acids and the term includes peptides, polypeptides,
proteins and peptidomimetics. A peptidomimetic is a compound
containing non-peptidic structural elements that is capable of
mimicking the biological action(s) of a natural parent peptide.
Some of the classical peptide characteristics such as enzymatically
scissile peptidic bonds are normally not present in a
peptidomimetic.
[0059] The term "amino acid" refers to a molecule which consists of
any one of the 20 naturally occurring amino acids, amino acids
which have been chemically modified or synthetic amino acids.
[0060] By "polysaccharide" is meant a linear or branched polymer
composed of covalently linked monosaccharides; glucose is the most
common monosaccharide and there are normally at least eight
monosaccharide units in a polysaccharide and usually many more.
Polysaccharides have a general formula of Cx(H2O)y where x is
usually a large number between 200 and 2500. Considering that the
repeating units in the polymer backbone are often six-carbon
monosaccharides, the general formula can also be represented as
(C6H10O5)n where 40.ltoreq.n.ltoreq.3000 i.e. there are normally
between 40 and 3000 monosaccharide units in a polysaccharide.
[0061] A "glycosaminoglycan" is a polysaccharide that contains
amino containing sugars.
[0062] Exemplary anionic therapeutic agents include polynucleotides
from various origins, and particularly from human, viral, animal,
eukaryotic or prokaryotic, plant, or synthetic origin, etc
including systems for therapeutic gene delivery. A polynucleotide
of interest may be of a variety of sizes, ranging from, for
example, a simple trace nucleotide to a gene fragment, or an entire
gene. It may be a viral gene or a plasmid. Exemplary
polynucleotides serving as therapeutic agents include specific DNA
sequences (e.g., coding genes), specific RNA sequences (e.g., RNA
aptamers, antisense RNA, short interfering RNA (siRNA) or a
specific inhibitory RNA (RNAi)), poly CPG, or poly I:C synthetic
polymers of polynucleotides.
[0063] Alternatively, the therapeutic agent can be a protein, such
as, for example, an enzyme, a hormone, an incretin, a proteoglycan,
a ribozyme, a cytokine, a peptide, an apolipoprotein, a growth
factor, a bioactive molecule, an antigen, or an antibody or
fragment(s) thereof, etc. The peptide can be a small peptide e.g.
from about 2 to about 40 amino acids, examples include
fibrinogen-receptor antagonists (RGD-containing peptides which are
tetrapeptides having an average molecular weight of about 600.
Exemplary peptides are somatostatin and analogs thereof e.g.
octreotide and lanreotide (Somatuline) which are both cyclic
octapeptides and pasireotide (SOM-230) which is a cyclic
hexapeptide (Weckbecker et al, 2002, Endocrinology 143(10)
4123-4130; Schmid, 2007, Molecular and Cellular Endocrinology 286,
69-74). Other exemplary peptides are glatiramer acetate
(Copaxone.RTM.) which is a tetrapeptide, terlipressin which is a 12
amino acid peptide analog (agonist) of lysine vasopressin (ADH) and
exenatide, a 39 amino acid peptide which is an incretin mimetic
agent, and other analogs of glucagon-like peptide-1 (GLP-1).
(Byetta.RTM. is the trade name for exenatide (Eli Lilly and
Company/Amylin Pharmaceuticals, Inc.). Other peptides include
dalargin which is a hexapeptide, and kyotorphin which is a
dipeptide. Peptides include growth hormone releasing peptides which
are peptides of about 12 amino acids or less; see for example
peptides disclosed in U.S. Pat. No. 4,411,890 (Momany) and U.S.
Pat. No. 4,839,344 (Bowers et al)
[0064] Examples of other peptides which can be used in the practice
of this invention are those disclosed in U.S. Pat. No. 4,589,881
(30 or more amino acid residues) of Pierschbacher et al; U.S. Pat.
No. 4,544,500 (20-30 residues) of Bittle et al; and EP0204480
(>34 residues) of Dimarchi et al and teriparatide. In some
embodiments, the therapeutic agent can include a polysaccharide,
such as a glycosaminoglycan. Exemplary glycosaminoglycans include
heparin, heparin derivatives, heparan sulfate, chondroitin sulfate,
dermatan sulfate, and hyaluronic acid. Examples of heparin
derivatives include, but are not limited to, low molecular weight
heparins such as enoxaparin, dalteparin and tinzaparin. A
therapeutic agent with a heparin-like effect is fondaparinux.
[0065] Other examples of therapeutic agents include, but are not
limited to hormones such as insulin, erythropoietin (EPO),
glucagon-like peptide 1 (GLP-1), melanocyte stimulating hormone
(alfa-MSH), parathyroid hormone (PTH), teriparatide, growth hormone
(GH), leuprolide, leuprolide acetate, factor VIII, growth hormone
releasing hormone (GHRH), peptide YY amino acids 3-36
(PYY.sub.(3-36)), calcitonin, somatotropin, somatostatin,
somatomedin, interleukins such as interleukin-2 (IL-2),
alfa-1-antirypsin, granulocyte/monocyte colony stimulating factor
(GM-CSF), granulocyte colony stimulating factor (G-CSF), T20,
testosterone, interferons such as interferon-alfa (IFN-.alpha.)
IFN-.beta. and IFN-.gamma., luteinizing hormone (LH),
follicle-stimulating hormone (FSH), human chorionic gonadotropin
(hCG), enkephalin, dalargin, kyotorphin, basic fibroblast growth
factor (bFGF), hirudin, hirulog, luteinizing hormone releasing
hormone (LHRH), gonadotropin releasing hormone (GnRH) analog,
brain-derived natriuretic peptide (BNP), tissue plasminogen
activator (TPA), oxytocin, and analogs and combinations
thereof.
[0066] Other examples of therapeutic agents include, but are not
limited to analgesic agents, anti-migraine agents, anti-coagulant
agents, anti-emetic agents, cardiovascular, anti-hypertensive and
vasodilator agents, sedatives, narcotic antagonists, chelating
agents, anti-diuretic agents and anti-neoplastic agents.
[0067] Analgesics include, but are not limited to, fentanyl,
sufentanil, butorphanol, buprenorphine, levorphanol, morphine,
hydromorphone, hydrocodeine, oxymorphone, methadone, lidocaine,
bupivacaine, diclofenac, naproxen, paverin, and analogs thereof.
Anti-migraine agents include, but are not limited to naratriptan,
naproxen, almotriptan, butalbital, frovatriptan, sumatriptan,
rizatriptan, acetaminophen, isometheptene, butorphanol,
dichloralphenazone, ergot alkaloids such as dihydroergotamine and
ergotamine, nonsteroidal anti-inflammatory drugs (NSAIDs) such as
ketoprofen and ketorolac, eletriptan, butorphanol, topiramate,
zolmitriptan, caffeine, aspirin and codeine, and analogs and
combinations thereof.
[0068] Anti-coagulant agents include, but are not limited to
heparin, hirudin, low molecular weight heparins and analogs thereof
and fondaparinux. Anti-emetic agents include but are not limited to
scopolamine, ondansetron, domperidone, etoclopramide, and analogs
thereof. Cardiovascular, anti-hypertensive and vasodilator agents
include, but are not limited to, diltiazem, clonidine, nifedipine,
verapamil, isosorbide-5-mononitrate, organic nitrates,
nitroglycerine and analogs thereof. Sedatives include, but are not
limited to, benzodiazeines, phenothiozines and analogs thereof.
Narcotic antagonists include, but are not limited to, naltrexone,
naloxone and analogs thereof. Chelating agents include, but are not
limited to deferoxamine and analogs thereof. Anti-diuretic agents
include, but are not limited to, desmopressin, vasopressin and
analogs (agonists) thereof such as terlipressin; the trade name of
terlipressin is glypressin.RTM.. Anti-neoplastic agents include,
but are not limited to, 5-fluorouracil, bleomycin, vincristine,
procarbazine, temezolamide, 6-thioguanine, hydroxyurea, cytarabine,
cyclophosphamide, doxorubicin, vinca alkaloid, epirubicin,
etoposide, ifosfamide, carboplatin and other platinum based
antineoplastic drugs (such as carboplatin (Paraplatin.RTM.,
tetraplatin, oxaliplatin, aroplatin and transplatin), vinblastine,
vinorelbine, chlorambucil, busulfan, mechlorethamine, mitomycin,
dacarbazine, thiotepa, daunorubicin, idarubicin, mitoxantrone,
esperamicin A1, dactinomycin, plicamycin, carmustine, lomustine
(CCNU), tauromustine, streptozocin, melphalan, dactinomycin,
procarbazine, dexamethasone, prednisone, 2-chlorodeoxyadenosine,
cytarabine, docetaxel, fludarabine, gemcitabine, herceptin,
hydroxyurea, irinotecan, methotrexate, rituxin, semustine, tomudex
and topotecan, taxol and taxol-like compounds and analogs and
combinations thereof.
[0069] Additional examples of therapeutic agents include, but are
not limited to coagulation factors and neurotrophic factors,
anti-TNF antibodies and fragments of TNF receptors.
[0070] Therapeutic agents also include pharmaceutically active
agents selected from the group consisting of vitamin B12, a
bisphosphonate (e.g., disodium pamidronate, alendronate,
etidronate, tiludronate, risedronate, zoledronic acid, sodium
clodronate, or ibandronic acid), taxol, caspofungin, or an
aminoglycoside antibiotic. Additional therapeutic agents include a
toxin, or an antipathogenic agent, such as an antibiotic (e.g.
vancomycin), an antiviral, an antifungal, or an anti-parasitic
agent. The therapeutic agent can itself be directly active or can
be activated in situ by the composition, by a distinct substance,
or by environmental conditions.
[0071] In some embodiments, the composition can include a plurality
of therapeutic agents (combination drugs). For example, the
composition can include Factor VIII and vWF, GLP-1 and PYY,
IFN-.alpha. and nucleotide analogues (i.e. ribavirin), and
alendronate or insulin and GLP-1.
[0072] In some embodiments, the composition can include a small
molecule and a peptide or protein. Exemplary combinations include a
combination of IFN-.alpha. and nucleotide analogues (i.e.
ribavirin) for the treatment of hepatitis C, teriparatide and
alendronate for treatment of bone disorders, a combination of GH
plus the medications for HIV therapy (e.g., HAART) to
simultaneously treat the viral infection and the accompanying HIV
lipodystrophy or AIDS wasting side effects. Combinations of two
small molecules can be used when one of them generally has poor
absorption or bioavailability even if the other generally has
effective absorption or bioavailability, such as some antibiotics
(e.g., a combination of vancomycin and an aminoglycoside such as
gentamicin. Exemplary combinations for the treatment and prevention
of metabolic disorders such as diabetes and obesity also include
combination of insulin and metformin, insulin and rosiglitazone,
GLP-1 (or exenatide) and metformin, and GLP-1 (or exenatide) and
rosiglitazone.
[0073] Indications and conditions which may be treated by
fondaparinux formulated as described herein include deep vein
thrombosis, hip or knee replacement, and bed-bound patients.
[0074] In some embodiments of the compositions described herein,
the composition includes a combination of a protein or peptide with
small molecules that either do or do not have good absorption or
bioavailability. For example, a composition can include at least
one therapeutic agent that may generally be characterized as poorly
absorbable or poorly bioavailable. The composition can also be used
for the administration of therapeutic agents that are absorbed in
the stomach and/or intestine, but cause irritation to the stomach
and/or intestine and therefore are difficult to tolerate. In such a
situation, a subject could benefit if the bioavailability of the
therapeutic agent were enhanced or if more of the therapeutic agent
were absorbed directly into the blood stream; if less therapeutic
agent is administered there will clearly be less chance of causing
irritation to the stomach and/or intestine. Thus compositions of
the invention are envisaged which comprises therein two or more
therapeutic agents.
[0075] In general, the composition may include from about 0.01% to
about 50% by weight of the therapeutic agent e.g. about 0.01, 0.02
0.05, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or
50% by weight. The maximum included in the composition is often in
the range of about 6%-33% by weight of the therapeutic agent.
[0076] In some embodiments of the compositions described herein,
the solid form including the therapeutic agent also includes a
stabilizer (e.g., a stabilizer of protein structure). Stabilizers
of protein structure are compounds that stabilize protein structure
under aqueous or non-aqueous conditions or can reduce or prevent
aggregation of the therapeutic agent, for example during a drying
process such as lyophilization or other processing step.
Stabilizers of structure can be polyanionic molecules, such as
phytic acid, polyvalent ions such as Ca, Zn or Mg, saccharides such
as a disaccharide (e.g., trehalose, maltose) or an oligo or
polysaccharide such as dextrin or dextran, or a sugar alcohol such
as mannitol, or an amino acid such as glycine, or polycationic
molecules, such as spermine, or surfactants such as Tween 80 or
Span 40 or pluronic acid. Uncharged polymers, such as methyl
cellulose and polyvinyl alcohol, are also suitable stabilizers.
Medium Chain Fatty Acid Salt:
[0077] The compositions described herein include the salt of a
medium chain fatty acid or a derivative thereof in a solid form.
For example, the salt of the medium chain fatty acid is in the form
of a particle such as a solid particle. In some embodiments, the
particle may be characterized as a granulated particle. In at least
some embodiments, the solid form may generally result from a spray
drying or evaporation process. In preferred embodiments, the salt
of the medium chain fatty acid is in the same particle as the
therapeutic agent. For example, the therapeutic agent and the salt
of the medium chain fatty acid can be prepared together by first
preparing a solution such as an aqueous solution comprising both
the therapeutic agent and the salt of the medium chain fatty acid
and co-lyophilizing the solution to provide a solid form or
particle that comprises both the therapeutic agent and the salt of
the medium chain fatty acid (and other ingredients). As described
above, the resulting solid particles are associated with a
hydrophobic medium. For example, the solid particles may be
suspended or immersed in a hydrophobic medium
[0078] In different embodiments of the compositions described
herein the medium chain fatty acid salt may be in the same particle
or in a different particle than that of the API. It was found that
bioavailability of a cargo compound was lower if the medium chain
fatty acid was in a different particle than the therapeutic agent
i.e. there was improved bioavailability if the medium chain fatty
acid salt and the cargo compound were dried after solubilization
together in the hydrophilic fraction. It is believed that if the
medium chain fatty acid salt and the cargo compound are dried after
solubilization together in the hydrophilic fraction then they are
in the same particle in the final powder.
[0079] Medium chain fatty acid salts include those having a carbon
chain length of from about 6 to about 14 carbon atoms. Examples of
fatty acid salts are sodium hexanoate, sodium heptanoate, sodium
octanoate (also termed sodium caprylate), sodium nonanoate, sodium
decanoate, sodium undecanoate, sodium dodecanoate, sodium
tridecanoate, and sodium tetradecanoate. In some embodiments, the
medium chain fatty acid salt contains a cation selected from the
group consisting of potassium, lithium, ammonium and other
monovalent cations e.g. the medium chain fatty acid salt is
selected from lithium octanoate or potassium octanoate or arginine
octanoate or other monovalent salts of the medium chain fatty
acids. The inventors found that raising the amount of medium chain
fatty acid salt increased the bioavailability of the resulting
formulation. In particular, raising the amount of medium chain
fatty acid salt, in particular sodium octanoate, above 10% to a
range of about 12% to 15% increased the bioavailability of the
therapeutic agents in the pharmaceutical compositions described
herein.
[0080] In general, the amount of medium chain fatty acid salt in
the compositions described herein may be from 10% up to about 50%
by weight of the bulk pharmaceutical composition. For example, the
medium chain fatty acid salt may be present at an amount of about
10%-50%, preferably about 11%-40% most preferably about 11%-28% by
weight for example at about 12%-13%, 13%-14%, 14%-15%, 15%-16%,
16%-17%, 17%-18%, 18%-19%, 19%-20%, 20%-21%, 21%-22%, 22%-23%,
23%-24%, 24%-25%, 25%-26%, 26%-27%, or 27%-28% by weight of the
bulk pharmaceutical composition. In other embodiments the medium
chain fatty acid salt may be present at an amount of at least about
11%, at least about 12%, at least about 13%, at least about 14%, at
least about 15% at least about 16%, at least about 17%, at least
about 18%, at least about 19%, at least about 20%, at least about
21%, at least about 22%, at least about 23%, at least about 24%, at
least about 25%, at least about 26%, at least about 27% or at least
about 28% by weight of the bulk pharmaceutical composition. In
specific embodiments the medium chain fatty acid salt (sodium,
potassium, lithium or ammonium salt or a mixture thereof) is
present at about 12%-21% by weight of the bulk pharmaceutical
composition preferably 11%-18% or about 11%-17% or 12%-16% or
12%-15% or 13%-16% or 13%-15% or 14%-16% or 14%-15% or 15%-16% or
most preferably 15% or 16%. In specific embodiments the medium
chain fatty acid salt (having a carbon chain length of from about 6
to about 14 carbon atoms particularly 8, 9 or 10 carbon atoms) is
present at about 12% -21% by weight of the bulk pharmaceutical
composition preferably 11%-18% about 11%-17% or 12%-16% or 12%-15%
or 13%-16% or 13%-15% or 14%-16% or 14%-15% or 15%-16% or most
preferably 15% or 16%. In specific embodiments the medium chain
fatty acid salt (for example salts of octanoic acid, salts of
suberic acid, salts of geranic acid) is present at about 12%-21% by
weight of the bulk pharmaceutical composition preferably 11%-18%
about 11%-17% or 12%-16% or 12%-15% or 13%-16% or 13%-15% or
14%-16% or 14%-15% or 15%-16% or most preferably 15% or 16%. In
certain embodiments the medium chain fatty acid salt is present in
the solid powder at an amount of 50% to 90%, preferably at an
amount of 70% to 80%.
[0081] One embodiment of the invention comprises a composition
comprising a suspension which consists essentially of an admixture
of a hydrophobic medium and a solid form wherein the solid form
comprises a therapeutically effective amount of a therapeutic agent
and at least one salt of a medium chain fatty acid, and wherein the
medium chain fatty acid salt is not a sodium salt. The salt may be
the salt of another cation e.g. lithium, potassium or ammonium; an
ammonium salt is preferred.
Matrix Forming Polymer:
[0082] In certain embodiments the composition of the invention
comprises a suspension which comprises an admixture of a
hydrophobic medium and a solid form wherein the solid form
comprises a therapeutically effective amount of a therapeutic
agent, at least one salt of a medium chain fatty acid and a matrix
forming polymer, and wherein the matrix forming polymer is present
in the composition at an amount of 3% or more by weight. In certain
embodiments the composition comprises a suspension which consists
essentially of an admixture of a hydrophobic medium and a solid
form wherein the solid form comprises a therapeutically effective
amount of a therapeutic agent, at least one salt of a medium chain
fatty acid and a matrix forming polymer, and wherein the matrix
forming polymer is present in the composition at an amount of 3% or
more by weight. In particular embodiments the matrix forming
polymer is dextran or polyvinylpyrrolidone (PVP). In particular
embodiments the polyvinylpyrrolidone is present in the composition
at an amount of about 2% to about 20% by weight, preferably at an
amount of about 3% to about 18% by weight, more preferably at an
amount of about 5% to about 15% by weight, most preferably at an
amount of about 10% by weight. In certain particular embodiments
the polyvinylpyrrolidone is PVP-12 and/or has a molecular weight of
about 3000. Other matrix forming polymers have a similar effect in
the compositions of the invention; such matrix forming polymers
include ionic polysaccharides (for example alginic acid and
alginates) or neutral polysaccharides (for example dextran and
HPMC), polyacrylic acid and poly methacrylic acid derivatives and
high molecular weight organic alcohols (for example polyvinyl
alcohol).
Protease Inhibitors:
[0083] It is generally accepted in the art of delivery of proteins,
polypeptides and peptides that protease inhibitors normally have to
be added to the formulation to prevent degradation of the API.
However in the formulations of the instant invention it is not
necessary to add protease inhibitors. The formulations of the
invention appear to confer stability of the therapeutic agent to
protease degradation within the time-frame of activity i.e. the
formulations of the invention are apparently environment inhibitory
for enzyme activity. Additionally, the inventors performed an
experiment wherein the protease inhibitor aprotinin was added to a
formulation and this had no beneficial effect on activity. A
similar experiment was performed where the protease inhibitor
.epsilon.-aminocaproic acid was added to a formulation and this too
had no beneficial effect on activity. Therefore, in some
embodiments, a pharmaceutical composition described herein is
substantially free of a protease inhibitor.
Hydrophilic Fraction:
[0084] In embodiments of the invention, the above compounds,
including the therapeutic agent and the medium chain fatty acid
salt are solubilized in an aqueous medium and then dried to produce
a powder. The drying process may be achieved for example by
lyophilization or granulation. The powder obtained is termed the
"hydrophilic fraction". In the hydrophilic fraction water is
normally present at an amount of less than 6%.
[0085] Lyophilization may be carried out as shown in the Examples
herein and by methods known in the art e.g. as described in
Lyophilization: Introduction and Basic Principles, Thomas Jennings,
published by Interpharm/CRC Press Ltd (1999, 2002) The lyophilizate
may optionally be milled (e.g. below 150 micron) or ground in a
mortar. During industrial production the lyophilizate is preferably
milled before mixing of the hydrophilic fraction and the
hydrophobic medium in order to produce batch-to-batch
reproducibility.
[0086] Granulation may be carried out as shown in the Examples
herein and by methods known in the art e.g. as described in
Granulation, Salman et al, eds, Elsevier (2006) and in Handbook of
Pharmaceutical Granulation Technology, 2.sup.nd edition, Dilip M.
Parikh, ed., (2005
[0087] Various binders may be used in the granulation process such
as celluloses (including microcrystalline celluloses), lactoses
(e.g. lactose monohydrate), dextroses, starch and mannitol and
other binders as described in the previous two references.
Hydrophobic Medium:
[0088] Oil: As described above, in the compositions of the
invention described herein the therapeutic agent and the medium
chain fatty acid salt are in intimate contact or association with a
hydrophobic medium. For example, one or both may be coated,
suspended, immersed or otherwise in association with a hydrophobic
medium. Suitable hydrophobic mediums can contain, for example,
aliphatic, cyclic or aromatic molecules. Examples of a suitable
aliphatic hydrophobic medium include, but are not limited to,
mineral oil, fatty acid monoglycerides, diglycerides,
triglycerides, ethers, esters, and combinations thereof. Examples
of a suitable fatty acid are octanoic acid, decanoic acid and
dodecanoic acid, also C7 and C9 fatty acids and di-acidic acids
such as sebacic acid and suberic acid, and derivatives thereof.
Examples of triglycerides include, but are not limited to, long
chain triglycerides, medium chain triglycerides, and short chain
triglycerides. For example, the long chain triglyceride can be
castor oil or coconut oil or olive oil, and the short chain
triglyceride can be glyceryl tributyrate and the medium chain
triglyceride can be glyceryl tricaprylate. Monoglycerides are
considered to be surfactants and are described below. Exemplary
esters include ethyl isovalerate and butyl acetate. Examples of a
suitable cyclic hydrophobic medium include, but are not limited to,
terpenoids, cholesterol, cholesterol derivatives (e.g., cholesterol
sulfate), and cholesterol esters of fatty acids. A non-limiting
example of an aromatic hydrophobic medium includes benzyl
benzoate.
[0089] In some embodiments of the compositions described herein, it
is desirable that the hydrophobic medium include a plurality of
hydrophobic molecules. In some embodiments of the compositions
described herein the hydrophobic medium also includes one or more
surfactants (see below).
[0090] In some embodiments of the compositions described herein,
the hydrophobic medium also includes one or more adhesive polymers
such as methylcellulose, ethylcellulose,
hydroxypropylmethylcellulose (HPMC), or poly(acrylate) derivative
Carbopol.RTM.934P (C934P). Such adhesive polymers may assist in the
consolidation of the formulation and/or help its adherence to
mucosal surfaces.
[0091] Surface Active Agents (surfactants): The compositions of
this invention described herein can further include a surface
active agent. For example, the surface active agent can be a
component of the hydrophobic medium as described above, and/or the
surface active agent can be a component of a solid form as
described above, for example in the solid form or particle that
includes the therapeutic agent.
[0092] Suitable surface active agents include ionic and non-ionic
surfactants. Examples of ionic surfactants are lecithin
(phosphatidyl choline), bile salts and detergents. Examples of
non-ionic surfactants include monoglycerides, cremophore, a
polyethylene glycol fatty alcohol ether, a sorbitan fatty acid
ester, a polyoxyethylene sorbitan fatty acid ester, Solutol HS15,
or a poloxamer or a combination thereof. Examples of monoglycerides
are glyceryl monocaprylate (also termed glyceryl monooctanoate),
glyceryl monodecanoate, glyceryl monolaurate, glyceryl
monomyristate, glyceryl monostearate, glyceryl monopalmitate, and
glyceryl monooleate. Examples of sorbitan fatty acid esters include
sorbitan monolaurate, sorbitan monooleate, and sorbitan
monopalmitate (Span 40), or a combination thereof. Examples of
polyoxyethylene sorbitan fatty acid esters include polyoxyethylene
sorbitan monooleate (Tween 80), polyoxyethylene sorbitan
monostearate, polyoxyethylene sorbitan monopalmitate or a
combination thereof. The commercial preparations of monoglycerides
that were used also contain various amounts of diglycerides and
triglycerides.
[0093] Compositions described herein including a surface active
agent generally include less than about 12% by weight of total
surface active agent (e.g., less than about 10%, less than about
8%, less than about 6%, less than about 4%, less than about 2%, or
less than about 1%). In particular embodiments of the invention the
total sum of all the surfactants is about 6%.
[0094] Methods of making pharmaceutical compositions and the
compositions produced: Also included in the invention are methods
of producing the compositions described herein. Thus one embodiment
of the invention is a process for producing a pharmaceutical
composition which comprises preparing a water-soluble composition
comprising a therapeutically effective amount of at least one
therapeutic agent and a medium chain fatty acid salt (as described
above), drying the water soluble composition to obtain a solid
powder, and suspending the solid powder in a hydrophobic medium, to
produce a suspension containing in solid form the therapeutic agent
and the medium chain fatty acid salt, thereby producing the
pharmaceutical composition, wherein the pharmaceutical composition
contains 10% or more by weight of medium chain fatty acid salt.
[0095] One embodiment is a process for producing a pharmaceutical
composition which comprises providing a solid powder of a
therapeutically effective amount of at least one therapeutic agent
and a solid powder comprising a medium chain fatty acid salt, and
suspending the solid powders in a hydrophobic medium, to produce a
suspension containing in solid form the therapeutic agent and the
medium chain fatty acid salt, thereby producing the pharmaceutical
composition, wherein the pharmaceutical composition contains 10% or
more by weight of medium chain fatty acid salt.
[0096] In one embodiment of the processes and compositions
described herein, the water-soluble composition is an aqueous
solution. In certain embodiments the drying of the water-soluble
composition is achieved by lyophilization or by granulation. In the
granulation process a binder may be added to the water soluble
composition before drying. In certain embodiments the drying step
removes sufficient water so that the water content in the
pharmaceutical composition is lower than about 6% by weight, about
5% by weight, about 4% by weight, about 3% or about 2% or about 1%
by weight. In certain embodiments of the processes and compositions
described herein the drying step removes an amount of water so that
the water content in the solid powder is lower than 6% or 5% or 4%
or 3% or preferably lower than 2% by weight. The water content is
normally low and the water may be adsorbed to the solid phase
during lyophilization i.e. the water may be retained by
intermolecular bonds. In certain embodiments the water soluble
composition additionally comprises a stabilizer for example methyl
cellulose. In preferred embodiments of the of the processes and
compositions described herein the hydrophobic medium is castor oil
or glyceryl tricaprylate or glyceryl tributyrate or a combination
thereof and may additionally contain octanoic acid; in certain
embodiments the hydrophobic medium comprises an aliphatic,
olefinic, cyclic or aromatic compound, a mineral oil, a paraffin, a
fatty acid such as octanoic acid, a monoglyceride, a diglyceride, a
triglyceride, an ether or an ester, or a combination thereof. In
certain embodiments of the processes and compositions described
herein the triglyceride is a long chain triglyceride, a medium
chain triglyceride preferably glyceryl tricaprylate or a short
chain triglyceride preferably glyceryl tributyrate, and the long
chain triglyceride is castor oil or coconut oil or a combination
thereof. In certain embodiments of the processes and compositions
described herein the hydrophobic medium comprises castor oil or
glyceryl tricaprylate or glyceryl tributyrate or a combination or
mixture thereof, and may additionally comprise octanoic acid. In
certain embodiments of the processes and compositions described
herein the hydrophobic medium comprises glyceryl tricaprylate or a
low molecular weight ester for example ethyl isovalerate or butyl
acetate. In certain embodiments of the processes and compositions
described herein the main component by weight of the hydrophobic
medium is castor oil and may additionally comprise glyceryl
tricaprylate. In certain embodiments of the processes and
compositions described herein the main component by weight of the
hydrophobic medium is glyceryl tricaprylate and may additionally
comprise castor oil.
[0097] A basic formulation is provided as an embodiment wherein the
hydrophobic medium consists essentially of castor oil, glyceryl
monooleate and glyceryl tributyrate; in a further embodiment of the
basic formulation the hydrophilic fraction consists essentially of
therapeutic agent, PVP-12 and sodium octanoate.
[0098] A particular formulation is provided as an embodiment
wherein the hydrophobic medium consists essentially of glyceryl
tricaprylate, castor oil, glyceryl monocaprylate, and Tween 80, and
the hydrophilic fraction consists essentially of therapeutic agent
(e.g. octreotide), PVP-12 and sodium octanoate. Another particular
formulation is provided as an embodiment wherein the hydrophobic
medium comprises glyceryl tricaprylate, castor oil, glyceryl
monocaprylate, and Tween 80, and the hydrophilic fraction comprises
therapeutic agent (e.g. octreotide), PVP-12 and sodium octanoate.
In certain embodiments the hydrophobic medium consists essentially
of glyceryl tricaprylate and in certain embodiments additionally
contains castor oil and/or glyceryl monocaprylate.
[0099] In certain embodiments the composition comprises a
suspension which consists essentially of an admixture of a
hydrophobic medium and a solid form wherein the solid form
comprises a therapeutically effective amount of a therapeutic agent
and at least one salt of a medium chain fatty acid, and wherein the
medium chain fatty acid salt is present in the composition at an
amount of 10% or more by weight. In certain embodiments the
hydrophobic medium consists essentially of castor oil, glyceryl
monooleate and glyceryl tributyrate; or the hydrophobic medium
consists essentially of glyceryl tricaprylate and glyceryl
monocaprylate; or the hydrophobic medium consists essentially of
castor oil, glyceryl tricaprylate and glyceryl monocaprylate. In
certain embodiments the hydrophobic medium comprises a triglyceride
and a monoglyceride and in certain particular embodiments the
monoglyceride has the same fatty acid radical as the triglyceride.
In certain of these embodiments the triglyceride is glyceryl
tricaprylate and the monoglyceride is glyceryl monocaprylate. In
certain embodiments the medium chain fatty acid salt in the
water-soluble composition has the same fatty acid radical as the
medium chain monoglyceride or as the medium chain triglyceride or a
combination thereof. In certain of these embodiments the medium
chain fatty acid salt is sodium caprylate (sodium octanoate) and
the monoglyceride is glyceryl monocaprylate and the triglyceride is
glyceryl tricaprylate.
[0100] Many of the compositions described herein comprise a
suspension which comprises an admixture of a hydrophobic medium and
a solid form wherein the solid form comprises a therapeutically
effective amount of a therapeutic agent and at least one salt of a
medium chain fatty acid, and wherein the medium chain fatty acid
salt is present in the composition at an amount of 10% or more by
weight. The solid form may be a particle (e.g., consist essentially
of particles, or consists of particles). The particle may be
produced by lyophilization or by granulation.
[0101] In a particular embodiment the formulation consists
essentially of a suspension which comprises an admixture of a
hydrophobic medium and a solid form wherein the solid form
comprises a therapeutically effective amount of a therapeutic agent
and about 10-20% preferably 15% medium chain fatty acid salt
preferably sodium octanoate, and about 5-10% preferably 10% PVP-12;
and wherein the hydrophobic medium comprises about 20-80%,
preferably 30-70% triglyceride preferably glyceryl tricaprylate or
glyceryl tributyrate or castor oil or a mixture thereof, about
3-10% surfactants, preferably about 6%, preferably glyceryl
monocaprylate and Tween 80 and about 1% water; in particular
embodiments the therapeutic agent is present at an amount of less
than 33%, or less than 25%, or less than 10%, or less than 1% or
less than 0.1%. The solid form may be a particle (e.g., consist
essentially of particles, or consists of particles). The particle
may be produced by lyophilization or by granulation. In a
particular embodiment the solid form may be a particle and may be
produced by lyophilization or by granulation.
[0102] In a further embodiment the formulation consists essentially
of a suspension which comprises an admixture of a hydrophobic
medium and a solid form wherein the solid form comprises a
therapeutically effective amount of a therapeutic agent and about
10-20% preferably 15% medium chain fatty acid salt preferably
sodium octanoate and about 5-10% preferably 10% PVP-12; and wherein
the hydrophobic medium comprises about 20-80%, preferably 30-70%
medium or short chain triglyceride preferably glyceryl tricaprylate
or glyceryl tributyrate, about 0-50% preferably 0-30% castor oil,
about 3-10% surfactants, preferably about 6%, preferably glyceryl
monocaprylate and Tween 80,and about 1% water; in particular
embodiments the therapeutic agent is present at an amount of less
than 33%, or less than 25%, or less than 10%, or less than 1% or
less than 0.1%.
[0103] In a particular embodiment the formulation consists
essentially of a suspension which comprises an admixture of a
hydrophobic medium and a solid form wherein the solid form
comprises a therapeutically effective amount of a therapeutic agent
and about 15% sodium octanoate and about 10% PVP-12; and wherein
the hydrophobic medium comprises about 41% glyceryl tricaprylate,
about 27% castor oil, about 4% glyceryl monocaprylate, about 2%
Tween 80, about 1% water and 1% or less therapeutic agent; when the
therapeutic agent is octreotide it is present at about 0.058%.
[0104] In another particular embodiment the formulation consists
essentially a suspension which comprises an admixture of a
hydrophobic medium and a solid form wherein the solid form
comprises a therapeutically effective amount of a therapeutic agent
and about 15% sodium octanoate and about 10% PVP-12; and wherein
the hydrophobic medium comprises about 68% glyceryl tricaprylate,
about 4% glyceryl monocaprylate, about 2% Tween 80, about 15%
sodium octanoate, about 10% PVP-12, about 1% water and less than 1%
therapeutic agent; when the therapeutic agent is octreotide it is
present at about 0.058%.
[0105] One embodiment is a composition comprising a suspension
which comprises an admixture of a hydrophobic medium and a solid
form wherein the solid form comprises a therapeutically effective
amount of octreotide and at least one salt of a medium chain fatty
acid; in a further embodiment the medium chain fatty acid salt is
present in the composition at an amount of 10% or more by weight,
preferably 15% by weight; in a further embodiment the solid form
additionally comprises a matrix-forming polymer. In a further
embodiment the matrix forming polymer is dextran or
polyvinylpyrrolidone (PVP). In a specific embodiment the matrix
forming polymer is polyvinylpyrrolidone and the
polyvinylpyrrolidone is present in the composition at an amount of
about 2% to about 20% by weight, preferably about 10% by weight. In
a specific embodiment the polyvinylpyrrolidone is PVP-12 and/or the
polyvinylpyrrolidone has a molecular weight of about 3000. In
specific embodiments the hydrophobic medium consists essentially of
glyceryl tricaprylate and the solid form additionally consists of
PVP-12 and sodium octanoate. In more specific embodiments the
hydrophobic medium additionally consists of castor oil or glyceryl
monocaprylate or a combination thereof and a surfactant. In further
specific embodiments the hydrophobic medium consists of glyceryl
tricaprylate, glyceryl monocaprylate, and polyoxyethylene sorbitan
monooleate (Tween 80). In a further embodiment the solid form
consists essentially of octreotide, PVP-12 and sodium octanoate. In
a particular embodiment the composition contains about 41% of
glyceryl tricaprylate, about 27% castor oil, about 4% glyceryl
monocaprylate, about 2% Tween 80, about 15% sodium octanoate, about
10% PVP-12, about 1% water and about 0.058% octreotide. In another
particular embodiment the composition contains about 68% of
glyceryl tricaprylate, about 4% glyceryl monocaprylate, about 2%
Tween 80, about 15% sodium octanoate, about 10% PVP-12, about 1%
water and about 0.058% octreotide.
[0106] In all the above formulations, the percentages recited are
weight/weight and the solid form may be a particle (e.g., consist
essentially of particles, or consists of particles). The particles
may be produced by lyophilization or by granulation.
[0107] Under normal storage conditions, the therapeutic agent
within the formulations of the invention is stable over an extended
period of time. The chemical and physical state of the formulation
is stable. Once administered to the intestine the therapeutic agent
is protected from damage by the GI environment since the
formulations are oil-based and therefore a separate local
environment is created in the intestine where the therapeutic agent
is contained in oil droplets, which confers stability in vivo.
[0108] In certain embodiments the process produces a composition
which consists essentially of a therapeutic agent and a medium
chain fatty acid salt and a hydrophobic medium. In embodiments of
the invention the solid powder (solid form) consists essentially of
a therapeutic agent and a medium chain fatty acid salt. Further
embodiments of the invention are pharmaceutical compositions
produced by the process describe herein. In certain pharmaceutical
compositions the therapeutic agent is a protein, a polypeptide, a
peptide, a glycosaminoglycan, a polysaccharide, a small molecule or
a polynucleotide and in particular embodiments the therapeutic
agent is insulin, growth hormone, parathyroid hormone,
teriparatide, interferon-alfa (IFN-.alpha.), a low molecular weight
heparin, leuprolide, fondaparinux, octreotide, exenatide,
terlipressin, vancomycin or gentamicin. Particular embodiments of
the invention comprise an oral dosage form comprising the
pharmaceutical composition, in particular an oral dosage form which
is enteric coated. Further embodiments of the invention comprise a
capsule containing the compositions of the invention, and in
various embodiments the capsule is a hard gel or a soft gel
capsule, and generally the capsule is enteric-coated. Other
embodiments of the invention comprise a rectal dosage form
comprising the pharmaceutical composition, in particular a
suppository, or a buccal dosage form. A kit comprising instructions
and the dosage form is also envisaged.
[0109] The therapeutic agent or medium chain fatty acid salt, or
any combination of therapeutic agent and other components, such as
protein stabilizers, can be prepared in a solution of a mixture
(e.g., forming an aqueous solution or mixture) which can be
lyophilized together and then suspended in a hydrophobic medium.
Other components of the composition can also be optionally
lyophilized or added during reconstitution of the solid
materials.
[0110] In some embodiments, the therapeutic agent is solubilized in
a mixture, for example, including one or more additional components
such as a medium chain fatty acid salt, a stabilizer and/or a
surface active agent, and the solvent is removed to provide a
resulting solid powder (solid form), which is suspended in a
hydrophobic medium. In some embodiments, the therapeutic agent
and/or the medium chain fatty acid salt may be formed into a
granulated particle that is then associated with the hydrophobic
medium (for example suspended in the hydrophobic medium or coated
with the hydrophobic medium). In general, the compositions
described herein are substantially free of "membrane fluidizing
agents" such as medium chain alcohols.
[0111] "Membrane fluidizing agents" are defined as medium chain
alcohols which have a carbon chain length of from 4 to 15 carbon
atoms (e.g., including 5 to 15, 5 to 12, 6, 7, 8, 9, 10, or 11
carbon atoms). For example, a membrane fluidizing agent can be a
linear (e.g., saturated or unsaturated), branched (e.g., saturated
or unsaturated), cyclical (e.g., saturated or unsaturated), or
aromatic alcohol. Examples of suitable linear alcohols include, but
are not limited to, butanol, pentanol, hexanol, heptanol, octanol,
nonanol, decanol, undecanol, dodecanol, tridecanol, tetradecanol,
and pentadecanol. Examples of branched alcohols include, but are
not limited to, geraniol, farnesol, rhodinol, citronellol. An
example of a cyclical alcohol includes, but is not limited to,
menthol, terpineol, myrtenol, perillyl and alcohol. Examples of
suitable aromatic alcohols include, but are not limited to, benzyl
alcohol, 4-hydroxycinnamic acid, thymol, styrene glycol, and
phenolic compounds. Examples of phenolic compounds include, but are
not limited to, phenol, m-cresol, and m-chlorocresol.
[0112] If desired, the pharmaceutical composition may also contain
minor amounts of non-toxic auxiliary substances such pH buffering
agents, and other substances such as for example, sodium acetate
and triethanolamine oleate.
[0113] In at least one embodiment, a therapeutic agent, such as a
protein, may be chemically modified to enhance its half-life in
circulation. For example, the therapeutic agent may undergo a
process such as pegylation.
[0114] In some embodiments the process for producing a
pharmaceutical composition comprises preparing a water-soluble
composition comprising a therapeutically effective amount of at
least one therapeutic agent and a medium chain fatty acid salt,
drying the water soluble composition to obtain a solid powder, and
dissolving the solid powder in a solution consisting essentially of
octanoic acid, thereby producing the pharmaceutical composition,
which is a solution. In some embodiments, the solid form may be a
particle (e.g., consist essentially of particles, or consists of
particles). In some embodiments, the particle may be produced by
lyophilization or by granulation. In some embodiments of this
process the octanoic acid is present in the composition at a level
of about 60% to about 90% or at a level of about 70 to about 85%
preferably about 78%. In some embodiments of this process the fatty
acid salt is sodium octanoate; in further embodiments of this
process the medium chain fatty acid salt is present in the
composition at an amount of about 11% to about 40% by weight or at
an amount of about 11% to about 28% by weight or at an amount of
about 15% by weight. In some embodiments of this process the
composition additionally comprises a matrix forming polymer and in
particular embodiments of this process the matrix forming polymer
is dextran or polyvinylpyrrolidone (PVP); in further embodiments of
this process the polyvinylpyrrolidone is present in the composition
at an amount of about 2% to about 20% by weight or at an amount of
about 5% to about 15% by weight, preferably at an amount of about
10% by weight. In certain embodiments of this process the
polyvinylpyrrolidone is PVP-12 and/or has a molecular weight of
about 3000. The composition may in addition include surfactants as
described above. The pharmaceutical products of these processes are
further embodiments of the invention e.g. a composition containing
octanoic acid at a level of about 60% to about 90% or at a level of
about 70 to about 85% preferably about 78%; fatty acid salt,
preferably sodium octanoate, present in the composition at an
amount of about 11% to about 40% by weight or at an amount of about
11% to about 28% by weight or at an amount of about 15% by weight;
matrix forming polymer e.g. polyvinylpyrrolidone, preferably
PVP-12, present in the composition at an amount of about 2% to
about 20% by weight or preferably an amount of about 5% to about
15% by weight, preferably at an amount of about 10% by weight; and
surfactants as described above. There also may be small quantities
of other hydrophobic constituents as described above.
[0115] Capsules: Preferred pharmaceutical compositions are oral
dosage forms or suppositories. Exemplary dosage forms include
gelatin or vegetarian capsules like starch
hydroxylpropyl-methylcellulose ("HPMC") capsules, enteric coated,
containing the bulk drug product. Capsules which may be used to
encapsulate the compositions of this invention are known in the art
and are described for example in Pharmaceutical Capsules edited by
Podczech and Jones, Pharmaceutical Press (2004) and in Hard gelatin
capsules today--and tomorrow, 2nd edition, Steggeman ed published
by Capsugel Library (2002).
[0116] Additional formulations: The compositions of the invention
may be formulated using additional methods known in the art, for
example as described in the following publications: Pharmaceutical
Dosage Forms Vols 1-3 ed. Lieberman, Lachman and Schwartz,
published by Marcel Dekker Inc, New York (1989); Water-insoluble
Drug Formulation 2.sup.nd edition, Liu, editor, published by CRC
Press, Taylor and Francis Group (2008); Therapeutic Peptides and
Proteins: Formulation, Processing and Delivery Systems, 2.sup.nd
edition by Ajay K. Banga (author) published by CRC Press, Taylor
and Francis Group (2006); Protein Formulation and Delivery,
2.sup.nd edition, McNally and Hasted eds, published by Informa
Healthcare USA Inc (2008); and Advanced Drug Formulation to
Optimize Therapeutic Outcomes, Williams et al eds, published by
Informa Healthcare USA (2008).
[0117] The compositions of the invention may be formulated using
microparticulate technology for example as described in
Microparticulate Oral Drug Delivery, Gerbre-Selassie ed., published
by Marcel Dekker Inc (1994) and in Dey et al, Multiparticulate Drug
Delivery Systems for Controlled Release, Tropical Journal of
Pharmaceutical Research, September 2008; 7 (3): 1067-1075.
[0118] Methods of treatment: The compositions described herein
exhibit effective, enteral delivery of an unaltered biologically
active substance (i.e. a therapeutic agent) and thus, have many
uses. For example, the compositions described herein can be used in
the treatment of diabetes.
[0119] In particular, insulin to treat and prevent subjects
(patients) suffering from Type II diabetes (prophylaxis of
diabetes), and to treat patients suffering from dysglycemia,
pre-diabetes and metabolic syndrome and other conditions, may be
administered in accordance with one or more embodiments of the
invention. Metabolic syndrome is a combination of medical disorders
that increase the risk of developing cardiovascular disease and
diabetes. Metabolic syndrome is a composite of different symptoms:
(1) fasting hyperglycemia (insulin resistance, type II diabetes,
etc); (2) decreased HDL cholesterol; (3) elevated triglycerides;
(4) high blood pressure; (5) central obesity; and (6)
proinflammatory state.
[0120] One embodiment of the invention is a method of treatment or
prevention of a subject suffering from the above conditions where
the amount of insulin sufficient to treat the condition is a low
dose of insulin formulated within the compositions of the
invention. Low dose insulin is provided by less than 300 or less
than 200 Units per capsule e.g. 40-200 Units per capsule.
[0121] Terlipressin (or other vasopressin analogs) to treat
subjects (patients) suffering from hepato-renal syndrome (HRS),
including HRS I and II, bleeding esophageal varices, portal
hypertension and other conditions may be administered in accordance
with one or more embodiments of the invention. Such terlipressin
formulations may also be used for primary and secondary prophylaxis
of variceal bleeding. A composition of the invention comprises a
suspension which comprises an admixture of a hydrophobic medium and
a solid form wherein the solid form comprises a therapeutically
effective amount of terlipressin (or other vasopressin analogues)
and at least one salt of a medium chain fatty acid.
[0122] Exenatide to improve glycemic control in subjects suffering
from Type II diabetes and to treat other conditions such as obesity
and for use in weight management may be administered in accordance
with one or more embodiments of the invention.
[0123] Interferon-alfa for the treatment of subjects suffering from
chronic hepatitis C and chronic hepatitis B and to treat other
conditions including cancer may be administered in accordance with
one or more embodiments of the invention.
[0124] Copaxone to treat subjects suffering from multiple sclerosis
and to treat other conditions including inflammatory diseases may
be administered in accordance with one or more embodiments of the
invention.
[0125] Desmopressin to treat subjects suffering from primary
nocturnal enuresis, central diabetes insipidus (DI) or bleeding
disorders (Von Willebrand Disease and Hemopilia A) may be
administered in accordance with one or more embodiments of the
invention. Oral desmopressin preparations known in the art suffer
from extremely low oral bioavailability.
[0126] Octreotide was first synthesized in 1979, and is an
octapeptide that mimics natural somatostatin pharmacologically,
though it is a more potent inhibitor of growth hormone, glucagon
and insulin than the natural hormone. Octreotide or other analogs
of somatostatin may be administered in accordance with one or more
embodiments of the invention for use in treating or preventing a
disease or disorder in a subject suffering from a disorder such as
acromegaly, abnormal GI motility, flushing episodes associated with
carcinoid syndrome, portal hypertension, an endocrine tumor (such
as carcinoids, VIPoma), gastroparesis, diarrhea, pancreatic leak or
a pancreatic pseudo-cyst. The diarrhea may result from radiotherapy
or may occur for example in subjects with vasoactive intestinal
peptide-secreting tumors (VIPomas). In addition, patients that
undergo pancreatic surgery may suffer from secretion of extrinsic
pancreas and are vulnerable to developing pancreatic leak or
pseudo-cysts which may be treated by octreotide products of the
invention. Some preferred embodiments are directed to a method of
treating a subject having a disorder such as acromegaly, abnormal
GI motility, flushing episodes associated with carcinoid syndrome,
portal hypertension, an endocrine tumor (such as carcinoids,
VIPoma), gastroparesis, diarrhea, pancreatic leak or a pancreatic
pseudo-cyst, which comprises administering to the subject a
composition of the invention, wherein the therapeutic agent is
octreotide, in an amount sufficient to treat the disorder.
Octreotide formulations of the invention may also be used for
primary and secondary prophylaxis of variceal bleeding, which may
be caused by portal hypertension; the varices may be gastric or
esophageal. Other uses of octreotide formulations of the invention
are in treatment of shock of hypovolemic (e.g. hemorrhagic) or
vasodilatory (e.g. septic) origin, hepatorenal syndrome (HRS),
cardiopulmonary resuscitation and anesthesia-induced hypotension.
Other analogs of somatostatin may be used in the methods and
compositions in which octreotide is used.
[0127] Vancomycin (molecular weight 1449 Da) is a glycopeptide
antibiotic used in the prophylaxis and treatment of infections
caused by Gram-positive bacteria. The original indication for
vancomycin was for the treatment of methycilin-resistant
Staphylococcus aureus (MRSA). Vancomycin never became first line
treatment for Staphylococcus aureus, one reason being that
vancomycin must be given intravenously. The prior art preparations
of vancomycin need to be given intravenously for systemic therapy,
since vancomycin does not cross through the intestinal lining. It
is a large hydrophilic molecule which partitions poorly across the
gastrointestinal mucosa. The only indication for oral vancomycin
therapy is in the treatment of pseudomembranous colitis where it
must be given orally to reach the site of infection in the colon.
Vancomycin for use in treating or preventing infection in a subject
may be administered orally to the subject in accordance with one or
more embodiments of the invention. Some preferred embodiments of
the invention are directed to a method of treating or preventing an
infection in a subject which comprises administering to the subject
a composition of the invention, wherein the therapeutic agent is
vancomycin, in an amount sufficient to treat or prevent the
infection.
[0128] Gentamicin (molecular weight=478) is an aminoglycoside
antibiotic, used to treat many types of bacterial infections,
particularly those caused by gram-negative bacteria. When
gentamicin is given orally in the prior art formulations, it is not
systemically active. This is because it is not absorbed to any
appreciable extent from the small intestine.
[0129] In addition, compositions of the invention also can be used
to treat conditions resulting from atherosclerosis and the
formation of thrombi and emboli such as myocardial infarction and
cerebrovascular accidents. Specifically, the compositions can be
used to deliver heparin or low molecular weight heparin or
fondaparinux across the mucosal epithelia.
[0130] The compositions of this invention can also be used to treat
hematological diseases and deficiency states such as anemia and
hypoxia that are amenable to administration of hematological growth
factors. The compositions of the invention can be used to deliver
vitamin B12 in a subject at high bioavailability wherein the
mucosal epithelia of the subject lacks sufficient intrinsic factor.
G-CSF may also be administered in accordance with various
embodiments. Additionally, the compositions of this invention can
be used to treat osteoporosis, such as through enteral
administration of PTH, teriparatide or calcitonin once or twice or
more daily.
[0131] Human growth hormone (hGH) to treat growth hormone
deficiency in particular in children may be administered in
accordance with one or more embodiments. In some preferred
embodiments, a composition described herein comprising growth
hormone can be administered to a subject to treat or prevent
metabolic and lipid-related disorders, e.g., obesity, abdominal
obesity, hyperlipidemia or hypercholestrolemia. For example a
composition of the invention comprising growth hormone can be
administered orally to a subject thereby treating obesity (e.g.,
abdominal obesity). In some preferred embodiments, a composition
described herein comprising growth hormone is administered to a
subject to treat or prevent HIV lipodistrophy (AIDS wasting) or to
treat Prader-Willi syndrome, growth disturbance due to insufficient
secretion of growth hormone (e.g. associated with gonadal
dysgenesis or Turner syndrome), growth disturbance in prepubertal
children with chronic renal insufficiency, and as replacement
therapy in adults with pronounced growth hormone deficiency.
Compositions of the invention comprising growth hormone can be
administered orally to a subject to promote wound healing and
attenuate catabolic responses in severe burns, sepsis, multiple
trauma, major operations, acute pancreatitis and intestinal
fistula. Many other conditions besides GH deficiency cause poor
growth, but growth benefits (height gains) are often poorer than
when GH deficiency is treated. Examples of other causes of
shortness which may be treated with compositions of the invention
comprising growth hormone are intrauterine growth retardation, and
severe idiopathic short stature. Other potential uses of
compositions of the invention comprising growth hormone include
treatment to reverse or prevent effects of aging in older adults,
to aid muscle-building and as treatment for fibromyalgia.
[0132] Some preferred embodiments are directed to a method of
treating a disorder such as obesity, HIV lipodistrophy, metabolic
disorder, or growth deficiency in a subject which comprises
administering to the subject a composition of the invention wherein
the therapeutic agent (the effector) is growth hormone, in an
amount sufficient to treat the disorder.
[0133] Some preferred embodiments are directed to a method of
treating a bone disorder in a subject which comprises administering
to the subject a composition of the invention, wherein the
therapeutic agent is teriparatide or parathyroid hormone, in an
amount sufficient to treat the bone disorder.
[0134] Some preferred embodiments are directed to a method of
treating or preventing a blood coagulative disorder in a subject
which comprises administering to the subject a composition of the
invention wherein the therapeutic agent is heparin or a heparin
derivative or fondaparinux, in an amount sufficient to treat or
prevent the blood coagulative disorder.
[0135] Leuprolide (GnRH agonist) formulated in an embodiment of the
invention may be delivered for treatment of female infertility
(e.g. once or twice daily dosage), prostate cancer and Alzheimer's
disease.
[0136] One embodiment of the invention relates to a method of
treating a subject suffering from a disease or disorder which
comprises administering to the subject a composition of the
invention in an amount sufficient to treat the condition. Another
embodiment of the invention relates to compositions of the
invention for use in treating a disease or disorder in a subject.
Another embodiment of the invention relates to the use of a
therapeutic agent in the manufacture of a medicament by the process
of the invention for the treatment of a disorder.
[0137] The dosage regimen utilizing the compounds is selected in
accordance with a variety of factors including type, species, age,
weight, sex and medical condition of the patient; the severity of
the condition to be treated; the route of administration; the renal
and hepatic function of the patient; and the particular compound or
salt thereof employed. 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. Oral dosages of the present invention,
when used for the indicated effects, may be provided in the form of
capsules containing 0.001, 0.0025, 0.005, 0.01, 0.025, 0.05, 0.1,
0.25, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0 or 100, 200, 300,
400, 500, 600, 700, 800 or 1000 mg of therapeutic agent.
[0138] Compounds of the present invention may be administered in a
single daily dose, or the total daily dosage may be administered in
divided doses of two, three, four, five or six times daily. In some
embodiments, the composition is administered at a daily dose of
from about 0.01 to about 5000 mg/day, e.g., administered once daily
(e.g., in the morning or before bedtime) or twice or more daily
(e.g. in the morning and before bedtime).
[0139] A representative product of the invention is an API-based
formulation orally administered as enteric coated-capsules: each
capsule contains API co-lyophilized with PVP-12 and sodium
octanoate, and suspended in a hydrophobic (lipophilic) medium
containing: glyceryl tricaprylate, glyceryl monocaprylate, and
Tween 80; in another representative product of the invention castor
oil is additionally present. The compositions described herein can
be administered to a subject i.e. a human or an animal, in order to
treat the subject with a pharmacologically or therapeutically
effective amount of a therapeutic agent described herein. The
animal may be a mammal e.g. a mouse, rat, pig horse, cow or sheep.
As used herein the term "pharmacologically or therapeutically
effective amount" means that amount of a drug or pharmaceutical
agent (the therapeutic agent) that will elicit the biological or
medical response of a tissue, system, animal or human that is being
sought by a researcher or clinician.
[0140] The formulations of the invention allow incorporation of the
therapeutic agent into the formulation without any chemical
modification of the therapeutic agent. Additionally, as shown
above, many different therapeutic agents have been successfully
formulated within the formulations of the invention, including
polypeptides, nucleotides, small molecules and even medium size
proteins. Furthermore, the formulations of the invention allow for
high flexibility in loading of the therapeutic agent. Loading
capacity is dependent on the therapeutic agent. To date, loading
capacity limits have not been reached; however loading of up to
1.5% wt/wt (polypeptides) and 6% wt/wt (small molecules) has been
achieved and higher loading up to 33% is envisaged. Finally, the
formulations of the invention protect the cargo compounds from
inactivation in the GI environment due to for example proteolytic
degradation and oxidation.
[0141] The function and advantages of these and other embodiments
will be more fully understood from the following examples. These
examples are intended to be illustrative in nature and are not to
be considered as limiting the scope of the systems and methods
discussed herein.
EXAMPLES
Example 1
Formulations
[0142] A. Composition of an Insulin Formulation
[0143] Table 1A presents an example of a composition in accordance
with one or more embodiments. More specifically, this composition
is an insulin formulation. Insulin was obtained from Diosynth
Biotechnology; sodium octanoate and NaOH from Merck; MgCl.sub.2,
MC400, Span40, lecithin and castor oil from Spectrum; PVP-12 from
BASF; ethyl isovalerate from Merck/Sigma; glyceryl tributyrate from
Acros/Penta; and glycerol monooleate from Abitec Corp.
TABLE-US-00001 TABLE 1A Ingredient % w/w Hydrophilic Insulin 0.417
Fraction NaOH 0.029 MgCl.sub.2 0.104 PVP-12 2.083 Sodium octanoate
3.125 Methyl cellulose 0.104 Hydrophobic Castor oil 52.858 Medium
Glyceryl tributyrate 28.466 Ethyl isovalerate 8.195 Glycerol
monooleate 1.779 Lecithin 1.893 Span-40 0.946
[0144] B. A formulation for leuprolide: Table 1B presents an
example of a composition for an API (Active Pharmaceutical
Ingredient) in accordance with one or more embodiments. More
specifically, this composition is a leuprolide formulation.
TABLE-US-00002 TABLE 1B Ingredient % w/w Hydrophilic Leuprolide
0.072 Fraction NaOH 0.038 MgCl.sub.2 0.137 PVP-12 2.740 Sodium
octanoate 12.002 Methyl cellulose 0.137 Water 0.605 Hydrophobic
Span-40 1.21 Medium Lecithin 2.43 Ethyl-isovalerate 10.52 Glycerol
monooleate 2.28 Glyceryl tributyrate 23.74 Castor Oil 44.09
[0145] C. A Formulation with Decreased Amount of Hydrophobic Medium
(50% of Hydrophobic Medium)
[0146] Table 1C presents an example of a composition for an API in
accordance with one or more embodiments. More specifically, this
composition is a formulation for dextran (FD4). The FD4 is
FITC-labeled dextran with a MW of 4.4 kDa (Sigma, FD4) and this is
the dextran which was used throughout the Examples unless stated
otherwise. This particular formulation contains coconut oil (Sigma)
instead of GTB.
TABLE-US-00003 TABLE 1C Ingredient % w/w Hydrophilic Dextran 0.939
Fraction NaOH 0.001 MgCl.sub.2 0.235 PVP-12 4.693 Sodium octanoate
20.662 Methyl cellulose 0.235 Water 1.071 Hydrophobic Span-40 1.04
Medium Lecithin 2.08 Ethyl-isovalerate 9.01 Glycerol-monooleate
1.95 Coconut oil 20.33 Castor oil 37.75
The above formulations are used for a wide variety of therapeutic
agents and give good bioavailability to the cargo compound in the
animal models described below. Note that the net amount of
therapeutic agent may vary as appropriate in any of the
formulations and there may be minor variations in the formulations;
for example NaOH is not always used; coconut oil may be used
instead of glyceryl tributyrate; MgCl.sub.2 is not always used
(e.g. with hGH it is not used); all ingredients may be substituted
as described above in the specification.
Example 2
Schematic Representation of Insulin Formulation Production
[0147] FIG. 1 illustrates a method of producing a composition in
accordance with one or more embodiments. For example, this method
may be implemented to make the compositions presented above in
Example 1.
Example 3
The Combination of Solid Particles Containing Sodium Octanoate and
Hydrophobic Medium is Critical for Permeation Activity
[0148] FIG. 2 presents data relating to serum insulin levels after
rectal administration to rats. Rats were anesthetized and were
administered 1000 .mu.L of bulk drug formulation containing an
insulin dose of 328 .mu.g/rat (9 IU/rat). Blood samples were
collected at 0, 3, 6, 10, 15, 25, 30, 40, 60 and 90 minutes post
administration and serum was prepared for determination of human
insulin by an immunoassay kit with no cross reactivity between rat
and human insulin.
[0149] Data is presented as MEAN.+-.SD, n=5. The left panel of FIG.
2 relates to administration of human insulin with sodium octanoate
(Na--C8) or solid hydrophilic fraction suspended in water (solid
particles in water). The right panel of FIG. 2 relates to
administration of full insulin formulation (solid particles in
hydrophobic medium). Table 2 below presents a summary of AUC values
calculated from the concentration vs. time curves.
TABLE-US-00004 TABLE 2 Test compound AUC.sub.(0-.infin.) Na--C8
5753 .+-. 3569 Solid particles in water 4083 .+-. 2569 Insulin in
formulation 280933 .+-. 78692 (Solid particles in hydrophobic
medium) Data are MEAN .+-. SD
[0150] The average exposure (expressed by AUC values) to insulin
after rectal administration of insulin-SCD was about 50-fold higher
than the exposure after administration without a hydrophobic
medium. Minimal exposure was detected in rats administered insulin
with sodium octanoate alone or as part of the solid particles of
the hydrophilic fraction (as listed in Example 1) suspended in
water. These data demonstrate synergy between solid sodium
octanoate and a hydrophobic medium.
Example 4
Intestinal Absorption of Insulin After GI Administration of Insulin
to Rats
[0151] FIG. 3 presents data relating to serum insulin levels and
blood glucose levels after rectal administration of insulin
solution and insulin in formulation to rats. Rats were anesthetized
and administered 100 .mu.L of test article (insulin in formulation
or insulin in PBS) containing an insulin dose of 328 .mu.g/rat (9
IU/rat). Blood samples were collected at 0, 3, 6, 10, 15, 25, 30,
40, 60 and 90 minutes post administration. Glucose level was
immediately determined with a glucometer and serum was prepared for
determination of human insulin by an immunoassay kit with no cross
reactivity between rat and human insulin.
[0152] Glucose levels are presented as the percentage form basal
levels measured before administration (time 0). The data of FIG. 3
is presented as MEAN.+-.SD, n=5.
[0153] Levels of insulin (left panel on FIG. 3) and glucose (right
panel of FIG. 3) after rectal administration of human insulin
solubilized in PBS (insulin solution) or incorporated in the
formulation are presented. Insulin levels rose rapidly in rat serum
after rectal administration of insulin in formulation. Maximal
levels were measured within 6 minutes post administration and a
gradual drop detected until reaching basal levels at about 90 min
post administration. This sharp and significant rise in insulin was
accompanied by a significant drop in glucose levels reaching an
average of 20% of the initial levels already at 30 min post
administration. By contrast, rectal administration of insulin in
PBS caused only a very slight glucose reduction, which is identical
to that observed following treatment with the PBS control
alone.
Example 5
Insulin Absorption After Rectal Administration of Insulin in
Formulation to Rats
[0154] FIG. 4 presents data relating to changes in blood glucose
and serum insulin concentrations following SC (subcutaneous)
administration of insulin solution (at 20 .mu.g/rat) and rectal
administration of insulin in formulation (at 328 .mu.g/rat). Blood
samples were collected at 0, 3, 6, 10, 15, 25, 30, 40, 60 and 90
minutes post rectal administration and at 0, 15, 30, 45, 60, 90
min, 2, 3, and 4 hours post SC administration. Glucose was
immediately determined with a glucometer and insulin by an
immunoassay kit. Glucose levels are presented as the percentage
form basal levels measured before administration (time 0). The data
of FIG. 4 is presented as MEAN.+-.SD, n=5.
[0155] The levels of insulin absorption from rat colon after
insulin in formulation administration were compared to the levels
of insulin absorbed after SC administration. Insulin exposure was
calculated from the area under the serum concentration versus time
curve (AUC) and the activity calculated as the relative
bioavailability (rBA) according to the following equation:
rBA=(rectal AUC.sub.(0-.infin.)/SC AUC.sub.(0-.infin.))*(SC
dose/rectal dose)
[0156] Insulin penetration into the bloodstream occurs during a
narrow window of time, generally within about 10 minutes of rectal
insulin in formulation administration. The rise in serum insulin is
paralleled by a fall in blood glucose levels.
[0157] In order to derive information about insulin bioavailability
when formulated insulin is presented into the colon,
AUC.sub.(0-.infin.) was determined for rectal and SC administration
and the rBA value of human insulin was 29.4.+-.3.4% with
coefficient of variance (CV)=11.4%.
[0158] Rectal administration of various insulin-containing
formulations was carried out on hundreds of animals. The assay was
further developed and qualified as a bioassay to support platform
development and batch release tests with a linear range of 10-200
.mu.g/rat, repeatability of 39% and intermediate precision of
33%.
[0159] The insulin formulation described herein was tested in five
different studies using a total of 25 rats. The rBA was
34.1.+-.12.6% with CV of 28.9%.
Example 6
Insulin Absorption After Intra-Jejunal Administration of Insulin in
Formulation to Rats
[0160] The absorption target site of the orally administered
platform of the invention is generally the small intestine. To test
the activity of insulin formulation in rat intestine, two major
obstacles were addressed: 1. Enteric-coated capsules for rats are
not available and therefore stomach bypass enabling direct
intra-jejunal administration is needed. 2. Insulin is extensively
metabolized by the liver; in humans 50-80% of endogenous insulin,
secreted by pancreatic .beta.-cells, is sequestered by the liver
and therefore can not be detected in the systemic circulation.
Insulin administered via the intestinal route (by way of insulin
formulation) mimics the endogenous route of insulin as the
intestinal blood flow is drained into the portal vein which leads
directly to the liver. Therefore to determine insulin absorbance,
blood samples must be drawn from the portal vein (portal
circulation, prior to the liver) as well as the jugular vein
(systemic circulation, after the liver).
[0161] A specialized rat model in which three different cannulas
are surgically implanted in anesthetized rats was developed: 1.
Jejunal cannula--stomach bypass, enables insulin formulation
administration, 2. Portal vein cannula--blood sampling prior to the
liver, determine insulin that cross the GI wall into the blood, and
3. Jugular vein cannula--to determine the systemic levels of
insulin. Using this model, the bioavailability of insulin in
formulation (rBA) was determined.
[0162] FIG. 5 presents data from a representative study relating to
insulin levels in the portal and systemic circulations after
intra-jejunal administration of insulin control and insulin
formulation to rats. Rats (8 rats per group) were anesthetized and
their jejunum exposed by abdominal surgery. The jejunum containing
intestinal loop was placed on gauze and kept moist and fully intact
throughout the entire study. A temporary cannula was inserted into
the jejunum and formulated insulin was administered. Blood was
collected from both portal and jugular veins at the same time
points, with approximately 4 time points per rat. The MEAN.+-.SD
value of each time point was used to create a plasma concentration
vs. time curve. AUC was determined and rBA was calculated.
[0163] Insulin levels in both the portal and systemic circulation
rose dramatically after intra-jejunum administration of insulin in
formulation. This is in contrast to the minimal insulin absorbance
detected when insulin control was administered. The window of
absorption was short and insulin levels peaked by 6 minutes. This
profile is similar to that seen after rectal administration of
formulated insulin (see above). Higher insulin levels were detected
in the portal compared to the systemic circulation, with rBA of
10.1% compared to 5.6%, respectively.
Example 7
Additional Formulations Comprising Various Cargo Compounds
[0164] Table 3A details the components of a range of dextran
formulations which were prepared as described in the following
Examples. The sodium caprate was obtained from Fluka/Sigma, the
olive oil from Fluka, the octanoic acid from Sigma and the mineral
oil from Acros.
TABLE-US-00005 TABLE 3A Cargo Dextran Formulation A B C D E F G H
Ingredient (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) (% w/w)
(% w/w) Hydrophilic Cargo 0.545 0.939 0.565 0.546 0.565 0.565 0.565
0.551 fraction NaOH 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001
MgCl.sub.2 0.136 0.235 0.141 0.156 0.141 0.141 0.141 0.138 PVP-12
2.726 4.693 2.823 3.117 2.823 2.823 2.823 2.754 Sodium 12.001
20.662 -- -- 9.002 9.002 9.002 12.125 octanoate Sodium -- -- 9.002
-- -- -- -- -- caprate MC 400 0.136 0.235 0.141 0.156 0.141 0.141
0.141 0.138 Water 0.622 1.071 0.507 0.159 0.507 0.507 0.507 0.661
Hydro- Span40 1.21 1.04 1.25 1.38 1.25 1.25 1.25 -- phobic Lecithin
2.42 2.08 2.50 2.76 2.50 2.50 2.50 -- medium Ethyliso- 10.46 9.01
10.83 11.96 10.83 10.83 10.83 11.23 valerate Glyceryl 2.27 1.95
2.35 2.60 2.35 2.35 2.35 -- monooleate Glyceryl 23.62 20.33 24.46
24.29 24.46 24.46 24.46 25.35 tributyrate Coconut oil -- -- -- --
-- -- -- -- Castor oil 43.86 37.75 45.42 45.07 45.42 -- -- 47.08
Octanoic acid -- -- -- 7.80 -- -- -- -- Mineral oil -- -- -- -- --
45.42 -- -- Olive oil -- -- -- -- -- -- 45.42 --
Table 3B details the components of a range of teriparatide acetate
and leuprolide formulations which were prepared as described in the
following Examples. Teriparatide was obtained from Novetide, and
leuprolide was obtained from Bambio.
TABLE-US-00006 TABLE 3B Cargo Teriparatide Leuprolide Formulation I
J K L Ingredient (% w/w) (% w/w) (% w/w) (% w/w) Cargo 0.118 0.118
0.050 0.050 NaOH -- -- 0.040 0.04 MgCl.sub.2 0.137 0.137 0.142 0.15
Hydro- PVP-12 2.740 2.740 2.838 2.99 philic Sodium 12.001 12.001
9.012 -- fraction octanoate Sodium -- -- -- 4.48 caprate MC 400
0.137 0.137 0.142 0.15 Water 0.605 0.605 0.489 0.33 Hydro- Span40
1.214 1.214 1.26 1.32 phobic Lecithin 2.428 2.428 2.52 2.65 medium
Ethyl-iso- 10.515 10.515 10.89 11.46 valerate Glyceryl 2.283 2.283
2.36 2.49 monooleate Glyceryl 23.740 -- 24.59 25.87 tributyrate
Coconut oil -- 23.740 -- -- Castor oil 44.082 44.082 45.66
48.04
Table 3C details the components of hGH formulations which were
prepared as described and the following Examples. The hGH was
obtained from PLR, Israel (GHP-24).
TABLE-US-00007 TABLE 3C Cargo hGH Formulation O P Ingredient (%
w/w) (% w/w) Hydrophilic Cargo 0.298 0.303 fraction NaOH -- --
MgCl.sub.2 -- -- PVP-12 2.836 2.738 Sodium 9.006 12.007 octanoate
Sodium -- -- caprate MC 400 0.142 0.137 Water 0.492 0.607
Hydrophobic Span40 1.257 1.213 medium Lecithin 2.514 2.427
Ethyl-iso- 10.885 10.508 valerate Glyceryl 2.363 2.281 monooleate
Glyceryl 24.575 23.725 tributyrate Coconut oil -- -- Castor oil
45.633 44.054
[0165] The production process for all these above formulations is
essentially as described in FIG. 1 and in Example 11.
Example 8
Effect of Dose of Sodium Octanoate Incorporated in Formulation on
Formulation Activity
[0166] The effect of increasing the amount of sodium octanoate
(Na--C8) in the formulation on the activity of the formulation was
tested using formulations containing dextran (average MW=4.4 kDa,
FITC labeled) as cargo compound and different doses of Na--C8
namely, formulation A in Table 3A (which contains 12% sodium
octanoate by weight) and similar dextran formulations containing
different Na--C8 doses: 9%, 6% and 3% respectively.
[0167] To test the activity of these formulations in the jejunum of
non-anesthetized rats, a rat model was established in which two
different cannulas are surgically implanted in male Sprague-Dowley
rats [0168] 1--Jejunal cannula to bypass the stomach and enable
direct formulation administration to the jejunum. [0169] 2--Jugular
vein cannula to determine the systematic levels of the administered
dextran following jejunal administration. Rats are allowed to
recover for 4 days before the study and are deprived of food for 18
hours before the start of the study.
[0170] FIG. 6 presents data from a study which determines
FITC-labeled dextran (4.4 kDa) bioavailability in non-anesthetized
rats following intra-jejunal administration of formulations
containing different amounts of Na--C8 or FITC-labeled dextran
solubilized with the Na--C8 in saline solution (control).
[0171] The bioavailability of the different dextran formulations
and the control was evaluated by administrating the different
formulations directly to the jejunum of non-anesthetized rats and
measuring plasma dextran levels at 3, 6, 10, 25, 60 and 90 minutes
post administration. Levels of plasma dextran following
administration of dextran in formulation or in saline were compared
to the levels of plasma dextran after intravenous administration.
Exposure values, AUC (0-90), were determined for jejunal and
intravenous administration and the absolute bioavailability (aBA)
was calculated according to the following equation:
aBA=(jejunal AUC(0-90))/(iv AUC (0-90))*(iv dose/jejunal dose).
Data are presented as Mean.+-.SD (n.gtoreq.5 rats per group).
The results show that increasing the amount of Na--C8 incorporated
in the formulation improves the bioavailability of the dextran in a
dose-responsive manner, reaching almost 30% aBA at the 12% (w/w)
dose. Dextran administered with Na--C8 at similar doses and
suspended in a saline solution (i.e. not formulated) showed much
lower bioavailability (.about.6% aBA). Further results
dose-response results are shown in Example 26.
Example 9
Effect of the Ratio of Hydrophilic Fraction/Hydrophobic Medium on
Formulation Activity
[0172] The effect on formulation activity of changing the ratio
(weight/weight) between the hydrophilic fraction and the
hydrophobic medium was tested using formulations containing dextran
(average MW=4.4 kDa, FITC labeled) as cargo (formulations A and B
in Table 3A). The in vivo non-anesthetized rat model described in
Example 8 was used in order to compare the activity of the
described formulations.
[0173] Table 4 presents bioavailability data following
intra-jejunal administration of formulations comprising a different
ratio of hydrophilic fraction to hydrophobic medium.
TABLE-US-00008 TABLE 4 Weight ratio between hydrophilic/ Route of
Formu- hydrophobic Animal adminis- % Cargo lation medium model
tration N aBA .+-. SD Dextran A 1/5.2 Rat Jejunal 17 28.0 .+-. 6.8
Non- anesthetized B 1/2.6 19 24.8 .+-. 25
[0174] Formulations A and B were administered directly to the
jejunum of non-anesthetized rats and plasma dextran levels were
measured at 3, 6, 10, 25, 60 and 90 minutes post formulation
administration. The levels of dextran absorption from rat jejunum
after administration of dextran in formulation were compared to the
levels of dextran absorbed after intravenous administration.
Exposure values, AUC (0-90), were determined for jejunal and
intravenous administration and the absolute bioavailability (aBA)
determined according to the following equation:
aBA=(jejunal AUC(0-90))/(iv AUC (0-90))*(iv dose/jejunal dose).
Data are presented as Mean.+-.SD (n.gtoreq.5 rats per group).
[0175] The results show that changing the ratio between the
hydrophilic fraction and the hydrophobic medium in these
formulations with a low % weight of therapeutic agent had no
significant effect on the bioavailability of the cargo which gives
a loading flexibility in devising additional formulations.
Example 10
Activity of Formulations Containing Different Cargo Compounds
[0176] In order to test the capability of the formulation platform,
the activity of formulations containing three different cargo
compounds (APIs) was tested in three different animal models:
jejunal administration to non-anesthetized rats, rectal
administration to anesthetized rats and jejunal administration to
non-anesthetized pigs. Table 5 summarizes the results of
representative experiments testing the bioavailability of
formulations containing different APIs in the three different
animal models described above.
TABLE-US-00009 TABLE 5 Route of Formu- Animal Adminis- % API lation
model tration N BA .+-.SD I Teriparatide I Rat Jejunal 5 14.0**
.+-.10.8 non- anesthe- tized II I Pig Jejunal 5 15.0** .+-.9.3 non-
anesthe- tized III Leuprolide K Rat Jejunal 4 10.1* .+-.7.5 non-
anesthe- tized IV hGH P Rat Rectal 5 17.9** .+-.3.9 anesthe- tized
*Absolute BA (compared to IV) **Relative BA (compared to SC)
[0177] A. Leuprolide Absorption After Jejunal Administration of
Leuprolide in Formulation to Rats
[0178] Table 5-III presents data from a representative study
relating to leuprolide % aBA following IV (intravenous)
administration of leuprolide solution (at 75 .mu.g/Kg) and jejunal
administration of leuprolide in formulation (at 450 .mu.g/Kg;
formulation K, Table 3B) to non-anesthetized rats, as previously
described in Example 8.
[0179] Blood samples were drawn from the jugular vein at 3, 6, 10,
15, 25, 40, 60 and 90 minutes post jejunal administration and at 3,
10, 25, 40, 90 min, 2, 3.3 and 5 hours post IV administration,
plasma was prepared and leuprolide levels were determined in each
sample. Leuprolide levels in systemic circulation rose dramatically
after jejunal administration of leuprolide in formulation.
Leuprolide blood levels peaked by 3 minutes post administration.
The average aBA achieved after jejunal administration of leuprolide
in formulation was calculated as described in the above Examples
and was 10.1%. In a control experiment, jejunal administration of
leuprolide in PBS demonstrated negligible penetration to the
bloodstream.
[0180] A similar leuprolide formulation containing 12% sodium
octanoate as described in Table 1B was prepared; it was tested in
the above model and showed bioavailability as follows:
rBA (compared to SC)=21.1%.+-.12.0 (CV=57%).
[0181] B. Teriparatide Absorption After Jejunal Administration of
Teriparatide in Formulation to Rats
[0182] Table 5-I presents data from a representative study relating
to plasma teriparatide concentration-time profiles following SC
administration of teriparatide solution (at 85 .mu.g/formulation
and jejunal administration of teriparatide (teriparatide) in
formulation (at 550 .mu.g/Kg; formulation I, Table 3B) to
non-anesthetized rats, as previously described in Example 8. Blood
samples were drawn from the jugular vein at 3, 6, 10, 25, 60 and 90
minutes post jejunal administration and at 3, 10, 30, 60, 90 min, 2
and 3 hours post SC administration, plasma was prepared and
teriparatide levels were determined in each sample. Teriparatide
levels in systemic circulation rose dramatically after jejunal
administration of teriparatide in formulation. Teriparatide levels
peaked by 3 minutes post-administration. The average rBA achieved
after jejunal administration of teriparatide in formulation was
calculated as described in the above Examples, and was 14.0%. In a
control experiment, jejunal administration of teriparatide in
saline demonstrated no penetration to the bloodstream.
[0183] C. Teriparatide Absorption after Jejunal Administration of
Teriparatide in Formulation to Pigs
[0184] Table 5-II presents data from a representative study
relating to plasma teriparatide concentration-time profiles
following SC administration of teriparatide solution (at 10.65
.mu.g/Kg) and jejunal administration of teriparatide in formulation
(at 100 .mu.g/Kg; formulation I, Table 3B) to non-anesthetized
pigs.
[0185] A pig model was established in which two different cannulas
were surgically permanently implanted in female domestic pigs:
[0186] 1--jejunal cannula to bypass the stomach and enable direct
formulation administration to the jejunum. [0187] 2--jugular vein
catheterization to determine the systematic levels of the
administered cargo following jejunal administration.
[0188] Pigs were allowed to recover for 7 days before the
experiment and deprived of food 18-20 hours before the start of the
experiment.
[0189] Blood samples were drawn from the jugular vein at 0, 3, 6,
10, 15, 25, 40, 60, 90 minutes, 2, 2.5 and 3 hours post jejunal
administration and at 0, 3, 6, 10, 15, 20, 30, 45, 60, 90 min, 2,
2.5, 3 and 4 hours post SC administration, plasma was prepared and
teriparatide levels were determined in each sample. Teriparatide
levels in systemic circulation rose dramatically after jejunal
administration of teriparatide in formulation. Teriparatide levels
peaked by 10 minutes post administration. The average rBA achieved
after jejunal administration of teriparatide in formulation was
calculated as described in the above Examples, and was 15.0%.
[0190] A similar pig experiment was performed using dextran (FD4,
formulation A in Table 3A) and it was determined that the average
bioavailability of dextran was 20% in pigs as compared to IV.
[0191] D. hGH Absorption after Rectal Administration of hGH in
Formulation to Rats
[0192] Table 5-IV presents data from a representative study
relating to plasma hGH concentration-time profiles following SC
administration of hGH solution (at 81 .mu.g/Kg) and rectal
administration of hGH in formulation (at 800 .mu.g/Kg; formulation
P, Table 3C), to anesthetized rats. Male Sprague-Dowley rats were
deprived of food for 18 hours before the start of the experiment.
Rats were anesthetized by a solution of ketamine: xylazine. The
formulation (100 .mu.L/rat) was administered rectally using a 14G
venflon. Blood samples were drawn from the jugular vein at 3, 6,
10, 15, 40, 60 and 90 minutes post rectal administration and at 15,
30, 45, 60, 90 min, 2, 3, and 4 hours post SC administration,
plasma was prepared and hGH levels were determined in each sample.
hGH levels in systemic circulation rose dramatically after rectal
administration of hGH in formulation. hGH levels peaked by 15
minutes. The average rBA achieved after rectal administration of
hGH in formulation was calculated as described in the above
Examples and was 17.9%. In a separate experiment hGH was
administered to the jejunum and the aBA was lower. In a control
experiment, rectal administration of hGH in PBS demonstrated no
penetration to the bloodstream.
[0193] Thus the results presented in Table 5 demonstrate that
substantial exposure was obtained for all cargo compounds tested in
all animal models tested.
[0194] The above results demonstrate that the formulations
described herein enable delivery of a wide range of different
macromolecules through the intestinal epithelium in different
animal models.
Example 11
Detailed Production Process of a formulation of Teriparatide
[0195] Production of the hydrophilic fraction: To 200 mL water the
following ingredients were slowly added one by one (with 2-3
minutes mixing between each ingredient): 172 mg of teriparatide,
200 mg of MgCl.sub.2, 4.0 g of PVP-12, 17.52 g of sodium octanoate
and 10.0 g of 2% MC-400 aqueous solution, prepared as follows: 1 g
of MC-400 powder was added to 50 mL water at 60.+-.2.degree. C.
while mixing. After 5 min of mixing, the beaker was transferred to
ice until a clear solution was obtained.
[0196] After addition of the MC-400 solution, the solution was
mixed for another 5 min and then lyophilized for about 24 h. This
procedure produced about 22 g of hydrophilic fraction.
[0197] Production of the hydrophobic medium: 2 g of Span 40, 4 g of
lecithin and 3.8 g of GMO were dissolved in 17.3 g of ethyl
isovalerate while mixing. To this solution were added 39.1 g of GTB
and 72.6 g of castor oil. This procedure produced about 136-138 g
of hydrophobic medium.
[0198] Production of the bulk drug product: Mixing of the
hydrophilic fraction and the hydrophobic medium was performed at
20.+-.2.degree. C.
[0199] 15.7 g of the hydrophilic fraction was slowly added during
mixing to 84.3 g of hydrophobic medium at 600.+-.50 RPM. After
addition of all the hydrophilic fraction, the mixing speed was
increased to 2000.+-.200 RPM for 2-10 min followed by 4-8 cycles of
15 min mixing at 600.+-.50 RPM and 2 min mixing at 2000.+-.200
RPM.
[0200] Degassing by vacuum was then applied as follows: 5 min at
600 mBar, 5 min at 500 mBar and 30-120 min at 400 mBar. The
resulting suspension was poured into a 100 mL dark bottle and
stored at 2-8.degree. C. This is the teriparatide formulation
designated "I" described in Table 3B.
[0201] All other formulations described herein were produced by
this method, varying ingredients and quantities according to the
details given in the relevant Tables (see e.g. Example 29). A
diagram of this method (with insulin as cargo) is shown in FIG.
1.
Example 12
Effect of the Oil Incorporated in the Formulation on Formulation
Activity
[0202] The effect of the type of oil incorporated in the
formulation (in the hydrophobic medium) on formulation activity was
tested. Formulations containing dextran (average MW=4.4 kDa, FITC
labeled) as cargo compound and different types of oils in the
hydrophobic medium (formulations E, F and G in Table 3A).were
tested in rats.
[0203] To test the activity of these formulations in the jejunum of
non-anesthetized rats, a rat model was established in which two
different cannulas are surgically implanted in male Sprague-Dowley
rats: [0204] 1--Jejunal cannula to bypass the stomach and enable
direct formulation administration to the jejunum. [0205] 2--Jugular
vein cannula to determine the systematic levels of the administered
dextran following jejunal administration. [0206] Rats are allowed
to recover for 4 days before the study and are deprived of food for
18 hours before the start of the study.
[0207] Table 6 presents data from a study in non-anesthetized rats
following intra jejunal administration of formulations containing
different oils in the hydrophobic medium.
TABLE-US-00010 TABLE 6 Cargo Formulation Oil N % aBA .+-. SD
Dextran E Castor oil + GTB 14 19.8 .+-. 5.5 F Mineral oil + 5 12.2
.+-. 5.0 GTB G Olive oil + GTB 5 12.0 .+-. 9.9
[0208] Formulations containing different oils were administered
directly to the jejunum of non-anesthetized rats and plasma dextran
levels were measured at 3, 6, 10, 25, 60 and 90 minutes post
formulation administration. The levels of dextran absorption from
rat jejunum after administration of dextran in formulation were
compared to the levels of dextran absorbed after intravenous
administration. Exposure values, AUC (0-90), were determined for
jejunal and intravenous administration and the absolute
Bioavailability (aBA) was determined according to the following
equation: aBA=(jejunal AUC(0-90))/(iv AUC(0-90))*(iv dose/jejunal
dose). Data are presented as Mean.+-.SD (n.gtoreq.5 rats per
group).
[0209] Similar bioavailability was achieved when dextran was
incorporated into formulations containing castor oil or coconut
oil. Good bioavailability was also obtained in rat jejunum when
teriparatide was used as cargo compound using formulations I and J;
these formulations contain castor oil and GTB, and castor oil and
coconut oil, respectively.
[0210] The results showed that formulations containing different
kinds of oils in their hydrophobic medium are active, enabling
penetration of the cargo (dextran, teriparatide) carried by the
formulation. Thus the data demonstrated that all tested oils enable
bioavailability of the cargo carried by the formulation. Castor oil
and coconut oil might be superior to the other tested oils.
Example 13
Preparation of a Formulation Using Granulation Instead of
Lyophilization
[0211] Production of the hydrophilic fraction: To a plastic bag,
the following ingredients were added: 1.00 g of PVP-30, 6.70 g of
sodium octanoate and 13.00 g of lactose monohydrate as binder.
After 5 min of mixing, all of the powder was transferred into a
mortar and pestle.
[0212] A dextran FD4 aqueous solution was prepared as followed:
0.42 g dextran was dissolved in 1.2 g of WFI. All of the dextran
solution was then added slowly to the powder while using a low
shear agitation in a mortar & pestle; the agitation took around
45 min. The mixture was then transferred into a lyophilization tray
and was oven-dried for about 20 h at 50.degree. C. This procedure
produced about 20 g of hydrophilic fraction, which was a fine
granulate.
[0213] Production of the hydrophobic medium: 2 g of Span 40, 4 g of
lecithin and 3.8 g of GMO were dissolved in 17.3 g of ethyl
isovalerate while mixing. To this solution were added 39.1 g of GTB
and 72.6 g of castor oil. This procedure produced about 136-138 g
of hydrophobic medium.
[0214] Production of the bulk drug product: Mixing of the
hydrophilic fraction and the hydrophobic medium was performed at
20.+-.2.degree. C.
[0215] 19.00 g (29.58% of the final BDP) of the hydrophilic
fraction was slowly added during mixing to 45.23 g (70.42% of the
final BDP) of hydrophobic medium at 600.+-.50 RPM. After addition
of all the hydrophilic fraction, the mixing speed was increased to
2000.+-.200 RPM for 2-10 min followed by 4-8 cycles of 15 min
mixing at 600.+-.50 RPM and 2 min mixing at 2000.+-.200 RPM.
[0216] Degassing by vacuum was then applied as follows: 5 min at
600 mBar, 5 min at 500 mBar and 30-120 min at 400 mBar. The
resulting suspension was poured into a 100 mL dark bottle and
stored at 2-8.degree. C.
[0217] Rat study: The above suspension was administered rectally to
rats as described above in the Examples and the results were as
follows: 35% BA, 12.9% SD. Another batch of suspension prepared by
granulation as described above was prepared and was administered to
the jejunum of rats as described above in the Examples, and the
results were as follows: 21.8% BA, 4.0% SD.A range of formulations
are prepared in a similar manner using granulation and
incorporating a selection of therapeutic agents and varying the
amount of sodium octanoate.
Example 14
Selection of Capsules
[0218] In vitro experiments were carried out using separately three
types of solutions: the hydrophobic medium as described in the
above Examples, ethyl isovalerate alone, and ethyl isovalerate
containing 5% of each of the following surfactants: lecithin, span
40 and glyceryl mono-oleate. 3 types of unsealed capsules, gelatin,
starch and HPMC, were each filled with each of these solutions. The
filled capsules were then maintained in vitro for 29 days at
22.+-.2.degree. C., 30-50% relative humidity. Gelatin and HPMC
capsules gave the best results, namely no deformation of the
capsule.
[0219] Similar experiments were carried out using the same three
solutions, and gelatin and HPMC capsules. The capsules were filled
with the solutions, sealed (bonded) and then were maintained for 8
days at 22.+-.2.degree. C., 30-50% relative humidity. Both types of
capsules showed stability to the solutions tested i.e. there was no
leakage and no deformation of the capsules.
Example 15
Effect of Varying the Cation in the Medium Chain Fatty Acid
Salt
[0220] Formulations were prepared with dextran (FD4) similar to
Formulation A of Table 3A except that 12% sodium octanoate (0.722M)
was replaced by an equal molarity of lithium octanoate or potassium
octanoate or arginine octanoate (the last as a model for an
ammonium salt). These formulations are shown below in Table 7A.
TABLE-US-00011 TABLE 7A Formulation, cargo = dextran K-octanoate
Li-octanoate Arg-octanoate Ingredient (% w/w) (% w/w) (% w/w)
Hydrophilic API 0.545 0.546 0.546 fraction MgCl2 0.134 0.136 0.124
PVP-12 2.673 2.722 2.475 Potassium 13.617 0.00 0.00 octanoate
Lithium 0.00 10.826 0.00 octanoate Arginine 0.00 0.00 22.989
octanoate MC 400 0.134 0.136 0.124 Water 0.684 0.627 0.919 Span40
1.185 1.206 1.097 Lecithin 2.369 2.412 2.193 Hydrophobic Ethyl
10.26 10.45 9.50 medium isovalerate Glyceryl 2.227 2.268 2.062
monooleate Glyceryl 23.16 23.58 21.44 tributyrate Castor oil 43.01
43.79 39.82
[0221] These formulations were each tested in the rat jejunal model
described in Example 8. The results were obtained and
bioavailability was calculated. The results are shown below in
Table 7B.
TABLE-US-00012 TABLE 7B Medium chain fatty acid salt in formulation
tested N % BA .+-. SD Sodium octanoate (Formulation A) 18 22.2 .+-.
10.8 Lithium octanoate 11 8.4 .+-. 3.8 Potassium octanoate 10 7.9
.+-. 6.4 Arginine octanoate 12 17.5 .+-. 7.4
[0222] The formulation A used in the above experiment was a
different batch to that used in Example 8, and so the BA results
given here for formulation A differ slightly from those recited in
Table 4.
[0223] The above results show that when 12% sodium octanoate was
replaced in the formulation by an equivalent molarity of lithium
octanoate or potassium octanoate, the formulation still had
bioavailability but at a lower level. The arginine octanoate
formulation had similar activity to the 12% sodium octanoate
formulation.
Example 16
Effect of Addition of Medium Chain Alcohols (Geraniol and Octanol)
to the Hydrophobic Medium
[0224] Formulations containing geraniol (BASF) and octanol
(Spectrum/MP) were prepared as described above, using the
ingredients shown below in Table 8. The sodium dodecanoate was
obtained from Spectrum/Acros).
[0225] Formulation Q-low % medium chain fatty acid salt: A dextran
(FD4) formulation was prepared essentially as described in Example
11, containing a total of 2.9% medium chain fatty acid
salt--(sodium octanoate 1.042%+sodium dodecanoate 1.869%)--and also
containing geraniol and octanol in the hydrophobic medium, all as
shown in Table 8 below.
[0226] Formulation R--over 10% medium chain fatty acid salt: A
dextran formulation was prepared essentially as described for
Formulation A except that geraniol and octanol were added to the
hydrophobic medium, all as shown in Table 8.
TABLE-US-00013 TABLE 8 Formulation, cargo Dextran Dextran Q R
Ingredient (% w/w) (% w/w) Hydrophilic API 0.545 0.456 fraction
NaOH 0.029 0.000 MgCl.sub.2 0.104 0.114 PVP-12 2.083 2.282 Sodium
octanoate 1.042 10.046 Sodium 1.869 -- dodecanoate MC 400 0.104
0.114 Water 0.231 0.521 Hydrophobic Geraniol 9.148 8.39 medium
Octanol 8.627 7.92 Span40 1.041 0.96 Lecithin 2.081 1.91 Ethyl
isovalerate 9.012 8.27 Glyceryl 1.956 1.80 monooleate Glyceryl
21.825 20.03 tributyrate Castor oil 40.532 37.20
[0227] Formulation Q (low % MCFA salt) was tested in the intra
jejunal rat model described above and the bioavailability was
calculated: aBA=4.4%, SD=3.8 (n=12). Formulation R (over 10% MCFA
salt) was tested in the intra jejunal rat model described above and
the bioavailability was calculated: aBA=22.7%. SD=1.6 (n=6). The BA
of these formulations do not differ significantly from similar
formulations, described in the above Examples, which do not contain
geraniol.
Example 17
Formulations for Gentamicin and for RNA
[0228] Formulations were prepared for gentamicin and for RNA
essentially as described in Example 11, with the ingredients of the
bulk drug product as shown below in Table 9. The gentamicin was
obtained from Applichem and the RNA was polyinosinic-polycytidylic
acid sodium salt (Sigma).
TABLE-US-00014 TABLE 9A Formulation, API Gentamicin RNA Ingredient
(% w/w) (% w/w) Hydrophilic API 6.000 0.100 fraction NaOH 0.670 --
MgCl.sub.2 0.127 0.137 PVP-12 2.545 2.741 Sodium octanoate 12.026
12.001 MC 400 0.127 0.137 Water 0.860 0.605 Hydrophobic Span40
1.119 1.214 medium Lecithin 2.238 2.429 Ethyl isovalerate 9.69
10.52 Glyceryl 2.103 2.283 monooleate Glyceryl 21.88 23.74
tributyrate Castor oil 40.62 44.09
[0229] The gentamicin formulation was tested in the rat jejunal
model described above and in the rat rectal model described above
(e.g. Examples 4 and 5). The gentamicin was assayed using an
immunoassay (ELISA). The results are shown in Table 9B below; % BA
is calculated compared comparing to IV administration. The
formulations were shown to provide bioavailability to the
gentamicin.
TABLE-US-00015 TABLE 9B Cargo Formulation ROA N % BA .+-. SD
Gentamicin As Table 9A jejunal 6 12.9 .+-. 4.5 As Table 9A rectal 5
50.1 .+-. 5.8
[0230] Similarly, the RNA formulation of Table 9A is tested in the
rat jejunal model and in the rat rectal model described above. The
RNA is assayed and the formulation is expected to provide
bioavailability to the RNA.
Example 18
Effect on Formulation Activity of the Surfactants in the
Hydrophobic Medium
[0231] The effect on formulation activity of withdrawing
surfactants from the hydrophobic medium was tested using
formulations containing dextran (average MW=4.4 kDa, FITC labeled)
as cargo (formulations A and H in Table 3A).
[0232] Table 10 presents data from a study in non-anesthetized rats
following intra-jejunal administration of formulations with or
without surfactants (e.g. Span40, lecithin, glyceryl monooleate) in
the hydrophobic medium.
TABLE-US-00016 TABLE 10 Surfactants in hydrophobic Cargo
Formulation medium N % aBA .+-. SD Dextran A + 17 28.0 .+-. 6.8 H -
4 11.1 .+-. 8.2
[0233] Formulations with or without surfactants in the hydrophobic
medium were administered directly to the jejunum of
non-anesthetized rats and plasma dextran levels were measured at 3,
6, 10, 25, 60 and 90 minutes post formulation administration. The
levels of dextran absorption from rat jejunum after administration
of dextran in formulation were compared to the levels of dextran
absorbed after intravenous administration.
[0234] Exposure values, AUC (0-90), were determined for jejunal and
intravenous administration and the absolute bioavailability (aBA)
was determined according to the following equation:aBA=(jejunal
AUC(0-90))/(iv AUC (0-90))*(iv dose/jejunal dose). Data are
presented as Mean aBA.+-.SD.
[0235] Lower bioavailability was achieved when dextran was
incorporated into a formulation not containing surfactants in the
hydrophobic medium (formulation H) as compared to a formulation
containing surfactants in the hydrophobic medium (formulation A).
The results demonstrate that withdrawing surfactants from the
hydrophobic medium adversely affects formulation activity.
Example 19
Effect on Formulation Activity of Withdrawing Medium Chain Fatty
Acids from the Hydrophilic Fraction
[0236] The effect on formulation activity of withdrawing medium
chain fatty acids (MCFA) from the hydrophilic fraction was tested
using formulations containing dextran (average MW=4.4 kDa, FITC
labeled) as cargo.
[0237] Table 11 presents data from a study in non-anesthetized rats
following intra jejunal administration of formulations with or
without sodium octanoate in the hydrophilic fraction (formulations
A and D in Table 3A, respectively).
TABLE-US-00017 TABLE 11 MCFA in hydrophilic Cargo Formulation
fraction N % aBA .+-. SD Dextran A + 17 28.0 .+-. 6.8 D - 5 0.6
.+-. 1.0
[0238] The formulations described above were administered directly
to the jejunum of non-anesthetized rats and plasma dextran levels
were measured at 3, 6, 10, 25, 60 and 90 minutes post formulation
administration. The levels of dextran absorption from rat jejunum
after administration of dextran in formulation were compared to the
levels of dextran absorbed after intravenous administration.
Exposure values, AUC (0-90), were determined for jejunal and
intravenous administration and the absolute bioavailability (aBA)
was determined according to the following equation: aBA=(jejunal
AUC(0-90))/(iv AUC (0-90))*(IV dose/jejunal dose). Data are
presented as Mean aBA.+-.SD.
[0239] Negligible penetration of dextran was achieved when dextran
was incorporated into a formulation lacking medium chain fatty
acids in the hydrophilic fraction (formulation D, % aBA=0.6.+-.1.0)
as compared to a formulation containing sodium octanoate at 12% w/w
in the hydrophilic fraction (formulation A, % aBA=28.0 .+-.6.8).
The results demonstrate that a formulation without medium chain
fatty acids in the hydrophilic fraction is not active.
[0240] A similar experiment was performed using octreotide as cargo
in the improved formulation (see below). The rBA was 0.11%
(CV=158%)
Example 20
Effect on Formulation Activity of Simplifying the Formulation
[0241] The effect on formulation activity of simplifying the
formulation was tested using formulations containing dextran
(average MW=4.4 kDa, FITC labeled) or octreotide (Novetide) as
cargo. The basic formulation described in the above Examples (e.g.
formulations designated A, I and P) was simplified by not adding
MgCl.sub.2, and MC 400 to the hydrophilic fraction and by not
adding span40, lecithin and ethyl iso-valerate to the hydrophobic
medium. There is a concomitant increase in the amounts of glyceryl
monooleate (surfactant) and glyceryl tributyrate added to the
hydrophobic medium. Such formulations are shown in Table 12A below.
These simplified formulations show no precipitation visually
although the particles are visible microscopically i.e. they are
stable suspensions.
TABLE-US-00018 TABLE 12A Formulation, API Dextran Octreotide
Simplified Simplified Ingredient (% w/w) (% w/w) Hydrophilic API
0.545 0.058 fraction NaOH 0.001 0.000 MgCl.sub.2 0.000 0.000 PVP-12
2.735 2.750 Sodium octanoate 12.000 12.019 MC 400 0.000 0.000 Water
0.611 0.593 Hydrophobic Span40 0.00 0.000 medium Lecithin 0.00
0.000 Ethyl isovalerate 0.00 0.000 Glyceryl monooleate 5.91 5.947
Glyceryl tributyrate 34.19 34.385 Castor oil 44.00 44.248
[0242] The production process for these above simplified
formulations is essentially as described in FIG. 1 and in Example
11 for the basic formulations.
[0243] The basic octreotide formulation is shown in Table 12B
below.
TABLE-US-00019 TABLE 12B Cargo Octreotide Basic Formulation M
Ingredient (% w/w) Hydrophilic Cargo 0.058 fraction NaOH 0.000
(HFP) MgCl.sub.2 0.137 PVP-12 2.742 Sodium 12.003 Octanoate MC 400
0.137 Water 0.603 Hydrophobic Span40 1.215 fraction Lecithin 2.430
(LFP) Ethyl-Iso- 10.522 valerate Glyceryl 2.284 Monooleate Glyceryl
23.756 Tributyrate Castor oil 44.113
[0244] Table 13 presents data from a study in non-anesthetized rats
following intra-jejunal administration of two different dextran
formulations--formulation A of Table 3A and the simplified
formulation shown in Table 12A.
TABLE-US-00020 TABLE 13 AUC (0-60 min)/dose/kg Cargo Formulation N
b.w. .+-. SD Dextran A(basic) 28 67062 .+-. 27368 Simplified 12
63897 .+-. 24210
[0245] The above results show that similar AUC values were achieved
when dextran was incorporated into a formulation containing the
basic formulation (formulation A) as compared to a simplified
formulation.
[0246] Table 14 below presents data from a study in
non-anesthetized rats following intra-jejunal administration of two
different octreotide formulations--the basic formulation shown in
Table 12B and the simplified formulation shown in Table 12A. The
levels of octreotide absorption from rat jejunum after
administration of octreotide in basic formulation and simplified
formulation were obtained. Exposure values, AUC (0-25), were
determined.
TABLE-US-00021 TABLE 14 AUC (0-25 min)/dose/kg Cargo Formulation N
b.w. .+-. SD Octreotide Basic 13 2.8 .+-. 1.4 Simplified 13 2.3
.+-. 0.8
[0247] The above results in Table 14 show that the AUC values were
slightly less when octreotide was incorporated into a simplified
formulation as compared to the full formulation.
Example 21
Effect on Formulation Activity of Replacing Castor Oil by Octanoic
Acid
[0248] The effect on formulation activity of replacing castor oil
(and glyceryl tributyrate and ethyl iso-valerate) by octanoic acid
(Aldritch) was tested using a formulation containing dextran as
cargo. This was done to maintain the C8 motif in the formulation
i.e. it was considered it might be advantageous to have C8 acid in
the hydrophobic medium in addition to the C8 salt in the
hydrophilic fraction.
[0249] The effect of adding ricinoleic acid (Spectrum) was also
tested by making a dextran formulation containing octanoic
acid/ricinoleic acid. Ricinoleic acid was chosen since the main
triglyceride component in castor oil is formed from ricinoleic
acid. Three formulations of dextran were prepared as shown in Table
15A below. The basic dextran formulation was prepared essentially
as described in the above Examples. The dextran octanoic
formulation was prepared essentially as described in the above
Examples but wherein castor oil, glyceryl tributyrate and ethyl
iso-valerate were replaced by octanoic acid. This formulation was
found to be a solution by visual analysis but true solubility
analysis was not performed. It seems that the octanoic acid at high
concentration (about 78% of this formulation) dissolves the solid
hydrophilic fraction, with the PVP and sodium octanoate being
soluble in octanoic acid at high concentration. The dextran
ricinoleic/octanoic acid formulation was prepared essentially as
described in the above Examples but wherein castor oil, glyceryl
tributyrate and ethyl iso-valerate were replaced by a mixture of
octanoic acid and ricinoleic acid. This formulation was a
suspension as is usual for most of the formulations of this
invention.
TABLE-US-00022 TABLE 15A Formulation, API Dextran Dextran
Ricinoleic/ Dextran Octanoic Octanoic basic acid acid Ingredient (%
w/w) (% w/w) (% w/w) Hydrophilic API 0.545 0.545 0.545 fraction
NaOH 0.001 0.001 0.001 MgCl.sub.2 0.136 0.136 0.136 PVP-12 2.726
2.726 2.726 Sodium octanoate 12.001 12.002 12.002 MC 400 0.136
0.136 0.136 Water 0.622 0.622 0.622 Hydrophobic Span40 1.208 1.207
1.207 medium Lecithin 2.416 2.414 2.414 Ethyl isovalerate 10.46
0.00 0.00 Glyceryl monooleate 2.271 2.272 2.272 Glyceryl
tributyrate 23.62 0.00 0.00 Castor oil 43.86 0.00 0.00 Octanoic
acid 0.000 77.94 23.38 Ricinoleic acid 0.000 0.00 46.76 Ethyl
Octanoate 0.000 0.00 7.80
[0250] The formulations described above in Table 15A were
administered directly to the jejunum of non-anesthetized rats, and
plasma dextran levels were measured post formulation
administration. Exposure values, AUC, were determined for the
different formulations. These results are shown below in Table
15B.
TABLE-US-00023 TABLE 15B AUC (0-60)/dose/kg Cargo Formulation N
b.w. .+-. SD Dextran Basic 12 72385 .+-. 37827 Octanoic acid 11
180824 .+-. 32778 Ricinoleic/Octanoic acid 11 113204 .+-. 33057
[0251] The results shown above in Table 15B demonstrate that the
absorption of dextran was much improved (over two-fold) in the
formulation containing octanoic acid. Additionally, the shape of
the graph was changed showing slower but longer release. This may
be advantageous since this allows the API to be longer-acting in
the body. The dextran ricinoleic/octanoic results showed less
activity than the octanoic acid formulation, but was still improved
over the basic formulation.
[0252] Since the octanoic acid and ricinoleic acid/octanoic acid
formulations showed high activity, similar formulations were
prepared with exenatide as cargo. Three formulations of exenatide
were produced as shown in Table 16A below. The basic exenatide
formulation was prepared essentially as described in the above
Examples. The exenatide/octanoic formulation was prepared
essentially as described in the above Examples but wherein castor
oil, glyceryl tributyrate and ethyl iso-valerate were replaced by
octanoic acid. This formulation containing about 78% octanoic acid
was found to be a solution by visual analysis, as was the similar
dextran formulation above. The exenatide ricinoleic/octanoic acid
formulation was prepared essentially as described in the above
Examples but wherein castor oil, glyceryl tributyrate and ethyl
iso-valerate were replaced by a mixture of octanoic acid and
ricinoleic acid.
TABLE-US-00024 TABLE 16A Formulation, API Exenatide Exenatide
Ricinoleic/ Exenatide Octanoic Octanoic basic acid acid Ingredient
(% w/w) (% w/w) (% w/w) Hydrophilic API 0.055 0.055 0.055 fraction
NaOH 0.000 0.000 0.000 MgCl.sub.2 0.137 0.137 0.137 PVP-12 2.742
2.742 2.742 Sodium octanoate 12.003 12.003 12.003 MC 400 0.137
0.137 0.137 Water 0.603 0.603 0.603 Hydrophobic Span40 1.213 1.214
1.214 medium Lecithin 2.434 2.429 2.429 Ethyl isovalerate 10.522
0.000 0.000 Glyceryl monooleate 2.283 2.285 2.285 Glyceryl
tributyrate 23.759 0.000 0.000 Castor oil 44.112 0.000 0.000
Octanoic acid 0.000 78.395 47.035 Ricinoleic acid 0.000 0.000
23.518 Ethyl Octanoate 0.000 0.000 7.842
[0253] The formulations described above in Table 16A were
administered directly to the jejunum of non-anesthetized rats, and
plasma exenatide levels were measured post formulation
administration. Exposure values, AUC, were determined for the
different formulations. These results are shown below in Table
16B.
TABLE-US-00025 TABLE 16B Cargo Formulation N AUC (0-90) .+-. SD %
BA .+-. SD Exenatide Basic 10 1961 .+-. 1791 8.8 .+-. 8.2 Octanoic
acid 11 612 .+-. 350 3.1 .+-. 1.8 [AUC (0-180) .+-. SD] Ricinoleic/
9 476 .+-. 321 2.2 .+-. 1.5 Octanoic acid
[0254] The results shown above in Table 16B demonstrate that the
exenatide formulation containing octanoic acid showed
bioavailability, but the absorption of exenatide was decreased
compared to the basic formulation. The shape of the graph was
changed showing slower but longer release as in the case of the
dextran octanoic acid formulation above; this prolonged PK profile
may be advantageous. Note that in the case of the octanoic acid
formulation, AUC 0-180 min was used for BA calculations due to the
prolonged PK profile. The exenatide ricinoleic/octanoic acid
formulation had even lower bioavailability than the octanoic acid
formulation.
Example 22
Dose Response for Octanoic Acid
[0255] A. Octreotide formulations: The effect on formulation
activity of varying the amount of octanoic acid was tested using
formulations containing octreotide as cargo. Four formulations of
octreotide were prepared using 0%, 5%, 10% or 15% octanoic acid as
shown in Table 17 below. The formulations are basic octreotide
formulations prepared essentially as described above wherein the
amount of octanoic acid varies as described and the amount of other
ingredients in the hydrophobic medium. (ethyl isovalerate and
glyceryl tributyrate) was concomitantly reduced. (In these
formulations the hydrophilic fraction was simplified to omit
MgCl.sub.2 and MC400.)
TABLE-US-00026 TABLE 17 Formulation, API Octreotide Octreotide
Octreotide Octreotide 0% Octanoic 5% Octanoic 10% Octanoic 15%
Octanoic Ingredient (% w/w) (% w/w) (% w/w) (% w/w) Hydrophilic API
0.058 0.057 0.057 0.057 fraction PVP-12 2.750 2.750 2.750 2.750
Sodium octanoate 12.019 12.034 12.034 12.034 Water 0.593 0.594
0.594 0.594 Hydrophobic Span40 1.217 1.219 1.219 1.219 medium
Lecithin 2.441 2.437 2.437 2.437 Ethyl isovalerate 10.554 0 0 0
Octanoic acid 0 5.053 10.553 15.021 Glyceryl monooleate 2.290 2.291
2.291 2.291 Glyceryl tributyrate 23.832 29.325 23.825 19.357 Castor
oil 44.246 44.241 44.241 44.241
[0256] B. Exenatide formulations: The effect on formulation
activity of varying the amount of octanoic acid was tested using
formulations containing exenatide as cargo. Five formulations of
exenatide were prepared using 0%, 10%, 15%, 20% or 35% octanoic
acid as shown in Table 18 below. The formulations are basic
exenatide formulations prepared essentially as described above
wherein the amount of octanoic acid varies as described and the
amount of other ingredients in the hydrophobic medium (ethyl
isovalerate and glyceryl tributyrate) was concomitantly
reduced.
TABLE-US-00027 TABLE 18 Exenatide Exenatide Exenatide Exenatide
Exenatide 0% 10% 15% 20% 35% Formulation, API Octanoic Octanoic
Octanoic Octanoic Octanoic Ingredient (% w/w) (% w/w) (% w/w) (%
w/w) (% w/w) Hydrophilic API 0.055 0.055 0.055 0.055 0.055 fraction
MgCl.sub.2 0.137 0.137 0.137 0.137 0.137 PVP-12 2.742 2.742 2.742
2.742 2.742 Sodium octanoate 12.003 12.003 12.003 12.003 12.003 MC
400 0.137 0.137 0.137 0.137 0.137 Water 0.603 0.603 0.603 0.603
0.603 Hydrophobic Span40 1.213 1.213 1.213 1.213 1.213 medium
Lecithin 2.434 2.434 2.434 2.434 2.434 Ethyl isovalerate 10.522 0 0
0 0 Octanoic acid 0 10.522 15.081 20.085 34.282 Glyceryl monooleate
2.283 2.283 2.283 2.283 2.283 Glyceryl tributyrate 23.759 23.759
19.201 14.197 0.000 Castor oil 44.112 44.112 44.112 44.112
44.112
[0257] The formulations described above in Tables 17 and 18 above
were administered directly to the jejunum of non-anesthetized rats,
and plasma octreotide or exenatide levels were measured post
formulation administration. Exposure values, AUC, were determined
for the different formulations. These results are shown below in
Table 19.
TABLE-US-00028 TABLE 19 AUC (0-60)/dose/kg Cargo Formulation N b.w.
.+-. SD Octreotide Basic 14 2.8 .+-. 1.0 Basic 12 2.7 .+-. 1.2 5%
Octanoic acid Basic) 12 3.2 .+-. 1.2 10% Octanoic acid Basic 12 4.5
.+-. 2.3 15% Octanoic acid Exenatide Basic 10 3.9 .+-. 3.8 Basic,
10% Octanoic acid 15 4.6 .+-. 2.8 Basic, 15% Octanoic acid 6 3.0
.+-. 1.8 Basic, 20% Octanoic acid 5 2.2 .+-. 0.5 Basic, 35%
Octanoic acid 6 1.9 .+-. 0.7
[0258] The results shown above in Table 19 demonstrate that the
octreotide formulation shows increased activity compared to the
basic formulation as the amount of octanoic acid is increased to
15% (the maximum amount tested). Additionally, the results shown
above in Table 19 demonstrate that the exenatide formulation shows
increased activity compared to the basic formulation as the amount
of octanoic acid is increased to 15% and the activity decreases at
higher levels of octanoic acid.
Example 23
Effect of Different Medium Chain Fatty Acid Salts
[0259] A. Sodium sebacate (disodium salt of decanedioic acid): The
effect on formulation activity of replacing sodium octanoate by
sodium sebacate (disodium C10 salt) in a dextran formulation was
tested. The sodium sebacate was prepared in situ from sebacic acid
(Aldrich) and sodium hydroxide. The formulation produced is
described in Table 20 below. The formulation was prepared
essentially as described above but 12% sodium octanoate was
replaced by sodium sebacate, at the same molar concentration as
sodium octanoate i.e. an equimolar amount of sodium sebacate was
used (viz., 0.72M).
TABLE-US-00029 TABLE 20 Formulation, API Dextran Na-Sebacate
Ingredient (% w/w) Hydrophilic API 0.545 fraction NaOH 0.000
MgCl.sub.2 0.129 PVP-12 2.589 Sodium Sebacate 16.190 MC 400 0.129
Water 0.783 Hydrophobic Span40 1.147 medium Lecithin 2.295 Ethyl
isovalerate 9.94 Glyceryl monooleate 2.157 Glyceryl tributyrate
22.44 Castor oil 41.66
[0260] The formulation described above in Table 20 was administered
directly to the jejunum of non-anesthetized rats, and plasma
dextran levels were measured post formulation administration.
Exposure value, AUC, was determined for the formulation and this is
compared with a similar formulation prepared with sodium octanoate.
These results are shown below in Table 21.
TABLE-US-00030 TABLE 21 AUC (0-60)/dose/kg Cargo Formulation N b.w.
.+-. SD Dextran With Na-octanoate 12 72385 .+-. 37827 With
Na-Sebacate 9 18691 .+-. 11887
[0261] The results shown in Table 21 demonstrate that the dextran
formulation containing sodium sebacate showed activity, but the
absorption of dextran was decreased compared to the formulation
containing an equimolar amount of sodium octanoate.
[0262] B. Mono-Sodium Suberate or Di-Sodium Suberate
[0263] Octreotide-containing formulations were prepared wherein 12%
sodium octanoate was replaced by an equimolar amount (0.72M) of
mono-sodium suberate or of di-sodium suberate, which are C8 salts.
These sodium salts were prepared in situ from suberic acid (Tokyo
Chemical Industry Co.) and sodium hydroxide.
TABLE-US-00031 TABLE 22A Formulation, API Octreotide Octreotide
mono-Sodium di-Sodium suberate suberate (0.72M) (0.72M) Ingredient
(% w/w) (% w/w) Hydrophilic API 0.058 0.059 fraction PVP-12 2.650
2.620 mono-Sodium Suberate 15.087 0 di-Sodium Suberate 0 15.996
Water 0.712 0.747 Hydrophobic Span40 1.173 1.159 medium Lecithin
2.352 2.325 Ethyl isovalerate 10.169 10.055 Glyceryl monooleate
2.206 2.181 Glyceryl tributyrate 22.962 22.704 Castor oil 42.632
42.152
[0264] The formulations described above in Table 22 are
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels are measured post formulation
administration. Exposure values, AUC, are determined for the
formulations and this is compared with a similar formulation
prepared with sodium octanoate.
[0265] C . Geranic Acid Salt
[0266] Two octreotide-containing formulations were prepared
essentially as described above wherein 12% sodium octanoate was
replaced by 18% geranic acid sodium salt (0.95M) and 14.6% (0.77M)
geranic acid sodium salt, which is 3,7-dimethyl-2,6-octadienoic
acid (obtained from SAFC.) . The formulations produced are
described in Table 22B below.
TABLE-US-00032 TABLE 22B Formulation, API Octreotide Octreotide
NaGeranate NaGeranate A B Ingredient (% w/w) (% w/w) Hydrophilic
API 0.057 0.057 fraction NaOH 0 0.543 PVP 12 10.006 9.833 Sodium
Geranate 18.053 14.625 Water 1.183 1.084 Hydrophobic Tween 80 2.001
1.970 medium Glyceryl monocaprylate 4.001 3.923 Glyceryl
tricaprylate 63.235 65.927 Castor oil 0.000 0
[0267] The formulations described above in Table 22B were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post formulation
administration. Exposure values, AUC, were determined for the
formulations and this was compared with a similar formulation
prepared with sodium octanoate. The results are shown below in
Table 22C and they demonstrate that the formulation with 18% sodium
geranate had similar activity as the 12% sodium octanoate
formulation, and the formulation with 14.6% sodium geranate had
increased activity.
TABLE-US-00033 TABLE 22C AUC (0-60)/dose/kg Cargo Formulation N
b.w. .+-. SD 15 Octreotide Sodium geranate A 9 4.48 .+-. 1.79
Sodium geranate B 9 6.33 .+-. 2.1 Improved 9 4.38 .+-. 1.66
Example 24
Effect of PVP (Polyvinylpyrrolidone) on Formulation Activity
[0268] The effect on formulation activity of replacing PVP-12 by
mannitol (Sigma) was tested using formulations containing exenatide
as cargo. It was understood in the art that PVP-12 is a stabilizer
and could be replaced in the formulation by another stabilizer such
as mannitol. The formulation shown in Table 23 below was prepared.
This formulation is a basic exenatide formulation prepared
essentially as described above, but wherein PVP-12 is replaced by
mannitol.
TABLE-US-00034 TABLE 23 Formulation, API Exenatide Mannitol
Ingredient (% w/w) Hydrophilic API 0.055 fraction MgCl.sub.2 0.137
Mannitol 2.742 Sodium octanoate 12.003 MC 400 0.137 Water 0.603
Hydrophobic Span40 1.213 medium Lecithin 2.434 Ethyl isovalerate
10.522 Glyceryl monooleate 2.283 Glyceryl tributyrate 23.759 Castor
oil 44.112
[0269] The formulation described above in Table 23 was administered
directly to the jejunum of non-anesthetized rats, and plasma
exenatide levels were measured post-formulation administration.
Exposure values, AUC, were determined for the formulation compared
to the basic formulation. These results are shown below in Table
24.
TABLE-US-00035 TABLE 24 AUC (0-60)/dose/kg Cargo Formulation N b.w.
.+-. SD Exenatide Basic 10 3.9 .+-. 3.8 Mannitol instead of PVP-12
6 1.6 .+-. 1.7
[0270] The results shown above in Table 24 demonstrate the
surprising and unexpected result that the exenatide formulation
without PVP-12 had significantly decreased activity compared to the
basic formulation. It was thus decided to investigate further the
effect of PVP on bioavailability.
[0271] The effect on formulation activity of varying the molecular
weight of PVP was tested using formulations containing exenatide as
cargo. Three formulations of exenatide were prepared using either
PVP-12, PVP-17 or PVP-25 (all obtained from BASF). PVP-12, PVP-17
and PVP-25 are all polyvinylpyrrolidone polymers; the average
molecular weights are about 2500-3000, 10000 and 30000
respectively. The formulations are basic exenatide formulations
prepared essentially as described above wherein the PVP varies as
described and wherein the hydrophilic fraction has been simplified
to omit MgCl.sub.2 and MC400.
TABLE-US-00036 TABLE 25 Formulation, API Exenatide PVP-12/17/25
Ingredient (% w/w) Hydrophilic API 0.022 fraction PVP 12/17/25
2.752 Sodium octanoate 12.005 Water 0.602 Hydrophobic Span40 1.218
medium Lecithin 2.442 Ethyl isovalerate 10.561 Glyceryl monooleate
2.291 Glyceryl tributyrate 23.846 Castor oil 44.272
[0272] The three formulations described above in Table 25 were
administered directly to the jejunum of non-anesthetized rats, and
plasma exenatide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the
formulations. The results are shown below in Table 26.
TABLE-US-00037 TABLE 26 AUC (0-60)/dose/kg Cargo Formulation N b.w.
.+-. SD (a) PVP-12 11 8.0 .+-. 7.7 Exenatide (b) PVP-17 6 3.4 .+-.
2.9 PVP-25 5 2.6 .+-. 2.3
[0273] The results shown above in Table 26 demonstrate that the
exenatide formulations containing PVP-12 showed much higher
activity than the exenatide formulations containing PVP-17 and
PVP-25. Thus the effect of PVP-12 only was investigated further,
and it was decided to perform a dose--response study using PVP-12.
The effect of increasing the amount of PVP-12 in the formulation on
the activity of the formulation was tested using formulations
containing octreotide as cargo compound and different doses of
PVP-12 as shown in Table 27 below. The PVP-12 doses tested were
2.75% (the standard dose used in the above formulations) and 5.0%,
7.5% and 10.0% PVP-12; the hydrophilic fraction has been simplified
to omit MgCl.sub.2 and MC400. The formulation containing 10% PVP
was semi-solid i.e. it was apparently a semi-solid suspension.
TABLE-US-00038 TABLE 27 Formulation, API Octreotide Octreotide
Octreotide Octreotide PVP 2.75% PVP 5.0% PVP 7.5% PVP 10.0%
Ingredient (% w/w) (% w/w) (% w/w) (% w/w) Hydrophilic API 0.058
0.057 0.057 0.057 fraction PVP-12 2.750 5.013 7.514 10.046 Sodium
octanoate 12.019 12.031 12.037 12.018 Water 0.593 0.684 0.784 0.885
Hydrophobic Span40 1.217 1.183 1.145 1.108 medium Lecithin 2.441
2.373 2.297 2.222 Ethyl isovalerate 10.554 10.259 9.934 9.608
Glyceryl monooleate 2.290 2.226 2.155 2.084 Glyceryl tributyrate
23.832 23.166 22.431 21.694 Castor oil 44.246 43.009 41.645
40.278
[0274] The formulations described above in Table 27 were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the four
different formulations. These results are shown below in Table
28A.
TABLE-US-00039 TABLE 28A AUC (0-60)/dose/kg Cargo Formulation N
b.w. .+-. SD Octreotide 2.75% PVP-12 14 2.8 .+-. 1.0 5.0% PVP-12 12
3.7 .+-. 1.6 7.5% PVP-12 12 4.2 .+-. 1.5 10.0% PVP-12 11 4.7 .+-.
1.4
[0275] The results shown above in Table 28A demonstrate that the
absorption of octreotide increased dramatically as the amount of
PVP in the formulation increased. The formulation containing 10%
PVP-12 had absorption of octreotide about 1.7 times greater that
the formulation containing 2.75% PVP-12. An improved octreotide
formulation in which there was 10 PVP-12 but no sodium octanoate
showed virtually no activity. The rBA was 0.11% (CV=158%) n=5.
[0276] It appears that the medium chain fatty acid salt acts as a
permeability enhancer (by facilitating or enhancing permeability
and/or absorption of the therapeutic agent), and that the PVP
serves to increase the effect of the permeability enhancer in a
synergistic manner since the PVP alone has virtually no effect. See
also Example 31.
[0277] A further experiment was performed to investigate if the 10%
PVP-12 could be replaced by dextran and still maintain activity of
the formulation. The dextran was manufactured by Fluka; the average
molecular weight is .about.6000. The formulations were prepared
essentially as described above wherein the PVP and dextran varies
as described and wherein the hydrophilic fraction has been
simplified to omit MgCl.sub.2 and MC400 and where the sodium
octanoate was increased to 15%; see Example 26.
TABLE-US-00040 TABLE 28B Formulation, API Octreotide Octreotide
Octreotide 10% PVP 10% Dextran no PVP 5% Dextran no PVP Ingredient
(% w/w) (% w/w) (% w/w) Hydrophilic API 0.058 0.058 0.058 fraction
PVP-12 10.011 0.0 0.0 Dextran 0.0 10.011 5.011 Sodium octanoate
15.008 15.008 15.015 Water 1.003 1.003 0.803 Hydrophobic Tween 80
2.027 2.027 2.169 medium Glyceryl 4.036 4.036 4.319 monocaprylate
Glyceryl tricaprylate 40.714 40.714 43.574 Castor oil 27.143 27.143
29.049
[0278] The three formulations described above in Table 28B were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the
formulations. The results are shown below in Table 28C.
TABLE-US-00041 TABLE 28C AUC (0-25)/dose/kg Cargo Formulation N
b.w. .+-. SD Octreotide 10% PVP 9 4.4 .+-. 1.7 10% Dextran; no PVP
5 3.3 .+-. 1.6 5% Dextran; no PVP 9 3.2 .+-. 1.5
[0279] The results shown above in Table 28C demonstrate that the
absorption of octreotide decreased when PVP in the formulation was
replaced by dextran but the activity was still significant. The
formulation containing 10% dextran had absorption of octreotide
about 75% of the formulation containing 10% PVP, and the
formulation containing 5% dextran had absorption of octreotide
about 73% of the formulation containing 10% PVP.
Example 25
A Comparative Study of C8, C9 and C10 Medium Chain Fatty Acid Salts
viz., Sodium Octanoate, Sodium Nonanoate and Sodium Decanoate
[0280] The effect on formulation activity of replacing sodium
octanoate with other medium chain fatty acid sodium salts was
tested using formulations containing octreotide as cargo. Three
formulations of octreotide were prepared, as shown in Table 29
below. These are all basic formulations prepared essentially as
described above where the hydrophilic fraction has been simplified
to omit MgCl.sub.2 and MC400 and wherein the medium chain fatty
acid salt is an equimolar amount of sodium octanoate, sodium
nonanoate or sodium decanoate.
TABLE-US-00042 TABLE 29 Formulation, API Octreotide Octreotide
Octreotide NaC8 12% NaC9 13% NaC10 14% (0.72M) (0.72M) (0.72M)
Ingredient (% w/w) (% w/w) (% w/w) Hydrophilic API 0.058 0.057
0.058 fraction PVP-12 2.750 2.718 2.685 Sodium octanoate 12.019 0 0
Sodium nonanoate 0 13.023 0 Sodium decanoate 0 0 14.019 Water 0.593
0.632 0.670 Hydrophobic Span40 1.217 1.203 1.188 medium Lecithin
2.441 2.412 2.383 Ethyl isovalerate 10.554 10.428 10.303 Glyceryl
monooleate 2.290 2.262 2.235 Glyceryl tributyrate 23.832 23.547
23.265 Castor oil 44.246 43.718 43.194
[0281] The formulations described above in Table 29 were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the
formulations. The results are shown below in Table 30.
TABLE-US-00043 TABLE 30 AUC (0-25)/dose/kg Cargo Formulation N b.w.
.+-. SD Octreotide sodium octanoate NaC8 9 2.1 .+-. 0.8 sodium
nonanoate NaC9 10 2.5 .+-. 0.4 sodium decanoate NaC10 10 1.7 .+-.
0.4
[0282] The results shown above in Table 30 demonstrate that when
sodium octanoate in the formulation is replaced by sodium nonanoate
or by sodium decanoate there is similar activity. Based on
statistical analysis, there is no difference in activity between
all three formulations.
Example 26
Dose Response of Sodium Octanoate
[0283] The dose response of sodium octanoate at 12%, 15% and 18%
was tested by making the formulations shown in Table 31. These are
all basic formulations prepared essentially as described above
where the hydrophilic fraction has been simplified to omit
MgCl.sub.2 and MC400 and the cargo compound was octreotide.
Additionally the formulation was corrected for viscosity i.e. the
same or similar viscosity was maintained for all three
formulations; this was achieved by varying the amounts of castor
oil and glyceryl tributyrate.
TABLE-US-00044 TABLE 31 Formulation, API Octreotide Octreotide
Octreotide NaC8 12% NaC8 15% NaC8 18% Ingredient (% w/w) (% w/w) (%
w/w) Hydrophilic API 0.058 0.058 0.058 fraction PVP-12 2.750 2.652
2.554 (simplified) Sodium octanoate 12.019 15.040 18.016 Water
0.593 0.710 0.825 Hydrophobic Span40 1.217 1.173 1.130 medium
Lecithin 2.441 2.353 2.267 Ethyl isovalerate 10.554 10.175 9.802
Glyceryl monooleate 2.290 2.207 2.126 Glyceryl tributyrate 23.832
32.816 41.090 Castor oil 44.246 32.816 22.132
[0284] The formulations described above in Table 31 were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the
formulations. The results are shown below in Table 32.
TABLE-US-00045 TABLE 32 AUC (0-60)/dose/kg Cargo Formulation N b.w.
.+-. SD Octreotide NaC8 12% 14 2.8 .+-. 1.0 NaC8 15% 12 4.1 .+-.
1.9 NaC8 18% 12 3.6 .+-. 1.1
[0285] The results shown above in Table 32 demonstrate that when
sodium octanoate in the formulation is increased from 12% to 15%
there is an increase in activity but a further increase of sodium
octanoate to 18% leads no higher activity than that obtained at
15%.Thus about 15% sodium octanoate appears to be the preferred
amount.
Example 27
Investigation of the Effect of Varying the Hydrophilic/Lipophilic
Balance of the Surfactants in the Formulation
[0286] Table 33 below describes various octreotide formulations.
The first column, formulation (a), is the basic formulation
prepared essentially as described above where the hydrophilic
fraction has been simplified to omit MgCl.sub.2 and MC400, and the
cargo compound is octreotide. The surfactants are Span 40, lecithin
and glyceryl monooleate, and by calculation the HLB is
approximately 5-6. In the other formulations (formulations b, c and
d) the HLB was changed as indicated (to 3.5, 6.7 and 14) by
replacing Span 40 and lecithin by differing amounts of Tween 80 and
by varying the amount of glyceryl monooleate.
TABLE-US-00046 TABLE 33 Octreotide Octreotide Octreotide Octreotide
Formulation, API HLB 5-6 [a] HLB 3.5[b] HLB 6.7[c] HLB 14[d]
Ingredient (% w/w) (% w/w) (% w/w) (% w/w) Hydrophilic API 0.058
0.057 0.057 0.057 fraction PVP-12 2.750 2.748 2.748 2.748 Sodium
octanoate 12.019 12.027 12.027 12.027 Water 0.593 0.594 0.594 0.594
Hydrophobic Span40 1.217 0 0 0 medium Lecithin 2.441 0 0 0 Ethyl
isovalerate 10.554 10.547 10.546 10.547 Tween 80 0 0.502 2.003
5.500 Glyceryl monooleate 2.290 5.500 4.002 0.502 Glyceryl
tributyrate 23.832 23.811 23.811 23.811 Castor oil 44.246 44.215
44.215 44.215
[0287] The formulations described above in Table 33 were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the
formulations. The results are shown below in Table 34.
TABLE-US-00047 TABLE 34 AUC (0-25)/dose/kg Cargo Formulation N b.w.
.+-. SD Octreotide HLB 5-6 - [a] 9 2.1 .+-. 0.8 HLB 3.5 - [b] 12
3.3 .+-. 0.9 HLB 6.7 - [c] 11 3.8 .+-. 0.9 HLB 14 - [d] 10 3.7 .+-.
0.9
[0288] The results shown above in Table 34 demonstrate that all the
three new formulations replacing Span 40 and lecithin with Tween 80
[b, c and d] had much better activity than the basic formulation
[a], even although the HLB in [b] was lower, in [c] was slightly
higher and in [d] was much higher than the HLB of the surfactants
in (a). Additionally, the activities of all the new formulations
[(b, c, and d] were statistically very similar. Thus the HLB alone
of the surfactants does not seem to affect activity but the
characteristics of the surfactants appear to play an important
role. In particular, replacing Span 40 and lecithin with Tween 80
is advantageous for activity in these octreotide formulations.
Example 28
Octreotide Formulations with Different Ratios of Glyceryl
Tricaprylate to Castor Oil
[0289] Based on the accumulation of results described above
including the PVP-12 dose response results, the sodium octanoate
dose response results and the surfactant results inter alia, a
series of octreotide formulations were prepared using 10% PVP-12
and 15% sodium octanoate, and varying the ratio of glyceryl
tricaprylate to castor oil. Additionally, glyceryl monooleate and
glyceryl tributyrate were replaced (if used) by glyceryl
monocaprylate and glyceryl tricaprylate (both supplied by Abitec).
This is to maintain the C8 motif within the formulation. Thus the
hydrophilic fraction contains a salt of a C8 acid (octanoate) and
the hydrophobic medium contains monoglycerides and triglycerides
incorporating the same C8 acid. The inventors believe that the use
of C-8 compounds in both the hydrophilic fraction and in the
hydrophobic medium may be advantageous for bioavailability. The
amounts of Tween 80 and glyceryl monocaprylate were also varied in
the formulations. The formulations were prepared are shown in Table
35A below. Formulations I, II, V and VI were semi-solid (apparently
suspensions) and formulations III and IV were the usual liquid
suspensions.
TABLE-US-00048 TABLE 35A Octreotide Octreotide Octreotide
Octreotide Octreotide Octreotide Formulation, API I II III IV V VI
Ingredient (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) (% w/w)
Hydrophilic API 0.058 0.058 0.058 0.058 0.058 0.058 fraction PVP-12
10.011 10.011 10.011 10.011 10.011 10.011 Sodium octanoate 15.008
15.008 15.008 15.008 15.008 15.008 Water 1.003 1.003 1.003 1.003
1.003 1.003 Hydrophobic Tween 80 2.027 2.027 2.027 2.027 6.063
6.062 medium Glyceryl 4.036 4.036 4.036 4.036 0 0 monocaprylate
Glyceryl 40.714 13.571 61.071 67.857 40.714 0 tricaprylate Castor
oil 27.143 54.286 6.786 0.000 27.143 67.857
[0290] The formulations described above in Table 35A were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the
formulations. The results are shown below in Table 35B.
TABLE-US-00049 TABLE 35B AUC (0-25)/dose/kg Cargo Formulation N
b.w. .+-. SD Octreo- Formulation I(GTC:castor oil 6:4) 9 4.4 .+-.
1.7 tide Formulation II(GTC:castor oil 2:8) 8 3.0 .+-. 1.7
Formulation III(GTC:castor oil 9:1) 9 3.1 .+-. 0.5 Formulation
IV(GTC:castor oil 10:0) 7 4.1 .+-. 2.1 Formulation V -without GMC 6
1.6 .+-. 1.0 (GTC:castor oil 6:4) Formulation VI -without
GMC>C 7 1.1 .+-. 0.6 (GTC:castor oil 0:10)
[0291] The results shown above in Table 35B demonstrate that
formulations 1 and IV have greatest activity. Since castor oil is
absent in formulation IV this demonstrates that castor oil is not
essential for activity. It seems that a high GTC: castor oil ratio
e.g. 6:4 is beneficial for activity. Additionally, since
formulation V (which has low activity) has the same GTC: castor oil
ratio as formulation I it appears that additionally GMC (or other
monoglyceride) is desirable for activity. Additionally a
formulation similar to formulation I of Table 36 was prepared but
sodium octanoate was omitted. This formulation showed virtually no
activity, rBA=0.1%.
[0292] Bulk drug product of formulation IV (improved, no castor
oil) was milled with a 150 micron screen, and then particle size
was determined using Malvern Laser
[0293] Diffraction technology. Preliminary results indicated that
90% (v/v) of the particles were below 130 microns, and 50% (v/v) of
the particles were below 45 microns.
[0294] Preliminary experiments using similar formulations to
formulation I, but with varying increased amounts of octreotide all
gave similar BA i.e. there was approximately linear exposure
independent of API loading. A preliminary experiment using a
similar formulation to formulation IV at even higher octreotide
loading--1.5% (wt/wt)--also gave similar BA.
[0295] A similar improved formulation to formulation I above was
prepared using FD4 as cargo instead of octreotide, and it was
compared to a basic formulation. These formulations are described
in Table 36A below.
TABLE-US-00050 TABLE 36A FD 4 Basic FD 4 Formulation, API (no Mg,
MC) Improved Ingredient (% w/w) (% w/w) Hydrophilic API 0.545 0.545
fraction NaOH 0.001 0 PVP-12 2.734 10.012 Sodium octanoate 12.036
15.009 Water 0.613 1.023 Hydrophobic Tween 80 0 2.013 medium
Glyceryl monocaprylate 0 4.008 Glyceryl tricaprylate 0 40.434
Span40 1.21 0 Lecithin 2.42 0 Ethyl-Iso-valerate 10.49 0 Glyceryl
mono-oleate 2.28 0 Glyceryl tributyrate 23.69 0 Castor oil 43.98
26.956
[0296] The formulations described above in Table 36A were
administered directly to the jejunum of non-anesthetized rats, and
plasma FD4 levels were measured post-formulation administration.
Exposure values, AUC, were determined for the formulations. The
results are shown below in Table 36B.
TABLE-US-00051 TABLE 36B AUC (0-90)/dose/kg Cargo Formulation N
b.w. .+-. SD FD 4 Basic 6 67448 .+-. 16977 (dextran) Improved 6
95374 .+-. 47490
[0297] The results shown above in Table 36B demonstrate that the
improved formulation has much greater activity than the basic
formulation.
Example 29
Detailed Production Process for a Selected (Improved) Octreotide
Formulation
[0298] The octreotide formulation in Example 28 (Table 6, first
column) was prepared essentially as described in the above
Examples. Below follows the detailed production process for this
formulation.
Production of the Hydrophilic Fraction:
[0299] To 150 mL water the following ingredients were slowly added
and mixed: 24.05 g of sodium octanoate, 16.04 g of PVP-12 and 92.4
g of 10 mg/mL aqueous octreotide solution. The resulting solution
was lyophilized.
Production of the Hydrophobic Medium:
[0300] 3.25 g Tween 80, 6.47 g of glyceryl monocaprylate, 65.25 g
of glyceryl tricaprylate and 43.50 g of castor oil were mixed
together.
Production of the Bulk Drug Product:
[0301] 26.08 g of the hydrophilic fraction was slowly added to
73.92 g of the hydrophobic medium at 20.+-.2.degree. C. while
mixing. After addition of the entire hydrophilic fraction, the
mixing speed was increased. Degassing by vacuum was then applied
and the resulting suspension was stored at 2-8.degree. C. [0302] To
enable larger amounts of octreotide to be dissolved the following
method was devised: [0303] 1. The amount of water of the
hydrophilic fraction preparation was the same as the calculated
volume of the final bulk drug product. [0304] 2. PVP-12 was
dissolved in half of the above amount of water. [0305] 3. Sodium
octanoate was dissolved in the second half amount of water. [0306]
4. Octreotide was dissolved in the PVP-12 solution (from paragraph
2). [0307] 5. The sodium octanoate solution was added to the
octreotide and PVP-12 solution.
[0308] At this stage there was some precipitation, but it became
soluble after mixing.
Example 30
Experiments in Pigs Using Capsules
[0309] In order to test the activity of the formulations of the
invention when administrated in capsules, an animal model allowing
capsule administration to pigs (domestic swine) was established. In
order to bypass the stomach and allow direct administration of
capsules to the small intestine of the pig, a well established
model in dogs ("Nipple Valve model"; Wilsson-Rahmberg & O.
Jonsson, Laboratory Animals (1997), 31, 231-240) was adapted to the
commercial pig.
[0310] The two octreotide formulations shown below in Table 37 were
prepared. The octreotide (x) formulation was prepared essentially
as described above for the basic formulation wherein the
hydrophilic fraction has been simplified to omit MgCl.sub.2 and
MC400. The octreotide (y) formulation was prepared essentially as
described above for the improved octreotide formulation. The
formulations were filled into gelatin capsules (from Capsugel),
basic formulation(x) at 0.42 mL/capsule and improved formulation
(y) at 0.44 mL/capsule, resulting in 5 mg net octreotide content in
both types of filled capsules. The capsules were not enteric-coated
i.e. they were uncoated.
TABLE-US-00052 TABLE 37 Octreotide (x) Octreotide(y) Formulation,
API basic improved Ingredient (% w/w) (% w/w) Hydrophilic API 1.357
1.277 fraction PVP-12 2.717 10.011 Sodium octanoate 12.011 15.008
Water 0.643 1.052 Hydrophobic Tween 80 0 1.992 medium Glyceryl
monocaprylate 0 3.967 Glyceryl tricaprylate 0 40.016 Castor oil
43.562 26.677 Span40 1.198 0 Lecithin 2.403 0 Ethyl isovalerate
10.391 0 Glyceryl monooleate 2.254 0 Glyceryl tributyrate 23.463
0
[0311] The formulations described above in Table 37 were
administered directly to the small intestine of the
non-anesthetized pigs via the gastric bypass described above, and
plasma octreotide levels were measured post-administration.
Exposure values, AUC were determined for the formulations. The % BA
was calculated compared to the exposure to octreotide after
subcutaneous administration. The results obtained are shown below
in Table 38.
TABLE-US-00053 TABLE 38 Cargo Formulation N AUC (0-240) .+-. SD %
BA .+-. SD Octreo- Octreotide(x) 4 896 .+-. 305 2.1 .+-. 0.7 tide
Octreotide(y) 4 2574 .+-. 889 6.2 .+-. 2.1
[0312] The above results in Table 38 show that there was
bioavailability in the pig model for encapsulated formulations, for
both the basic and improved formulations. Octreotide
bioavailability of the improved formulation was about three times
the level of bioavailability of the basic formulation.
[0313] The results given here for bioavailability are
underestimated because sampling time was not sufficient for
octreotide levels to go back to baseline (0 ng/mL). This was due to
the unexpectedly longer exposure time in pigs as compared to what
had been previously measured in rats. The shape of the graph was
changed compared to the rat results showing longer time to reach
maximal peak levels and extended time in which octreotide is
resident in the blood. This may be advantageous since this allows
the octreotide to be longer-acting in the body. Thus the actual
bioavailability in pigs must be higher than the numbers given.
[0314] Based on the results in rats, the level of bioavailability
in pigs of octreotide administered in aqueous solution is
extrapolated to be about 0.1%. This level of bioavailability is
below the level of sensitivity of the bioassay used for pigs.
Example 31
Dose-Response Results for PVP in the Improved Formulation
[0315] Further to the PVP results in Example 24, the effect on
activity of increasing the amount of PVP-12 in the improved
formulation was studied. The improved formulations, made
essentially as described above, contained octreotide as cargo
compound and different doses of PVP-12 as shown in Table 39 below.
The PVP-12 doses tested were 7.5%, 10.0% and 15.0% PVP-12. The
formulations containing 10% and 15.0% PVP were semi-solid i.e. they
were apparently semi-solid suspensions--and the formulation
containing 7.5% PVP was a viscous suspension.
TABLE-US-00054 TABLE 39 Octreotide Octreotide Octreotide
Formulation, API PVP 7.5% PVP 10.0% PVP 15.0% Ingredient (% w/w) (%
w/w) (% w/w) Hydrophilic API 0.058 0.058 0.058 fraction PVP- 12
7.506 10.011 15.009 Sodium octanoate 15.012 15.008 15.009 Water
0.903 1.003 1.203 Hydrophobic Tween 80 2.098 2.027 1.884 medium
Glyceryl 4.178 4.036 3.752 monocaprylate Glyceryl 42.147 40.714
37.851 tricaprylate Castor oil 28.098 27.143 22.234
[0316] The formulations described above in Table 39 were
administered directly to the jejunum of non-anesthetized rats, and
plasma octreotide levels were measured post-formulation
administration. Exposure values, AUC, were determined for the three
formulations. These results are shown below in Table 40.
TABLE-US-00055 TABLE 40 T AUC (0-25)/dose/kg Cargo Formulation N
b.w. .+-. SD Octreotide 7.5% PVP-12 7 2.9 .+-. 2.2 10.0% PVP-12 9
4.4 .+-. 1.7 .sup. 15% PVP-12 10 2.1 .+-. 1.2
[0317] The results shown above in Table 40 demonstrate that the
absorption of octreotide was greatest when PVP in the formulation
was 10%, and increasing the amount to 15% results in significant
decrease in activity. This confirms the choice of 10% PVP in the
improved formulation.
Experiment 32: Activity of API Packed in Formulation Compared to
API Administered Concomitant to Formulation
[0318] Three different basic formulations of three different cargo
compounds were prepared (dextran, gentamicin and exenatide),
essentially as described above (wherein the basic formulation is
the basic non-simplified hydrophilic fraction). Each of these three
formulations was administered directly to the jejunum of
non-anesthetized rats, and plasma cargo levels were measured
post-formulation administration. Exposure values, AUC, were
determined for the formulations. Additionally, a similar
formulation was prepared with a non-relevant cargo compound (a mock
formulation). Separately, the mock formulation was administered
concomitantly with dextran, gentamicin or exenatide in aqueous
solution and exposure values, AUC, were determined. Concomitant
administration was achieved by administrating cargo in aqueous
solution immediately followed by mock formulation administration
via a jejunal--implanted cannula (gastric bypass). For each
compound, exposure after administration of the formulated cargo was
compared to exposure after administration of the unformulated cargo
(concomitant). The comparative results are shown below in Table 41.
The results show that there is higher activity (bioavailability)
when the cargo is formulated compared to unformulated (concomitant)
in all three cases, and that exenatide showed by far the greatest
increase in activity due to formulation. Note that dextran and
gentamicin are compounds that are not sensitive to protease
degradation, whereas exenatide being a peptide is subject to
degradation by intestinal enzymes. The large difference in activity
between the formulated exenatide compared to unformulated exenatide
may be due to the protective effect of the formulation against
degradation.
TABLE-US-00056 TABLE 41 Formulated versus unformulated API/cargo
(fold activity) Dextran 1.7 Gentamicin 1.5 Exenatide 4.4
Example 33
Intestinal Hyperpermeability Evaluation
[0319] A. Size limitation: The technology and formulations
described above are intended to enhance the permeability of the
intestine, allowing specific delivery of proteins, peptides and
other otherwise impermeable molecules across this barrier. A
certain degree of non-specific penetration of intestinal content
may result as a side-effect of this enhancement of specific
permeability. The size of molecules which could possibly penetrate
the intestine in a non-specific manner was evaluated using
different molecular size markers.
[0320] In order to evaluate the molecular size limit of increased
GI permeability, five different FITC-labeled dextrans of different
molecular weight were chosen to serve as molecular markers to test
increased intestinal permeability; the average molecular weight of
the five dextrans was 4.4, 10, 20, 40 and 70 kDa, equivalent to a
radius of 14, 23, 33, 45 and 60 .ANG. respectively. These different
size markers were administered directly to the jejunum of
non-anaesthetized rats, through an intestinal implanted cannula,
and showed virtually no basal intestinal penetration when tested
alone. Each of these markers was then administered directly to the
jejunum of non-anesthetized rats together with 300 .mu.L of basic
formulation, and the degree of its penetration was evaluated by
testing dextran levels in blood.
[0321] The plasma dextran levels were measured pre-dosing and at
3', 6', 10', 25', 60', 90' minutes post formulation administration.
Exposure values, AUC (0-90), were determined and the results are
shown in FIG. 7. Data is presented as MEAN.+-.SD, n.gtoreq.4.
[0322] The results show that while the smallest molecular marker
tested (dextran of average MW=4.4 kDa), penetrates the intestine
when administered concomitantly with a formulation, as the molecule
size increases, penetration extent decreases: a marker molecule of
10 kDa penetrates to a smaller extent and a 20 kDa marker to an
even smaller extent. A marker molecule of 40 kDa shows minimal
penetration, while a marker molecule of 70 kDa shows no penetration
at all (basal penetration). These results indicate that 40-70 kDa
is a cutoff size for non-specific permeability enhancement by
formulations of the invention. Thus administration of a large
volume of formulation (300 .mu.L) to the jejunum of rats resulted
in permeability enhancement of the intestinal barrier, and this
enhanced permeability is restricted by molecular size, showing a
cutoff size of 40-70 kDa and minimal penetration at 40 kDa.
[0323] Published values of the size of hazardous molecules
(molecular weight and radius) which could potentially be present in
the intestine are shown below in Table 42.
TABLE-US-00057 TABLE 42 MW (kDa) Radius (.ANG.) Macromolecules
>4 14 or larger LPS >100 Short-100 Long-1000 Enterobacterial
70-900 -- Toxins Viruses -- 600-1000 Bacteria -- 10,000 or
larger
[0324] Table 42 demonstrates that potentially hazardous molecules
present in the intestine are above the cutoff size of permeability
enhancement by the tested formulations, as shown above. Thus these
results suggest that the tested formulations will not facilitate
penetration of hazardous molecules through the intestinal barrier
and these formulations can therefore be considered as safe. Other
formulations of the invention give similar results.
[0325] B. Formulation repeated dosing: In order to investigate if
repeated dosing of formulation affects intestinal permeability, the
octreotide improved formulation (12% sodium octanoate with castor
oil) was dosed to rats for 14 sequential days using the above in
vivo model (rat implanted with two cannulas in the jejunum). At
days 1, 7 and 14 of administration, a dextran permeability marker
(FITC-dextran of 4.4 kDa MW; FD4), was administered 60 minutes post
formulation administration. This was to assess the permeability of
the intestine by the penetration of the FD4 from the intestine to
blood. No significant difference in FD4 exposure following 14 days
of formulation repeated dosing was found. These results suggest
there is no increase in intestinal permeability following this
period of repeated dosing of formulation, and intestinal enhanced
permeability remains a reversible process during this period.
[0326] The results suggest that the formulation causes no damage to
the intestinal tissue, but acts by specifically opening the
intestinal barrier, showing no additive permeability enhancing
effect.
Example 34
Intestinal Hyperpermeability Evaluation: Time-Course and
Reversibility
[0327] Further to the study in the above Example, a study was
designed in order to define the time-course of increased intestinal
permeability due to the formulations of the invention, and the
reversibility of this process, using dextran as a permeability
marker.
[0328] In order to define the time window of increased intestinal
permeability, an in vivo model was developed in rats, in which one
or two cannulas are implanted in the jejunum of the rats.
FITC-labeled dextran (average molecular weight 4.4 kDa, FD4), which
has virtually no basal intestinal penetration, served as a
molecular marker to test intestinal permeability. An experiment was
designed in which the dextran marker was administered concomitant
to the formulation (by a jejunal implanted cannula), or at
different time intervals from the formulation administration (by a
second separate jejunal implanted cannula). Intestinal permeability
was evaluated by testing FD4 penetration to blood. Rats were
administered a basic formulation concomitant with the dextran
marker, or the basic formulation and then the dextran marker at
different intervals of time (10, 30 and 60 minutes). Blood samples
were analyzed for dextran concentration pre-administration and at
3, 6, 10, 25, 60 and 90 min following dextran administration. The
results are shown in FIG. 8. Data is presented as Mean.+-.SD,
n.gtoreq.5.
[0329] FIG. 8 demonstrates that the dextran marker penetrates the
intestine to the highest extent when administered together with the
formulation. An interval of 10 minutes between administration of
the formulation and administration of the dextran marker results in
significantly decreased amount of marker penetration, and
increasing the interval further results in exponential reduction of
marker penetration.
[0330] These results show that while there is some degree of
non-specific permeability enhancing by the formulation, it is
restricted to a short period of time following administration of
the formulation. The permeability of the intestine decreases
sharply with time, and 60 minutes from administration of the
formulation there is no more marker penetration. Thus
administration of the formulation to the rat intestine results in a
very short period of hyperpermeability of the intestinal barrier.
Other formulations of the invention gave similar results.
Example 35
Oral Administration of Octreotide to Monkeys
[0331] In order to test the pharmacokinetics of octreotide
following oral administration of formulated octreotide to monkeys,
five Cynomologus monkeys were orally dosed with capsules containing
an improved castor oil formulation of octreotide (similar to
formulation I of Table 35--but with higher load of octreotide). The
capsules used were size 1 gelatin capsules coated with 6.7%
Acryl-EZE.RTM. enteric coating; this coating prevents capsule
disintegration in the stomach and allows opening of the capsules in
the small intestine of the dosed animals. The octreotide dose used
was 5 mg/capsule.
[0332] Monkeys were fasted overnight prior to capsule
administration. Following oral administration, blood samples were
withdrawn over a period of 9.75 hours, processed for plasma and
analyzed for octreotide content by the LC/MS/MS method: see FIG. 9.
Similar experiments were performed with the improved no castor
oil/GTC formulation (similar to formulation IV of Table 35 but with
higher load of API) and similar results were obtained. Similar
experiments were also performed with several different enteric
coatings and similar results were obtained.
[0333] In order to compare the pharmacokinetics of octreotide
following administration of the improved octreotide formulation, to
the pharmacokinetics of injected octreotide, octreotide acetate
solution (0.1 mg/monkey) was administered subcutaneously to two
monkeys from the above group to serve as a reference. Blood samples
were withdrawn over a period of four hours, processed for plasma
and analyzed for octreotide content by the LC/MS/MS method.
[0334] The pharmacokinetics of octreotide following oral octreotide
and subcutaneous injected octreotide solution were compared (see
FIGS. 9 and 10). The results of the oral formulation showed
absorption over a period of a few hours. The shape of the graph was
changed compared to subcutaneous, showing slower but longer release
of octreotide into the blood. This may be advantageous since this
allows the persistence of octreotide for a longer time in the blood
potentially prolonging the activity window.
[0335] An approved dose for injected octreotide acetate in humans
is 0.1 mg/patient. The above results in the monkeys suggest that
the improved formulation containing about 10 mg octreotide per dose
will generate therapeutic exposure in humans.
Example 36
Stability Data
[0336] Basic and improved octreotide formulations of the invention
were maintained both at 4.degree. C. and at 25.degree. C. and were
tested for octreotide content periodically. Both formulations were
found to be stable.
Example 37
Formulations Incorporating Vancomycin, Interferon-Alfa and
Terlipressin
[0337] A. Vancomycin: Table 43 below describes a vancomycin
improved formulation, containing 10% PVP and 15% sodium octanoate
in the hydrophilic fraction, and containing glyceryl tricaprylate
as the main constituent of the hydrophobic medium. The vancomycin
was obtained from Gold Biotechnology.
TABLE-US-00058 [0337] TABLE 43 Formulation, API Vancomycin
Ingredient (% w/w) Hydrophilic API 6.267 fraction NaOH 0.082 PVP-12
10.005 Sodium octanoate 15.016 Water 1.216 Hydrophobic Tween 80
2.004 medium Glyceryl monocaprylate 4.008 Glyceryl tricaprylate
61.400 Castor oil 0.000
[0338] In a preliminary experiment, the formulation described above
in Table 43 was administered directly to the jejunum of
non-anesthetized rats, and plasma vancomycin levels were measured
post-formulation administration. Exposure value, AUC, was
determined for the formulation. The results are that the absolute
BA is around 5% (comparative to IV, n=6). When vancomycin in saline
solution was administered to the jejunum of non-anesthetized rats
no BA was detected.
[0339] Interferon-alfa: Table 44 below describes an interferon-alfa
improved formulation, containing 10% PVP and 15% sodium octanoate
in the hydrophilic fraction, and containing glyceryl tricaprylate
as the main constituent of the hydrophobic medium. The
interferon-alfa is supplied in a buffer (from Intas
Biopharmaceuticals) and the ingredients of the interferon-alfa
buffer in the formulation are marked by an asterisk (*).
TABLE-US-00059 TABLE 44 Formulation, API IFN-.alpha. Ingredient (%
w/w) Hydrophilic API 0.050 fraction *Na.sub.2HPO.sub.4 0.032
*NaH.sub.2PO.sub.4 0.030 *Polysorbate (Tween) 80 0.002 *Disodium
EDTA 0.002 PVP- 12 10.026 Sodium Octanoate 14.997 Water 1.006
Hydrophobic Tween 80 2.005 medium Glyceryl monocaprylate 4.005
Glyceryl tricaprylate 67.84 Castor oil 0
[0340] The formulation described above in Table 44 is administered
directly to the jejunum of non-anesthetized rats. Plasma
interferon-alfa levels are measured post-formulation
administration.
[0341] C. Terlipressin: Table 45 below describes a terlipressin
basic formulation and a terlipressin improved formulation
containing 10% PVP and 15% sodium octanoate in the hydrophilic
fraction, and containing glyceryl tricaprylate as the main
constituent of the hydrophobic medium. The terlipressin was
obtained from Bambio. The basic formulation was prepared
essentially as described above and the improved formulation is also
prepared essentially as described above.
TABLE-US-00060 TABLE 45 Terlipressin Terlipressin Formulation, API
basic improved Ingredient (% w/w) (% w/w) Hydrophilic API 0.235
0.235 fraction MgCl.sub.2 0.137 0.000 PVP 12 2.736 10.004 Sodium
octanoate 12.004 15.015 MC 400 0.137 0.000 Water 0.610 1.010
Hydrophobic Span40 1.211 0.000 medium Lecithin 2.428 0.000 Ethyl
isovalerate 10.500 0.000 Glyceryl monooleate 2.278 0.000 Glyceryl
tributyrate 23.708 0.000 Castor oil 44.016 0.000 Tween 80 0.000
2.002 GMC 0.000 4.004 GTC 0.000 67.731
[0342] The formulations described above in Table 45 are
administered directly to the jejunum of non-anesthetized rats.
Plasma terlipressin levels are measured post-formulation
administration.
Example 38
Inhibition of Growth Hormone In Vivo by Octreotide
[0343] One of the best characterized effects of octreotide is the
inhibition of growth hormone release. In order to test for the
efficacy of an octreotide formulation of the invention on growth
hormone inhibition, a rat model was used in which endogenous rat
growth hormone (rGH) levels were monitored following octreotide
formulation administration to the jejunum of the non-anesthetized
rat model (described above). Administration of a basic octreotide
formulation (containing 12% sodium octanoate) to the jejunum of
rats was shown to reduce rGH levels by 87.4% compared to
administration of a saline control. This result demonstrates that
the octreotide formulations described herein enable delivery of
octreotide in its active form from the intestinal lumen into the
blood stream.
Example 39
Toxicology Studies
[0344] A 28-day toxicity administration study of formulation
control (excipients only, no cargo) was performed in Wistar rats.
The animals in the test group were daily administered rectally with
the maximal feasible dose of formulation (100 .mu.L/animal/day) for
28 consecutive days. The test group was compared to two control
groups: a naive group (non-treated) and a saline administered
group, (n=15/group).
[0345] General clinical observations were made twice daily, and
detailed clinical observations were performed weekly. Body weight
and food consumption were measured weekly. Clinical pathology and
gross pathology were conducted one day after the last treatment. A
histological examination was performed on rectum, colon, liver and
kidneys, and no toxic effects were detected. There was clean
histopathology with no local GI or systemic findings, no
formulation related clinical findings, no changes in hematological
and blood chemistry parameters, no macroscopic findings at necropsy
and no mortality. In conclusion, this experiment demonstrated that
there was no observed toxicity during a daily rectal dosing of
formulation to rats for 28 consecutive days.
[0346] Having thus described several aspects of at least one
embodiment, it is to be appreciated that various alterations,
modifications, and improvements will readily occur to those skilled
in the art. Such alterations, modifications, and improvements are
intended to be part of this disclosure and are intended to be
within the scope of the invention. Accordingly, the foregoing
description and drawings are by way of example only, and the scope
of the invention should be determined from proper construction of
the appended claims, and their equivalents.
* * * * *