Sustained release of biotherapeutic from biodegradable polymer particles

Abstract

Disclosed herein are particles, methods of forming particles, and formulations comprising particles. The particles are particles comprising a biodegradable polymer, a biotherapeutic, and vitamin E. The biodegradable polymer is a polyesteramide (PEA) of defined structure. The particles may exhibit desirable release properties in vivo.

Description

[0001] SUSTAINED RELEASE OF BIOTHERAPEUTIC FROM BIODEGRADABLE POLYMER PARTICLES FIELD OF THE INVENTION

[0002]

[0001] Aspects of the invention relate to particles of polyesteramide polymers encapsulating water- soluble biotherapeutic and more particularly those exhibiting advantageous in vivo performance characteristics and other properties that render the particles suitable, according to specific embodiments, for use in administering biotherapeutics with desirable release profiles.

[0003] BACKGROUND

[0004]

[0002] Biotherapeutics are medicinal products that are peptides, polypeptides, proteins, nucleic acids or nucleic acid based, monoclonal antibodies, vaccines, or formulations thereof. Biotherapeutics are manufactured in, extracted from, or semi-synthesized from biological sources.

[0005]

[0003] Biotherapeutics are administered to patients for myriad medical reasons, often by injection or infusion. Due to limited plasma lifetime, administration of biotherapeutics may be required at frequent intervals. Certain biotherapeutics may have their properties modified to allow administration at weekly intervals, but longer intervals between injections is still desired.

[0006]

[0004] Sustained release or pulsatile release of the biotherapeutic after a single injection or implantation would reduce the number of injections required to achieve the desired therapeutic effect. Sustained release systems may release the biotherapeutic over the course of a few days to several months. Typically, limited burst release is desired so as to maintain efficient therapeutic concentration of the biotherapeutic.

[0007]

[0005] A pulsatile drug delivery system may involve the simultaneous administration of an immediate release component and a modified release component. The immediate release component may act as a first dose or provide an immediately felt impact on the patient, while the modified release component may act as a second dose and / or provide a longer term benefit to the patient.

[0008]

[0006] One type of biotherapeutic is a glucagon-like peptide-1 receptor agonist (GLP-1 RA). GLP-1 RA treatments have become extremely popular for treating Type 2 diabetes and obesity. Examples of GLP-1 RA treatments are exenatide, tirzepatide, liraglutide, semaglutide, and dulaglutide. GLP-1 RA treatments require frequent subcutaneous injections. Such frequent injections are uncomfortable for patients and inhibit patient compliance.2025P00125WG

[0009]

[0007] For example, tirzepatide requires weekly subcutaneous injections. The approved label for tirzepatide requires initial dosing for four weeks using a once weekly 2.5mg dose, increasing in 2.5 mg increments after at least 4 weeks until the recommended weekly maintenance dose of 10 mg or 15 mg is reached. Accordingly, several months of treatment may be required to achieve the desired dose for a particular patient. This can be prolonged if a dose is missed. Improved patient outcomes could be achieved if the desired dose could be achieved quicker and / or if less frequent subcutaneous injections were required for efficacy.

[0010]

[0008] US2009 / 0169632 discloses a sustained release composition comprising a polymer and manufacturing method thereof. The sustained release composition comprises a polymer, a bioactive agent, and a release rate determined agent, wherein the release rate determined agent is dispersed in the sustained release composition to control the release rate of the bioactive agent. The method comprises providing an oil phase comprising a bioactive agent, a polymer, and a release rate determined agent; providing an aqueous phase comprising a surfactant; mixing the oil phase with the aqueous phase to form the sustained release composition having a controlled release effect.

[0011]

[0009] US2014 / 0120170 describes particles comprising polyesteramide co-polymers that may provide a sustained release of bioactive agents and degrade hydrolytically at physiological conditions via bulk erosion mechanism in contrast with the polyesteramide co-polymers previously known in the prior art that degrade only in presence of certain classes of enzymes by surface erosion.

[0012]

[0010] WO2024 / 134493 describes biocompatible polymeric drug delivery systems for long-acting or controlled release of glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide- 1 (GLP-1) dual agonist, or pharmaceutically acceptable salts thereof comprising a polymeric material to form a polymer matrix; and GIP and GLP-1 dual agonist is homogeneously dispersed within the polymer matrix, wherein the polymer matrix comprises a biodegradable polymer and pharmaceutically acceptable excipients. The biocompatible polymeric drug delivery system can be administered into a patient to release for long-acting or controlled or sustained period.

[0013]

[0011] Drug delivery systems that are able to achieve the therapeutic effect of a biotherapeutic with a reduced number of injections would be desirable.SUMMARY

[0014]

[0012] One desired aspect of a drug delivery system is that an initial “burst” of biotherapeutic upon injection should usually be maintained in a safe range. For sustained release, it is desired to rather release the biotherapeutic slowly over time and over the course of a few weeks to several months. For a pulsatile system, it is desired to release the dose, whether as a burst or gradually, sometime after initial injection. For example, after 25-30 days the majority of the delayed dose is released over ten days or less.

[0015]

[0013] In contrast to the objectives pursued in certain prior art sustained-release systems, such as those described in US2009 / 0169632, the present invention is not directed to eliminating or shortening an initial lag phase in drug release. Rather, the present invention is specifically concerned with achieving prolonged, controlled release over an extended duration while minimizing or avoiding an initial burst release of the active agent. The prior art emphasizes release profiles designed to prevent a lag phase following administration, often prioritizing early availability of the bioactive agent. By contrast, the present invention recognizes that, for certain therapeutic applications, excessive early release may be undesirable and may compromise safety, tolerability, or dosing control. Accordingly, the present invention focuses on formulations and material selections that suppress burst release and sustain delivery over time, thereby addressing a fundamentally different technical problem than that addressed by the prior art.

[0016]

[0014] Achieving sustained release profiles in an injectable formulation while ensuring sufficiently high loading of biotherapeutic is challenging. The administered formulation must be injectable, typically requiring particles of sufficiently small size. Increased biotherapeutic payload may negatively affect particle formation generally as there is naturally less of the other components of the formulation needed to form particles as the amount of biotherapeutic increases. Achieving particles that are sufficiently stable once formed is also challenging, as is achieving a desired release profile once sufficient particles are formed.

[0017]

[0015] Aspects of the invention are associated with the discovery that certain formulations comprising a water soluble biotherapeutic and certain biodegradable polymers yield surprising sustained release properties. Benefits of the disclosed inventions may include, alone or in combination, an improved release duration, limited burst release, improved initial release over the first seven days, a quicker achievement of a desired dose, reduced injection frequency, an improved dosing regimen, higher drug loading, patient comfort, patient compliance, reduced side effects,reduced gastrointestinal effect or discomfort, manufacturing efficiency or repeatability, and improved product stability.

[0018]

[0016] These and other embodiments, aspects, and advantages relating to the present invention are apparent from the following Detailed Description.

[0019] BRIEF DESCRIPTION OF THE DRAWINGS

[0020]

[0017] A more complete understanding of exemplary embodiments of the invention and the advantages thereof may be acquired by referring to the following description in consideration of the accompanying figures, which serve to illustrate various features and certain principles involved.

[0021]

[0018] Fig. 1 is a chart of particle size distribution of four particle formulations, each employing a different polyesteramide polymer.

[0022]

[0019] Fig. 2 is a chart of average cumulative release of tirzepatide over time from four particle formulations, each employing a different polyesteramide polymer.

[0023]

[0020] Fig. 3 is a chart of average cumulative release of tirzepatide over time for five particle formulations of varying composition.

[0024]

[0021] Fig. 4 is a chart of average cumulative release from two particle formulations of varying composition.

[0025]

[0022] Fig. 5 is a chart of average cumulative release from four particle formulations of varying composition.

[0026] DETAILED DESCRIPTION

[0027]

[0023] The disclosures herein relate to particles comprising a water-soluble biotherapeutics encapsulated by certain biodegradable polymers, so-called encapsulated particles. The biodegradable polymers are polyesteramides (PEAs) that are random copolymers comprising certain “units” as hereinafter described. The particles comprise a water-soluble biotherapeutic, the biodegradable polymer, and vitamin E.

[0028]

[0024] As used herein, “vitamin E” means a group of compounds known collectively as vitamin E in the art, including both fat-soluble and water-soluble forms and derivatives thereof. Fat-soluble vitamin E compounds include tocopherols and tocotrienols, such as alpha-tocopherol, beta-tocopherol, gamma-tocopherol, delta-tocopherol, alpha-tocotrienol, beta-tocotrienol,gamma-tocotrienol, and delta-tocotrienol, as well as esters thereof, including acetate, succinate, and phosphate esters. Water-soluble vitamin E derivatives include vitamin E conjugates modified to increase hydrophilicity, such as tocopheryl polyethylene glycol succinate (vitamin E-TPGS), tocopheryl phosphate salts, and other polymer- or surfactant-modified tocopherol derivatives. Unless otherwise indicated, the term “vitamin E” encompasses any of the foregoing compounds and derivatives, whether naturally occurring or synthetic. In a preferred embodiment, the Vitamin E comprises alpha-tocopherol. In a preferred embodiment, the Vitamin E consists of alpha-tocopherol. In an embodiment, the vitamin E comprises a fatsoluble form of vitamin E. In an embodiment, the vitamin E consists of one or more fat-soluble forms of vitamin E. In an embodiment, the vitamin E comprises a water-soluble form of vitamin E. In an embodiment, the vitamin E consists of one or more water-soluble forms of vitamin E.

[0029]

[0025] As used herein, “tirzepatide” means a GIP / GLP1 dual agonist peptide as described in US 9,474,780 and described by CAS Registry Number: 2023788-19-2. US9474780 describes and claims tirzepatide. More specifically, tirzepatide is described in Example 1 of US 9,474,780, with the following sequence: YX1EGTFTSDYSIX2LDKIAQKAFVQWLIAGGPSSGAPPPS wherein Xi is Aib; X2 is Aib; K at position 20 is chemically modified through conjugation to the epsilon-amino group of the K side-chain with (2-[2-(2-Amino-ethoxy)-ethoxy]-acetyl)2- (yGlu)i-CO-(CH2) 18-CO2H; and the C-terminal amino acid is amidated as a C-terminal primary amide (SEQ ID NO: 1).

[0030]

[0026] As used throughout this description, the biotherapeutic is a water-soluble peptide. In an embodiment, the biotherapeutic is a GLP-1 receptor agonist. In an embodiment, the peptide is a glucose-dependent insulinotropic polypeptide (GIP) agonist.

[0031]

[0027] In an embodiment, the biotherapeutic comprises tirzepatide. Tirzepatide is a GIP / GLP1 dual agonist peptide as described in US 9,474,780 and described by CAS Registry Number: 2023788-19-2. More specifically, tirzepatide is described in Example 1 of US 9,474,780, with the following sequence: YX1EGTFTSDYSIX2LDKIAQKAFVQWLIAGGPSSGAPPPS wherein Xi is Aib; X2 is Aib; K at position 20 is chemically modified through conjugation to the epsilon-amino group of the K side-chain with (2-[2-(2-Amino-ethoxy)-ethoxy]-acetyl)2- (yGlu)i-CO-(CH2) 18-CO2H; and the C-terminal amino acid is amidated as a C-terminal primary amide (SEQ ID NO: 1).

[0028] In an embodiment, the biotherapeutic comprises from 15 to 100 amino acid residues. In an embodiment, the biotherapeutic comprises at least 15, 20, 25, or 30 amino acid residues. In an embodiment, the biotherapeutic comprises at most 100, 90, 80, 70, 60, 50 or 40 amino acid residues.

[0032]

[0029] In addition to amino acid residues, the biotherapeutic may comprise certain modifications to enhance cell uptake, binding to albumin, or stability. In an embodiment, the biotherapeutic comprises a peptide coupled to a fatty acid moiety, such as eicosanedioic acid.

[0033]

[0030] In an embodiment, the biotherapeutic has a molecular weight of from 3000 to 80,000 g / mol. In an embodiment, the biotherapeutic has a molecular weight of from 3000 to 6000 g / mol, preferably 3500 to 5500 g / mol. In an embodiment, the biotherapeutic has a molecular weight of from 4500 to 5200 g / mol.

[0034]

[0031] In an embodiment, the water-soluble biotherapeutic is present in an amount of from 3 wt% to 12 wt%, based on the total weight of the particles, preferably 4 to 11 wt%, and more preferably from 5 to 10 wt%. In an embodiment, the water-soluble biotherapeutic is present in an amount of from 3 wt%, 4 wt%, 5 wt%, 6 wt%, or 7 wt%, based on the total weight of the particles. In an embodiment, the water-soluble biotherapeutic is present in an amount of from 12 wt%, 11 wt%, 10 wt%, 9 wt%, or 8 wt%, based on the total weight of the particles.

[0035]

[0032] In an embodiment, the biodegradable polymer is present in an amount of from 78 wt% to 92 wt%, based on the total weight of the particles, preferably 80 to 92 wt%. In an embodiment, the biodegradable polymer is present in an amount of from 78 wt%, 79 wt%, 80 wt%, 81 wt%, 82 wt%, 83 wt%, 84 wt%, or 85 wt%, based on the total weight of the particles. In an embodiment, the biodegradable polymer is present in an amount of from 92 wt%, 91 wt%, 90 wt%, 89 wt%, or 88 wt%, based on the total weight of the particles.

[0036]

[0033] In an embodiment, the vitamin E is present in an amount of from 1 wt% to 10 wt%, based on the total weight of the particles, preferably 2 to 8 wt%. In an embodiment, the vitamin E is present in an amount of from 2 wt%, 3 wt%, 4 wt%, or 5 wt%, based on the total weight of the particles. In an embodiment, the vitamin E is present in an amount of from 9 wt%, 8 wt%, 7 wt%, or 6 wt%.

[0037]

[0034] In an embodiment, the particles further comprise an additive. In an embodiment, the particles further comprise a surfactant, plasticizer, antioxidant, chelating agent, buffer, or osmotic agent.In an embodiment, the particles consists of the biodegradable polymer, the biotherapeutic, and the vitamin E.

[0038]

[0035] In an embodiment, the average particle size (d50) of the particles is from 1 to 500 pm, preferably from 5 to 200 pm. In an embodiment, the particles have a d50 particle size of from 5, 10, 15, 20, 25, or 30 pm. In an embodiment, the particles have a d50 particle size of at most 200, 150, 120, 100, 90, or 80 pm. d50 particle size is measured in Milli Q® water by static light laser scattering using a Malvern Mastersizer equipped with an aqueous medium sample dispersion chamber.

[0039]

[0036] In an embodiment, the particles are formed by a so-called water in oil in water (W / O / W) technique. In this technique, an oil phase is first formed by dissolving the PEA and vitamin E in an organic solvent that is suitable to both dissolve the PEA and form an emulsion with the water phase. In an embodiment, the organic solvent comprises dichloromethane. A water phase is formed by dissolving the water-soluble biotherapeutic in water. Next, an emulsion is formed by adding the water phase to the oil phase and emulsifying. Then, the emulsion is injected into water and mixed. Lastly, the emulsion in water is added into a larger water bath to dissolve the remaining organic solvent and further harden the particles. The particles are then sieved and dried.

[0040]

[0037] The particles are preferably delivered to the patient in an injectable formulation. The formulation may comprise the particles along with a suitable pharmaceutically acceptable carrier, such as saline or an oily vehicle.

[0041]

[0038] In an embodiment, the formulation may further comprise second particles. The second particles may comprise essentially the same components as the (first) particles but may differ in a desired attribute. For example, the second particles may differ from the first particles in particle size, biotherapeutic loading, biodegradable polymer formulation, comprise additional biodegradable polymers, shell thickness, or comprise a further shell.

[0042]

[0039] In an embodiment, the formulation further comprises unencapsulated water-soluble biotherapeutic. In this way, an immediate dose of biotherapeutic may be delivered with a delayed dose being delivered via the first particles. In an embodiment, the unencapsulated biotherapeutic is present as particles. In an embodiment, such unencapsulated biotherapeutic is the same as the water-soluble biotherapeutic in the first particles.

[0040] In an embodiment, less than 20% of the water-soluble biotherapeutic is released from the particles after twenty-four hours in phosphate buffer at pH 7.4 containing 137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPO4 and 1.8 mM KH2PO4 at 37 °C. In an embodiment, less than 25% of the water-soluble biotherapeutic is released from the particles after 7 days in phosphate buffer at pH 7.4 containing 137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPO4 and 1.8 mM KH2PO4 at 37 °C.

[0043]

[0041] An injectable formulation comprising the particles may be administered to a patient by injecting the formulation. In an embodiment, less than 20% of the water-soluble biotherapeutic is released from the particles after twenty-four hours. In an embodiment, less than 25% of the water-soluble biotherapeutic is released from the particles after 24 hours.

[0044]

[0042] In addition to use of the encapsulated particles in an injected formulation, the encapsulated particles may be processed with another degradable or non-degradable biomaterial. In an embodiment, the particles are dispersed within a degradable or non-degradable polymer to become a (component of) a drug-eluting implant. In such an implant, the particles may be dispersed into another biomaterial to control the initial burst and drug release kinetics. In an embodiment, the particles may form the interior of an implant comprising an external degradable polymer layer or coating. In another embodiment, a melt is formed comprising a degradable or non-degradable polymer and the encapsulated particles, and a (component of) a medical implant is formed. In an embodiment the degradable or non-degradable polymer is a degradable polyester, such as PLA, PLLA, PLGA, or a combination thereof. In an embodiment the degradable or non-degradable polymer is ethylene-vinyl acetate (EVA) or a thermoplastic polyurethane (TPU). In the case of a degradable polymer, upon degradation of the external layer or body of the component, the encapsulated particles may be released. The implant may be formed by a hot-melt process, injection molding, or extrusion. Such an implant is typically a solid or a semi-solid.

[0045]

[0043] In a further embodiment, a plurality of encapsulated particles is further encapsulated via a solid- oil-in-water emulsification process to form an injectable particles formulation. In this embodiment the primary particles are suspended into an encapsulating polymer-containing oil phase which is added to an aqueous phase under the conditions of an emulsification process to yield injectable particles of average particle size (d50) of from 40 to 250 pm. In such an embodiment, each particle comprises a shell comprising a degradable encapsulating polymer encapsulating a plurality of the encapsulated particles.

[0044] The PEA has a structure that includes, or that may consist of, n units, each unit having residues, for example 2, 3, or 4 different types of residues, as defined below, which are present in each unit at respective molar equivalents that are designated by m, p, q, and x, in which m+p+q+x=l. The n units may be characterized as “repeating” units to the extent that the defined values or ranges of m, p, q, and x are consistent from one unit to the next, although it is not required for each unit to be identical in structure, as long as the defined values or ranges are met. As is apparent from the structures of residues given below, they are linked by amide bonds [(- C=O)NH-)], whereas certain of these residues include ester bonds [(-C=O)O-)] that help confer water permeability. Given the teachings herein, those skilled in the art will be able to appreciate the structural modifications and corresponding adjustments in synthesis procedures that influence certain properties described herein.

[0046]

[0045] For ease of understanding, dashed lines are used in structures of the individual residues, and in other structures, to emphasize that they represent points of attachment (bonding) and not methyl groups. To the extent that structures may include chiral carbon atoms, the lack of an express illustration of stereochemistry is meant to convey that all stereoisomers are intended. However, those skilled in organic chemistry, given the teachings herein including the specific synthesis procedures, will be able to ascertain particular stereochemical configurations that are representative of PEA polymers in preferred embodiments.

[0047]

[0046] More specifically, the n units in the in the structure of the PEA may have:

[0048] m molar equivalents of a first residue having the formula

[0049]

[0050] p molar equivalents of a second residue having the formula

[0051] O O H O O H

[0052] - C - R1- C - N - C - C - O - R6- O - C - C - N -

[0053]

[0054] H R4R4H q molar equivalents of a third residue having the formulaO O H H

[0055] - C - R1- C - N - C - R8- N - H C - O - R7

[0056]

[0057] 0, and

[0058] x molar equivalents of a fourth residue having the formula

[0059] O O H H

[0060] - C - R1- C - N - C - R8- N - H C - O - H

[0061]

[0062] O

[0063]

[0047] The values m, p, q, and x represent molar equivalents of the corresponding first, second, third, and fourth residues in the units of the random copolymer, which molar equivalents are scaled such that m+p+q+x=l, thereby indicating relative molar equivalents among the residues.

[0064]

[0048] According to a first embodiment, m is from 0 to 0.8, p is from 0 to 0.95, m+p is from is from 0.5 to 1, q is from 0.02 to 0.25, and x is from 0.05 to 0.25; n is from 5 to 300; R1is (C2- C2o)alkylene; R3and R4are selected from the group consisting of hydrogen, (Ci-C6)alkyl, (C2- C6)alkenyl, (C2-C6)alkynyl, (C6-Cio)aiyl, -CH2SH, -(CH2)2S(CH3), -CH2OH, -CH(OH)CH3, -(CH2)4NH3+, -(CH2)3NHC(=NH2+)NH2, -CH2COOH, -CH2-CO-NH2, -CH2CH2-CO-NH2, -CH2CH2COOH, CH3-CH2-CH(CH3)-, (CH3)2CH-CH2-, H2N-(CH2)4-, Ph-CH2-, CH=C-CH2- , (CH3)2CH-, Ph-NH-, NH-(CH2)3-C-, or NH-CH=N-CH=C-CH2-, wherein R3and R4are the 1 _ 1 1 - 1

[0065] same or different; R5is (C2-C2o)alkylene, optionally having one or more alkylene carbon atoms (-C-) replaced by an oxygen atom (-O-), and / or optionally having 5 or 6 consecutive alkylene carbon atoms together form a respective 5- or 6-membered carbocyclic ring;

[0066] R6is

[0067]

[0068] R7is (C6-Cio)aryl(Ci-C6)alkylene; and R8is (C3-Cs)alkylene. In such PEA polymers, higher values of p, i.e. , increased proportions of the second residue, can lead to a higher glass transition temperature, and more particularly a higher glass transition temperature of the polymer in its hydrated state (wet Tg).

[0069]

[0049] As used herein, the term “alkyl” means a monovalent straight or branched chain hydrocarbon group including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-hexyl, and the like, with more specific alkyl groups being limited according to ranges of carbon atom numbers, as indicated in parentheses, preceding “alkyl.”

[0070]

[0050] As used herein, the term “alkylene” means a divalent straight or branched chain hydrocarbon group such as -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CIfc)?-, and the like, with more specific alkyl groups being limited according to ranges of carbon atom numbers, as indicated in parentheses, preceding “alkylene.”

[0071]

[0051] As used herein, the term “alkenyl” means a monovalent straight or branched chain hydrocarbon group having at least one carbon-carbon double bond in the main chain or in a side chain, with more specific alkenyl groups being limited according to ranges of carbon atom numbers, as indicated in parentheses, preceding “alkenyl.”

[0072]

[0052] As used herein, the term “alkenylene” means a divalent straight or branched chain hydrocarbon group having at least one carbon-carbon double bond in the main chain or in a side chain, with more specific alkenylene groups being limited according to ranges of carbon atom numbers, as indicated in parentheses, preceding “alkenylene.”

[0073]

[0053] As used herein, “alkynyl” means a monovalent straight or branched chain hydrocarbon group having at least one carbon-carbon triple bond in the main chain or in a side chain, with more specific alkynyl groups being limited according to ranges of carbon atom numbers, as indicated in parentheses, preceding “alkynyl.”

[0054] As used herein, “aryl” means an unsubstituted or optionally substituted phenyl radical or an unsubstituted or optionally substituted ortho-fused bicyclic carbocyclic radical having nine or ten ring atoms, in which at least one ring is aromatic. Examples of aryl include, but are not limited to, phenyl, naphthyl, and nitrophenyl.

[0074]

[0055] As used herein, “biodegradable" means a material which is capable of being completely or substantially degraded or eroded when exposed to an in vivo environment. A polymer is capable of being degraded or eroded when it can be gradually broken down, resorbed, absorbed, and / or eliminated by, for example, hydrolysis, enzymolysis, oxidation, metabolic processes, bulk or surface erosion, and the like. As noted above, in preferred embodiments, PEA polymers may be characterized as being biodegradable. This leads to advantages in the use of such polymers in delivery of biotherapeutics.

[0075]

[0056] According to the PEA structure as defined above and including n units, each having residues, for example 2, 3, or 4 different types of residues, in the case of a “random copolymer,” the residues, present in each unit at respective molar equivalents that are designated by m, p, q, and x, are distributed randomly throughout the units and throughout the copolymer.

[0076]

[0057] Given the structures of the monomers above, it can be appreciated that, in any of the embodiments as defined herein, the PEA may have the following structure:

[0077] O O H O O H O O H O O H II , .5II „ II I II „ II I — C— R1-C— N— C— C-O— R5-O— C— C— N C— R1-C— N— C— C-O— R6-O— C— C— N I I I I H R3R3H R4R4H

[0078]

[0079] wherein n, m, p, q, and x, as well as the substituents R1, R3, R4, R5, R6, R7, and R8are as defined above.

[0080]

[0058] In more specific embodiments, the PEA is defined as according to the first embodiment, wherein m may be from 0, 0.10, 0.15, 0.20, or 0.25 to 0.75, 0.70, 0.65, 0.60, 0.55, 0.50, 0.45,0.40, or 0.35. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein p may be from 0.10, 0.20, 0.30, 0.35, or 0.40 to 0.95, 0.90, 0.85, 0.75, 0.70, 0.65, 0.60, 0.55, 0.50, or 0.45. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein p may be greater than or equal to m. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein m and p may be both greater than zero. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein m:p may be from 2:1, 1 : 1, or 2:3 to 1:5, 1:4, 1 :3, or 1:2. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein q is from 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, or 0.20 to 0.25, 0.23, 0.21, 0.20, 0.19, 0.18, 0.17, 0.16, 0.15, 0.11, 0.10, 0.09, 0.08, or 0.07. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein x is from 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, or 0.20 to 0.30, 0.25, 0.20, 0.19, 0.18, 0.17, 0.16, 0.15, 0.14, 0.13, 0.125, 0.12, 0.11, or 0.10. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein the ratio q:x is from 50:50, 45:55, 40:60, or 35:65 to 20:80, 25:75, 30:70, or 35:65. In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein m is about 0.3, p is about 0.45, q is about 0.10, and x is about 0.15 (X60). In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein m is about 0.3, p is about 0.45, q is about 0.09, and x is about 0.16 (X65). In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein m is about 0.3, p is about 0.45, q is about 0.08, and x is about 0.18 (X70). In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein m is about 0.3, p is about 0.45, q is about 0.06, and x is about 0.19 (X75). In other more specific embodiments, the PEA is defined as according to the first embodiment, wherein n is from 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, or 75 to 300, 290, 280, 270, 260, 250, 240, 230, 220, 210, 200, 190, 180, 170, 160, or 150. In other more specific embodiments, the PEA is defined as according to the first embodiment and has a number average molecular weight (Mn) of at least 15,000 g / mol, at least 20,000 g / mol, at least 25,000 g / mol, at least 30,000 g / mol, or at least 35,000 g / mol. In other more specific embodiments, the PEA is defined as according to the first embodiment and has an Mn of at most 250,000 g / mol, at most 225,000 g / mol, at most 200,000 g / mol, at most 175,000 g / mol, at most 150,000 g / mol, at most 125,000 g / mol, at most 100,000 g / mol, or at most 75,000 g / mol. Mn is measured via gel permeation chromatography (GPC) in tetrahydrofuran (THF) with polystyrene as standard.

[0059] In yet further specific embodiments, the PEA is defined as according to the first embodiment, and may have any one or more of: ranges of m as defined above; ranges of p as defined above; p being greater than m; m and p being both greater than zero; ranges of m:p as defined above; ranges of q as defined above; ranges of x as defined above; ranges of q:x as defined above; values of m, p, q, and x as defined above; ranges of n as defined above; lower limits of Mn as defined above; and / or upper limits of Mn as defined above.

[0081]

[0060] In yet further specific embodiments, the PEA is defined as according to the first embodiment, and may be more particularly characterized by any one or more of: R3is selected from the group consisting of hydrogen, (Ci-C6)alkyl, CH3-CH2-CH(CH3)-, (CEE CH-CEE-, PI1-CH2-, and (CHshCH-; R4is selected from the group consisting of hydrogen, (Ci-C6)alkyl, CH3-CH2- CH(CH3)-, (CH3)2CH-CH2-, Ph-CH2-, and (CH3)2CH-; R3and R4are the same; R5is (C2- C2o)alkylene; R7is (C6)aryl-CH2- (z.e. benzyl or phenylmethyl); and / or R8is -(CH2)4-.

[0082]

[0061] In yet further specific embodiments, the PEA is defined as according to the first embodiment, and may be more particularly characterized by any one or more of: p is from 0 to 0.8 and m+p is from 0.5 to 0.9; q is from 0.05 to 0.15; x is from 0.10 to 0.25; q:x is from 50:50 to 20:80, or more particularly from 45:55 to 25:75, or more particularly from 40:60 to 30:70; and / or m is 0.

[0083]

[0062] In yet further specific embodiments, the PEA is defined as according to the first embodiment, and may be more particularly characterized by any one or more of: m is from 0.1 to 0.5, p is from 0.1 to 0.75, m+p is from 0.5 to 0.8, q is from 0.05 to 0.15, and x is from 0.10 to 0.25; R1is (C2-Cio)alkylene; R3and R4are (Ci-C6)alkyl; R5is (C2-Cio)alkylene;

[0084] R6is

[0085]

[0086] '» ; R7is Ph-(Ci-C6)alkylene; and R8is (C3-C6)alkylene. For example, the PEA may be characterized by one or more of these definitions of m, p, m+p, q, x, R1, R3, R4, R5, R6, R7, and R8, such as in the case of the PEA being characterized by all of these definitions.

[0063] Polyesteramide random copolymers may be synthesized by adapting a procedure known in the art from Katsarava et al. (WON J POLYM SCI A: POLYM CHEM 1999:37 : 391-407). Briefly, the polymers are prepared via solution polycondensation of di-p-toluenesulfonic or hydrochloric acid salts of bis-(a-amino acid) a,co-diol diesters, lysine benzyl ester, lysine, and / or di-N- hydroxysuccinimide ester of sebacic acid in anhydrous DMSO. Typically, the salts are converted to free amines by addition of triethylamine and these amines are further reacted with the di-acid derivative. The usage of pre-activated acid in the reaction allows polymerization at relatively low temperature, such as 65°C, affording side-product free polycondensates and predictable degradation products. Subsequently, the obtained reaction mixture is purified via a water precipitation followed by an organic precipitation and filtration. Drying under reduced pressure yields the polyesteramide random copolymer.

[0087]

[0064] For example, such polymers may be prepared by reacting lysine, lysine benzyl ester, and hexahydrofuro[3,2-b]furan-3,6-diyl bis(2-amino-4-methylpentanoate) with di-N- hydroxysuccinimide ester activated sebacic acid in DMSO for 24 hours. The polymer is then isolated from the reaction mixture in two precipitation steps and characterized by means of proton NMR and THF-based GPC relative to polystyrene standards.

[0088] EXAMPLES

[0089]

[0065] The following examples are set forth as representative of the present invention. These examples are not to be construed as limiting the scope of the invention as other equivalent embodiments will be apparent in view of the present disclosure and appended claims.

[0090] Preparation of Copolymers

[0091] The PEAs are random copolymer within the scope of PEA polymers as described herein and have the structure:O O H O O H O O H O O H II < II I IIKH I IIdII I II . II I — C— R1-C— I N— C— C-O— R5-O— C— C— N C— R1-C— N— C— C-O— R6-O— C— C— N A A= I I I I R3H R4R4H

[0092] O O H H O O H H

[0093] C— R1-C— N— C— R8-N C— R1-C— N— C— R8-N I I I H C-O— R H C-O— H II II

[0094] O o

[0095]

[0096] This PEA polymer therefore has residues with the structures as described generally herein, in which m is 0.30; p is 0.45; R1is octylene [(Cs)alkylene] ; R3and R4are isobutyl; R5is hexylene [(Ce)alkylene] ;

[0097] R

[0098]

[0099] 6is '» ;

[0100] R7is benzyl [Ph-(Ci)alkylene, or phenylmethyl]; and R8is butylene [(C4)alkylene] .

[0101]

[0066] The four prepared PEAs differ in the ratio of unit q and unit x. This is reported in Table 1, below:Table 1: PEA Description

[0102] PEA X

[0103] Name q

[0104] PEA-X25 0.1875 0.0625

[0105] PEA-X50 0.125 0.125

[0106] PEA-X65 0.0875 0.1625

[0107] PEA-X75 0.0625 0.1875

[0108]

[0109]

[0067] PEA-X65 and PEA-X75 were prepared as follows. Triethylamine and DMSO were added to a mixture of di-N-hydroxysuccinimide ester of sebacic acid (Di-NHS-sebacic acid), L-leucine- (DAS)-2TosOH, L-leucine(6)-2TosOH, L-lysine-2HCl and L-lysine(Bz)-2TosOH in a nitrogen flushed 1000 ml round bottomed flask equipped with an overhead stirrer at room temperature. The subsequent mixture was heated to 60°C to allow the reaction to proceed, with monitoring by GPC analysis in THF. After 36 hours, a stable molecular weight was obtained. The reaction mixture was diluted with 500 ml DMSO and was allowed to cool to room temperature. At room temperature, acetic anhydride was added to acylate the amino functional end groups of the polymer. Next, the mixture was stirred at room temperature for 24 hours.

[0110]

[0068] PEA-X25 and PEA-X50 were prepared as follows. Triethylamine and DMSO were added to a mixture of di-N-hydroxysuccinimide ester of sebacic acid (Di-NHS-sebacic acid), L-leucine- (DAS)-2TosOH, L-leucine(6)-2TosOH, L-lysine-2HCl and L-lysine(Bz)-2TosOH in a nitrogen flushed 500 ml round bottomed flask equipped with an overhead stirrer at room temperature. The subsequent mixture was heated to 60°C to allow the reaction to proceed, with monitoring by GPC analysis in THF. After 36 hours, a stable molecular weight was obtained. The reaction mixture was diluted with 250 ml DMSO and was allowed to cool to room temperature. At room temperature, acetic anhydride was added to acylate the amino functional end groups of the polymer. Next, the mixture was stirred at room temperature for 24 hours.

[0111]

[0069] The amount of the ingredients used to prepare each PEA is shown in Table 2, below.Table 2: PEA Ingredients

[0112] Di-NHS- L-leucine- L- L- TriethylL- leuci Acetic Name DMSO sebacic (DAS)- ne(6)- lysine(Bz)- amine 2 lysine-2HCl anhydride acid TosOH 2TosOH 2TosOH

[0113] PEA- 31 ml, 54 ml, 40.398g, 32.876 g, 21.062 g, 1.396 g, 11.097 g, 1.89 ml, X25 0.222 mol 0.76 mol 0.101 mol 0.046 mol 0.030 mol 0.006 mol 0.019 mol 0.0199 mol PEA- 31 ml, 54 ml, 39.805 g, 32.390 g, 20.753 g, 2.750 g, 7.289 g, 1.89 ml, X50 0.222 mol 0.76 mol 0.100 mol 0.045 mol 0.030 mol 0.013 mol 0.013 mol 0.0199 mol PEA- 64 ml, 110 ml, 82.398 g, 66.975 g, 42.920 g, 7.402 g, 10.555 g, 3.78 ml, X65 0.458 mol 1.55 mol 0.208 mol 0.093 mol 0.062 mol 0.034 mol 0.018 mol 0.0398 mol PEA- 65 mL, 110 ml, 84.429 g, 68.704 g, 42.021 g, 8.751 g, 7.727 g, 3.86 ml, X75 0.465 mol 1.55 mol 0.213 mol 0.096 mol 0.064 mol 0.040 mol 0.013 mol 0.0406 mol

[0114]

[0115]

[0070] The obtained crude polymer mixture was precipitated in water at a 10:1 ratio (water: reaction mixture). The polymer was collected and dissolved in ethanol (500 ml, 8.57 mole) and then precipitated a second time. The polymer was again dissolved in ethanol (500 ml, 8.57 mole) and precipitated in ethylacetate (5000 ml, 50.91 mole) by dropwise addition to a stirring solution. The precipitated polymer was washed with ethylacetate (100 ml, 1.00 mole), the supernatant was removed, and the precipitate was washed again with ethylacetate (100 ml, 1.00 mole). After the removal of the supernatant, the precipitate was dried and dissolved in ethanol (500 ml, 8.57 mole), and filtered over a 0.2 pm PTFE membrane filter. The filtered polymer solution was dried under reduced pressure at 65°C. From this synthesis procedure, a typical yield is 75%, and the number average molecular weight (Mn) is normally in the range of 40- 70 kDa, measured via gel permeation chromatography (GPC) using tetrahydrofuran (THF) as the mobile phase on dried samples, and determined relative to polystyrene standards.

[0116] UPLC analysis for loading determination and in-vitro release studies

[0117]

[0071] The loading of the biotherapeutic was determined via an analysis on an Acquity H class apparatus, equipped with a diode array UV-VIS detector operating at 214 nm. The chromatographic column was Acquity Premiers BEH Cl 8 column (1.7 pm particle size, 2.1 mm x 50 mm). A binary gradient was used, with a mobile phase A = 0.1% TFA + MilliQ® water and a mobile phase B = 0.1% TFA + Acetonitrile. The gradient used is specified in Table 3, below.Table 3: UPLC Gradient

[0118] Time (minutes) Flow rate (mL / min) A (%) B (%)

[0119] 0 0.4 65 35

[0120] 4 0.4 30 70

[0121] 4.1 0.4 5 95

[0122] 5 0.4 5 95

[0123] 5.5 0.4 65 35

[0124]

[0125] 7.5 0.4 65 35

[0126] The flow rate is maintained at 0.4 mL / min, the column’s temperature is set at 40°C and the injection volume for loading and extraction is set at 3 pL, while the injection volume for release is set at 5 pL.

[0127] Biotherapeutic Dissolution Assay

[0128]

[0072] Release experiments are executed in triplicates. Around 10 mg of microparticles are weighed in lobind Eppendorf tubes. 1 mL of phosphate buffer solution (PBS) containing 0.05% NaNs is added to a concentration of 10 mg / mL. The samples are incubated at 37°C under shaking at 100 rpm at typical sampling points, in this case on days 1, 2, 3, 4, 7, 9, 11, 14, 17, and 21. Weekly or biweekly, 800 pL of buffer is transferred to HPLC vials and measured using the UPLC method above, and 800 pL of fresh buffer solution is added.

[0129] Encapsulation Efficiency Calculation

[0130]

[0073] The encapsulation efficiency is calculated as a ratio of the measured biotherapeutic load and the theoretical biotherapeutic load, using the following equation:

[0131] (BiotherapeuticTm (Bioth.erapeutic)m EE(%) = f—— - - - • 100 = - - - - - 100

[0132] Biotherapeutic) th > "Lbtp _

[0133]

[0134] mbtp+ + mPEA 1+ mPEA 2where EE is the encapsulation efficiency in percent, (biotherapeutic)m is the measured biotherapeutic load (wt.%), (biotherapeutic)th is the theoretical biotherapeutic load (wt.%), mbtp is the amount of biotherapeutic (g), madd is the amount of additive (g), mpEA,i is the amount of PEA polymer 1 (g), and mpEA,2 is the amount of PEA polymer 2 (g).

[0135] Particle Formation

[0136]

[0074] Particles of biotherapeutic encapsulated by PEA are formed as follows.

[0137]

[0075] Preparation of the Oil Phase (O)PEA and vitamin E (DL-alpha-Tocopherol, Thermo Scientific Chemicals, Batch 10188932, Cat# A17039.0B; appearance: clear yellowish brown viscous liquid), if present, were dissolved in dichloromethane and 1-butanol (DCM:BuOH w / w between 13 and 15.6, average 14.3) using aNalgene™ LDPE Sample Vials with cap. The sample vials were placed on a shaker overnight at room temperature to ensure complete dissolution.

[0138]

[0076] Preparation of the outer water phase (W2 and hardening bath)

[0139] Per particle batch, 400 mL of outer water phase phase was made by dissolving 1 w / w% PVA and 2.5 w / w% NaCl in 400 mL MilliQ® water.

[0140]

[0077] Preparation of the inner water phase (Wl)

[0141] Tirzepatide (BOC Sciences, CAT No-006246, purity >98%) was used as the biotherapeutic. Tirzepatide powder was dissolved in MilliQ® water to form the inner water phase.

[0142]

[0078] Forming the W / O emulsion

[0143] Walls of the sample vial were rinsed with oil phase before adding the inner water phase to the oil phase using a pipette. The phases were emulsified using the ultrasonic probe with the ultrasonic at 75% intensity turning on and off for 1 second each time with a total “on” time of 10 seconds. A white, viscous emulsion was formed.

[0144]

[0079] Forming the W / O / W emulsion

[0145] The primary (W / O) emulsion is injected through a needle into a glass vial containing 20 mL W2 with a glass syringe and mixed with the ultraturrax for 3 minutes at 4000 rpm. After mixing, the dispersion head was rinsed with hardening phase and added to the 400 mL beaker containing 375 mL hardening phase and a magnetic stirrer bar. The solutions were left at room temperature overnight, with stirring at 400 rpm to allow the dichloromethane to evaporate.

[0146]

[0080] Sieving and drying

[0147] The next day, the particle sizes of the crude mixtures were analysed on the Mastersizer 2000. The microparticles were sieved with a 125-pm sieve and a 20-pm sieve. The beaker and sieves were washed three times with 0.04% Tween 80 in MilliQ® water solution using 50 mL each wash. The collected microparticles were added to a 10 mL vial, which was pre- weighed (together with a closing cap), covered with a dust-free tissue and dried in a freeze dryer at - 50°C under vacuum for 3 days. After drying, the vials were weighed, and the yield wasdetermined. The dried microparticles were analysed again on the Mastersizer 2000 for final particle size measurements. The particles were redispersed in MilliQ® water before analysis.

[0148] Effect of PEA Formula on Tirzepatide Release

[0149]

[0081] Biotherapeutic-loaded microparticles that consist of a PEA polymer and tirzepatide were formed according to the above procedures. Targeted loading of biotherapeutic was 10 wt%. The PEA polymer in each batch of particles was varied among PEA-X25, PEA-X50, PEA- X65, and PEA-75. The particle size distribution of each is shown in Fig. 1.

[0150]

[0082] Average cumulative release was tested over 30 days according to the above procedure for biotherapeutic dissolution assay. The results are shown in Fig. 2. It was observed that the release from PEA-X25 and PEA-X50 was insufficient and the release from PEA-X65 and PEA- X75 was too fast.

[0151] Effect of modified Formulations on Tirzepatide Release

[0152]

[0083] Attempts were made to modify the release observed in the prior experiment. Biotherapeutic- loaded microparticles were formed according to the above procedures. The particles have the formulations, measured particle sizes, and measured biotherapeutic loadings as shown in Table 5. Tirzepatide load was measured by UPLC and the amount of PEA(s) and vitamin E was then calculated based on the ratios used when forming the oil phase. Stated wt% is based on the total weight of the particles.

[0153] Table 4: Biotherapeutic-loaded particle formulations

[0154] PEAIIIX50 PEAIIIX65 TZP VitE

[0155] Sample Name

[0156] (wt%) (wt%) (wt%) (wt%)

[0157] PEAIIIX50 90.98% 0 9.02 0

[0158] PEAIIIX65 0 91.65 8.35 0 PEAIIIX65 / X50 blend 15.93 75.1 8.97 0 PEAIIIX65&Vit E 0 86.77 8.29 4.94 PEAIIIX65&higher

[0159] 0 87.04 12.96 0

[0160] TZP load

[0161]

[0162] Average cumulative release was tested according to the above procedures. The results are shown in Fig. 3. It was observed that the addition of vitamin E has a desirable effect on limiting release over the first seven days, which was further investigated in subsequent experiments.Effect of Vitamin E on Tirzepatide Release

[0163]

[0084] Biotherapeutic-loaded microparticles are formed according to the above procedures. The particles have the formulations, measured particle sizes, and measured biotherapeutic loadings as shown in Table 5.

[0164] Table 5: Characteristics of biotherapeutic-loaded microparticle samples Sample PEA DCM 1-butanol Vitamin BioMilliQ D50 No. (g) (g) (g) E (g) therapeutic (g) water (g) (gm)

[0165] 1 0.306 5.220 0.367 - 0.0363 0.960 40 2 0.296 5.220 0.361 0.0081 0.0335 0.960 29 3 0.299 5.190 0.360 0.0132 0.0335 0.960 29

[0166]

[0167] 4 0.165 2.659 0.180 0.0136 0.0191 0.480 65

[0168]

[0085] The release of 10 wt.% biotherapeutic from the microparticle samples of Table 5 is studied, and a comparison is made between microparticles with 5 wt.% vitamin E and microparticles without vitamin E in Figure 4. Further results are displayed in Figure 5 for four ranges of vitamin E loading: (1) no vitamin E added, (2) addition of 2.5 wt.% vitamin E, (3) addition of 5 wt.% vitamin E, and (4) addition of 7.5 wt.% vitamin E.

[0169]

[0086] The addition of vitamin E has an advantageous effect on the microparticles’ sustained release of biotherapeutic. Figure 4 shows that the initial release is controlled to about 25% in 5 days from an initial release of about 80% in 5 days. Figure 5 shows that the addition of different amounts of vitamin E leads to more controlled releases over the first seven days. The addition of 7.5% of vitamin E especially leads to very low initial release, leading to a properly sustained release of biotherapeutic overtime.

[0170]

[0087] Those skilled in the art having knowledge of the present disclosure will recognize that various changes can be made to these polymers and their applications, to attain these and other advantages, without departing from the scope of the present disclosure. As such, it should be understood that the features of the disclosure are susceptible to modifications and / or substitutions without departing from the scope of the invention. The specific embodiments illustrated and described herein are for illustrative purposes only, and not limiting of the invention as set forth in the appended claims.EXEMPLARY DESCRIPTION OF CERTAIN EMBODIMENTS

[0171] Particles comprising a water-soluble biotherapeutic, a biodegradable polymer, and vitamin E, wherein the biodegradable polymer comprises a random copolymer having a structure including n units of:

[0172] m molar equivalents of a first residue having the formula

[0173] O O H O O H

[0174] - C - R1- C - N - C - C - O - R5- O - C - C - N - I I I I

[0175]

[0176] H R3R3H p molar equivalents of a second residue having the formula

[0177] O O H O O H

[0178]

[0179] H R4R4H q molar equivalents of a third residue having the formula

[0180] O O H H

[0181] - C - R1- C - N - C - R8- N - H C - O - R7

[0182]

[0183] 0, and

[0184] x molar equivalents of a fourth residue having the formula

[0185] O O H H

[0186] - C - R1- C - N - C - R8- N - H C - O - H

[0187]

[0188] O

[0189] wherein m+p+q+x = 1 ;

[0190] m is from 0 to 0.5, p is from 0.1 to 0.95, m+p is from 0.5 to 0.95, q is from 0.02 to 0.25, and x is from 0.05 to 0.25;n is from 5 to 300;

[0191] the ratio q:x is from 50:50 to 20:80;

[0192] R1is (C2-C2o)alkylene;

[0193] R3and R4are selected from the group consisting of hydrogen, (Ci-C6)alkyl, (C2-Ce)alkenyl, (C2-C6)alkynyl, (C6-Cio)aryl, -CH2SH, -(CH2)2S(CH3), -CH2OH, -CH(OH)CH3, -(CH2)4NH3+, -(CH2)3NHC(=NH2+)NH2, -CH2COOH, -CH2-CO-NH2, -CH2CH2-CO-NH2, -CH2CH2COOH, CH3-CH2-CH(CH3)-, (CH3)2CH-CH2-, H2N-(CH2)4-, Ph-CH2-, CH=C-CH2-, (CH3)2CH-, Ph-NH-, NH-(CH2)3-C-, or NH-CH=N-CH=C-CH2-, wherein R3

[0194]

[0195] I - 1 I _ I and R4are the same or different;

[0196] R5is (C2-C2o)alkylene, optionally having one or more alkylene carbon atoms (-C-) replaced by an oxygen atom (-O-), and / or optionally having 5 or 6 consecutive alkylene carbon atoms together form a respective 5- or 6-membered carbocyclic ring;

[0197] R6is

[0198]

[0199] R7is (C6-Cio)aryl(Ci-C6)alkylene; and

[0200] R8is (C3-Cs)alkylene.

[0201] The particles of the previous exemplary embodiment, wherein the biodegradable polymer has the following formula:O O H O O H O O H O O H II < II I II c H I II JI I II > II I — C— R1-c— I N— C— C-O— R5-O— C— C— N C— R1-C— N— C-C-O— R6-O-C— C— N A A= I I I I R3H R4R4H

[0202]

[0203] wherein the units m, p, q, and x are randomly distributed throughout the biodegradable polymer.

[0204] The particles of any one of the previous exemplary embodiments, wherein, in the structure of the biodegradable polymer, p is from 0.3 to 0.8 and m+p is from is from 0.6 to 0.9.

[0205] The particles of any one of the previous exemplary embodiments, wherein, in the structure of the biodegradable polymer, q is from 0.05 to 0.15.

[0206] The particles of any one of the previous exemplary embodiments, wherein, in the structure of the biodegradable polymer, x is from 0.08 to 0.25.

[0207] The particles of any one of the previous exemplary embodiments, wherein, in the structure of the biodegradable polymer, x is from 0.10 to 0.20.

[0208] The particles of any one of the previous exemplary embodiments, wherein, in the structure of the biodegradable polymer, q:x is from 40:60 to 25:75.

[0209] The particles of any one of the previous exemplary embodiments, wherein, in the structure of the biodegradable polymer, q:x is from 40:60 to 30:70.

[0210] The particles of any one of the previous exemplary embodiments, wherein, in the structure of the biodegradable polymer, m is 0.The particles of any one of the previous exemplary embodiments, wherein m is from 0, 0.10, 0.15, 0.20, or 0.25 to 0.75, 0.70, 0.65, 0.60, 0.55, 0.50, 0.45, 0.40, or 0.35.

[0211] The particles of any one of the previous exemplary embodiments, wherein p is from 0.10, 0.20, 0.30, 0.35, or 0.40 to 0.95, 0.90, 0.85, 0.75, 0.70, 0.65, 0.60, 0.55, 0.50, or 0.45.

[0212] The particles of any one of the previous exemplary embodiments, wherein m:p is from 2:1, 1:1, or 2:3 to 1:5, 1:4, 1:3, or 1:2.

[0213] The particles of any one of the previous exemplary embodiments, wherein q is from 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, or 0.20 to 0.25, 0.23, 0.21, 0.20, 0.19, 0.18, 0.17, 0.16, 0.15, 0.11, 0.10, 0.09, 0.08, or 0.07.

[0214] The particles of any one of the previous exemplary embodiments, wherein x is from 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, or 0.20 to 0.30, 0.25, 0.20, 0.19, 0.18, 0.17, 0.16, 0.15, 0.14, 0.13, 0.125, 0.12, 0.11, or 0.10.

[0215] The particles of any one of the previous exemplary embodiments, wherein the ratio q:x is from 50:50 to 35:65.

[0216] The particles of any one of the previous exemplary embodiments, wherein the ratio q:x is from 50:50, 45:55, 40:60, or 35:65 to 20:80, 25:75, 30:70, or 35:65.

[0217] The particles of any one of the previous exemplary embodiments, wherein m is about 0.3, p is about 0.45, q is about 0.10, and x is about 0.15.

[0218] The particles of any one of the previous exemplary embodiments, wherein m is about 0.3, p is about 0.45, q is about 0.09, and x is about 0.16.

[0219] The particles of any one of the previous exemplary embodiments, wherein m is about 0.3, p is about 0.45, q is about 0.08, and x is about 0.18.

[0220] The particles of any one of the previous exemplary embodiments, wherein m is about 0.3, p is about 0.45, q is about 0.06, and x is about 0.19.The particles of any one of the previous exemplary embodiments, wherein m is from 0.1 to 0.5, p is from 0.1 to 0.75, m+p is from 0.5 to 0.8, q is from 0.05 to 0.15, and x is from x is from 0.10 to 0.25.

[0221] The particles of any one of the previous exemplary embodiments, wherein n is from 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, or 75 to 300, 290, 280, 270, 260, 250, 240, 230, 220, 210, 200, 190, 180, 170, 160, or 150.

[0222] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer has a number average molecular weight (Mn) of from 15,000 g / mol to 250,000 g / mol.

[0223] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer has a number average molecular weight (Mn) of at least 15,000 g / mol, at least 20,000 g / mol, at least 25,000 g / mol, at least 30,000 g / mol, or at least 35,000 g / mol.

[0224] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer has a Mn of at most 250,000 g / mol, at most 225,000 g / mol, at most 200,000 g / mol, at most 175,000 g / mol, at most 150,000 g / mol, at most 125,000 g / mol, at most 100,000 g / mol, or at most 75,000 g / mol.

[0225] The particles of any one of the previous exemplary embodiments, wherein m is from 0.1 to 0.5, p is from 0.1 to 0.75, m+p is from 0.5 to 0.8, q is from 0.05 to 0.15, and x is from 0.10 to 0.25;

[0226] R1is (C2-Cio)alkylene;

[0227] R3and R4are (Ci-Ce)alkyl;

[0228] R5is (C2-Cio)alkylene;

[0229] R6is

[0230]

[0231] R7is Ph-(Ci-C6)alkylene; and

[0232] R8is (C3-C6)alkylene.

[0233] The particles of any one of the previous exemplary embodiments, wherein the structure of the biodegradable polymer is further characterized by one or more of the following: m is from 0.1 to 0.5, p is from 0.1 to 0.75, m+p is from is from 0.5 to 0.8, q is from 0.05 to 0.15, and x is from 0.10 to 0.25;

[0234] R1is (C2-Cio)alkylene;

[0235] R3and R4are (Ci-C6)alkyl or (CHs CH-CTh-;

[0236] R5is (C2-Cio)alkylene;

[0237] R6is

[0238]

[0239] R7is Ph-(Ci-C6)alkylene; and

[0240] R8is (C3-C6)alkylene.

[0241] The particles of any one of the previous exemplary embodiments, wherein the structure of the biodegradable polymer is further characterized by: m is 0.2-0.4, p is 0.3-0.6, q is 0.05-0.15, and x is 0.10-0.25;

[0242] The particles of any one of the previous exemplary embodiments, wherein the structure of the biodegradable polymer is further characterized by one or more of the following:m is 0.3, p is 0.45, q is 0.05-0.15, and x is 0.10-0.25;

[0243] R1is octylene;

[0244] R3and R4are isobutyl;

[0245] R5is hexylene;

[0246] R6is

[0247]

[0248] R7is benzyl; and

[0249] R8is butylene.

[0250] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer has a glass transition temperature in the hydrated state of less than about 37 °C.

[0251] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer is amorphous.

[0252] The particles of any one of the previous exemplary embodiments, wherein the particles further comprise a second biodegradable polymer.

[0253] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic comprises a peptide.

[0254] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic comprises from 15 to 100 amino acid residues.

[0255] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic comprises at least 15, 20, 25, or 30 amino acid residues.The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic comprises at most 100, 90, 80, 70, 60, 50 or 40 amino acid residues.

[0256] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic comprises a peptide coupled to a fatty acid moiety.

[0257] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic has a molecular weight of from 3000 to 80,000 g / mol.

[0258] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic has a molecular weight of from 3000 to 6000 g / mol, preferably 3500 to 5500 g / mol.

[0259] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic has a molecular weight of from 4500 to 5200 g / mol.

[0260] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic is a GLP-1 receptor agonist.

[0261] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic is tirzepatide.

[0262] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic is present in an amount of from 3 to 12 wt%, based on the total weight of the particles.

[0263] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic is present in an amount of from 3 wt%, 4 wt%, 5 wt%, 6 wt%, or 7 wt%, based on the total weight of the particles.

[0264] The particles of any one of the previous exemplary embodiments, wherein the water-soluble biotherapeutic is present in an amount of from 12 wt%, 11 wt%, 10 wt%, 9 wt%, or 8 wt%, based on the total weight of the particles.The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer is present in an amount of from 78 wt% to 92 wt%, based on the total weight of the particles.

[0265] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer is present in an amount of from 80 wt% to 92 wt%, based on the total weight of the particles.

[0266] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer is present in an amount of from 78 wt%, 79 wt%, 80 wt%, 81 wt%, 82 wt%, 83 wt%, 84 wt%, or 85 wt%, based on the total weight of the particles.

[0267] The particles of any one of the previous exemplary embodiments, wherein the biodegradable polymer is present in an amount of from 92 wt%, 91 wt%, 90 wt%, 89 wt%, or 88 wt%, based on the total weight of the particles.

[0268] The particles of any one of the previous exemplary embodiments, wherein the vitamin E is present in an amount of from 1 wt% to 10 wt%, based on the total weight of the particles, preferably 2 to 8 wt%.

[0269] The particles of any one of the previous exemplary embodiments, wherein the vitamin E is present in an amount of from 2 wt%, 3 wt%, 4 wt%, or 5 wt%, based on the total weight of the particles.

[0270] The particles of any one of the previous exemplary embodiments, wherein the vitamin E is present in an amount of from 9 wt%, 8 wt%, 7 wt%, or 6 wt%, based on the total weight of the particles.

[0271] The particles of any one of the previous exemplary embodiments, wherein the vitamin E comprises alpha-tocopherol, beta-tocopherol, gamma-tocopherol, or delta-tocopherol.

[0272] The particles of any one of the previous exemplary embodiments, wherein the vitamin E comprises alpha-tocopherol.The particles of any one of the previous exemplary embodiments, wherein the vitamin E consists of alpha-tocopherol.

[0273] The particles of any one of the previous exemplary embodiments, wherein the vitamin E comprises a fat-soluble form of vitamin E.

[0274] The particles of any one of the previous exemplary embodiments, wherein the vitamin E consists of one or more fat-soluble forms of vitamin E.

[0275] The particles of any one of the previous exemplary embodiments, wherein the vitamin E comprises a water-soluble form of vitamin E.

[0276] The particles of any one of the previous exemplary embodiments, wherein the vitamin E consists of one or more water-soluble forms of vitamin E.

[0277] The particles of any one of the previous exemplary embodiments, wherein the particles further comprise an additive.

[0278] The particles of any one of the previous exemplary embodiments, wherein the particles further comprise a surfactant, plasticizer, antioxidant, chelating agent, buffer, or osmotic agent.

[0279] The particles of any one of the previous exemplary embodiments, wherein the particles consist of the biodegradable polymer, the water-soluble biotherapeutic, and vitamin E.

[0280] The particles of any one of the previous exemplary embodiments, wherein the average particle size (d50) of the particles is from 1 to 500 pm.

[0281] The particles of any one of the previous exemplary embodiments, wherein the average particle size (d50) of the particles is from 5 to 200 pm.

[0282] The particles of any one of the previous exemplary embodiments, wherein the particles have a d50 particle size of from 5, 10, 15, 20, 25, or 30 pm.The particles of any one of the previous exemplary embodiments, wherein the average particle size (d50) of the particles have a d50 particle size of at most 200, 150, 120, 100, 90, or 80 pm.

[0283] The particles of any one of the previous exemplary embodiments, wherein the particles have an average particles size (d50) of from 40 to 250 pm.

[0284] The particles of any one of the previous exemplary embodiments, wherein less than 20% of the water-soluble biotherapeutic is released from the particles after twenty -four hours in phosphate buffer at pH 7.4 containing 137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPO4 and 1.8 mM KH2PO4at37 °C.

[0285] The particles of any one of the previous exemplary embodiments, wherein less than 25% of the water-soluble biotherapeutic is released from the particles after 7 days in phosphate buffer at pH 7.4 containing 137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPO4 and 1.8 mM KH2PO4 at 37 °C.

[0286] The particles of any one of the previous exemplary embodiments, wherein less than 25% of the water-soluble biotherapeutic is released from the particles after 5 days in phosphate buffer at pH 7.4 containing 137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPO4 and 1.8 mM KH2PO4 at 37 °C.

[0287] The particles of any one of the previous exemplary embodiments, wherein less than 10% of the water-soluble biotherapeutic is released from the particles after 7 days in phosphate buffer at pH 7.4 containing 137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPO4 and 1.8 mM KH2PO4 at 37 °C.

[0288] A method of forming particles comprising the steps of:

[0289] a. forming an oil-phase by dissolving a biodegradable polymer and Vitamin E in an organic solvent;

[0290] b. forming a water phase by dissolving a water-soluble biotherapeutic in water; c. adding the water phase to the oil phase and emulsifying, thereby forming an emulsion; and

[0291] d. dispersing the emulsion in water.The method of forming particles of the previous exemplary embodiment, wherein the biodegradable polymer, vitamin E, and / or the water-soluble biotherapeutic are as described in any of the previous exemplary embodiments.

[0292] Particles formed according to the method of any one of the previous exemplary embodiments.

[0293] An injectable formulation comprising the particles of any one of the previous exemplary embodiments.

[0294] The injectable formulation of the previous exemplary embodiment, further comprising unencapsulated water-soluble biotherapeutic.

[0295] The injectable formulation of any one of the previous exemplary embodiments, further comprising unencapsulated biotherapeutic, wherein the unencapsulated biotherapeutic is present as particles.

[0296] The injectable formulation of any one of the previous exemplary embodiments, further comprising unencapsulated water-soluble biotherapeutic, wherein the unencapsulated biotherapeutic is the same as in the particles.

[0297] A method for administering a biotherapeutic to a patient comprising the step of injecting the particles according to any one of the previous exemplary embodiments, wherein less than 20% of the water-soluble biotherapeutic is released from the particles after twenty-four hours.

[0298] A method for administering a biotherapeutic to a patient comprising the step of injecting the particles according to any one of the previous exemplary embodiments, wherein less than 25% of the water-soluble biotherapeutic is released from the particles after 24 hours.

[0299] An implant comprising particles dispersed within a degradable or non-degradable polymer, wherein the particles comprise the particles present in any one of the previous exemplary embodiments or formed by the method of forming particles of any one of the previous exemplary embodiments, and wherein the implant is solid or semi-solid.82. A pre-loaded syringe comprising the particles according to any one of the previous exemplary embodiments.

Claims

CLAIMS:

1. Particles comprising a water-soluble biotherapeutic, a biodegradable polymer, and vitamin E, wherein the biodegradable polymer comprises a random copolymer having a structure including n units of:m molar equivalents of a first residue having the formulaO O H O O H- C - R1- C - N - C - C - O - R5- O - C - C - N - I I I IH R3R3H p molar equivalents of a second residue having the formulaO O H O O HH R4R4H q molar equivalents of a third residue having the formulaO O H H- C - R1- C - N - C - R8- N - H C - O - R70, andx molar equivalents of a fourth residue having the formulaO O H H- C - R1- C - N - C - R8- N - H C - O - HOwherein m+p+q+x = 1 ;m is from 0 to 0.5, p is from 0.1 to 0.95, m+p is from 0.5 to 0.95, q is from 0.02 to 0.25, and x is from 0.05 to 0.25;n is from 5 to 300;the ratio q:x is from 50:50 to 20:80;R1is (C2-C2o)alkylene;R3and R4are selected from the group consisting of hydrogen, (Ci-C6)alkyl, (C2-Ce)alkenyl, (C2-C6)alkynyl, (C6-Cio)aryl, -CH2SH, -(CH2)2S(CH3), -CH2OH, -CH(OH)CH3, -(CH2)4NH3+, -(CH2)3NHC(=NH2+)NH2, -CH2COOH, -CH2-CO-NH2, -CH2CH2-CO-NH2, -CH2CH2COOH, CH3-CH2-CH(CH3)-, (CH3)2CH-CH2-, H2N-(CH2)4-, Ph-CH2-, CH=C-CH2-, (CH3)2CH-, Ph-NH-, NH-(CH2)3-C-, or NH-CH=N-CH=C-CH2-, wherein R3I - 1 I _ I and R4are the same or different;R5is (C2-C2o)alkylene, optionally having one or more alkylene carbon atoms (-C-) replaced by an oxygen atom (-O-), and / or optionally having 5 or 6 consecutive alkylene carbon atoms together form a respective 5- or 6-membered carbocyclic ring;R6isR7is (C6-Cio)aryl(Ci-C6)alkylene; andR8is (C3-Cs)alkylene.

2. The particles of claim 1, wherein the biodegradable polymer has the following formula:37O O H O O H O O H O O H II < II I II c H I II JI I II > II I — C— R1-c— I N— C— C-O— R5-O— C— C— N C— R1-C— N— C-C-O— R6-O— C— C— N A A= I I I I R3H R4R4HO O H H O O H HC— R1-C— N— C— R8-N C— R1-C— N— C— R8-N I I H C-O— R IIOwherein the units m, p, q, and x are randomly distributed throughout the biodegradable polymer.

3. The particles of any one of the previous claims, wherein, in the structure of the biodegradable polymer, p is from 0.3 to 0.8 and m+p is from is from 0.6 to 0.9.

4. The particles of any one of the previous claims, wherein the ratio q:x is from q:x is from 40:60 to 25:75.

5. The particles of any one of the previous claims, wherein the structure of the biodegradable polymer is further characterized by one or more of the following:m is from 0.1 to 0.5, p is from 0.1 to 0.75, m+p is from is from 0.5 to 0.8, q is from 0.05 to 0.15, and x is from 0.10 to 0.25;R1is (C2-Cio)alkylene;R3and R4are (Ci-C6)alkyl or (CHs CH-CTh-;R5is (C2-Cio)alkylene;R6isR7is Ph-(Ci-C6)alkylene; andR8is (C3-C6)alkylene.

6. The particles of any one of the previous claims, wherein the water-soluble biotherapeutic comprises a peptide comprising from 15 to 100 amino acid residues.

7. The particles of any one of the previous claims, wherein the water-soluble biotherapeutic has a molecular weight of from 3000 to 6000 g / mol, preferably 3500 to 5500 g / mol.

8. The particles of any one of the previous claims, wherein the water-soluble biotherapeutic is a GLP-1 receptor agonist.

9. The particles of any one of the previous claims, wherein the water-soluble biotherapeutic comprises tirzepatide.

10. The particles of any one of the previous claims, wherein the water-soluble biotherapeutic is present in an amount of from 3 to 12 wt%, based on the total weight of the particles.

11. The particles of any one of the previous claims, wherein the biodegradable polymer is present in an amount of from 78 wt% to 92 wt%, based on the total weight of the particles.

12. The particles of any one of the previous claims, wherein the vitamin E is present in an amount of from 1 wt% to 10 wt%, based on the total weight of the particles, preferably 2 to 8 wt%.

13. The particles of any one of the previous claims, wherein less than 25% of the water-soluble biotherapeutic is released from the particles after 5 days in phosphate buffer at pH 7.4 containing 137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPO4 and 1.8 mM KH2PO4at 37 °C.

14. A method of forming particles comprising the steps of:a. forming an oil-phase by dissolving a biodegradable polymer and Vitamin E in an organic solvent;b. forming a water phase by dissolving a water-soluble biotherapeutic in water; c. adding the water phase to the oil phase and emulsifying, thereby forming an emulsion; andd. dispersing the emulsion in water,wherein the biodegradable polymer comprises a random copolymer having a structure including n units of:m molar equivalents of a first residue having the formulap molar equivalents of a second residue having the formulaq molar equivalents of a third residue having the formulaO O H H- C - R1- C - N - C - R8- N - H C - O - R70, andx molar equivalents of a fourth residue having the formulaO O H H- C - R1- C - N - C - R8- N - H C - O - HOwherein m+p+q+x = 1 ;m is from 0 to 0.5, p is from 0.1 to 0.95, m+p is from 0.5 to 0.95, q is from 0.02 to 0.25, and x is from 0.05 to 0.25;n is from 5 to 300;the ratio q:x is from 50:50 to 20:80;R1is (C2-C2o)alkylene;R3and R4are selected from the group consisting of hydrogen, (Ci-C6)alkyl, (C2-Ce)alkenyl, (C2-C6)alkynyl, (C6-Cio)aryl, -CH2SH, -(CH2)2S(CH3), -CH2OH, -CH(OH)CH3, -(CH2)4NH3+, -(CH2)3NHC(=NH2+)NH2, -CH2COOH, -CH2-CO-NH2, -CH2CH2-CO-NH2, -CH2CH2COOH, CH3-CH2-CH(CH3)-, (CH3)2CH-CH2-, H2N-(CH2)4-, Ph-CH2-, CH=C-CH2-, (CH3)2CH-, Ph-NH-, NH-(CH2)3-C-, or NH-CH=N-CH=C-CH2-, wherein R3I - 1 I _ I and R4are the same or different;R5is (C2-C2o)alkylene, optionally having one or more alkylene carbon atoms (-C-) replaced by an oxygen atom (-O-), and / or optionally having 5 or 6 consecutive alkylene carbon atoms together form a respective 5- or 6-membered carbocyclic ring;R6isR7is (C6-Cio)aryl(Ci-C6)alkylene; andR8is (C3-Cs)alkylene.4115. Particles formed according to the method of the previous claim.

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