U.S. patent application number 14/524521 was filed with the patent office on 2015-02-26 for sustained release formulations using non-aqueous carriers.
This patent application is currently assigned to AstraZeneca Pharmaceuticals LP. The applicant listed for this patent is Amylin Pharmaceuticals, LLC, AstraZeneca Pharmaceuticals LP. Invention is credited to Scott H. COLEMAN, Mary L. HOUCHIN, Robert N. JENNINGS, Robin H. LEE, Greg OEHRTMAN, Hong QI.
Application Number | 20150056285 14/524521 |
Document ID | / |
Family ID | 41797509 |
Filed Date | 2015-02-26 |
United States Patent
Application |
20150056285 |
Kind Code |
A1 |
HOUCHIN; Mary L. ; et
al. |
February 26, 2015 |
SUSTAINED RELEASE FORMULATIONS USING NON-AQUEOUS CARRIERS
Abstract
The disclosure provides one-component, injectable, sustained
release formulations which comprise microspheres containing active
pharmaceutical ingredients (e.g., exenatide), wherein the
microspheres are suspended in a non-aqueous carrier. The
non-aqueous carrier can be an oil, a fractionated oil,
triglycerides, diglycerides, monoglycerides, propylene glycol fatty
acid diesters, and the like. The formulations offer distinct
advantages of long shelf life for the stability and potency of the
formulation and sustained release of active pharmaceutical
ingredients to reduce the frequency of medication dosing and to
increase patient compliance.
Inventors: |
HOUCHIN; Mary L.;
(Rockville, MD) ; LEE; Robin H.; (San Diego,
CA) ; QI; Hong; (San Diego, CA) ; OEHRTMAN;
Greg; (San Diego, CA) ; JENNINGS; Robert N.;
(San Diego, CA) ; COLEMAN; Scott H.; (San Diego,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Amylin Pharmaceuticals, LLC
AstraZeneca Pharmaceuticals LP |
San Diego
Wilmington |
CA
DE |
US
US |
|
|
Assignee: |
AstraZeneca Pharmaceuticals
LP
Amylin Pharmaceuticals, LLC
|
Family ID: |
41797509 |
Appl. No.: |
14/524521 |
Filed: |
October 27, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13060225 |
May 16, 2011 |
8895033 |
|
|
PCT/US09/56058 |
Sep 4, 2009 |
|
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14524521 |
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61094381 |
Sep 4, 2008 |
|
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Current U.S.
Class: |
424/489 ;
514/11.7; 514/5.3; 514/6.7; 514/7.2 |
Current CPC
Class: |
A61K 9/1647 20130101;
A61P 3/00 20180101; A61K 45/06 20130101; A61K 9/5153 20130101; A61K
47/14 20130101; A61P 1/16 20180101; A61K 31/60 20130101; A61K
9/1617 20130101; A61P 3/08 20180101; A61P 5/50 20180101; A61K 38/26
20130101; A61K 9/0019 20130101; A61K 9/1623 20130101; A61K 38/2278
20130101; A61P 3/10 20180101; A61K 31/65 20130101; A61P 3/04
20180101; A61K 31/00 20130101; A61P 43/00 20180101; A61K 9/10
20130101; A61K 38/28 20130101; A61P 3/06 20180101; A61K 47/44
20130101; A61K 31/60 20130101; A61K 2300/00 20130101; A61K 31/65
20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/489 ;
514/11.7; 514/7.2; 514/6.7; 514/5.3 |
International
Class: |
A61K 9/16 20060101
A61K009/16; A61K 38/26 20060101 A61K038/26 |
Claims
1. (canceled)
2. A manufactured pre-mixed formulation comprising a suspension of
(i) a pharmaceutically acceptable non-aqueous carrier comprising
one or more glycerol or propylene glycol esters of fatty acids,
wherein the one or more esters comprise 0 to 2 wt %, C.sub.6 fatty
acid, 50 to 65 wt % C.sub.8 fatty acid, 30 to 45 wt % C.sub.10
fatty acid, 0 to 2 wt % C.sub.12 fatty acid, and 0 to 2 wt %
C.sub.14 fatty acid based on the total fatty acid content of the
one or more esters; and (ii) microspheres comprising a
biocompatible, biodegradable polymer and an active pharmaceutical
ingredient.
3-11. (canceled)
12. The formulation of claim 2, wherein the one or more glycerol
esters of the fatty acids are one or more monoglycerides, one or
more diglycerides, one or more triglycerides, or a combination of
two or more thereof.
13. The formulation of claim 2, wherein the one or more propylene
glycol esters of the fatty acids are propylene glycol diesters.
14. The formulation of claim 2, wherein the fatty acids are
saturated fatty acids.
15. The formulation of claim 12, wherein the one or more glycerol
esters of the fatty acids are one or more triglycerides which
comprise esters of C.sub.6 to C.sub.12 fatty acids.
16-18. (canceled)
19. The formulation of claim 15, wherein the triglycerides further
comprise ester of C.sub.14 fatty acid.
20. (canceled)
21. The formulation of claim 2, further comprising a
pharmaceutically acceptable excipient.
22. The formulation of claim 21, wherein the pharmaceutically
acceptable excipient is a sugar or a sugar alcohol.
23. The formulation of claim 21, wherein the pharmaceutically
acceptable excipient is sucrose, glucose, dextrose, galactose,
maltose, trehalose, fructose, maltodextrin, glycol, glycerol,
erythritol, threitol, arabitol, ribitol, sorbitol, dulcitol,
iditol, isomalt, maltitol, lactitol, mannitol, xylitol, benzoic
acid, sorbic acid, meta cresol, sodium benzoate, potassium sorbate,
methylparaben, propylparaben, butylparaben, benzalkonium chloride,
sodium metabisulfite, butylated hydroxy anisole, butylated hydroxy
toluene, sodium sulfite, tocopherol thymol, ascorbate, propyl
gallate, or a combination of two or more thereof.
24. The formulation of claim 23, wherein the pharmaceutically
acceptable excipient is sucrose.
25. The formulation of claim 2, wherein the biocompatible,
biodegradable polymer is a polylactide, a copolymer of a
polylactide, a polyglycolide, a copolymer of a polyglycolide, a
poly(lactide-co-glycolide) copolymer, a polylactic acid, a
copolymer of a polylactic acid, a polyglycolic acid, a copolymer of
a polyglycolic acid, a poly(lactic acid-co-glycolic acid)
copolymer, a polycaprolactone, a copolymer of a polycaprolactone, a
polycarbonate, a copolymer of a polycarbonate, a polyesteramide, a
copolymer of a polyesteramide, a polyanhydride, a copolymer of a
polyanhydride, a polyamino acid, a copolymer of a polyamino acid, a
polyorthoester, a copolymer of a polyorthoester, a
polycyanoacrylate, a copolymer of a polycyanoacrylate, a
poly(p-dioxanone), a copolymer of a poly(p-dioxanone), a
polyalkylene oxalate, a copolymer of a polyalkylene oxalate, a
polyurethane, a copolymer of a polyurethane, or a combination of
two or more thereof.
26. The formulation of claim 2, wherein the biocompatible,
biodegradable polymer is a poly(lactide-co-glycolide)
copolymer.
27-29. (canceled)
30. The formulation of claim 2, wherein the active pharmaceutical
ingredient is a GLP-1 receptor agonist.
31. The formulation of claim 2, wherein the active pharmaceutical
ingredient is exenatide.
32. The formulation of claim 2, wherein the active pharmaceutical
ingredient is GLP-1 (7-37) or GLP-1(7-36)-NH.sub.2.
33. The formulation of claim 2, wherein the active pharmaceutical
ingredient is pramlintide, davalintide, Val.sup.27-davalintide,
metreleptin, insulin, a glucagon agonist or antagonist, a chimera
of a GLP-1 receptor agonist and a glucagon agonist, bovine serum
albumin, sodium salicylate, salicylic acid, minocycline HCl,
insulin, or a small molecule organic compound.
34-46. (canceled)
47. The formulation of claim 2, wherein the microspheres are
present in the formulation at a concentration of from 10 mg/ml to
500 mg/ml.
48. The formulation of claim 31, wherein the exenatide is present
in the microspheres in an amount of about 5 wt %.
49. The formulation of claim 21, wherein the pharmaceutically
acceptable excipient is sucrose, the biocompatible, biodegradable
polymer is a poly(lactide-co-glycolide) copolymer, and the active
pharmaceutical ingredient is exenatide.
50-66. (canceled)
67. A method for treating diabetes, stimulating insulin release;
lowering plasma glucagon; reducing food intake; reducing appetite;
decreasing gastric motility; delaying gastric emptying: lowering
plasma lipid levels; treating impaired glucose tolerance; treating
hyperglycemia: treating obesity: treating overweight; treating
fatty liver disease: or treating non-alcoholic steatohepatitis in a
patient in need thereof, the method comprising administering to the
patient the formulation of claim 2.
68-89. (canceled)
90. The kit of claim 88, wherein the container is a single-dose or
multi-dose pen injector, a single-dose or multi-dose vial, or a
single-dose or multi-dose cartridge.
91-118. (canceled)
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 13/060,225 filed on Feb. 22, 2011, which is a .sctn.371
national stage entry of International Application No.
PCT/US09/56058 filed on Sep. 4, 2009, which claims priority to U.S.
Application No. 61/094,381 filed Sep. 4, 2008; the disclosures of
all of which are herein incorporated by reference in their
entirety.
BACKGROUND
[0002] Injectable sustained release formulations offer the
opportunity to provide therapeutic amounts of active pharmaceutical
ingredients over an extended period of time from a single
injection, thus eliminating the need for once or twice daily
injections. Presently available injectable sustained release
formulations utilizing, for example, microspheres and an aqueous
carrier, carry several disadvantages. The formulations do not offer
long term stability in the aqueous carrier, thus necessitating
separate packaging and storage for the microspheres and aqueous
carrier, and the patient must take several steps to combine the
microspheres and aqueous carrier before administering the
injection.
[0003] Another disadvantage of presently available injectable
microsphere formulations is a large burst release following
injection, which causes an undesirable in vivo release of active
pharmaceutical ingredient in a single burst. When medications have
toxic or deleterious side effects, this is undesirable.
[0004] There is a need for formulations and methods of safely
administering sustained release pharmaceutical formulations to
patients so that the active ingredient will be released in vivo
over an extended period of time and without an unacceptable initial
burst release. Ideally the active ingredient is released so as to
maintain levels within the therapeutic window, i.e., in the
concentration range above that needed to cause the desired clinical
effect, but below that where undesirable side effects outweigh the
benefits of the drug. It is also necessary that this active
pharmaceutical ingredient be provided in a manner that is easy and
convenient for the patient to self-administer and that is provided
in a formulation that maintains stability for a long period of time
in a liquid state. The disclosure is directed to these as well as
other important ends.
SUMMARY
[0005] The disclosure provides formulations comprising microspheres
that contain active pharmaceutical ingredients, where the
microspheres are suspended in a non-aqueous pharmaceutically
acceptable carrier. The formulations are one-component injectable
microsphere formulations, such that they do not require the patient
to mix the formulation with a pharmaceutically acceptable carrier
prior to injection. The disclosure offers distinct advantages over
prior two-component formulations by providing for a long shelf life
of the composition in the carrier, sustained release of the active
pharmaceutical ingredient, a less complex carrier, a more easily
manufactured carrier, a less complex injection-delivery apparatus,
a kit with less components, and ease of use by patients.
[0006] The disclosure provides sustained release formulations
comprising a pharmaceutically acceptable carrier which consists
essentially of one or more triglycerides which comprise
C.sub.6-C.sub.12 fatty acids; and microspheres which consist
essentially of a poly(lactide-co-glycolide) polymer having
dispersed therein about 1% to 10% (w/w) exenatide and about 0.1% to
5% (w/w) of a sugar; wherein the ratio of lactide:glycolide in the
polymer is about 70:30 to 30:70, or about 1:1. In one embodiment,
the exenatide is present in an amount of 1% to 5% (w/w) or 5% (w/w)
and the sugar is present in an amount of 2% (w/w). The sugar may
be, e.g., glucose, dextrose, galactose, maltose, fructose, mannose,
sucrose, lactose, trehalose, raffinose, acarbose, glycol, glycerol,
erythritol, threitol, arabitol, ribitol, sorbitol, dulcitol,
iditol, isomalt, maltitol, lactitol, mannitol, xylitol, or a
combination of two or more thereof. In one embodiment, the sugar is
sucrose. The formulation is a suspension whereby the microspheres
are suspended in the carrier. In one embodiment, the total pore
volume of the microspheres is about 0.1 mL/g or less, as determined
using mercury intrusion porosimetry, to provide a release profile
having a ratio of maximum serum concentration of exenatide during
the period of release (C.sub.max) to average serum concentration of
exenatide during the period of release (C.sub.ave) of about 3 or
less. Further, although the microspheres are formulated in oil
(i.e. a carrier as disclosed herein), the microspheres do not
necessarily have oil contained within the interior spaces or pores,
or within a substantial number of interior spaces or pores, of the
microspheres, and yet can achieve the surprising properties
disclosed herein.
[0007] The disclosure provides sustained release formulations
comprising a pharmaceutically acceptable non-aqueous carrier and
microspheres which comprise a biocompatible, biodegradable polymer
and an active pharmaceutical ingredient. In one embodiment, the
total pore volume of the microspheres is about 0.1 mL/g or less, as
determined using mercury intrusion porosimetry, to provide a
release profile having a ratio of maximum serum concentration of
the active pharmaceutical ingredient during the period of release
(C.sub.max) to average serum concentration of the active
pharmaceutical ingredient during the period of release (C.sub.ave)
of about 3 or less. Further, although the microspheres are
formulated in oil (i.e. a carrier as disclosed herein), in some
embodiments the microspheres do not have oil contained within the
interior spaces or pores, or do not have oil within a substantial
number of interior spaces or pores of the microspheres, and yet can
achieve the surprising properties disclosed herein. The formulation
is a suspension whereby the microspheres are suspended in the
carrier. The non-aqueous carrier may be an oil, such as
fractionated oils, triglycerides, diglycerides, monoglycerides,
propylene glycol fatty acid diesters, and the like.
[0008] In one embodiment the active ingredient is not soluble in
the carrier. In various other embodiments the active ingredient has
a solubility in the carrier of less than 0.01 mg/ml, or less than
0.05 mg/ml or less than 0.1 mg/ml or less than 0.5 mg/ml or less
than 1 mg/ml. In still other embodiments the active pharmaceutical
ingredient has a solubility in the carrier such that less than 10%
of the active ingredient in the formulation is contained within the
carrier with the remaining 90% contained within the microparticles.
In further embodiments less than 5% or less than 2% or less than 1%
or less than 0.5% of the active ingredient is contained in the
carrier. In still further embodiments where it is desirable to have
some active ingredient immediately available, it may also be
directly incorporated into the carrier in a pharmaceutically
effective amount.
[0009] The disclosure provides a kit, available to a patient or
medical service provider. The kit contains a container having a
formulation of the invention, and instructions for use. In one
embodiment the container is a pen injector. The pen injector can be
a single-dose pen injector or a multi-dose pen injector. In one
embodiment the container is a vial, which can be either a
single-dose vial or a multi-dose vial. In another embodiment the
container is a cartridge, such as a cartridge for use in a
injection apparatus. The cartridge can be either a single-dose or a
multi-dose cartridge. In different embodiments the kit contains 1,
2, 3, 4, or even 5 or more such containers carrying a formulation
of the invention. One further advantage of the formulations is that
in one embodiment the container is provided preservative free. But
in other embodiments a preservative can be soluble in the selected
carrier and provided in the formulation.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] For each of FIGS. 1-6, the microspheres comprise a
poly(lactide-co-glycolide) copolymer having exenatide dispersed
therein, as described in Example 1. For each of FIGS. 2-6, the oil
carrier is a medium chain triglyceride (MCT) commercially available
as MIGLYOL.RTM. 812 (Sasol Germany GmbH, Witten, Germany).
[0011] FIG. 1 provides a comparison of the pharmacokinetics of four
different formulations of microspheres. In three formulations, the
carrier was an oil (e.g., sesame oil; MIGLYOL.RTM. 812; ethyl
oleate). In the comparative formulation, the carrier was an aqueous
diluent.
[0012] FIG. 2 is a graphical simulation (i.e., nanoparametric
superposition) of data extrapolated from FIG. 1 of the plasma
exenatide concentration over time for the microsphere formulation
comprising the oil carrier and the microsphere formulation
comprising the aqueous carrier in male Sprague Dawley Rats. The
plasma concentration plateau of exenatide may be reached after
about 5 dosings.
[0013] FIG. 3 illustrates the in vitro release for a formulation
comprising microspheres in an oil carrier compared to formulations
comprising microspheres in an aqueous carrier.
[0014] FIG. 4 illustrates the in vivo release profile in rats over
10 hours for a formulation comprising microspheres in an oil
carrier and a formulation comprising microspheres in an aqueous
carrier.
[0015] FIGS. 5A and B illustrate the purity of exenatide over 9
months at temperatures of 5.degree. C. and 6 months at 25.degree.
C. when stored in the formulations comprising the microspheres of
Example 1 with an oil carrier as compared to the purity of
exenatide that was stored in dry microspheres of Example 1. In FIG.
5A, the purity of exenatide was determined by strong cation
exchange HPLC. In FIG. 5B, the purity of exenatide was determined
by reverse-phase HPLC.
[0016] FIG. 6 illustrates the stability/potency of exenatide in a
formulation where the microspheres are suspended in an oil carrier,
where one formulation is stored at 5.degree. C. and one formulation
is stored at 25.degree. C.
DETAILED DESCRIPTION
[0017] The disclosure provides sustained release compositions
provided in pharmaceutically acceptable carriers, for the sustained
release of an active pharmaceutical ingredient (API). The
formulations may comprise microspheres comprised of a
biocompatible, biodegradable polymer having an active
pharmaceutical ingredient dispersed therein, where the microspheres
are suspended in a non-aqueous carrier. The formulations are
one-component injectable formulations, compared to two-component
formulations which require that the microspheres be stored dry in
one container while the liquid carrier can be stored in a separate
container, such that the patient must mix the two together prior to
injection. The formulations offer the convenience of long term
stability of a pharmaceutical composition in a non-aqueous liquid
carrier, thus eliminating any need for the patient to add a
pharmaceutically acceptable carrier to the pharmaceutical
composition prior to injection. The formulations are provided in a
single container for easy use by the patient, who only need to
lightly agitate the formulation before injecting it from the same
container. When the container provided is also an injection device,
even the step of syringing the formulation is eliminated. The
formulations described herein offer the additional important
advantage of substantially reducing burst release of the active
pharmaceutical ingredient. Thus, even active pharmaceutical
ingredients that have a toxic effect at higher concentrations can
be safely administered using the formulations described herein.
[0018] The term "patient" refers to mammals, including humans,
animal pets, farm animals, zoo animals, and the like. In one
embodiment, the patient is a human.
[0019] The terms "treating" or "treatment" refer to the
administration of one or more active pharmaceutical ingredients to
a patient who has a condition or disorder or a predisposition
toward a condition or disorder, with the purpose to alleviate,
relieve, remedy, ameliorate, improve, slow or stop the progression
or worsening of the disease, or at least one symptom of the
disease, condition or disorder, or the predisposition toward the
condition or disorder.
[0020] "Exenatide" has the same meaning and amino acid sequence as
exendin-4. More particularly, exenatide is a synthetic peptide with
the same amino acid sequence as exendin-4, which is a peptide
isolated from the venom of the Gila monster.
One Component Formulation
[0021] Previous injectable formulations contained at least two
components. The first component may be dry microspheres and the
second component may be an aqueous pharmaceutically acceptable
carrier. The first component and second component are stored in
separate sealed containers (e.g., vials, injection pen chambers).
The patient receives the two-component formulation, and the patient
or pharmacist must physically mix the two components together prior
to injection. In the case of an injection pen, the two components
are mixed together immediately prior to injection into the patient.
Two-component formulations typically are administered to the
patient within a short time after being mixed with the
pharmaceutically acceptable carrier. For example, the microsphere
component and the pharmaceutically acceptable aqueous carrier are
mixed together and then the formulation is administered to the
patient within about 30 or 60 minutes.
[0022] The formulations described herein are one component
injectable formulations. A one component injectable formulation
refers to a formulation that contains both the microspheres and the
pharmaceutically acceptable carrier provided in the same container,
and that may be administered to the patient without the need to
first combine the microspheres and the pharmaceutically acceptable
carrier. Accordingly, the one component formulation is manufactured
as a pre-mixed formulation for injection. A one-component
formulation provides significant convenience for manufacturing,
transport, storage, and patient use.
[0023] In another embodiment the one-component formulation
described herein is provided in a sealed container. A "sealed
container" is a container that has not been opened, punctured, or
had anything introduced into it since its time of completion of
manufacture. The time of completion of manufacture is the time when
the container holding the formulation is initially sealed.
Containers may include vials (single use or multi-use), syringes,
injection pens (e.g., single use or multi-use), and the like.
Carrier
[0024] "Carrier" (or vehicle) refers to a pharmaceutically
acceptable non-aqueous liquid material. The carrier is
substantially inert so that it does not interact with the
microspheres described herein and is non-toxic so that it does not
negatively impact the patient. The carrier is preferably approved
by or is awaiting approval by a regulatory agency of the Federal or
a state government or listed in the U.S. Pharmacopoeia or other
generally recognized pharmacopoeia for use in mammals, such as
humans. The term "carrier" may include one or more compounds. The
carrier is a non-solubilizing carrier, in that the carrier does not
solubilize the polymer(s) that forms the microspheres. In a further
embodiment, the carrier does not solubilize the active
pharmaceutical ingredient(s) within the microspheres. For example,
the carrier will not solubilize exenatide or other water-soluble
therapeutic peptides or proteins.
[0025] The term "non-aqueous" does not exclude trace amounts of
residual water that do not have a demonstrated negative impact on
the stability of the sustained release compositions. Thus, a
composition may have about 0.1% (w/v) water or even about 0.25%
water or less than 0.1% (w/v) water or less than 0.25% (w/v) water
and still be considered non-aqueous. The carrier does not
solubilize the microspheres to the extent of having a demonstrated
negative impact on the stability of the microspheres or
demonstrated loss of burst release control. In one embodiment, the
carrier does not enter or permeate the biocompatible, biodegradable
polymer and is not dispersed within the biocompatible,
biodegradable polymer. The carrier also does not cause swelling of
the microspheres to an extent that has a demonstrated negative
impact on the stability of the microspheres. For example swelling
may occur to a degree of less than 1% and still be considered a
non-aqueous carrier that is non-swelling of the microspheres.
[0026] In one embodiment, the non-aqueous carrier is a
pharmaceutically acceptable oil. An oil is a substance that is in a
viscous liquid state at ambient temperatures or slightly warmer,
and is both hydrophobic (immiscible with water) and lipophilic
(miscible with other oils, literally). Exemplary pharmaceutically
acceptable oil carriers include vegetable oils and volatile
essential oils. Exemplary pharmaceutically acceptable oil carriers
include coconut oil, palm oil, palm kernel oil, sesame oil, soybean
oil, almond oil, rapeseed oil, corn oil, sunflower oil, peanut oil,
olive oil, castor oil, soybean oil, safflower oil, cottonseed oil,
ethyl oleate, and the like. The carrier may comprise one oil or a
combination of two or more oils.
[0027] In one embodiment, the carrier is a fractionated oil or a
combination of two or more fractionated oils. Exemplary
pharmaceutically acceptable oil carriers include fractionated
coconut oil, fractionated palm oil, fractionated palm kernel oil,
fractionated sesame oil, fractionated soybean oil, fractionated
almond oil, fractionated rapeseed oil, fractionated corn oil,
fractionated sunflower oil, fractionated peanut oil, fractionated
olive oil, fractionated castor oil, fractionated soybean oil,
fractionated safflower oil, fractionated cottonseed oil, and the
like. In one embodiment, the carrier is fractionated coconut oil.
In one embodiment, the carrier is fractionated palm kernel oil. In
one embodiment, the carrier is a combination of fractionated
coconut oil and fractionated palm kernel oil.
[0028] As used herein, fractionation is a process whereby long
chain fatty acids are removed from the oil, such that the resulting
fractionated oil substantially comprises medium chain
triglycerides. The skilled artisan will appreciate that some
long-chain fatty acids may remain in the fractionated oil, but
generally in amounts less than 5 wt % or less than 2 wt % of the
total fatty acid content of the fractionated oil.
[0029] In one embodiment, the carrier is a long chain triglyceride,
a medium chain triglyceride, a diglyceride, a monoglyceride, a
propylene glycol fatty acid diester, or a combination of two or
more thereof.
[0030] In one embodiment, the carrier is a medium chain
triglyceride. The medium chain triglyceride may be synthetic or
natural (e.g., produced from fractionated oils, such as coconut oil
and/or palm kernel oil). "Medium chain triglyceride" refers to
esters of glycerol having three C.sub.6 to C.sub.12 fatty acid
chains, where the three fatty acid chains may be the same or
different. Medium chain triglycerides are represented by the
compound of Formula (I):
##STR00001##
wherein each x is independently 4, 6, 8, or 10. When x is 4, the
chain is referred to as a C.sub.6 fatty acid. When x is 6, the
chain is referred to as a C.sub.8 fatty acid. When x is 8, the
chain is referred to as a C.sub.10 fatty acid. When x is 10, the
chain is referred to as a C.sub.12 fatty acid. In various
embodiments, each x is the same integer, two x are the same integer
and one x is a different integer, or each x is a different
integer.
[0031] In various embodiment, the medium chain triglyceride
comprises esters of (i) three C.sub.8 fatty acids; (ii) three
C.sub.10 fatty acids; (iii) two C.sub.8 fatty acids and one
C.sub.10 fatty acid; (iv) two C.sub.10 fatty acids and one C.sub.8
fatty acid; (v) two C.sub.8 fatty acids and one C.sub.6 fatty acid;
(vi) two C.sub.10 fatty acids and one C.sub.6 fatty acid; (vii) one
C.sub.8 fatty acid, one C.sub.10 fatty acid, and one C.sub.6 fatty
acid; or (viii) any other combination of C.sub.6, C.sub.8,
C.sub.10, and C.sub.12 fatty acids. In one embodiment, the medium
chain triglyceride comprises two C.sub.8 fatty acids and one
C.sub.10 fatty acid. In one embodiment, the medium chain
triglyceride comprises two C.sub.10 fatty acids and one C.sub.8
fatty acid.
[0032] The skilled artisan will appreciate that a mixture of medium
chain triglycerides may result from any process (e.g.,
fractionation, hydrogenation) used to prepare medium chain
triglycerides. For example, substantially all of the medium chain
triglycerides obtained from fractionated coconut oil may comprise
C.sub.8 and/or C.sub.10 fatty acids; however, there may be some
medium chain triglycerides containing C.sub.6 and/or C.sub.12 fatty
acids.
[0033] In one embodiment, the medium chain triglycerides comprise
esters of (i) 0 to 2 wt % C.sub.6 fatty acid, 65 to 80 wt % C.sub.8
fatty acid, 20 to 35 wt % C.sub.10 fatty acid, and 0 to 2 wt %
C.sub.12 fatty acid; (ii) 0 to 2 wt % C.sub.6 fatty acid, 50 to 65
wt % C.sub.8 fatty acid, 30 to 45 wt % C.sub.10 fatty acid, and 0
to 2 wt % C.sub.12 fatty acid; (iii) 0 to 2 wt % C.sub.6 fatty
acid, 45 to 65 wt % C.sub.8 fatty acid, 30 to 45 wt % C.sub.10
fatty acid, 0 to 3 wt % C.sub.12 fatty acid; and 0 to 5 wt %
linoleic acid; or (iv) 0 to 2 wt % C.sub.6 fatty acid, 45 to 55 wt
% C.sub.8 fatty acid, 30 to 40 wt % C.sub.10 fatty acid, 0 to 3 wt
% C.sub.12 fatty acid, and 10 to 20 succinic. In one embodiment,
the medium chain triglyceride comprises 0 to 2 wt % C.sub.6 fatty
acid, 50 to 65 wt % C.sub.8 fatty acid, 30 to 45 wt % C.sub.10
fatty acid, and 0 to 2 wt % C.sub.12 fatty acid, and which is
commercially available as MIGLYOL.RTM. 812 (Sasol Germany GmbH,
Witten, Germany) The weight % is based of the total fatty acid
content of the triglycerides. In one embodiment, the medium chain
triglycerides may comprise up to 2% C.sub.14 fatty acids.
[0034] The carrier may comprise one, two, three, four or more
different medium chain triglycerides. In one embodiment, the
carrier comprises a medium chain triglyceride comprising esters of
two C.sub.8 fatty acids and one C.sub.10 fatty acid. In one
embodiment, the carrier comprises a medium chain triglyceride
comprising esters of one C.sub.8 fatty acid and two C.sub.10 fatty
acids. In one embodiment, the carrier comprises two different
medium chain triglycerides, where a first medium chain triglyceride
comprises esters of two C.sub.8 fatty acids and one C.sub.10 fatty
acid and a second medium chain triglyceride comprises esters of one
C.sub.8 fatty acid and two C.sub.10 fatty acids. In one embodiment,
the carrier comprises a medium chain triglyceride which comprises 0
to 2 wt % C.sub.6 fatty acid, 50 to 65 wt % C.sub.8 fatty acid, 30
to 45 wt % C.sub.10 fatty acid, 0 to 2 wt % C.sub.12 fatty acid,
based on the total fatty acid content of the medium chain
triglyceride.
[0035] The triglycerides may be prepared by methods known in the
art and are commercially available as MIGLYOL.RTM. 810, 812, 818,
829 (Sasol Germany GmbH, Witten, Germany) or NEOBEE.RTM. 1053, 895,
M-5 (Stepan Company, Northfield, Ill.).
[0036] In another embodiment the carrier is a propylene glycol
diester of saturated vegetable fatty acids with chain lengths of
C.sub.8 and C.sub.10 (caprylic and capric acid). An example of one
such commercially available carrier is MIGLYOL.RTM. 840 (Sasol
Germany GmbH, Witten, Germany). The pharmaceutically acceptable,
non-aqueous carrier may optionally comprise other pharmaceutically
acceptable excipients. Exemplary excipients include sugars (e.g.,
sucrose, glucose, dextrose, galactose, maltose, trehalose,
fructose, maltodextrin); sugar alcohols (e.g., glycol, glycerol,
erythritol, threitol, arabitol, ribitol, sorbitol, dulcitol,
iditol, isomalt, maltitol, lactitol, mannitol, xylitol);
preservatives (e.g., benzoic acid, sorbic acid, meta cresol, sodium
benzoate, potassium sorbate, methylparaben, propylparaben,
butylparaben, benzalkonium chloride, and the like, generally
oil-soluble, with some solubility in the selected carrier); and
antioxidants (e.g., sodium metabisulfite, butylated hydroxy
anisole, butylated hydroxy toluene, sodium sulfite, tocopherol,
thymol, ascorbate, propyl gallate, and the like). In one
embodiment, the carrier optionally comprises mannitol,
maltodextrin, sorbitol, or a combination of two or more
thereof.
[0037] The pharmaceutically acceptable carrier may contain a
gel-forming agent; however, the gel-forming agent may only be
present in an amount that does not cause a gel-depot to form at the
site of in vivo administration of the formulation. In one
embodiment, the pharmaceutically acceptable carrier does not
contain a gel-forming agent. Exemplary gel-forming agents include
cellulose derivatives (e.g., hydroxypropyl cellulose, carboxymethyl
cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose,
methylcellulose); polyoxyethylene and polyoxypropylene polymers or
co-polymers (poloxamers); chitosan acid, and the like. The skilled
artisan will understand that the formation of gels in vivo can be
determined by methods known in the art, such as the use of
histological sections and colored dyes.
[0038] In certain embodiments the non-aqueous, non-solubilizing
carrier has a viscosity of from 5 cP to 200 cP or from 10 cP to 90
cP. In other embodiments the viscosity of the non-aqueous,
non-solubilizing carrier is from 20 cP to 80 cP or from 30 cP to 70
cP. Thus, with reference to this disclosure the person of ordinary
skill will be able to identify other oils, triglycerides, or
non-aqueous compounds that also can be present in the non-aqueous,
non-solubilizing carrier.
Microspheres
[0039] The term "microspheres" includes microspheres,
microparticles, nanoparticles, pellets, cylinders, rods, discs, and
the like. A microsphere can have a spherical, non-spherical or
irregular shape. The microsphere will be of a size suitable for
injection. A typical size range for microspheres is 1000 microns or
less. In a particular embodiment, the microsphere ranges from about
one to about 180 microns in diameter. In yet further embodiments
suitable release profiles are obtained when microspheres range from
about 1 to 100 microns, from about 30 to 90 microns, or from about
50 to 70 microns. In one embodiment the mean microsphere size is
not less than or is equal to about 50, 60 or 70 microns, and
preferably less than about 80, 90, or 100 microns. At larger sizes,
microsphere are preferably substantially non-aggregated to allow
passage through a 25 gauge needle, or a 27 gauge needle, or a 30
gauge needle, or a 31 gauge needle.
[0040] Consistent and superior release profiles are obtained by
controlling size distribution. In one embodiment a mean microsphere
size is about 50 microns and the lower and upper range of
microsphere are about 30 and 90 microns, respectively. Distribution
of microspheres can be described using a mean diameter of the
volume. Mean diameter of the volume distribution represents the
center of gravity of the distribution and is a type of "average
particle size." In various embodiments, the microspheres have a
mean diameter of the volume distribution of about 50 to 70 microns,
about 50 to 60 microns or about 50, 60 or 70 microns, with a
Distribution of Volume (DV) of less than or about 5%, 10%, or 15%
at 30 microns and a DV of greater than or about 80%, 85%, 90% or
95% at 90 microns. In one embodiment, the microspheres have a mean
diameter of the volume distribution of about 60 microns, with a
Distribution of Volume (DV) of less than or about 10% at 30 microns
and a DV of greater than or about 90% at 90 microns.
[0041] Microspheres may be prepared by processes known in the art
and described, e.g., in U.S. Pat. Nos. 7,563,871, 7,456,254,
7,223,440, 6,824,822, 6,667,061, 6,495,164, and 6,479,065, the
disclosures of which are incorporated by reference herein.
[0042] In a further embodiment, the microspheres have a less porous
outer layer, and further can have a non-porous outer layer.
Accordingly, in the formulations disclosed herein the oil does not
have access to the interior spaces or pores or even to a
substantial portion of the interior spaces or pores. It is
specifically, contemplated that for each of the formulations
disclosed herein the microspheres can additionally lack oil (or a
carrier as disclosed herein) in the interior spaces of the
microspheres. Thus, the advantages of the present formulations can
be achieved without the presence of oil in the interior spaces of
the microspheres when formulated.
Polymers The microspheres comprise biocompatible, biodegradable
polymers. A polymer is biocompatible if the polymer and any
degradation products of the polymer are non-toxic to the patient at
administered levels and also possess no demonstrated deleterious or
untoward effects on the patient's body, for example a substantial
immunological reaction at the injection site. Biodegradable means
the polymer will degrade or erode in vivo to form smaller units or
chemical species. Degradation can result, for example, by
enzymatic, chemical and physical processes.
[0043] Exemplary biocompatible, biodegradable polymers include, for
example, polylactides, polyglycolides, poly(lactide-co-glycolides),
polylactic acids, polyglycolic acids, poly(lactic acid-co-glycolic
acid)s, polycaprolactones, polycarbonates, polyesteramides,
polyanhydrides, polyamino acids, polyorthoesters,
polycyanoacrylates, poly(p-dioxanone), polyalkylene oxalates,
biodegradable polyurethanes, blends thereof and copolymers thereof.
Acceptable molecular weights for the biocompatible, biodegradable
polymers can be determined by a person of ordinary skill in the art
taking into consideration factors such as the desired polymer
degradation rate, physical properties such as mechanical strength,
end group chemistry and rate of dissolution of polymer. Typically,
an acceptable range of molecular weight is of about 2,000 Daltons
to about 2,000,000 Daltons. The biocompatible, biodegradable
polymer can also be selected based upon the polymer's inherent
viscosity. Suitable inherent viscosities are about 0.06 to 1.0
dL/g; about 0.2 to 0.6 dL/g; or about 0.3 to 0.5 dL/g.
[0044] In one embodiment, the biocompatible, biodegradable polymer
is a poly(lactide-co-glycolide) copolymer (also referred to as
"PLGA") having a lactide:glycolide ratio from 70:30 to 30:70, or
from 60:40 to 40:60 or about 50:50. The molecular weight of the
poly(lactide-co-glycolide) copolymer is about 10,000 Daltons to
about 90,000 Daltons. In another embodiment, the molecular weight
of the poly(lactide-co-glycolide) copolymer is about 30,000 Daltons
to about 70,000, or from about 50,000 to about 60,000 Daltons.
[0045] The formulation may contain microspheres at a concentration
of from 1 mg/ml to 500 mg/ml; from 25 mg/ml to 300 mg/ml; or from
50 mg/ml to 200 mg/ml.
Active Pharmaceutical Ingredient
[0046] An active pharmaceutical ingredient is a biologically active
compound that has a therapeutic, prophylactic, or other beneficial
pharmacological and/or physiological effect on the patient. The
active pharmaceutical ingredient can also be a mixture of two or
more compounds. The term "peptide" refers to any compound having
two or more consecutive amino acids. As used herein, the term
"peptide" is synonymous with peptide, polypeptide, and protein. In
one embodiment, the peptide has a molecular weight of from 500 Da
to 100 kDa; from 1 kDa to 80 kDa; from 1 kDa to 50 kDa; from 1 kDa
to 30 kDa; or from 1 kDa to 20 kDa. In one embodiment, the peptide
comprises 2 to 500 amino acid residues; 2 to 250 amino acid
residues; 5 to 100 amino acid residues; or 5 to 50 amino acid
residues.
[0047] In one embodiment, the active pharmaceutical ingredient is a
GLP-1 receptor agonist compound, such as an exendin, an exendin
analog, GLP-1(7-37), a GLP-1(7-37) analog, and the like. Exemplary
GLP-1 receptor agonist compounds include exendin-3, exenatide,
GLP-1(1-37), GLP-1(7-37)-NH.sub.2, GLP-1(7-36),
GLP-1(7-36)-NH.sub.2, Leu.sup.14-exendin-4, Leu.sup.14,
Phe.sup.25-exendin-4, exendin-4(1-28), Leu.sup.14-exendin-4(1-28),
Leu.sup.14, Phe.sup.25-exendin-4(1-28), exendin-4(1-30),
Leu.sup.14-exendin-4(1-30), Leu.sup.14, Phe.sup.25-exendin-4(1-30),
liraglutide, and the compounds described in, e.g., U.S. Pat. No.
7,157,555, U.S. Pat. No. 7,220,721, U.S. Pat. No. 7,223,725, and WO
2007/139941, the disclosures of which are incorporated herein by
reference.
[0048] Other peptides known in the art can be used as the active
pharmaceutical ingredient in the formulations described herein.
Exemplary peptides include amylin, amylin agonists (e.g.,
pramlintide, davalintide, Val.sup.27-davalintide); leptin, leptin
agonists (e.g., metreleptin); PYY(3-36) and agonist analogs
thereof; glucagon, glucagon agonists, glucagon antagonists, peptide
chimera of GLP-1 receptor agonists and glucagon agonists, peptide
chimera of human amylin and salmon calcitonin, insulin, heparin,
low-molecular weight heparin, angiotensin, argipressin, argireline,
atosiban, bivalirudin, cetrorelix, desmopressin, enfuvirtide,
deptifibatide, GHRP-2, GHRP-6, gonadorelin, leuprolide,
lysipressin, melanotan, nesiritide, octreotide, oxytocin, PT141,
calcitonin, sermorelin, somatostatin, terlipressin, thymopentin,
thymosin al, triptorelin, vapreotide, elcatonin, ziconotide,
ghrelin, nafarelin, BNP-32, and the like.
[0049] The active pharmaceutical ingredient can also be a small
molecule. A "small molecule" is an organic molecule. Exemplary
small molecules include metformin, sulfonylureas, TZDs, statins
(e.g., atorvastatin, cerivastatin, fluvastatin, Lovastatin,
mevastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin);
non-selective beta blockers and/or alpha-1 blockers (e.g.,
carvedilol, dilatrend, eucardic, carloc); PDE3 inhibitors (e.g.,
cilostazol); antiplatelet drugs, antithrombotic drugs,
anticoagulant drugs, glycoprotein IIb/IIIa inhibitors (e.g.,
abciximab, eptifibatide, tirofiban); antibacterial drugs (e.g.,
ciprofloxacin, norfloxacin, levofloxacin, moxifloxacin,
sparfloxacin, gemifloxacin, ecinofloxacin, delafloxacin); Factor Xa
inhibitors (e.g., glycosaminoglycans, oligosaccharides,
heparinoid); direct Xa inhibitors (e.g., xabans); direct thrombin
(II) inhibitors (e.g., hirudin, argatroban, dabigatran, melagatran,
ximelagatran, defibrotide, ramatroban, antithrombin III, protein
C); thrombolytic drugs (e.g., plasminogen activators, urokinase,
streptokinase, serine endopiptidases); ACE inhibitors (e.g.,
lisinopril, aceon, acertil, armix, coverene, coverex, coversum,
prestarium, prexanil, Prexum, procaptan); ADP receptor/P2Y 12
inhibitors (e.g., clopidogrel, ticlopidine, prasugrel);
prostaglandin analogs (e.g., beraprost, prostacyclin, iloprost,
treprostinil); anticoagulants (e.g., coumarin, coumatetralyl,
dicoumarol, ethyl biscoumacetate, phenprocoumon, warfarin,
clorindione, diphenadione, phenindione, tioclomarol); diuretics
(e.g., hydrochlorothiazide); macrolides (e.g., azithromycin,
clarithromycin, dirithromycin, erythromycin, roxithromycin,
telithromycin); NSAIDs and COX-3 inhibitors (e.g., celecoxib,
etoricoxib, parecoxib); sulphonanilides (e.g., nimesulide), and the
like.
[0050] The skilled artisan will appreciate that the formulations
described herein may contain two or more peptides; two or more
small molecules; or a combination of small molecules and peptides.
For example, the formulation may comprise two different sets of
microspheres, where one set of microspheres contain one peptide
(e.g., pramlintide) and another set of microspheres contain a
different peptide (e.g., metreleptin). In one embodiment, 1 to 99%
of the microspheres comprise one active pharmaceutical ingredient
and 99 to 1% of the microspheres comprise a different active
pharmaceutical ingredient. In another embodiment 30 to 70% of the
microspheres comprise one active pharmaceutical ingredient and 70
to 30% of the microspheres comprise a different active
pharmaceutical ingredient. The skilled artisan will appreciate that
the percentage of each type of peptide in the formulation will be
determined by the relative potency of the peptides. This
formulation advantageously allows high potency peptides to be
combined with low potency peptides for simultaneous delivery to a
patient because the low potency peptides can be provided in more
microspheres and the high potency peptides can be provided in fewer
microspheres in the same formulation. Exemplary combinations of
peptides and/or small molecules that can be administered in
different sets of microspheres and in the same formulation include:
pramlintide and insulin; pramlintide and metreleptin; davalintide
and metreleptin; exenatide and metreleptin; lovastatin and niacin;
atorvastatin and amlodipine; simvastatin and ezetimibe; exenatide
and metformin; and the like.
[0051] The formulations generally contain from about 0.01% (w/w) to
about 50% (w/w) of the active pharmaceutical ingredient (based on
the total weight of the composition). For example, the amount of
active pharmaceutical ingredient can be from about 0.1% (w/w) to
about 30% (w/w) of the total weight of the composition. The amount
of active pharmaceutical ingredient will vary depending upon the
desired effect, potency of the agent, the planned release levels,
and the time span over which the peptide will be released. In
certain embodiments, the range of loading is between about 0.1%
(w/w) to about 10% (w/w), for example, from 0.5% (w/w) to about 5%
(w/w), or from 1% to 5% (w/w). When the active pharmaceutical
ingredient is a GLP-1 receptor agonist, suitable release profiles
can be obtained when the active pharmaceutical ingredient, for
example exenatide, is loaded at about 2% w/w to about 7% w/w,
including at about 2% w/w, about 3% w/w, about 4% w/w, about 5%
w/w, about 6% w/w, or about 7% w/w.
Sugars
[0052] The microspheres may also comprise one or more sugars. A
sugar is a monosaccharide, disaccharide or oligosaccharide or a
derivative thereof. Sugar alcohols of monosaccharides are suitable
derivatives of sugar. Monosaccharides include, but are not limited
to, glucose, fructose and mannose. A disaccharide, as further
defined herein, is a compound which upon hydrolysis yields two
molecules of a monosaccharide. Suitable disaccharides include, but
are not limited to, sucrose, lactose and trehalose. Suitable
oligosaccharides include, but are not limited to, raffinose and
acarbose. The microspheres may further comprise glucose, dextrose,
galactose, maltose, fructose, mannose, sucrose, lactose, trehalose,
raffinose, acarbose, glycol, glycerol, erythritol, threitol,
arabitol, ribitol, sorbitol, dulcitol, iditol, isomalt, maltitol,
lactitol, mannitol, xylitol, or a combination of two or more
thereof. In one embodiment, the sugar is sucrose, glucose, mannose,
or fructose. In one embodiment, the sugar is sucrose.
[0053] The amount of sugar present in the microspheres can range
from about 0.01% (w/w) to about 50% (w/w), such as from about 0.01%
(w/w) to about 10% (w/w), such as from about 0.1% (w/w) to about 5%
(w/w) of the total weight of the composition. In one embodiment,
about 2% (w/w) sucrose is used.
[0054] Alternatively, the amount of sugar present in the
microspheres can be referred to on a weight ratio with the active
pharmaceutical ingredient. For example, the active pharmaceutical
ingredient and sugar can be present in a ratio from about 10:1 to
about 1:10 weight:weight. In particularly preferred embodiments,
the ratio of active pharmaceutical ingredient (e.g., exenatide) to
sugar (e.g., sucrose) is about 3:2 (w/w), 4:2 (w/w), or 5:2 (w/w).
Combinations of two or more sugars can also be used. The amount of
sugar, when a combination is employed, is the same as the ranges
recited above.
Sustained Release
[0055] The compositions are sustained release compositions, meaning
that the active pharmaceutical ingredient contained in the
compositions will be released into the patient over an extended
period of time such as, for example, a period of two days, or three
days, or at least two days, or at least three days, or over a
period of one week, two weeks, one month, three months, or one
year. The release of the active pharmaceutical ingredient is
considered complete when there is no longer a therapeutic level of
active pharmaceutical ingredient in the patient's body, as
determined by the medical judgment of those of ordinary skill in
the art.
[0056] Cmax as used herein is the maximum serum concentration of
drug which occurs during the period of release which is monitored.
Cave as used herein, is the average serum concentration of drug
derived by dividing the area under the curve (AUC) of the release
profile by the duration of the release.
[0057] In one embodiment the ratio of Cmax to Cave is about 3 or
less. This profile is particularly desirable for anti-diabetic or
glucoregulatory polypeptides, such as those described herein. A
ratio of about 3 or less can provide a Cave in a therapeutic window
while avoiding adverse drug side effects which can result from
higher ratios. Further by controlling the physical aspects of the
sustained release composition, as described herein, a superior
desired release profile can be achieved and controlled, for
example, by appropriate selection of carrier properties, such as
viscosity. There is thus provided a reduced burst (i.e. initial
release; e.g., Cmax at 0-1 day). In other embodiments the Cmax to
Cave ratio is from about 1 to about 3, or from 1 to 3, or from
about 2 to about 3, or from 2 to 3. Further, a Cmax, if present,
can be shifted from the burst or initial release period into the
"sustained phase" of release. In one embodiment the Cmax can occur
at at least 7, 14, 21, 28, 35 or 42 days post administration and
can occur at any integer day in between. In a further embodiment
the Cmax occurs at about 21 to 35 days after administration, and in
yet another embodiment is at about 28 to 31 days, and further at
about 28 days after administration. In a further embodiment the
maximal concentration of drug (e.g. plasma concentration) occurs at
at least 7, 14, 21, 28, 35 or 42 days post administration and can
occur at any integer day in between. In yet a further embodiment
the maximal concentration of drug occurs at about 21 to 35 days
after administration, particularly in the case of glucoregulatory
agents such as exendin-4, GLP-1, GIP or their analogs.
Longer Shelf Life
[0058] One advantage offered by the present formulations is a
longer shelf life for the formulation. It was discovered
unexpectedly that sustained release compositions retain remarkable
stability when stored in a non-aqueous carrier as described herein.
In one embodiment the formulation has a shelf life of at least 6
months. In other embodiments the formulation has a shelf life of at
least 1 year, or at least 18 months, or at least 2 years. By "shelf
life" is meant the formulation can be stored or maintained for that
period of time under appropriate environmental conditions while
retaining at least 90% of the desired activity of the active
pharmaceutical ingredient relative to the activity at initial
formulation (as 100%). In another embodiment the active
pharmaceutical ingredient retains at least 95%, or at least 98% or
at least 99% of its desired activity as compared to its activity
immediately before storage. When the formulation contains
microspheres, shelf life also refers to the retention of particle
size and/or morphology of the microspheres. Retention of size
morphology can be determined by microscopic examination, the use of
which is known to persons of ordinary skill in the art.
[0059] When formulated as disclosed herein a peptide or protein as
active ingredient is less susceptible to oxidation and to
hydrolysis, either chemical or proteolytic, both during storage and
during its sustained release period after injection. The addition
of an anti-oxidant or other stabilizer is not required in these
formulations, particularly those where the carrier is a medium
chain triglyceride.
Reduced Burst Release
[0060] Another advantage of the present formulations is that
formulations according to the present disclosure offer a
significantly reduced burst release rate compared with other
formulations. When previously available injectable sustained
release formulations are injected into a patient there is often a
"burst" of active ingredient or agent associated with the
injection. Without wanting to be bound by any specific theory, it
is believed this burst is caused by that quantity of active
pharmaceutical ingredient in the formulation that is not retained
within the polymer that is released over time. By "burst release"
is meant that quantity of active pharmaceutical ingredient released
within the first 24 hours after injection. In other embodiments it
is that quantity of active that is release over 1 hour, or 2 hours,
or 4 hours, or 8 hours, or 12 hours after injection. In various
embodiments the formulation of the invention has a burst release
after injection of less than 10% or less than 5%, or less than 3%,
or less than 2.5%, or less than 2%, or less than 1% or less than
0.75% or less than 0.5% or less than 0.25% or less than 0.1%.
Percentages refer to the percentage of the total amount of active
pharmaceutical ingredient in the injected formulation. Following
injection of the formulation in the patient, the burst release may
occur at any time up to about 24 hours, thereafter there may be a
lag time where substantially no active pharmaceutical ingredient is
released from the microspheres, and then the polymeric microspheres
begin degrading and releasing the active pharmaceutical ingredient.
The skilled artisan will appreciate that the time period when the
burst release occurs may vary from patient to patient.
[0061] Burst can be assessed by measuring the proportion of the
total area under the curve for a particular time period following
administration of a drug. Area under the curve (AUC) is a well
established measurement in the pharmaceutical sciences and measures
the amount of drug or active ingredient that reaches the
bloodstream in a set period of time. As is well known in the art,
the period of time selected will varying depending on the time
period the concentration of the drug in the blood is expected to be
detectable or within the drug's therapeutic window. AUC is
calculated by plotting the concentration of the drug in the blood,
for example plasma concentrations, at various times during the
selected time period and then calculating the total area under the
curve obtained. In one exemplary embodiment, the area under the
curve is measured for a 42 day period and using the formulations
described herein, the release or burst as measured within the first
24 hours is 5% or less, 2% or less, 1.5% or less, 1% or less, or
0.5% or less of the total AUC. In another embodiment, the
formulations described herein result in a burst or proportion of
the AUC that is 20% or less, 15% or less, 10% or less, 5% or less,
or 2% or less than that obtained when the sustained release
composition is contained in a carrier in which the active
pharmaceutical ingredient is soluble.
[0062] In another embodiment, the formulations described herein
limit initial burst such that the upper limit of the therapeutic
window for the active pharmaceutical ingredient is not exceeded.
The therapeutic window is the range of concentration of active
pharmaceutical ingredient in the circulation, above which the
active pharmaceutical ingredient has its desired effect, but below
the concentration at which the adverse effects associated with the
active pharmaceutical ingredient outweigh the benefits as would be
generally accepted among physicians. In one exemplary embodiment,
the active pharmaceutical ingredient is an exendin, for example
exenatide, or agonist analogue thereof, and administration of the
formulations described do not result in a circulating level of
active pharmaceutical ingredient exceeding 400 pg/ml during the
first 24 hours following administration. In another exemplary
embodiment the active pharmaceutical ingredient is an exendin, for
example exenatide, or agonist analogue thereof, and administration
of the formulations described does not result in a circulating
level of active pharmaceutical ingredient exceeding 350 pg/ml
during the first 24 hours following administration.
[0063] Initial burst can also be assessed by comparing circulating
concentrations of the active pharmaceutical ingredient in a time
period immediately following administration of the formulation with
the circulating concentration of the drug in a second time period
that immediately follows the first. In one embodiment, use of the
formulations of the present disclosure result in circulating
concentrations of active pharmaceutical ingredient during the first
24 hours following administration that do not exceed the
circulating concentration during the next 24 hour period. In
another embodiment, use of the formulations of the present
disclosure result in average circulating concentration of active
pharmaceutical ingredient during the first 24 hours following
administration do not exceed the average circulating concentration
during the next 24 hour period.
Methods of Storing
[0064] Another aspect provides methods of storing the sustained
release formulations described herein. The methods of storing the
formulations described herein may also be referred to as methods of
preventing the degradation of the microspheres. By "storing" is
meant that the formulation is retained for a period of time within
its container without adding any additional component to the
container and without removing the formulation from the container
(e.g., in the manufacturing facility, during transport, in the
pharmacy). The storage time will typically be the amount of time
between packaging of the formulation and its use by the patient.
After the storage time the formulation is administered to the
patient in need thereof. "Administering" to the patient includes
self-administration. The methods involve storing the sustained
release formulations for a period of at least 1 week, at least 2
weeks, at least 1 month, at least 3 months, at least 1 year, at
least 18 months, or at least 2 years. In some embodiments, the
formulations can be stored at 5.degree. C. or 25.degree. C. There
is minimal degradation of the microspheres when the formulations
are stored for such extended periods of time.
[0065] In another embodiment the invention provides methods of
maintaining the potency of (e.g., preventing the loss of biological
activity) and/or purity (e.g. preventing chemical changes in the
molecule) an active pharmaceutical ingredient. Thus, a peptide or
protein or other API that has undergone a chemical change (e.g.
oxidation) may result in a loss of purity, but may still retain its
potency. The methods involve storing a microsphere comprising a
active pharmaceutical ingredient in a non-aqueous carrier as
described herein for a period of time, whereby the potency and/or
purity of the active pharmaceutical ingredient is maintained by the
microspheres and the non-aqueous carrier. In the formulations
described herein, at least 80%, at least 90%; at least 95%; at
least 98%; or at least 99% of the potency and/or purity of the
active pharmaceutical ingredient is retained for a period of time
of at least 1 week, at least 2 weeks, at least 1 month, at least 3
months, at least 1 year, at least 18 months, or at least 2
years.
Methods of Administering/Treatment
[0066] In another aspect the present invention provides methods of
administering an active pharmaceutical ingredient to a patient in
need thereof. The methods involve administering to the patient a
formulation or composition as described herein. Any of the
formulations described herein can be administered by parenteral
administration, using any of the methods described herein. For
example, the formulations can be administered by subcutaneous,
intra-muscular, intra-peritoneal, intra-abdominal, intravenous, or
any suitable manner of administration. In one embodiment, the
formulations described herein are administered subcutaneously. In
one embodiment the methods involve injecting the formulation
without the patient performing a prior step of combining the
sustained release composition with a second carrier.
[0067] In one embodiment the administration does not comprise a
mixing step. A mixing step is a step where the microspheres are
combined with a carrier prior to injection. In various embodiments
the mixing step is a step where the microspheres are combined with
a carrier within the 1 week period prior to injection in the
patient. The carrier can be a non-aqueous carrier, such as those
described herein. Administration of the formulation refers to the
complete process of the user interacting with the formulation,
including mixing, combining any ingredients forming the
formulation, and the actual injection or other form of providing
the formulation to the patient.
[0068] The frequency of administration can vary depending on any
one or a combination of factors such as the amount of the
formulation administered, the release profile of the formulation,
the amount of active pharmaceutical ingredient in the formulation,
and the circulating level of active pharmaceutical ingredient to be
achieved. In particular embodiments, the formulations described
herein can be administered once daily, once per week, once every
two weeks, once a month, once every two months, once every three
months, once every four months, once every six months or once per
year. In one embodiment, the formulation is administered once a
week. In another embodiment, the formulation is administered once a
month.
[0069] When the formulations comprise a GLP-1 receptor agonist,
such as GLP-1 or an analog thereof, or an exendin (e.g., exenatide)
or an analog thereof, they can be used to treat numerous diseases,
such as diabetes (e.g., Type 1 diabetes, Type II diabetes,
gestational diabetes), impaired glucose tolerance, hyperglycemia
(e.g., fasting and postprandial), obesity, overweight,
non-alcoholic fatty liver disease, non-alcoholic steatohepatitis
(NASH), and the like. The formulations comprising a GLP-1 receptor
agonist (e.g., exenatide) will also be useful to stimulate insulin
release; lower plasma glucagon; reduce food intake, reduce
appetite, decrease gastric motility, delay gastric emptying, lower
plasma lipid (e.g., triglycerides, cholesterol) levels, and the
like. These methods of treatment are described, for example, in
U.S. Pat. No. 5,424,286, U.S. Pat. No. 6,858,576, U.S. Pat. No.
6,872,700, U.S. Pat. No. 6,956,025, and U.S. Pat. No. 6,956,025,
and WO 2007/022518, the disclosures of which are incorporated by
reference herein.
[0070] In certain embodiments, administration of any of the
formulations provided herein comprising a glucoregulatory peptide
such as an exendin, e.g. exenatide, result in a 2 hour plasma
glucose of less than 300 mg/dl, less than 275 mg/dl, less than 250
mg/dl, or less than 225 mg/dl. In a particular embodiment
administration of any of the formulations provided herein
comprising a glucoregulatory peptide such as an exendin, e.g.
exenatide, results in a 2 hour plasma glucose of less than 200
mg/dl. In other embodiments, administration of any of the
formulations provided herein comprising a glucoregulatory peptide
such as an exendin, e.g. exenatide, results in a 2 hour plasma
glucose of less than 190 mg/dl, less than 180 mg/dl, less than 170
mg/dl, less than 160 mg/dl, or less than 150 mg/dl. In certain
embodiments, administration of any of the formulations provided
herein comprising a glucoregulatory peptide such as an exendin,
e.g. exenatide, results in a 2 hour plasma glucose less than 140
mg/dl. In further embodiments, administration of any of the
formulations provided herein comprising a glucoregulatory peptide
such as an exendin, e.g. exenatide, results in a venous or
capillary fasting blood glucose (FBG) level of less than 200 mg/dl,
less than 175 mg/dl, less than 150 mg/dl, less than 140 mg/dl, less
than 130 mg/dl, less than 120 mg/dl, or less than 115 mg/dl. In one
embodiment, a FBG level of less than 110 mg/dl is achieved, while
in another embodiment a FBG level of less than 100 mg/dl is
achieved.
[0071] In additional embodiments, administration of any of the
formulations provided herein comprising a glucoregulatory peptide
such as an exendin, e.g. exenatide, results in a 2 hour venous or
capillary blood glucose level of less than 300 mg/dl, less than 275
mg/dl, less than 250 mg/dl, less than 225 mg/dl, or less than 200
mg/dl. In a particular embodiment administration of any of the
formulations provided herein comprising a glucoregulatory peptide
such as an exendin, e.g. exenatide, results in a 2 hour blood
glucose level of less than 180 mg/dl. In further embodiments,
administration of any of the formulations provided herein
comprising a glucoregulatory peptide such as an exendin, e.g.
exenatide, result in blood glucose levels of less than 170 mg/dl,
less than 160 mg/dl, less than 150 mg/dl, less than 140 mg/dl, less
than 130 mg/dl, or less than 120 mg/dl. In particular embodiments,
administration of any of the formulations provided herein
comprising a glucoregulatory peptide such as an exendin, e.g.
exenatide, result in a 2 hour venous blood glucose level of less
than 120 mg/dl, while in other embodiments, a 2 hour capillary
blood glucose level of less than 140 mg/dl is achieved.
[0072] In one embodiment, glucose levels are average glucose levels
calculated over a chosen time period. Specific examples include,
but are not limited to, daily average glucose levels, weekly
average glucose levels, monthly average glucose levels or yearly
average glucose levels. Two hour circulating glucose levels are
determined after an oral glucose tolerance test (OGTT). In the
standard test, 75 g of anhydrous glucose is dissolved in 250-300 ml
of water and administered over 5 minutes. In children, glucose is
administered at a rate of 1.75 g/kg body weight up to a maximum of
75 grams of glucose. A baseline glucose level is obtained prior to
ingestion and then typically every 30 minutes for 2 hours. For
gestational diabetes, a 100 g, 3 hour test is often used.
[0073] Because glucose freely crosses the cell membrane of red
blood cells, erythrocyte hemoglobin undergoes a nonenzymatic
glycosylation at the amine residues. Hemoglobin Alc (HbAlc) refers
to the percentage of hemoglobin molecules with glucose moieties
attached to the N-terminal valines of each of the two beta-chains.
Glycohemoglobin includes HbAlc along with other forms of hemoglobin
where glycosylation has occurred at other amino acids. The
percentage of hemoglobin molecules undergoing glycosylation is
proportional to the average ambient glucose concentrations during
the previous during the previous 60-90 days. HbAlc is a commonly
used measure to assess the state of glycemic control in patients
with diabetes.
[0074] In one embodiment, administration of any of the formulations
provided herein comprising a glucoregulatory peptide such as an
exendin, e.g. exenatide, results in a reduction to, maintenance of,
or both of HbAlc levels of less than 8%. In another embodiment
HbAlc levels are reduced to, maintained at, or both to less than
7.5%, while in yet another embodiment, HbAlc levels are reduced to,
maintained at, or both at less than 7%. In further embodiments,
administration of any of the formulations provided herein
comprising a glucoregulatory peptide such as an exendin, e.g.
exenatide, results in a reduction to or maintenance of, or both of
HbAlc levels to less than 6.5%, less than 6%, less than 5.5%, less
than 5% less than 4.5% or less than 4%. Thus, the compositions
disclosed herein are useful in a method of reducing or maintaining
HbAlc levels in the blood, the methods comprising administering a
composition disclosed herein. In another embodiment, administration
of any of the formulations provided herein comprising a
glucoregulatory peptide such as an exendin, e.g. exenatide, results
in a reduction to, maintenance of, or both of glycosylated
hemoglobin levels of less than 10%. In another embodiment,
glycosylated hemoglobin levels are reduced to, maintained at, or
both to less than 9.5%; while in yet another embodiment,
glycosylated hemoglobin levels are reduced to, maintained at, or
both at less than 9%. In further embodiments administration of any
of the formulations provided herein comprising a glucoregulatory
peptide such as an exendin, e.g. exenatide, results in a reduction
to, or maintenance of, or both of glycosylated hemoglobin levels to
less than 8.5%, less than 8%, less than 7.5%, less than 7% less
than 6.5%, less than 6%, less than 5.5%, less than 5%, less than
4.5% or less than 4%. In other aspects administration of any of the
formulations provided herein comprising a glucoregulatory peptide
such as an exendin, e.g. exenatide, results in a lower of HbAlc by
at least 0.2%, at least 0.4%, at least 0.6%, at least 0.8%, at
least 1%, at least 1.2%, at least 1.4%, at least 1.6%, at least
1.8%, or at least 2%. Thus, the invention provides methods of
reducing or maintaining glycosylated hemoglobin levels in the
blood, the methods involving administering a composition described
herein.
[0075] It should be realized that a subject in need of lowering of
blood glucose is not limited to patients having diabetes mellitus,
but may include any subject suffering from hyperglycemia for
whatever reason including, but not limited to, injury, trauma,
surgery, stroke and myocardial infarction. The amount of glucose
lowering will vary with the subject in question and depend on
factors such as the severity of the hyperglycemia and the severity
of the disease, disorder or condition in question.
EXAMPLES
[0076] The following non-limiting examples provide further
illustrations of making and using the formulations described
herein, and are not intended to limit the scope of the appended
claims. With respect to the Examples herein, MCT oil refers to
medium chain triglyceride oil which is commercially available as
MIGLYOL.RTM. 812 (Sasol Germany GmbH, Witten, Germany).
Example 1
[0077] Microspheres may be prepared by processes known in the art
and described, e.g., in U.S. Pat. No. 7,563,871 and U.S. Pat. No.
7,456,254. Microspheres comprising a poly(lactide-co-glycolide)
copolymer having dispersed therein 5% (w/w) exenatide and 2% (w/w)
sucrose were obtained. The poly(lactide-co-glycolide) copolymer had
a ratio of lactide:glycolide of 1:1. These microspheres are
currently being developed by Amylin Pharmaceuticals, Inc. (San
Diego, Calif.), Alkermes, Inc. (Cambridge, Mass.), and Eli Lilly
and Company (Indianapolis, Ind.) for a once-weekly formulation for
treating diabetes. Gedulin et al, Diabetologia, 48:1380-1385
(2004).
Example 2
[0078] The stability of the microspheres from Example 1 was
investigated to determine their stability over an extended period
of time while stored in a non-aqueous carrier. Microspheres from
Example 1 were stored for a period of 6 months at 5.degree. C. in a
formulation comprising a non-aqueous carrier (i.e., sesame oil; MCT
oil; and ethyl oleate, which is a monoglyceride). The control was
an aqueous formulation comprising the microspheres from Example 1
in an aqueous carrier containing carboxymethylcellulose and a
surfactant.
[0079] The stability of the microspheres was determined by
morphology and particle size via examination under a microscope.
Exenatide purity, potency (by HPLC evaluation), and in vitro
release were also determined. As shown in Table 1, after 6 months
of storage the physical structure (i.e., size, morphology) of the
microspheres did not change.
[0080] As shown in Table 2, the microspheres stored in a MCT oil
showed no change in the purity of exenatide based on an HPLC
analysis. Impurities might also be referred to as degradation
products from the peptide. High purity means relatively little
degradation of the peptide. The purity is relative to the
formulation at time zero. The microspheres stored in sesame oil and
ethyl oleate showed a slight decrease in the purity of exenatide.
The impurities did not appear to be oil or
poly(lactide-co-glycolide) polymer related (based on retention
times), but appeared to be related to the stability of exenatide
itself.
[0081] Table 3 shows that the potency of exenatide did not
significantly decrease over the 6 month period regardless of the
non-aqueous carrier that was used.
TABLE-US-00001 TABLE 1 Particle size and morphology using
microscope size (.mu.m) (standard deviation (.mu.m)) morphology T =
0 1 month 6 months 0 to 6 months sesame oil 64 (22) 63 (23) 64 (12)
no change MCT oil 65 (19) 60 (22) 61 (17) no change ethyl oleate 64
(16) 62 (16) 59 (13) no change
TABLE-US-00002 TABLE 2 Change in Purity of Exenatide Containing
Formulation % purity of exenatide 1 % % 6 % t = 0 month change* 3
month change* month change* sesame 95.93 95.68 -0.25 94.55 -1.38
95.00 -0.93 oil MCT 95.63 95.56 -0.07 94.67 -0.96 95.50 -0.13 oil
ethyl 95.60 95.80 0.20 93.67 -1.93 94.70 -0.90 oleate *Changes less
than 0.5% are considered to be insignificant
TABLE-US-00003 TABLE 3 Change in Potency of Exenatide Based on
Carrier in Formulation time carrier zero 1 month 3 months 6 months
sesame oil 97 104 98 98 MCT oil 94 108 99 99 ethyl oleate 95 98 99
100
Example 3
[0082] The pharmacokinetics of the formulations in Example 2 were
determined, except that 2% (w/w) lecithin was added to the ethyl
oleate carrier. Single injections with a dose of 53 mg/ml of
microspheres per ml of non-aqueous carrier were administered to 6
rats with a 21G needle. In the study, a comparison was also made to
the microspheres from Example 1 that were mixed with an aqueous
carrier just before injection.
[0083] FIG. 1 provides a comparison of the pharmacokinetics of the
four different formulations of microspheres containing exenatide.
In three formulations, the carrier is an oil (e.g., sesame oil; MCT
oil; ethyl oleate). In one comparative formulation, the carrier is
an aqueous diluent. As can be seen from the data, the formulations
having an oil carrier had reduced burst when compared to the
formulation having an aqueous carrier.
[0084] FIG. 2 is a graphical simulation of data extrapolated from
FIG. 1 of the plasma exenatide concentration over time of the
formulation comprising the MCT oil carrier and the comparative
formulation comprising the aqueous carrier. The plasma
concentration plateau of exenatide may be reached after about 5
dosings.
Example 4
[0085] A formulation comprising the microspheres of Example 1 in an
aqueous carrier and a formulation comprising the microspheres of
Example 1 in an MCT carrier were prepared. The burst release was
evaluated by adding about 0.75 mL of the formulations to a 10 mM
HEPES release buffer. The mixture was agitated to ensure that the
microspheres achieved full contact with the HEPES release buffer.
After incubation at 37.degree. C. for one hour, the mixture was
centrifuged and the aqueous phase was analyzed by HPLC to determine
the burst release. The concentration of the dose tested for release
was 150 mg/mL.
[0086] FIG. 3 shows the lower burst release of the formulation
having the oil carrier compared to the formulations having an
aqueous carrier. The graph shows that with an aqueous carrier,
about 0.6% of exenatide was released in the burst. With the
formulation having the MCT oil carrier, less than 0.1% of exenatide
was released in the burst.
[0087] FIG. 4 illustrates the in vivo release profile in rats over
10 hours for the formulation of Example 1 in MCT oil compared to a
formulation comprising the same microspheres in an aqueous (saline)
carrier. In the time period following sub-cutaneous administration
of the formulation, the entrance of exenatide into the plasma was
markedly lower than the same microspheres administered in the
aqueous carrier. The formulation of the invention shows no burst
release, and a markedly more gradual entrance into the blood plasma
versus the aqueous formulation. In contrast, the aqueous
formulation showed a burst release followed by a sharper entrance
into the blood plasma.
Example 5
[0088] Microparticles were prepared in a manner similar to that
described in the examples in U.S. Pat. No. 5,439,688, the
disclosure of which is incorporated by reference herein. Eight
samples were prepared by briefly mixing an active pharmaceutical
ingredient (i.e., davalintide, pramlintide, metreleptin, bovine
serum albumin, sodium salicylate, salicylic acid, minocycline HCl,
insulin) and polymer (i.e, poly(lactide-co-glycolide) copolymer or
polycaprolactone/PLGA copolymer) and then the mixture was placed in
a grinder to obtain a well-homogenized powder. Mixtures ranged from
2% to 10% w/w of the active pharmaceutical ingredient. The mixed
powder was transferred to an extruder where the temperature was
adjusted according to the chosen polymer. Some polymers needed
higher temperatures to produce a melt with good flow properties.
The extruder contained twin screws that moved clockwise to produce
efficient mixing. The material was extruded through a 1.5 mm
orifice, collected, cooled at room temperature, and cut into short
strands about 1-2 inches long. These strands were then fed into a
12-tooth rotor mill, followed by a sieving step to produce
microparticles of about 20 to 100 microns. The microparticles were
collected and stored at 5.degree. C. until further use.
[0089] Experimental samples were prepared by dispersing about 50 mg
of the microparticles into 0.75 mL of a MCT oil carrier. The
samples were stored at 5.degree. C. and 25.degree. C. for two days,
two weeks, or one month, at which times representative samples were
tested. The fraction of drug that remained in the microparticles
and the fraction of drug that partitioned into the MCT oil carrier
were determined. Briefly, the samples were centrifuged to separate
the microparticles from the MCT oil carrier. Each portion was
treated independently to determine the amount of drug it contained.
Results are reported on the basis of the percent residing in each
independent portion.
TABLE-US-00004 TABLE 4 PLGA copolymer; 2 Days Storage at 5.degree.
C. Compound Microparticles MCT Carrier davalintide 99.8% 0.2%
pramlintide 100.0% 0.0% metreleptin 100.0% 0.0% bovine serum
albumin 100.0% 0.0% sodium salicylate 99.5% 0.5% salicylic acid
98.9% 1.1% minocycline 99.1% 0.9%
TABLE-US-00005 TABLE 5 PLGA copolymer; 1 Month Storage at 5.degree.
C. Compound Microparticles MCT Carrier davalintide 99.4% 0.6%
pramlintide 99.7% 0.3% metreleptin 100.0% 0.0% bovine serum albumin
100.0% 0.0% sodium salicylate 98.7% 1.3% salicylic acid 99.9% 0.1%
minocycline 99.9% 0.1% insulin 99.5% 0.5%
TABLE-US-00006 TABLE 6 PLGA copolymer; 2 Days Storage at 25.degree.
C. Compound Microparticles MCT Carrier davalintide 100.0% 0.0%
pramlintide 100.0% 0.0% metreleptin 100.0% 0.0% bovine serum
albumin 100.0% 0.0% sodium salicylate 97.7% 2.3% salicylic acid
99.1% 0.9% minocycline 99.4% 0.6%
TABLE-US-00007 TABLE 7 PLGA copolymer; 1 Month Storage at
25.degree. C. PLGA Polymer; 1 Month Storage at 25.degree. C.
Compound Microparticles MCT Carrier davalintide 100.0% 0.0%
pramlintide 100.0% 0.0% metreleptin 100.0% 0.0% bovine serum
albumin 100.0% 0.0% sodium salicylate 98.5% 1.5% salicylic acid
99.8% 0.2% minocycline 99.6% 0.4% insulin 99.3% 0.7%
TABLE-US-00008 TABLE 8 polycaprolactone/PLGA copolymer; Two Weeks
Storage 25.degree. C. 5.degree. C. MCT Compound Microparticles MCT
Carrier Microparticles Carrier pramlintide 100.0% 0.0% 100.0%
0.0%
[0090] The data in Tables 4-8 illustrate the broad applicability of
the sustained release formulations described herein to a variety of
different active pharmaceutical ingredients, including peptides and
small molecules. The compositions have been successfully produced
using a variety of peptides, bovine serum albumin, and even a
selection of small molecules. Surprisingly salicylic acid, which is
oil soluble, did not migrate into the MCT carrier oil, despite that
its solubility in the MCT oil is greater than 30 mg/ml. Thus, the
microparticles remain intact upon storage in MCT even when the
active pharmaceutical ingredient is soluble in MCT. The data
further illustrate that the compositions can be successfully
produced even using other polymer mixtures in the
microparticles.
Example 6
[0091] The percentage purity of exenatide was measured by HPLC at
one month intervals over a 9 month period in the following four
formulations: (i) a formulation comprising the microspheres of
Example 1 stored in an oil MCT oil carrier at 5.degree. C.; (ii) a
formulation comprising the microspheres of Example 1 stored in an
MCT oil carrier at 25.degree. C.; (iii) dry microspheres of Example
1 that had been stored in a container for 9 months at 5.degree. C.
without a liquid carrier, and that were then admixed with an
aqueous carrier immediately prior to the study; and (iv) dry
microspheres of Example 1 that had been stored in a container for 9
months at 25.degree. C. without a liquid carrier, and that were
then admixed with an aqueous carrier immediately prior to the
study.
[0092] FIGS. 5A and B show the following: (i) exenatide had a
purity greater than 93% at 6 months and 9 months in the formulation
with the oil carrier at a temperature of 5.degree. C.; (ii)
exenatide had a purity greater than 86% at 6 months and 9 months in
the formulation with the oil carrier at a temperature of 25.degree.
C.; (iii) exenatide had a purity of greater than 94% at 6 months
where the microspheres had been stored dry at 5 OC; and (iv)
exenatide had a purity of greater than 90% at 6 months in the
formulation where the microspheres had been stored dry at a
temperature of 25.degree. C. In FIG. 5A, the purity of exenatide
was determined by strong cation exchange HPLC. In FIG. 5B, the
purity of exenatide was determined by reverse-phase HPLC.
Example 7
[0093] Formulations containing the microspheres from Example 1 and
an MCT oil carrier were stored at 50 and the potency of exenatide
was measured at monthly intervals for 9 months. Additionally,
formulations containing the microspheres from Example 1 and an MCT
oil carrier were stored at 250 and the potency of exenatide was
measured at monthly intervals for 6 months. FIG. 6 presents the
results which show that the potency of exenatide was preserved for
at least 9 months.
Example 8
[0094] The physical integrity of a formulation containing the
microspheres from Example 1 in an MCT oil carrier was analyzed.
After storage for a period of 6 months at 5.degree. C., the
molecular weight of the poly(lactide-co-glycolide) copolymer did
not change relative to time zero. After storage for a period of 6
months at 25.degree. C., the molecular weight of the
poly(lactide-co-glycolide) copolymer decreased by 6 kDaltons, which
was comparable to the molecular weight change of dry microspheres
(i.e., microspheres stored for 6 months at 25.degree. C. not in any
carrier). The mean diameter of the microspheres was measured after
storage at 3, 6, and 9 months at either 5.degree. C. or 25.degree.
C., and no change in mean diameter was detected relative to time
zero.
Example 9
[0095] The ratio of lactide/glycolide for the microparticles was
also investigated for use with various APIs. The Table below
provides the various lactide/glycolide ratios used.
TABLE-US-00009 Approx polymer MW Lactide/Glycolide Polymer Drug
(kDa) ratio for PLGA PLGA davalintide 10 50/50 PLGA pramlintide 10
50/50 PLGA Leptin 10 75/25 PLGA BSA 25 50/50 PLGA Na Salicylate 25
50/50 PLGA Salicylic acid 25 50/50 PLGA Minocycline 10 75/25 PLGA
Insulin 25 50/50 1.1:1 PCL/PLGA pramlintide PCL = 150 50/50 PLGA =
10
[0096] All publications and patents are incorporated by reference
herein. The foregoing has been described in detail, and the skilled
artisan will recognize that modifications may be made without
departing from the spirit or scope of the disclosure or appended
claims.
* * * * *