Sustained release formulations using non-aqueous carriers.

(54) Title: SUSTAINED RELEASE FORMULATIONS USING NON-AQUEOUS CARRIERS.

(54) Title: SUSTAINED RELEASE FORMULATIONS USING NON-AQUEOUS CARRIERS.

(57) Summary

The present invention relates to one component injectable sustained release formulations which comprise microspheres containing active pharmaceutical ingredients (eg, 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 extended shelf life benefits for formulation stability and potency and sustained release of active pharmaceutical ingredients to reduce medication dosing frequency and increase patient compliance.

(57) Abstract

The disclosure provides one-component, injectable, sustained release formulations which comprise microspheres containing active pharmaceutical ingredient (eg, 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 Ufe for the stability and potency of the formulation and sustained release of active pharmaceutical ingredient to reduce the frequency of medication dosing and to increaee patient compliance.

<img file="MX352189B_D0001.tif" />

PATENT TITLE No. 352189

Headlines):

AMYLIN PHARMACEUTICALS, LLC; ASTRAZENECA PHARMACEUTICALS LP

Address: 9373 Towne Center Orive, San Diego, California, 92121, USA; 1800 Concord Pike,

Wilmington, Delaware, USA

D nomination: SUSTAINED RELEASE FORMULATIONS USING NO CARRIERS

AQUEOUS.

CIP:

Classification:

CPC &lt;

Inventor (s)

<img file="MX352189B_D0002.tif" />

0; * e20

1994

0M&gt; 02.

ndustrial nanas (DOF 12/15/1999. amended on 02/04/2000, 07/29/2004.

A61K47 A6

A61K '1K9 / 1647; A61 cse &lt;:¹marV l. j ^ Nning

Number:

MX / a / 20T1 / Q0239á Validity: VBÜ ^ Mños »4,

Date of Ve Date of Exp n: 10 of

The reference patent In accordance with the article from the filing date

Whoever subscribes this title lo h (Official Gazette of the Federation (01/25/2006. 05/06/2009, 06/01/2010, 06/18 / Regulations of the Mexican Institute of articles 1, 3^or, 4th, 5th fraction V subsection a), 12/27/1999, amended on 10/10/2002, 07/29/2004, 08/04 General Deputy, Coordinator, Divisional Directors, Departmental Titles and other subordinates of the Institute Mexican from the

04/08/2004 and 09/13/2007).

^ 61K9 / 16; A61K9 / 51; A61K31 / 40;

A61K9 / 16 17; A61K9 / 1623;

G OEHftTMAN; ROBERT N.

le £ 009. ^ pM Industrial.

Non-extendable, counted under the Rights of the Industrial Property Law 5/1999, 01/26/2004, 06/16/2005, subsection a), 4th and 12th sections I and III of 07/15/2004, 07/28 / 2004 and 7/09/2007);

Mexican Law of Industrial Property (DO F. Agreement that delegates powers to the Eglonal Directors. Divisional Deputy Directors, Coordinators

This official letter is signed with an advanced electronic signature (FIEL), based on articles 7 BIS 2 of the Industrial Property Law; 3 of its Regulations, and 1 section III, 2 section V, 26 BIS and 26 TER of the Agreement establishing the guidelines for the use of the Payment and Electronic Services Portal (PASE) of the Mexican Institute of Industrial Property, in the procedures indicated.

THE DIVISIONAL DIRECTOR OF PATENTS

NAHANNY CANAL REYES

Ξ Original Chain:

“NAHANNY MARISOL CANAL REYES | 00001000000403252793 | Administration Service: Tax | 1695 || MX / 2017/92840 | MX / a / 2011/002398 | PCT patent title | 1223 | GAGV | Page (s)

2 | APFNcLMZ9iRo3ml8NyEULIIW + 7l =

<img file="MX352189B_D0003.tif" />

Digital stamp:

xm4DsP1cFSMP9XRS3DxEHSNBBzEoRjNRoTwOVe8yUmDDEm2lkjhjKNhy1 JGAZcBVrfY46WDIqzs / YA4alDlrwFGN2 m9m2wvLGxAzqTUk5zFNS¡4ILiaJN4RBiWYWN3emrR4zXEtb6CM + JVz4rso9O1i9HeCU769n6GwO2jUt3ztsFHoZiTo DsfZJA0QEfmVRzyGpOEiq0tRafR2OajlEOPUgddCx3BCsy7ba + 8h2FIUafg9z8iRuSZupRmKwT5k9z5bntXw2h / 2WP IOUfhAcmlxFc7XxjrOaeZOHdp2gkQQqx896l2A + FEmgzT61 pjtY4RmHbYEiPdMli68cBXkzw == * Additional information on the back

Arenal No 550. Floor 1. Pueblo Santa María Tepepan. Xochimíco, 16020 Mexico City (55) 53340700 VAwgob.mx/irnpi

<img file="MX352189B_D0004.tif" />

Αχ / i. ..a _ IMPI

SUSTAINED RELEASE FORMULATIONS WgaMBQs ^

INDUSTRIAL

<img file="MX352189B_D0005.tif" />

NON-AQUEOUS CARRIERS

CROSS REFERENCE TO RELATED REQUESTS

This application claims priority to US Application No. 61 / 094,371 filed September 4, 2008, the disclosure of which is incorporated by reference herein.

i, '

BACKGROUND OF THE INVENTION

Injectable sustained-release formulations offer the

Opportunity to provide therapeutic amounts of active pharmaceutical ingredients for extended periods of time from a single injection, thus eliminating the need for one or two injections: daily. Currently injectable sustained-release formulations¹ Available ones use, for example, microspheres and an aqueous carrier, which have several disadvantages. The formulations do not offer long-term stability in the aqueous carrier, thus it is required to separate the packaging and storage for the microspheres and aqueous carrier, and the patient must take: various measures to combine the microspheres and aqueous carrier before administering them to the injection.

* IMPI \

INSTITUTO MSXICANO, Dt LA PROPIEDAD ¹ INDUSTRIAL

Another disadvantage of currently available injectable microsphere formulations is a high-burst release after injection, which causes undesirable in vivo release of the ingredient. active pharmaceutical in a single blast. When medications have 5 toxic or harmful side effects, this is undesirable.

There is a need for formulations and methods to 'safely administer sustained release pharmaceutical formulations' to patients so that the active ingredient is released in vivo over a prolonged period of time and without an unacceptable initial burst release. Ideally, the active ingredient is released in order to maintain levels within the therapeutic window, that is, in a range of: concentration above that necessary to cause the desired clinical effect, I.

but below that where the undesirable side effects are outweighed by the benefits of the drug. It is also necessary that this 15: active pharmaceutical ingredient be provided in a way that is easy and convenient for the patient to self-administer and that it is provided in a formulation that maintains stability for a prolonged period of time in a liquid state. The description addresses these as well as other important purposes.

<img file="MX352189B_D0006.tif" />

IMPI

MEXICAN INSTITUTE

Say THE INDUSTRIAL PROPERTY

BRIEF DESCRIPTION OF THE INVENTION

The disclosure provides formulations comprising microspheres containing active pharmaceutical ingredients, wherein the microspheres are suspended in a non-aqueous pharmaceutically acceptable carrier. The formulations are one-component injectable microsphere formulations, so the patient is not required to mix the formulation with a pharmaceutically acceptable carrier prior to injection. The description offers distinct advantages over the previous two-component formulations by providing a shelf life of the composition in the carrier, sustained release of the active pharmaceutical ingredient, a less complex carrier, an easier manufactured carrier, a less complex injection delivery apparatus, a kit with fewer components and ease of use by the patient.

The disclosure provides sustained release formulations comprising a pharmaceutically acceptable carrier which consists essentially of one or more triglycerides which comprise C6-C12 fatty acids; and microspheres which consist essentially of a poly (lactide-co-glycolide) polymer that has dispersed in it approximately 1% to 10% (w / w) of exenatide and approximately 0.1% to 5% (w / w) of a sugar , wherein the lactide: glycolide ratio in the polymer is about 70:30 to 30:70 or about 1: 1. In one embodiment, exenatide is present in an amount of 1% to 5%.⁴ IMPIAS • «UTUTO MUICANO D6 LA PWOF1BDAO INDUSTRIAL (w / w) or 5% (w / w) and the sugar is present in an amount of 2% (w / w). The sugar can be, for example, glucose, dextrose, galactose, maltose, fructose, manny, sucrose, lactose, trehalose, raffinose, acarbose, glycol, glycerol, erythritol, treitol, 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 approximately 0.1 mL / g or less, as determined using mercury intrusion porositometry, to provide a release profile that has a maximum serum concentration ratio of exenatide during the period release (C_max) at average serum concentration of exenatide during the release period (C_prom) of approximately 3 or less. Furthermore, although microspheres are formulated in oil (i.e., a carrier as described herein), microspheres do not necessarily have oil contained within interior spaces or pores, or within a substantial number of interior spaces or pores. , of the microspheres, and the amazing properties described herein can still be achieved.

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 approximately 0.1 ml / g or

Δ J_t «b

IMPI ^

ΙΝΓΠΤυτο MEXICANO PE THE minor industrial PROPERTY, as determined using mercury intrusion porositometry, J «Z __ to provide a release profile that has a maximum serum concentration ratio of the active pharmaceutical ingredient during the release period (C_max) at average serum concentration of the active pharmaceutical ingredient during the release period (C_prom) of approximately 3 or less. Also, although the microspheres are formulated in oil (i.e., a carrier as described herein), in some embodiments the microspheres have no oil contained within the interior spaces or pores, or have no oil within a number of substantial space or internal pores of the microspheres, and can still achieve the amazing properties described herein. The formulation is a suspension, thereby the microspheres are suspended in the carrier. The non-aqueous carrier can be an oil, such as fractionated oils, triglycerides, diglycerides, monoglycerides, propylene glycol fatty acid diesters, and the like.

In one embodiment, the active ingredient is not soluble in the carrier. In various other embodiments, the active ingredient has a potency solubility 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 additional modalities, less than 5% or

<img file="MX352189B_D0007.tif" />

IMPI

MgXCANO HSTITUTE OF FINDUAL PROPERTY 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 can also be directly incorporated into the carrier in a pharmaceutically effective amount.

The description provides a kit, available to a patient or medical service provider. The kit contains a container that has a formulation of the invention, and instructions for use. In some embodiment, the container is a pen-shaped injector. The pen-shaped injector may be a single-dose pen-shaped injector or a multi-dose pen-shaped injector. In one embodiment, the container is a vial, which can be either a single dose vial or a multiple dose vial. In another embodiment, the container is a cartridge, such that it is a cartridge for use in an injection apparatus. The cartridge can be either a single dose cartridge or a multiple dose cartridge. In different embodiments, the kit contains 1, 2, 3, 4 or even 5 or more of the containers that carry a formulation of the invention. A further advantage of the formulations is that in one embodiment, the container is provided preservative free. But in other embodiments, a preservative may be soluble in the carrier selected and provided in the formulation.

<img file="MX352189B_D0008.tif" />

, IMPI

BRIEF DESCRIPTION OF THE FIGURES tNcusnuAt

For each of Figures 1-6, the microspheres comprise a poly (lactide-co-glycolide) copolymer having exenatide dispersed therein, as described in Example 1. For each of Figures 2-6, the carrier The oil is a medium chain triglyceride (MCT) commercially available as MIGLYOL® 812 (Sasol Germany GmbH, Witten, Germany).

Figure 1 provides a comparison of the pharmacokinetics of four different microsphere formulations. In three formulations, the carrier is an oil (eg, sesame oil; MIGLYOL® 812; ethyl oleate). In the comparative formulation, the carrier is an aqueous diluent.

Figure 2 is a graphical simulation (i.e. nanoparametric overlay) of extrapolated data from Figure 1 of the plasma exenatide concentration over time for the microsphere formulation comprising the oily carrier and the microsphere formulation comprising the carrier Aqueous in Sprague Dawley Rats. The plateau of exenatide plasma concentration can be reached after approximately 5 dosages.

Figure 3 illustrates the in vitro release for a formulation comprising microspheres in an oily carrier compared to formulations comprising microspheres in an aqueous carrier.

IMPI

INSTITUTO M1XCANO HEARD INDUSTRIAL NEWS

<img file="MX352189B_D0009.tif" />

Figure 4 illustrates the in vivo release profile in rats over hours for a formulation comprising microspheres in an oily carrier and a formulation comprising microspheres in an aqueous carrier.

Figures 5A and 5B illustrate the purity of exenatide for 9 months at temperatures of 5C and 6 months at 25 ° C when stored in the formulations comprising the microspheres of Example 1 with an oily carrier compared to the purity of exenatide that is stored. on dry microspheres from Example 1. In Figure 5A, the purity of exenatide is determined by strong cation exchange HPLC. In Figure 5B, the purity of exenatide is determined by reverse phase HPLC.

Figure 6 illustrates the stability / potency of exenatide in a formulation where the microspheres are suspended in an oily carrier, where a formulation is stored at 5 ° C and a formulation is stored at 15 to 25 ° C.

DETAILED DESCRIPTION OF THE INVENTION

The description 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 a pharmaceutical ingredient

<img file="MX352189B_D0010.tif" />

ΙΜΡΙ &lt;

'Πτυτο Mexican'

OF THE PROPERTY active dispersed in this, where the microspheres are suSp ^ flüfen non-aqueous carrier. The formulations are one-component injectable formulations, compared to two-component formulations which require microspheres to be stored in one container while a liquid carrier can be stored in a separate container, so that the patient can mix the two. gaskets prior to injection. The formulations offer the long-acting stability convenience 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 ease of use by the patient, who only needs to lightly shake the formulation before injecting it from the same container. When the container also provides an injection device, even the step of syringing the formulation is eliminated. The formulations described herein offer the additional important advantage of substantially reducing the burst release of the active pharmaceutical ingredient. In this way, even pharmaceutical ingredients that have a toxic effect at higher concentrations can be safely administered using the formulations described herein.

The term "patient" refers to mammals, including humans, pet animals, farm animals, zoo animals, and the like. In one embodiment, the patient is a human.

<img file="MX352189B_D0011.tif" />

The terms

IMPI, Mexican institute • E LA monedad

INDUSTRIAL treat "or" treatment "refers to the administration of one or more active pharmaceutical ingredients to a patient who has a condition or disorder or a predisposition towards a condition or disease, for the purpose of alleviating, diminishing, remedying, mitigating, improving, slowing or stopping the progress or worsening of the disease, or at least one symptom of the disease, condition or disorder, or the predisposition towards the condition or disorder.

"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 Gila monster venom.

Formulation of a Component

Previous injectable formulations contained in at least two components. The first component can be dry microspheres and the second component can be an aqueous pharmaceutically acceptable carrier. The first component and the second component are stored in separate sealed containers (eg vials, pen chamber for injection). The patient receives the two-component formulation, and the patient or pharmacist can physically mix the two components together prior to injection. In the case of an injection pen, the two components are mixed together immediately before injecting the patient. Two component formulations are typically administered

ΙΜΡΙ £&gt;

INSTITUTO MSXfCANO,,. obla wtoptiüAD VAZz ^ Br to the patient within a short time after being mixed «l ^^TOcorS ^ _ ^ pharmaceutically acceptable carrier. For example, the microsphere component and the pharmaceutically acceptable aqueous carrier are mixed together and the formulation is then administered to the patient within about 30 or 60 minutes.

The formulations described herein are one component injectable formulations. A one-component injectable formulation refers to a formulation that contains both microspheres and the pharmaceutically acceptable carrier provided in the same container, and that can be administered to the patient without the need to first combine the microspheres and the pharmaceutically acceptable carrier. Therefore, the one-component formulation is manufactured as a premixed formulation for injection. A one-component formulation provides significant convenience for manufacture, transportation, storage, and use by the patient.

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 nothing has been inserted since its time of manufacture completion. The manufacturing completion time is the time when the container holds the formulation and is initially sealed. Containers can include (single or multiple use), syringes, injection pens (eg, single use or multiple use), and the like.

<img file="MX352189B_D0012.tif" />

Carrier

IMPI

INSTITUTO MÉKCANO • e la pftomoAo INDUSTRIAL “Carrier” (or vehicle) refers to a liquid, non-aqueous, pharmaceutically acceptable 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 awaiting approval by the Federal regulatory agency or a state government or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia for use in animals, such as humans. The term "carrier" can include one or more compounds. The carrier is a non-solubilizing carrier, in the art the carrier does not solubilize the polymers that form the microspheres. In a further embodiment, the carrier does not solubilize the active pharmaceutical ingredients within the microspheres. For example, the carrier cannot solubilize exenatide or other water soluble peptides or therapeutic proteins.

The term "non-aqueous" does not exclude residual amounts of wastewater that have no demonstrated negative impact on the stability of sustained release compositions. In this way, a composition can have approximately 0.1% (w / v) water or even approximately 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 that it has a demonstrated negative impact on the stability of the microspheres or demonstrates loss of release control

<img file="MX352189B_D0013.tif" />

IMPI INSTITUTO MEXICANO DE LA PROPERTY Explosion. In one embodiment, the carrier does not enter or peiTR ^^ I biocompatible, biodegradable and does not disperse within 361 biocompatible, biodegradable polymer. The carrier also does not cause swelling of the microspheres as the negative impact on the stability of the microspheres has been demonstrated. For example, swelling can occur to a degree of less than 1% and still be considered a non-aqueous carrier that the microspheres do not swell.

In one embodiment, the non-aqueous carrier is a pharmaceutically acceptable oil. An oil is a substance that is in an aqueous liquid state at slightly warm or ambient temperatures 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, castor oil, soybean oil, safflower oil, cottonseed oil, ethyl oleate, and the like. The carrier can comprise an oil or a combination of two or more oils.

In one embodiment, the carrier is a fractionated oil or a combination of two or more fractionated oils. Exemplary pharmaceutically acceptable oily carriers include coconut oil

IMPI nSTITUTO MÉXICANO fractionated, fractionated palm oil, fractionated alrriártfl ^^ oil, fractionated sesame oil, soyq ft ^ cinnated oil, fractionated almond oil, fractionated rapeseed oil, corn oil

<img file="MX352189B_D0014.tif" />

fractionated, 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 poop oil. In one embodiment, the carrier is fractionated palm kernel oil. In one embodiment, the carrier is a combination of coconut oil and fractionated palm kernel oil.

As used herein, fractionation is a process by which long chain fatty acids are removed from oil, such as the resulting fractionated oil substantially comprises medium chain triglycerides. The person skilled in the art will appreciate that some long chain fatty acids can remain in the fractionated oil, but generally in amounts less than 5% by weight or less than 2% by weight of the total fatty acid content of the fractionated oil.

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.

In one embodiment, the carrier is a medium chain triglyceride. Medium chain triglyceride can be synthetic or natural (for example, it is produced from fractionated oils, such as coconut oil and / or

IMPI

INSTITUTO MIRICA NO DB THE PROPERTY

<img file="MX352189B_D0015.tif" />

palm kernel oil). 'Medium chain triglyceride'^Nl^ "Fefie esters of glycerol having three fatty acid chains Ce to Ci2&gt; where the three fatty acid chains can be the same or different. Medium chain triglycerides are represented by the compound of Formula (I):

<img file="MX352189B_D0016.tif" />

where each X is independently 4, 6, 8, or 10. When x is 4, the chain refers to a Ce fatty acid. When x is 6, the chain refers to a C fatty acid₈. When x is 8, the chain refers to a C fatty acid₁₀. When x is 10, the chain refers to a C12 fatty acid. In various embodiments, each x is the same integer; two x's are the same integer and one x is a different integer; or each x is a different whole number.

In various embodiments, the esters comprise medium chain triglycerides of (i) three C fatty acids₈; (i¡) three fatty acids Ci₀; (iii) two C fatty acids₈ and a fatty acid C₁₀; (iv) two C10 fatty acids and one C fatty acid₈¡(V) two C fatty acids₈ and a fatty acid C₆; (vi) two C10 fatty acids and one C fatty acid₆; (vii) a C fatty acid₈, a C10 fatty acid and a C fatty acid₈; or (viii) any other combination of C fatty acids₆, C₈, C10 and

IMPI ^

INSTITUTE MiXICANC

Ci2 · In another embodiment, the medium chain triglyceride compreno ^ Swácroe? Fatty Ce and a Fatty Acid Ci₀. In one embodiment, the triglyceride of Cade ici media comprises two fatty acids C₁₀ and a fatty acid Ce.

One skilled in the art will appreciate that a mixture of medium chain triglycerides can result from any process (eg, fractionation, hydrogenation) used to prepare medium chain triglycerides. For example, substantially all of the medium chain triglycerides obtained from fractionated coconut oil can comprise C8 and / or C10 fatty acids; however, there may be some medium chain triglycerides that contain C fatty acids₆ and / or C- |₂.

In one embodiment, the medium chain triglycerides comprise esters of (i) 0 to 2% by weight of Ce fatty acid, 65 to 80% by weight of Ce fatty acid, 20 to 35% by weight of Cio fatty acid, and 0 to 2% by weight of fatty acid Ci₂; (ii) 0 to 2% by weight of fatty acid C₆, 50 to 65% by weight of Ce fatty acid, 30 to 45% by weight of Cio fatty acid, and 0 to 2% by weight of Ci fatty acid₂; (iii) 0 to 2% by weight of fatty acid C₆, 45 to 65% by weight of fatty acid C₈, 30 to 45% by weight of Cio fatty acid, 0 to 3% by weight of C fatty acid₁₂And 0 to 5% by weight of linoleic acid; or (iv) 0 to 2% by weight of fatty acid C₆, 45 to 55% by weight of fatty acid C_8&gt; 30 to 40% by weight of Cw fatty acid, 0 to 3% by weight of Ci fatty acid₂, and 10 to 20 succinic. In one embodiment, the medium chain triglyceride comprises 0 to 2% by weight of fatty acid C₈, 50 to 65% by weight of fatty acid C₈, 30 to 45% by weight of Cio fatty acid, and 0 to 2% by weight of C fatty acid₁₂, and which is

IMPI tMSTlTUTO MEXICANO commercially available as MIGLYOL® 812 (Sasol Ge ^ áS ^ rl ^

<img file="MX352189B_D0017.tif" />

Witten, Germany) Weight% is based on total acid content or triglycerides. In one embodiment, the medium chain triglycerides can comprise up to 2% C14 fatty acids.

The carrier can 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 fatty acids.₈ and a C10 fatty acid. In one embodiment, the carrier comprises a medium chain triglyceride comprising esters of a C fatty acid₈ and two C10 fatty acids. In one embodiment, the carrier comprises two different medium chain triglycerides, wherein a first medium chain triglyceride comprises esters of two C fatty acids.₈ and a C10 fatty acid and a second medium chain triglyceride comprise esters of a C fatty acid₈ and two fatty acids Cw. In one embodiment, the carrier comprises a medium chain triglyceride which comprises 0 to 2% by weight of C fatty acid.₆, 50 to 65% by weight of fatty acid C₈, 30 to 45% by weight of fatty acid C₁₀0 to 2% by weight of C12 fatty acid, based on the total fatty acid content of the medium chain triglyceride.

Triglycerides can be prepared by methods known in the art and are commercially available as MIGLYOL® 810, 812, 818, 829 (Sasol Germany GmbH, Witten, Germany) or NEOBEE® 1053, 895, M-5 (Stepan Company, Northfield, IL).

IMPI ^

In another embodiment the carrier is a diester

INDUSTRIAL saturated vegetable fatty acids with long chains of Ce and Cw (capric and caprylic acid). An example of such a commercially available carrier is MIGLYOL® 840 (Sasol Germany GmbH, Witten, Germany). The non-aqueous, pharmaceutically acceptable carrier may optionally comprise other pharmaceutically acceptable excipients. Exemplary excipients include sugars (eg, sucrose, glucose, dextrose, galactose, maltose, trehalose, fructose, maltodextrin); sugar alcohols (eg, glycol, glycerol, erythritol, treitol, arabitol, ribitol, sorbitol, dulcitol, iditol, somali, maltitol, lactitol, mannitol, xylitol); preservatives (eg, benzoic acid, sorbic acid, meta cresol, sodium benzoate, potassium sorbate, methyl paraben, propyl paraben, butyl paraben, benzalkonium chloride, and the like, generally soluble in oil, with some solubility in the selected carrier); and antioxidants (eg, sodium metabisulfite, butylated hydroxy anisole, butylated hydroxytoluene, sodium sulfite, tocopherol, thymol, ascorbate, propylgalate, and the like). In one embodiment, the carrier optionally comprises mannitol, maltodextrin, sorbitol, or a combination of two or more thereof.

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 deposit to form at the in vivo administration site of the formulation. In one embodiment, the pharmaceutically acceptable carrier does not contain a gel-forming agent.

IMPI ^ mtTtTuTo Mexican

Exemplary gel-forming agents include derivatives^M"FewSS ^ os" 5I ^^ example, hydroxypropyl cellulose, carboxymethyl cellulose, "hydroxyctyl ooluloca, hydroxypropyl methyl cellulose, methyl cellulose); polymers or copolymers of polyoxyethylene and polyoxypropylene (poloxamers); chitosan acid, and the like. One of ordinary skill in the art will understand that in vivo gel formation can be determined by methods known in the art, such as the use of colored stains and histological sections.

In certain embodiments the non-solubilizing, non-aqueous 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-solubilizing, non-aqueous carrier is from 20 cP to 80 cP or from 30 cP to 70 cP. Thus, with reference to this description the person of ordinary skill will be able to identify other oils, triglycerides, or nonaqueous compounds that may also be present in the nonaqueous, nonsolubilizing carrier.

Microspheres

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 for microspheres is 1000 microns or less. In one particular embodiment, the microsphere ranges from about one to about 180 microns in diameter. In still additional embodiments, adequate release profiles are obtained when microesl

DI THE INDUSTRIAL FROFIBILITY

<img file="MX352189B_D0018.tif" />

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 equal to about

50, 60, or 70 microns, and preferably less than about 80, 90, or 100 microns. At larger sizes, the microspheres 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.

Superior and consistent release profiles are obtained by controlling the size distribution. In one embodiment a median microsphere size is approximately 50 microns and the lower and upper ranges of the microsphere are approximately 30 and 90 microns, respectively. Microsphere distribution can be described using a volume mean diameter. The 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 volume distribution diameter of about 50 to 70 microns, about 50 to 60 microns, or about 50, 60, or 70 microns, with a volume distribution (DV) of less than or about 5%. , 10%, or 15% at 30 microns and a DV greater than or approximately 80%, 85%, 90%, or 95% at 90 microns. In one embodiment, the microspheres have a volume distribution mean diameter of

IMPI

ΚβΤΠνίΌ * &lt;£ I «CAWX1&gt;

oeiA MOísetMij approximately 60 microns, with a voliyñWt ^ Dvf ^^ cremenos distribution of or approximately 10% at 30 microns and a greater uv dt ^ o —— approximately 90% at 90 microns.

Microspheres can be prepared by processes known in the art and are described, for example, in U.S. Patent Nos. 7,563,871, 7,456,254, 7,223,440, 6,824,822, 6,667,061, 6,495,164, and 6,479,065, the descriptions of which are incorporated by reference into the Present.

In a further embodiment, the microspheres have a less porous outer layer, and may also have a non-porous outer layer. Accordingly, in the formulations described herein, the oil does not have access to the interior spaces or pores or even a substantial portion of the interior spaces or pores. Specifically, it is contemplated that for each of the formulations described herein, the microspheres may additionally be oil-free (or a carrier as described 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 they are formulated.

Polymers

The microspheres comprise biocompatible, biodegradable polymers. A polymer is biocompatible if the polymer and any of the polymer's degradation products are non-toxic to the patient at levels

INSTITUTO MUUCANI OE LA MOKEtMC administered and also possess undesired or 'déíStéreo ^ -Tfo ^⁹^ Demonstrated in the patient's body, for example Substantial immunological damage at the injection site. Biodegradable means that 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.

Exemplary biocompatible, biodegradable polymers include, for example, polylactides, polyglycolides, poly (lactide-co-glycolides), polylactic acids, polyglycolic acids, poly (lactic acid-co-glycolic acid), polycaprolactones, polycarbonates, polyesteramides, polyanhydrides, polyamino acid, polyorthoesters, polycyanoacrylates, poly (p-dioxanone), polyalkylene oxalates, biodegradable polyurethanes, mixtures thereof, and copolymers thereof. Acceptable molecular weights for biocompatible, biodegradable polymers can be determined by one of ordinary skill in the art taking into consideration factors such as the rate of degradation of the desired polymer, physical properties such as mechanical intensity, end group chemistry, and dissolution rate. of the polymer. Typically, an acceptable molecular weight range is from about 2,000 Daltons to about 2,000,000 Daltons. The biocompatible, biodegradable polymer can also be selected based on the inherent viscosity of the polymer. Suitable inherent viscosities are about 0.06 to 1.0 dL / g; approximately 0.2 to 0.6 dL / g; or about 0.3 to 0.5 dL / g.

’ -4 ²³

INSTITUTO MEXJCaMC 'DE LA PtOFIEOAO INDUSTRIAL LS * »

In one embodiment, the biocompatible, biodegradable polymer is a poly (lactide-co-glycolide) copolymer (also referred to as "PLGA") having a lactide: glycolide ratio of from 70:30 to 30:70, or from 60:40 to 40:60 or approximately 50:50. The molecular weight of the poly (lactide-coglycolide) copolymer is about 10,000 Daltons to about 90,000 Daltons. In another embodiment, the molecular weight of the poly (lactide-coglycolide) copolymer is about 30,000 Daltons to about 70,000, or from about 50,000 to about 60,000 Daltons.

The formulation can contain microspheres at a concentration 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

An active pharmaceutical ingredient is a biologically active compound that has a beneficial therapeutic, prophylactic or other pharmacological and / or physiological effect in the patient. The active pharmaceutical ingredient can also be a mixture of two or more compounds. The term "peptide" refers to any compound that has 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

<img file="MX352189B_D0019.tif" />

IMPI «« nruTOMexicANo

Μ LA MOMSDAD industrial one embodiment, the peptide comprises 2 to 500 amino acid residues; 2nd

250 amino acid residues; 5 to 100 amino acid residues; or 5 to 50 amino acid residues.

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) -NH₂, GLP-1 (7-36), GLP-1 (7-36) -NH₂, Leu¹⁴exendin-4, Leu¹⁴, Phe²⁵-exendin-4, exendin-4 (1-28), Leu¹⁴-exendina-4 (128), Leu¹⁴, Phe²⁵-exendin-4 (1-28), exendin-4 (1-30), Leu¹⁴-exendina-4 (1-30), Leu¹⁴, Phe²⁵-exendin-4 (1-30), liratuglide, and the compounds described in, for example, US Patent No. 7,157,555, US Patent No. 7,220,721, US Patent No. 7,223,725, and WO 2007/139941, the descriptions of which are incorporated herein by reference.

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 (eg, pramlintide, davalintide, Val²⁷-davalintida); leptin, leptin agonists (eg, metreleptin); PYY (3-36) and agonist analogs thereof; glucagon, glucagon agonists, glucagon antagonists, peptide chimera of GLP-1 receptor agonists and glucagon agonists, human amylin peptide chimera and salmon calcite, insulin, heparin,

<img file="MX352189B_D0020.tif" />

IMPI

INSTITUTO MEXICANO DE LA PROPIEDAD INDUSTRIAL low molecular weight heparin, angiotensin, argipressin, argireline, atosiban, bivalirudin, cetrorelix, desmopressin, enfuvirtide, deptiphin,

GHRP-2, GHRP-6, Gonadorelin, Leuprolide, Lysipressin, Melanotan, Nesiritide, Octreotide, Oxytocin, PT141, Calcitonin, Sermorelin, Somatostatin, Terlipressin, Thymopentin, Thymosin A1, Triptorelin, Vapreotide, Elcatonin, BNP, Zicon -32, and the like.

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 (eg, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin); non-selective beta blockers and / or alpha-1 blockers (eg, carvedilol, dilatrend, eucardic, carloc); PDE inhibitors (eg, cilostazol); antiplatelet drugs, antithrombotic drugs, anticoagulant drugs, llb / llla glycoprotein inhibitors (eg, abciximab, eptifibatide, tirofiban); antibacterial drugs (eg, ciprofloxacin, norfloxacin, levofloxacin, moxifloxacin, sparfloxacin, gemifloxacin, ecinofloxacin, delafloxacin); Factor Xa inhibitors (eg, glycosaminoglycans, oligosaccharides, heparinoid); direct Xa inhibitors (eg, xaban); direct thrombin (II) inhibitors (eg, hirudin, argatroban, dabigatran, melagatran, ximelagatran, defibrotide, ramatroban, antithrombin III, protein C); thrombolytic drugs (eg, plasminogen activators, urokinase, streptokinase, serine endopipidases);

²⁶ IMPI ^

INSTITUTO MEXICANO DE LA PROPIEDAD INDUSTRIAL ACE inhibitors (eg, lisinopril, aceon, acertil, armix, coveren, coverex, coversum, prestaium, prexanil, Prexum, procaptan); ADP / P2Y12 receptor inhibitors (eg, clopidogrel, ticlopidine, prasugrel); prostaglandin analogs (eg, beraprost, prostacyclin, iloprost, treprostinil); anticoagulants (eg, coumarin, coumatetralyl, dicoumarol, ethyl biscoumacetate, fenprocoumon, warfarin, chlorindione, diphenadione, phenindione, thioclomarol); diuretics (eg, hydrochlorothiazide); macrolides (eg, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, telithromycin); NSAIDs and COX-3 inhibitors (eg, celecoxib, etoricoxib, parecoxib); sulfonanilides (eg, nimesulide), and the like.

One skilled in the art 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 contains one peptide (eg, pramlintide) and another set of microspheres contains a different peptide (eg, 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. One skilled in the art will appreciate that the

IMPI

INSTITUTO MEXICANO Dt LA PROPERTY

<img file="MX352189B_D0021.tif" />

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 low-potency peptides can be provided in more microspheres and high-potency peptides can be provided in fewer microspheres therein. formulation. Exemplary combinations of peptides and / or small molecules that can be delivered in different series of microspheres and in the same array include: pramlintide and insulin; pramlintide and metreleptin; davalintide and metreleptin; exenatide and metreleptin; lovastatin and niacin; atorvastatin and amlodipine; simvastatin and ezetimíb; exenatide and metformin; and the like.

Formulations in general 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 the 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 the active pharmaceutical ingredient will vary depending on the desired effect, potency of the agent, planned release levels, and the elapsed time during which the peptide will be released. In certain modes, the loading range 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 ingredient

<img file="MX352189B_D0022.tif" />

IMPI

MMfCANO INSTITUTE

DF LA MtOPIIDAü INDUSTRIAL active pharmaceutical 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

The microspheres can also comprise one or more sugars. A sugar is a monosaccharide, disaccharide, or oligosaccharide or a derivative thereof. Monosaccharide sugar alcohols 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 provides 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, rafmosa and acarbose. The microspheres can also comprise glucose, dextrose, galactose, maltose, fructose, manny, sucrose, lactose, trehalose, raphmose, acarbose, glycol, glycerol, erythritol, treitol, arabitol, ribitol, sorbitol, dulcitol, iditol, maltol, isomalt , 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.

<img file="MX352189B_D0023.tif" />

IMPI

MEXICAN INSTITUTE OF PROPERTY

The amount of sugar present in the microns will vary from about 0.01% (w / w) to about 50% - ⁷ (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.

Alternatively, the amount of sugar present in the microspheres can be related to a weight ratio with the active pharmaceutical ingredient. For example, the active pharmaceutical ingredient and sugar can be present in a ratio of from about 10: 1 to about 1:10 weight: weight. In particularly preferred embodiments, the ratio of the active pharmaceutical ingredient (eg, exenatide) to sugar (eg, sucrose) is about 3: 2 (w / w), 4: 2 (w / w), or 5: 2 ( p / p). Combinations of two or more sugars can also be used. The amount of sugar, when a combination is used, is the same as the ranges mentioned above.

Sustained Release

The compositions are sustained release compositions, which means that the active pharmaceutical ingredient contained in the compositions will be released into the patient over a prolonged 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,

IMPI

INSTITUTO MÉXIGANC two weeks, one month, three months, or one year. The release WWT ^ SK

<img file="MX352189B_D0024.tif" />

Active pharmaceutical is considered complete when there is no longer a therapeutic level of the active pharmaceutical ingredient in the patient's body, as determined by the medical judgment of those of ordinary skill in the art.

Cmax as used herein is the maximum serum concentration of the drug which occurs during the release period which is monitored. Cave as used herein is the average serum concentration of the drug delivered by dividing the area under the curve (AUC) of the release profile by the duration of release. In one embodiment, the ratio of Cmax to Cave is about 3 or less. This profile is particularly desirable for antidiabetic or glycoregulatory 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. By further controlling the physical aspects of the sustained release composition, as described herein, a desired superior release profile can be achieved and controlled, for example, by appropriate selection of carrier properties, such as viscosity. This provides a reduced burst (ie, initial release; eg, 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. Furthermore, ³ ΙΜΡΙ ^%

INSTITUTO MEXICANO U € LA MONEDAD a Cmax, if present, can be changed from the initial release period to the “sustained phase” of release. In one mode the

Cmax can occur in at least 7, 14, 21, 28, 35 or 42 days after administration and can occur on any whole day in between. In a further embodiment the Cmax occurs at about 21 to 35 days after administration, and in still another embodiment it is at about 28 to 31 days, and furthermore at about 28 days after administration. In a further embodiment, the peak drug concentration (eg, plasma concentration) occurs at least 7, 14, 21, 28, 35, or 42 days after administration and can occur on any full day in between. In yet a further embodiment the maximum drug concentration occurs at approximately 21 to 35 days after administration, particularly in the case of glycoregulatory agents such as exendin4, GLP-1, GIP or their analogues.

Long shelf life

One advantage offered by the present formulations is a long shelf life for the formulation. Sustained release compositions were unexpectedly found to retain remarkable stability when stored in a nonaqueous carrier as described herein. In one embodiment the formulation has a shelf life of at least 6 months. In other modalities the formulation has a shelf life of at least 1 year, or at least 18 months, or at least 2 years. By "shelf life" it means

IMPI

ΙΝΓΠΤυΤΟ MEXICANO DI LA PROPERTY that the formulation can be stored or maintained ροΓϊίΓρβηο

<img file="MX352189B_D0025.tif" />

time under appropriate environmental conditions while retaining at least

90% of the desired activity of the active pharmaceutical ingredient relative to the activity of an initial formulation (such 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 compared to its activity immediately prior to storage. When the formulation contains microspheres, shelf life also refers to retention of the particle size and / or morphology of the microspheres. The retention of size morphology can be determined by microscopic examination, the use of which is known to persons of ordinary skill in the art. When formulated as described herein, a peptide or protein as an active ingredient is less susceptible to oxidation and hydrolysis, either chemical or proteolytic, both during storage and during its period of sustained release 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

Another advantage of the present formulations is that the formulations according to the present description offer a significantly reduced burst release ratio compared to other formulations. When sustained release formulations

<img file="MX352189B_D0026.tif" />

IMPI

INSTITUTO MEXICANO DE LA PROPERTY INDUSTRIAL Previously available injectables are injected into a patient is often ¹ II ww * · V u - "a" burst "of the active ingredient or agent associated with the injection. Without wishing to be limited by any specific theory, it is believed that this burst is caused by the amount of the active pharmaceutical ingredient in the formulation that does not it is retained within the polymer that is released over time. By "burst release" it is meant that the amount of the active pharmaceutical ingredient is released within the first 24 hours after injection. In other embodiments it is the amount of active that is released during 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 referred to the percentage of the total of the active pharmaceutical ingredient in the injected formulation. After injection of the formulation into the patient, burst release can occur at any time up to about 24 hours, subsequently there may be a lag time where substantially no active pharmaceutical ingredient is released from the microspheres, and then the Polymeric microspheres begin to degrade and release the active pharmaceutical ingredient. One skilled in the art will appreciate that the period of time when burst release occurs may vary from patient to patient.

IMPI

ΙΝΓΠΤυΤΟ MUtlCAN

The burst can be assessed by measuring the pro- ^ tyflSSf ^

<img file="MX352189B_D0027.tif" />

total under the curve for a period of time will start. »^the&lt;; after the administration of a drug. The area under the curve (AUC) is a well-established measurement in 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 selected period of time will vary depending on the period of time the drug concentration is expected to be detectable in the blood or within the therapeutic window of the drug. The AUC is calculated by plotting the concentration of the drug in the blood, eg plasma concentrations, at various times during the selected time period and then calculating the total area under the obtained curve. In an exemplary embodiment, the area under the curve is measured for a period of 42 days and the formulations described herein are used, the release or burst already 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.

In another embodiment, the formulations described herein limit the initial burst so that the upper limit of the therapeutic window for the active pharmaceutical ingredient is not exceeded. Window

IMPI

INSTITUTO MEXICANO DI LA PROPERTY Y ** - —i INDUSTRIAL therapeutic is the concentration range of the active pharmaceutical ingredient in circulation, above which the active pharmaceutical ingredient has its desired effect, but below the concentration at which adverse effects associated with the active pharmaceutical ingredient outweigh the benefits as might be generally accepted among specialists. In an exemplary embodiment, the active pharmaceutical ingredient is an exendin, for example exenatide, or agonist analog thereof, and the administration of the disclosed formulations does not result in a circulating level of the active pharmaceutical ingredient that exceeds 400 pg / ml during the first few 24 hours after administration. In another exemplary embodiment the active pharmaceutical ingredient is an exendin, for example exenatide, or agonist analog thereof, and administration of the disclosed formulations will not result in a circulating level of the active pharmaceutical ingredient that exceeds 350 pg / ml during the first 24 hours after administration.

The initial burst can also be assessed by comparing circulating concentrations of the active pharmaceutical ingredient in a period of time immediately after administration of the formulation with the circulating concentration of the drug in a second period of time immediately following the first. In one embodiment, use of the formulations of the present disclosure results in circulating concentrations of the active pharmaceutical ingredient during the first 24 hours after administration that do not exceed the

<img file="MX352189B_D0028.tif" />

circulating concentration over the next 24 hour period. In another embodiment, use of the formulations of the present disclosure results in the average circulating concentration of the active pharmaceutical ingredient during the first 24 hours after administration that does not exceed the average circulating concentration during the next 24 hour period.

Classification Methods

Another aspect provides methods for classifying the sustained release formulations described herein. The methods for classifying the formulations described herein can also be referred to as methods for preventing microsphere degradation. By "sort" it means that the formulation is retained for a period of time within its container without adding any additional components to the container and without removing the formulation from the container (for example, at the manufacturing facility, during transportation, at the pharmacy ). 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

IMPI

INSTITUTO MBXICANC modalities, the formulations can be stored at

<img file="MX352189B_D0029.tif" />

minimal microsphere degradation when the formulations are stored for such long periods of time.

In another embodiment the invention provides methods of maintaining the potency of (eg, preventing loss of biological activity) and / or purity (eg, preventing chemical changes in the molecule) an active pharmaceutical ingredient. Thus, a peptide or protein or other API that has undergone a chemical change (eg, oxidation) may result in a loss of purity, but may still retain its potency. The methods involve storing a microsphere comprising an active pharmaceutical ingredient in a non-aqueous carrier as described herein for a period of time, thereby the potency and / or purity of the active pharmaceutical ingredient is maintained by the microspheres and the carrier is not. aqueous. 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 at least 18 months, or at least 2 years.

Methods of administration / treatment

In another aspect the present invention provides methods for administering an active pharmaceutical ingredient to a patient in need of the

<img file="MX352189B_D0030.tif" />

IMPI Mexican inequtttuto DS LA ntOMEDAD INDUSTRIAL itself. 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 subcutaneous, intra-muscular, intra-peritoneal, intra-abdominal, intravenous, or any suitable mode 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 previous step of combining the sustained release composition with a second carrier.

In one embodiment the administration does not comprise a mixing step. A mixing stage is a stage where the microspheres are combined with a carrier prior to injection. In various embodiments the mixing stage is a stage where the microspheres are combined with a carrier within a period of 1 week prior to injection into the patient. The carrier can be a non-aqueous carrier, such as those described herein. Formulation administration refers to the entire process of user interaction with the formulation, which includes mixing, combining any of the ingredients that make up the formulation, and the actual injection or other way of providing the formulation to the patient.

IMPI ^

INSTITUTO MEXICANO DI LA PROPERTY INDUSTRIAL

The frequency of administration may vary depending on any or a combination of factors such as the amount of the formulation administered, the release profile of the formulation, the amount of the active pharmaceutical ingredient in the formulation, and the circulation level of the active pharmaceutical ingredient. to achieve. In particular modalities, the formulations described herein can be administered once daily, once a 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 a year. In one embodiment, the formulation is administered once a week. In another embodiment, the formulation is administered once a month.

When the formulations comprise a GLP-1 receptor agonist, such as GLP-1 or an analog thereof, or an exendin (eg, 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, nonalcoholic steatohepatitis (NASH), and the like. Formulations comprising a GLP-1 receptor agonist (eg, exenatide) will also be useful to stimulate insulin release; decrease plasma glucagon; reduce food absorption, reduce appetite, reduce gastric motility, retard gastric vacuum, lower plasma lipid levels (e.g. triglycerides,

<img file="MX352189B_D0031.tif" />

IMPI INSTITUTO MEXICANO DE LA PtOHEDAD industrial cholesterol), and the like. These treatment methods are described, for example, in US Patent No. 5,424,286, US Patent No.

US Patent No. 6,858,576, US Patent No. 6,872,700, the

US Patent No. 6,956,025, and US Patent No. 6,956,025, and WO 2007/022518, the disclosures of which are incorporated by reference herein.

In certain embodiments, administration of any of the formulations provided herein comprising a glycoregulatory peptide such as an exendin, eg, exenatide, results 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 the administration of any of the formulations provided herein comprising a glycoregulatory peptide such as an exendin, eg 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 glycoregulatory peptide such as an exendin, eg, 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 glycoregulatory peptide such as an exendin, eg, exenatide, results in a 2 hour plasma glucose of less than 140 mg / dl. In additional modalities, the

IMPI χ '_ι I ·. I χ „I · INDUSTRIAL administration of any of the formulations provided

MEXICAN INSTITUTE OF PROPERTY

<img file="MX352189B_D0032.tif" />

present that comprise a tai glycoregulatory peptide such as an exendin, for example exenatide, results in a capillary or venous 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 modality, a FBG level of less than 110 mg / dl is achieved, while in another modality a FBG level of less than 100 mg / dl is achieved.

In additional embodiments, administration of any of the formulations provided herein comprising a glycoregulatory peptide such as an exendin, eg, exenatide, results in a 2-hour capillary or venous 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 the administration of any of the formulations provided herein comprising a glycoregulatory peptide such as an exendin, eg exenatide, results in a 2 hour blood glucose level of less than 180 mg / dl. In additional embodiments, administration of any of the formulations provided herein comprising a glycoregulatory peptide such as an exendin, eg, exenatide, results in blood glucose levels of less than 170 mg / dl, less than 160 mg / dl. dl, less than 150 mg / dl, less than 140 mg / dl, less than 130 mg / dl, or less than 120 mg / dl. In particular modalities, the administration of any of the

<img file="MX352189B_D0033.tif" />

IMPI

INSTITUTO MEXICANO DE LA PROPIEDAD INDUSTRIAL Formulations provided herein comprising a glycoregulatory peptide such as an exendin, e.g., exenatide, result in a 2-hour blood glucose level of less than 120 mg / dl, while in other modalities, a capillary blood glucose level of 2 of less than 140 mg / dl is achieved.

In one embodiment, glucose levels are average glucose levels calculated over a selected period of time. 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 ratio of 1.75 g / kg of 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 3-hour, 100-g test is often used.

Because glucose freely crosses the cell membrane of red blood cells, erythrocyte hemoglobin undergoes nonenzymatic glycosylation at amine residues. Hemoglobin A1c (HbA1c) refers to the percentage of hemoglobin molecules with glucose portions attached to the N-terminal valines of each of the two chains

IMPI ^ beta. Glycohemoglobin includes HbA1c along with other forms13§ &lt;WI || ^^ where glycosylation to another amino acid has occurred. The percentage of hemoglobin molecules that undergo glycosylation is proportional to the average ambient glucose concentrations during the previous 60-90 days. HbA1c is a commonly used measure to assess the status of glycemic control in patients with diabetes.

In one embodiment, administration of any of the formulations provided herein comprising a glycoregulatory peptide such as an exendin, eg, exenatide, results in a reduction to, maintenance of, or both of HbA1c levels of less than 8 %. In another modality HbA1c levels are reduced to, maintained at, or both less than 7.5%, while in still another modality, HbA1c levels are reduced to, maintained at, or both less than 7%. In additional embodiments, administration of any of the formulations provided herein that comprise a glycoregulatory peptide such as an exendin, eg, exenatide, results in a reduction to or maintenance of, or both, HbA1c 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 described herein are useful in a method for reducing or maintaining HbA1c levels in the blood, the methods comprise administering a composition described herein. In another embodiment, the administration of any of the formulations provided herein comprising such a glycoregulatory peptide

IMPI

INSTITUTO M &amp; KJCAHO of INDUSTUAL nonecAD as an exendin, eg, 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 less than 9.5%; while in yet another embodiment, glycosylated hemoglobin levels are reduced to, maintained at, or both less than 9%. In further embodiments the administration of any of the formulations provided herein comprising a glycoregulatory peptide such as an exendin, eg, exenatide, results in a reduction to, or maintenance of, or both of the 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 that comprise a glycoregulatory peptide such as an exendin, eg, exenatide, results in a decrease in HbA1c 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 for reducing or maintaining glycosylated hemoglobin levels in the blood, the methods involving administering a composition described herein.

It should be noted that a subject in need of lowering blood glucose is not limited to patients who have diabetes mellitus, but may include any subject suffering from

<img file="MX352189B_D0034.tif" />

IMPI

INSTITUTO MEXICANO DE LA PROPIEDAD INDUSTRIAL hyperglycemia for any reason, including but not limited to injury, ~ 111 «i ILMMUJ1.

trauma, surgery, stroke and myocardial infarction. The amount of glucose decreased will vary with the subject in question and depends on factors such as the severity of the hyperglycemia and the severity of the disease, disorder or condition in question.

EXAMPLES

The following non-limiting examples provide additional illustrations for making and using the formulations described herein, and are 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® 812 (Sasol Germany GmbH, Witten, Germany).

EXAMPLE 1

Microspheres can be prepared by processes known in the art and are described, for example, in US Patent No. 7,563,871 and US Patent No. 7,456,254. Microspheres are obtained comprising a poly (lactide-co-glycolide) copolymer having dispersed there 5% (w / w) of exenatide and 2% (w / w) sucrose. Poly (lactide-co-glycolide) copolymer has a lactide: glycolide ratio

Ή

IMPI

<img file="MX352189B_D0035.tif" />

Mexican tNrrrnrro DI LA reoMEDAD, 1: 1. These microspheres are currently being developed ^^ Á

Pharmaceuticals, Inc. (San Diego, CA), Alkermes, Inc. (uambrioge, MA), and Eh

Lilly and Company (Indianapolis, IN) for a once weekly formulation for treating diabetes. Gedulin et al, Diabetologia, 48: 1380-1385 (2004).

EXAMPLE 2

The stability of the microspheres of Example 1 was investigated for their stability over an extended period of time while stored in a non-aqueous carrier. The microspheres of Example 1 were stored for a period of 6 months at 5 ° 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 of Example 1 in an aqueous carrier containing carboxymethylcellulose and a surfactant.

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 (ie size, morphology) of the microspheres did not change.

<img file="MX352189B_D0036.tif" />

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

As shown in Table 2, microspheres stored in MCT oil showed no change in exenatide purity based on HPLC analysis. Impurities could also be referred to as peptide degradation products. High purity means relatively little degradation of the peptide. Purity is relative to zero time formulation. Microspheres stored in sesame oil and ethyl oleate showed a slight reduction in purity of exenatide. The impurities do not appear to be oil or poly (lactide-coglycolide) polymer (based on retention times), but appear to be related to the stability of exenatide itself.

Table 3 shows that the potency of exenatide is not significantly reduced during the 6-month period relative to the non-aqueous carrier used.

TABLE 1

Particle size and morphology using microscope

<td></td><td colspan="3">size (pm) (standard deviation (pm))</td><td>morphology</td>

<td></td><td>T = 0</td><td>1 month</td><td>6 months</td><td>0 to 6 months</td>

<td>Sesame oil</td><td> 64 (22)</td><td> 63 (23)</td><td> 64 (12)</td><td>without changes</td>

<td>MCT oil</td><td> 65 (19)</td><td> 60 (22)</td><td> 61 (17)</td><td>without changes</td>

<td>ethyl oleate</td><td> 64 (16)</td><td> 62 (16)</td><td> 59 (13)</td><td>without changes</td>

<img file="MX352189B_D0037.tif" />

TABLE 2

IMPI iwCTtTUTO MEXICANO ^W ntlLA nongj® INDUSTRIAL

Change in purity of exenatide containing formulation

<td colspan="5">% purity exena</td><td colspan="3">tida</td>

<td></td><td>t = 0</td><td>1 month</td><td>% exchange *</td><td>3 months</td><td>% change ★</td><td>6 months</td><td>% exchange*</td>

<td>Sesame oil</td><td> 95.93</td><td> 95.68</td><td> -0.25</td><td> 94.55</td><td> -1.38</td><td> 95.00</td><td> -0.93</td>

<td>MCT oil</td><td> 95.63</td><td> 95.56</td><td> -0.07</td><td> 94.67</td><td> -0.96</td><td> 95.50</td><td> -0.13</td>

<td>ethyl oleate</td><td> 95.60</td><td> 95.80</td><td> 0.20</td><td> 93.67</td><td> -1.93</td><td> 94.70</td><td> -0.90</td>

'Changes of less than 0.5% are considered to be insignificant

TABLE 3

Change in potency of exenatide based on carrier in formulation

<td>carrier</td><td>size zero</td><td>1 month</td><td>3 months</td><td>6 months</td>

<td>Sesame oil</td><td> 97</td><td> 104</td><td> 98</td><td> 98</td>

<td>MCT oil</td><td> 94</td><td> 108</td><td> 99</td><td> 99</td>

<td>ethyl oleate</td><td> 95</td><td> 98</td><td> 99</td><td> 100</td>

<td></td><td></td><td></td><td></td><td></td>

EXAMPLE 3

The pharmacokinetics of the formulations in Example 2 were determined, except that lecithin (w / w) 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 with microspheres of Example 1 that were mixed with an aqueous carrier only prior to injection.

<img file="MX352189B_D0038.tif" />

IMPI

MEXICAN INSTITUTE

BE THE INDUSTRIAL PROPERTY

Figure 1 provides a comparison of the pharmacokinetics of the four different exenatide-containing microsphere formulations. In three formulations, the carrier is an oil (eg, sesame oil; MCT oil; ethyl oleate). In a comparative formulation, the carrier is an aqueous diluent. As can be seen from the data, formulations having an oily carrier have reduced popping when compared to formulation having an aqueous carrier.

Figure 2 is a graphical simulation of extrapolated data from Figure 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 can be reached after approximately 5 dosages.

EXAMPLE 4

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. Burst of release was evaluated by adding approximately 0.75 ml of the formulations at 10 mM of a HEPES release buffer. The mixture was agitated to ensure that the microspheres made full contact with the buffer.

<img file="MX352189B_D0039.tif" />

IMPI

CTrtVTO MWCMNO I HEARD HtOHHMC) moumiAi HEPES release. After incubation at 37 ° C for one hour, the mixture was centrifuged and the aqueous phase was analyzed by HPLC to determine the burst of release. The concentration of the dose tested for release was 150 mg / ml.

Figure 3 shows the decrease in burst release of the formulation having the oil carrier compared to the formulations having an aqueous carrier. The graph showed that with the aqueous carrier, approximately 0.6% exenatide was released on burst. With the formulation having the MCT oil carrier, less than 0.1% exenatide was released on the burst.

Figure 4 illustrates the in vivo release profile in rats over 10 hours for the formulation of Example 1 in the oily MCT compared to a formulation comprising the same microspheres in an aqueous carrier (saline). In the period of time after subcutaneous administration of the formulation, the entry of exenatide into the plasma was markedly lower than the same microspheres administered in the aqueous carrier. The formulation of the invention showed no burst of release, and a markedly more gradual entry into blood plasma versus the aqueous formulation. In contrast, the aqueous formulations showed a burst of release followed by a more defined entry into the blood plasma.

<img file="MX352189B_D0040.tif" />

EXAMPLE 5

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

Microparticles were prepared in a manner similar to those described in the Examples in US Patent No. 5,439,688, the disclosure which is incorporated herein by reference. Eight samples were prepared by briefly mixing an active pharmaceutical ingredient (i.e., davalintide, pramlintide, metreleptin, bovine serum albumin, sodium salicylate, salicylic acid, minocycline HCI, insulin) and polymer (i.e., poly (lactide-co- copolymer). glycolide) or polycaprolactone / PLGA copolymer) and then the mixture was placed in a grinder to obtain a well homogenized powder. The mixtures range 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 require high temperatures to produce a melt with good flow properties. The extruder contained double screws that stir like clockwise to produce efficient mixing. The material was extruded through a 1.5 mm hole, collected, cooled to room temperature, and cut into short strands approximately 1-2 inches long. These strands were then fed into a 12 tooth rotor mill, followed by a sieving step to produce microparticles of approximately 20 to 100 microns. The microparticles were collected and stored at 5 ° C until further use.

<img file="MX352189B_D0041.tif" />

IMPI

INSTITUTO MEXICANO DI LA FROPTIDAD INDUSTRIAL

Experimental samples were prepared by dispersing approximately 50 mg of the microparticles in 0.75 ml of an MCT oil carrier. The samples were stored at 5 ° C and 25 ° C for two days, two weeks or one month, times in which representative samples were tested. The fraction of the drug that remained in the microparticles and the fraction of the drug that cleaved in the MCT oil carrier were determined. Briefly, the samples were centrifuged to separate the microparticles from the MCT oil carrier. Each portion was independently treated to determine the amount of drug contained. The 10 results were reported on the basis of the percentage resident in each independent portion.

TABLE 4

PLGA copolymer; 2 Days of storage at 5 ° C

<td>Compound</td><td>Microparticles</td><td>MCT carrier</td>

<td>Davalintida</td><td> 99.8%</td><td> 0.2%</td>

<td>pramlintide</td><td> 100.0%</td><td> 0.0%</td>

<td>Metreleptin</td><td> 100.0%</td><td> 0.0%</td>

<td>Bovine serum albumin</td><td> 100.0%</td><td> 0.0%</td>

<td>Sodium salicylate</td><td> 99.5%</td><td> 0.5%</td>

<td>Salicylic acid</td><td> 98.9%</td><td> 1.1%</td>

<td>minocycline</td><td> 99.1%</td><td> 0.9%</td>

<img file="MX352189B_D0042.tif" />

IMPI

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

TABLE 5

PLGA copolymer; 1 month storage at 5 ° C

<td>Compound</td><td>Microparticles</td><td>MCT carrier</td>

<td>Davalintida</td><td> 99.4%</td><td> 0.6%</td>

<td>Pramlintide</td><td> 99.7%</td><td> 0.3%</td>

<td>Metreleptin</td><td> 100.0%</td><td> 0.0%</td>

<td>Bovine serum albumin</td><td> 100.0%</td><td> 0.0%</td>

<td>Sodium salicylate</td><td> 98.7%</td><td> 1.3%</td>

<td>Salicylic acid</td><td> 99.9%·</td><td> 0.1%</td>

<td>minocycline</td><td> 99.9%</td><td> 0.1%</td>

<td>insulin</td><td> 99.5%</td><td> 0.5%</td>

TABLE 6

PLGA copolymer; 2 Days of Storage at 25 ° C

<td>Compound</td><td>Microparticles</td><td>MCT carrier</td>

<td>Davalintida</td><td> 100.0%</td><td> 0.0%</td>

<td>Pramlintide</td><td> 100.0%</td><td> 0.0%</td>

<td>Metreleptin</td><td> 100.0%</td><td> 0.0%</td>

<td>Bovine serum albumin</td><td> 100.0%</td><td> 0.0%</td>

<td>Sodium salicylate</td><td> 97.7%</td><td> 2.3%</td>

<td>Salicylic acid</td><td> 99.1%</td><td> 0.9%</td>

<td>minocycline</td><td> 99.4%</td><td> 0.6%</td>

TABLE 7

PLGA copolymer; 1 Month Storage at 25 ° C

<td colspan="3">PLGA polymer; 1 Month Storage at 25 ° C</td>

<td>Compound</td><td>Microparticles</td><td>MCT carrier</td>

<td>Davalintida</td><td> 100.0%</td><td> 0.0%</td>

<td>Pramlintide</td><td> 100.0%</td><td> 0.0%</td>

<td>Metreleptin</td><td> 100.0%</td><td> 0.0%</td>

<td>Bovine serum albumin</td><td> 100.0%</td><td> 0.0%</td>

<td>Sodium salicylate</td><td> 98.5%</td><td> 1.5%</td>

<td>Salicylic acid</td><td> 99.8%</td><td> 0.2%</td>

<td>minocycline</td><td> 99.6%</td><td> 0.4%</td>

<td>insulin</td><td> 99.3%</td><td> 0.7%</td>

IMPI

<img file="MX352189B_D0043.tif" />

TABLE 8

INSTITUTO MEXICANO Di IA PtOPIIDAO

INDUSTRIAL

Polycaprolactone / PLGA copolymer; Two Weeks of Storage

<td></td><td colspan="2">5 ° C</td><td colspan="2">25 ° C</td>

<td>Compound</td><td>Microparticles</td><td>MCT carrier</td><td>Microparticles</td><td>MCT carrier</td>

<td>Pramlintide</td><td> 100.0%</td><td> 0.0%</td><td> 100.0%</td><td> 0.0%</td>

The data in Tables 4-8 illustrates 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 a soluble oil, did not migrate in the MCT oil carrier, even though its solubility in MCT oil is greater than 30 mg / ml. In this way, the microparticles remain intact in storage in MCT even when the active pharmaceutical ingredient is soluble in MCT. The data further illustrates that the compositions can be successfully produced even using other polymer mixtures in the microparticles.

EXAMPLE 6

The percent purity of exenatide was measured by HPLC at one-month intervals over a 9-month period in the next four

<img file="MX352189B_D0044.tif" />

IMPI

MEXICAN INSTITUTE

OF INDUSTRIAL PROPERTY formulations: (i) a formulation comprising loo miorocaferaa dei^{, mm}~ Example 1 stored in a MCT oil carrier at 5 ° C; (Ii) a formulation comprising the microspheres of Example 1 stored in an MCT oil carrier at 25 ° C; (iii) dry microspheres from Example 1 that have been stored in a container for 9 months at 5 ° C without a liquid carrier, and where they were mixed with an aqueous carrier immediately prior to study; and (iv) dried microspheres from Example 1 that have been stored in a container for 9 months at 25 ° C without a liquid carrier, and that were mixed with an aqueous carrier immediately prior to study.

Figures 5A and 5B show the following: (i) exenatide has a purity greater than 93% in 6 months and 9 months in the formulation with the oil carrier at a temperature of 5 ° C; (ii) exenatide has a purity greater than 86% in 6 months and 9 months in the formulation with the oil carrier at a temperature of 25 ° C; (iii) exenatide has a purity greater than 94% in 6 months where the microspheres have been stored at 5 ° C; and (iv) exenatide has a purity greater than 90% in 6 months in the formulation where the microspheres have been stored dry at a temperature of 25 ° C. In Figure 5A, the purity of exenatide was determined by strong cation exchange HPLC. In Figure 5B, the purity of exenatide was determined by reverse phase HPLC.

<img file="MX352189B_D0045.tif" />

IMPI

Mexican Institute of Industrial Property

EXAMPLE 7

Formulations containing the microspheres of Example 1 and an MCT oil carrier were stored at 5 ° C and the potency of exenatide was measured at monthly intervals for 9 months. Additionally, the formulations containing the microspheres of Example 1 and an MCT oil carrier were stored at 25 ° C and the potency of exenatide was measured at monthly intervals for 6 months. Figure 6 presents the results which show that the potency of exenatide was preserved for at least 9 months.

EXAMPLE 8

A formulation containing the microspheres of Example 1 in an MCT oil carrier was tested for physical integrity. After storage for a period of 6 months at 5 ° C, the molar 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 ° C, the molecular weight of the poly (lactide-co-glycolide) copolymer decreased by 6 kDaltons, which is comparable to the change in molecular weight of dry microspheres (i.e. microspheres stored for 6 months at 25 ° C not in any carrier). The mean diameter of the microspheres was measured after storage at 3, 6 and 9 months either at 5 ° C or 25 ° C, and no change in mean diameter was detected in

<img file="MX352189B_D0046.tif" />

IMPI

MIXICaw INSTITUTE

DÉ LA rHOmr? AU INDUSTRIA !, relation to zero time.

EXAMPLE 9

The lactide / glycolide ratio for microparticles for use with various APIs was also investigated. The table below provides the varied lactide / glycolide ratios used.

<td>Polymer</td><td>Drug</td><td>Polymer MW approx. (kDa)</td><td>Lactide / Glycolide Ratio</td>

<td></td><td></td><td></td><td>for PLGA</td>

<td>PLGA</td><td>Davalintida</td><td> 10</td><td> 50/50</td>

<td>10 PLGA</td><td>Pramlintide</td><td> 10</td><td> 50/50</td>

<td>PLGA</td><td>Leptin</td><td> 10</td><td> 75/25</td>

<td>PLGA</td><td>BSA</td><td> 25</td><td> 50/50</td>

<td>PLGA</td><td>Na salicylate</td><td> 25</td><td> 50/50</td>

<td>PLGA</td><td>Salicylic acid</td><td> 25</td><td> 50/50</td>

<td>PLGA</td><td>Minocycline</td><td> 10</td><td> 75/25</td>

<td>PLGA</td><td>insulin</td><td> 25</td><td> 50/50</td>

<td>1.1: 1 PLC / PLGA</td><td>pramlintide</td><td>PCL = 150</td><td> 50/50</td>

PLGA = 10

All publications and patents are incorporated by reference herein. The foregoing has been described in detail and one skilled in the art will recognize that modifications may be made without departing from the spirit or scope of the appended description or claims.

<img file="MX352189B_D0047.tif" />