EA045374B1 - PHARMACEUTICAL COMPOSITION - Google Patents

Description

Field of technology

The present invention relates to controlled release drug delivery or pharmaceutical compositions, in particular to pharmaceutical compositions suitable for in-situ depot formation. In particular, the present invention relates to a pharmaceutical composition comprising a biodegradable multi-branched copolymer containing at least three polyester arms attached to a central polyester-containing core, wherein said multi-branched copolymer is substantially insoluble in an aqueous solution, wherein said composition additionally contains at least one pharmaceutically active ingredient.

State of the art

WO 2012/090070 A describes a technology for the production of in situ formed depots (ISFD) by solvent exchange, comprising dissolving a mixture of linear polyesters (m)PEG in a biocompatible organic solvent. Upon injection, the solvent diffuses and water-insoluble polymers precipitate and form a depot that can entrap the active pharmaceutical ingredient (API). The drug substance is released from this depot over a long period of time.

There remains a need to develop improved technology to provide sustained release. The use of branched PEG polyester block copolymers has been proposed as a potential improvement to currently used technologies, which have disadvantages such as high viscosity, high injectability index values and slow degradation kinetics.

PEG polyester star copolymers are branched structures consisting of several (three or more) linear chains connected to a central core. Star copolymers can be divided into two categories: star homopolymers or star copolymers.

Star-shaped homopolymers consist of a symmetrical structure containing radiating arms with the same chemical composition and the same molecular weight. Star copolymers consist of a symmetrical structure containing radiating arms with the same molecular weight but formed by at least two different monomers.

Star copolymers (also known as multi-armed or multi-branched) are described in Cameron et al., Chemical Society Reviews, 40, 1761, 2011 and Burke et al., Biomacromolecules, 18, 728, 2017).

Hiemstra et al., Biomacromolecules, 7, 2790, 2006; Buwalda et al., Biomacromolecules, 11, 224, 2010; Calucci et al., Langmuir, 26, 12890, 2010; Mayadunne et al., US 2016/0058698 A1, 2016, describe aqueous systems that form a gel when temperature changes.

Application EP 1404294 B1 describes the use of branched (co)polymers for the production of in situ formed depots (ISFD) by solvent replacement.

Accordingly, there is a need to develop new ISFD formulations based on solvent exchange star copolymers that have lower viscosity, improved needle permeability upon administration, improved or different API release characteristics, or depot degradation kinetics.

Brief description of the invention

The present invention provides a pharmaceutical composition comprising a biodegradable multi-branched copolymer comprising at least three polyester arms attached to a central polyester-containing core, wherein the multi-branched copolymer is substantially insoluble in an aqueous solution, wherein said composition further comprises at least at least one pharmaceutically active ingredient.

Typically, the molecular weight of the polyether is 10 kDa or less, preferably 5 kDa or less, 4 kDa or less, 3 kDa or less, or 2 kDa or less, or 1 kDa or less, or 0.5 kDa or less.

The present invention also provides a pharmaceutical composition comprising a biodegradable multi-branched polyester copolymer comprising at least three polyester arms attached to a central polyester-containing core, wherein the molecular weight of said polyester is 10 kDa or less, preferably 5 kDa or less, 4 kDa or less, 3 kDa or less, or 2 kDa or less, or 1 kDa or less, or 0.5 kDa or less, wherein the composition further contains at least one pharmaceutically active ingredient.

The above compositions are suitable for the formation of in-situ depots.

The present inventors have surprisingly discovered that formulations based on multiply branched or star copolymers have lower viscosity and improved ability to pass through a needle upon administration, while providing improved or different release profiles of the active ingredient compared to formulations containing only linear analogues.

- 1 045374 copolymers. In addition, depots obtained from compositions based on multiply branched or star-shaped components decompose faster than depots obtained from analogue linear copolymers.

Typically, the multi-branched copolymer is substantially insoluble in aqueous solution.

In a preferred embodiment, the multi-branched copolymer has a solubility in aqueous solution of less than 15 mg/ml, optionally less than 10 mg/ml, less than 5 mg/ml, less than 2 mg/ml, or less than 1 mg/ml. Typically, solubility is measured at 37°C.

Typically, the multi-branched copolymer is represented by the formula A(B)n, where A is the central core and B is the polyester arms, and n is an integer equal to at least 3. In embodiments of the present invention, n is at least at least 4, or at least 6, or at least 8; n can be 3, 4, 6 or 8. Preferably n is 4.

In one embodiment, the central core comprises a multi-branched polyether poly(ethylene glycol) (PEG) polyol. Typically, a polyol contains at least three hydroxyl groups. A polyol is typically a hydrocarbon functionalized with at least three hydroxyl groups, optionally 3, 4, 5, 6 or 8 hydroxyl groups. According to some embodiments of the present invention, the polyol further contains one or more ether groups. Preferably the polyol is pentaerythritol (PE), dipentaerythritol (DPE), trimethylolpropane (TMP), glycerol, hexaglycerol, erythritol, xylitol, di(trimethylolpropane) (diTMP), sorbitol or inositol.

In preferred embodiments, the multi-branched polyether is characterized by Formula 1 or Formula 2 or Formula 3 or Formula 4

Formula 1 where m is an integer from 2 to 150.

R represents H, or alkyl, or PEG. Preferably R is PEG.

Formula 2 where m is an integer from 2 to 150 and p is 6,

Formula 3 where m is an integer from 2 to 150.

- 2 045374

Formula 4 where m is an integer from 2 to 150.

Typically, the polyester is or is formed from at least one polymer or copolymer selected from the group consisting of poly(lactic acid) (PLA), poly(glycolic acid) (PGA), poly(epsilon-caprolactone) (PCL), poly(ethylene adipate) (PEA), copolymer of lactic acid and glycolic acid (PLGA) and poly(hydroxyalkanoate) (PGA) or mixtures thereof. Polyester arms are typically formed by reacting a polyester precursor or monomer with a polyester core. For example, when forming arms based on PLA, polyester is reacted with D,L-lactide.

In preferred embodiments, each branch of the multi-branched polyester contains a reactive end group capable of reacting with the polyester or a monomer or precursor thereof. Typically, the terminal reactive group is a hydroxyl group or an amino group, but preferably a hydroxyl group.

In one embodiment, the polyester is a homopolymer.

In one embodiment, the polyester is derived from more than one monomer. When a polyester is derived from more than one monomer, the polyester may be a random copolymer or a block copolymer.

In a preferred embodiment, the polyester is or contains PLA.

In a preferred embodiment, the multi-branched copolymer is prepared by reacting the multi-branched polyether as defined above with D,L lactide. The multi-branched copolymer can be prepared by ring-opening polymerization of D,L-lactide initiated by said multi-branched polyether.

In a preferred embodiment, the multi-branched copolymer is characterized by Formula 5, or Formula 6, Formula 7, or Formula 8

where u is an integer from 4 to 200 and m is an integer from 2 to 150.

Formula 6

- 3 045374 where u is an integer from 4 to 200, m is an integer from 2 to 150 and p is 6.

Formula 7 where u is an integer from 4 to 200 and m is an integer from 2 to 150.

Formula 8 where u is an integer from 4 to 200 and m is an integer from 2 to 150.

In preferred embodiments of the above multi-branched copolymer of Formula 5, the polyester core (i.e., the compound of Formula 5 without the PLA arms) has a molecular weight of 2 kDa, wherein the molar ratio of ester repeat unit to ethylene oxide is 2, 3, or 6.

In one embodiment, the number of repeating ester units in each arm independently ranges from 4 to 200.

In a preferred embodiment, the molecular weight of the polyether is 0.5 to 10 kDa, optionally 1 to 10 kDa, preferably 2 to 10 kDa, most preferably 2 to 5 kDa. In a further embodiment, the molecular weight of the polyester is in the range of 0.5 to 2 kDa.

In preferred embodiments, the molar ratio of repeating ester unit to ethylene oxide for the multi-branched copolymer in the composition is from 1 to 10, preferably from 2 to 6.

In a further embodiment, the composition of the present invention may comprise one or more additional biodegradable multi-branched copolymers as defined above. A composition containing two or more multi-branched copolymers provides an additional means of modulating the release of a pharmaceutically active agent. Various combinations of multi-branched copolymers are also provided, and the relative amounts of two or more multi-branched copolymers can be varied. This allows the release profile of the pharmaceutically active agent to be controlled.

In one embodiment, the composition comprises a first biodegradable multi-branched copolymer as defined above and a second distinct biodegradable multi-branched copolymer as defined above, optionally the content of the first biodegradable multi-branched copolymer being from 15 to 25% w/w. , optional from 18 to

20% w/w, and the content of the second biodegradable multi-branched copolymer is from 15 to 25% w/w, optionally from 18 to 20% w/w, based on the total weight of the composition.

In one embodiment, each of the first and second biodegradable multi-branched copolymers has a structure of formula 5

where u is an integer from 4 to 200 and m is an integer from 2 to 150;

and wherein the first biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 2;

and wherein the second biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 6; or the first biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 5 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 2; and wherein the second biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 6.

According to an alternative embodiment of the present invention, the composition further comprises a biodegradable triblock copolymer characterized by the formula

Av-Bw-Ax, where A is a polyester, B is polyethylene glycol, v and x are the number of repeat units ranging from 1 to 3000 and w is the number of repeat units ranging from 3 to 300, wherein v= x or v^x.

The combination of a multi-branched copolymer with a linear triblock copolymer provides an additional option for modulating the release of a pharmaceutically active ingredient.

Typically, the weight of the polyethylene glycol chain in the triblock copolymer is in the range of 180 Da to 12 kDa, or 194 Da to 12 kDa, or 200 Da to 12 kDa, or 100 Da to 4 kDa, preferably 1 to 2 kDa .

Typically, the molar ratio of ester repeating unit to ethylene oxide repeating unit in the triblock copolymer is from 0.5 to 22.3, optionally from 0.5 to 10, preferably from 0.5 to 3.5.

In a preferred embodiment, the triblock copolymer contains a polyethylene glycol chain having a mass of 1 kDa, and the molar ratio of ester repeat unit to ethylene oxide repeat unit is 4, and the multi-branched copolymers have the structure of formula 5

where u is an integer from 4 to 200 and m is an integer from 2 to 150; containing a polyether core having a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 3; or wherein the triblock copolymer contains a polyethylene glycol chain having a mass of 2 kDa, and the molar ratio of the ester repeating unit and the ethylene oxide repeating unit is 2, wherein the multi-branched copolymers have a structure of formula 5 containing a polyether core having a molecular mass of 2 kDa, with

- 5 045374 In this case, the molar ratio of the ester repeating unit and the ethylene oxide repeating unit is 2; or wherein the triblock copolymer contains a polyethylene glycol chain having a mass of 2 kDa, and the molar ratio of the ester repeat unit and the ethylene oxide repeat unit is 2, wherein the multi-branched copolymers have a structure of formula 5 containing a polyether core having a molecular weight of 2 kDa, wherein the molar ratio of the ester repeating unit and the ethylene oxide repeating unit is 6;

optionally the multi-branched copolymer content is from 15 to 25% w/w, optionally from 18 to 20% w/w. based on the total weight of the composition, and the triblock copolymer content is from 15 to 25% wt./wt., optionally from 18 to 20% wt./wt. from the total mass of the composition.

According to a further alternative embodiment, the composition also contains a biodegradable diblock copolymer characterized by the formula

Cy-Az, wherein A is a polyester and C is a capped polyethylene glycol, and y and z are the number of repeat units, with y ranging from 2 to 250 and z ranging from 1 to 3000.

The combination of a multi-branched copolymer with a linear diblock copolymer provides an additional option for modulating the release of a pharmaceutically active ingredient.

In one embodiment, the molecular weight of the diblock copolymer-terminated polyethylene glycol chain is 100 Da to 10 kDa, or 164 Da to 2 kDa, preferably 1 to 2 kDa.

In one embodiment, the molar ratio of ester repeat unit to ethylene oxide repeat unit in the diblock copolymer is from 0.8 to 15, optionally from 1 to 10.

Typically, the polyester A in the triblock or diblock copolymer is selected from the group consisting of polylactic acid (PLA), polyglycolic acid, polycaprolactone, polyethylene adipate, polyhydroxyalkanoate, ε-caprolactone-lactide copolymer (PCLA), lactic acid-glycolic acid copolymer ( PMCG) and mixtures thereof, and optionally the capping polyethylene glycol is methoxy polyethylene glycol. Preferably, polyester A is polylactic acid.

According to one embodiment of the present invention, the composition further comprises a pharmaceutically acceptable carrier, wherein optionally the pharmaceutically acceptable carrier is an organic solvent, wherein optionally the organic solvent is a biocompatible organic solvent, wherein optionally the amount of said carrier is at least 25% or at least 35% % w/w from the total mass of the composition. Preferably, the pharmaceutically acceptable carrier is selected from the group: benzyl alcohol, benzyl benzoate, dimethyl isosorbide (DMI), dimethyl sulfoxide (DMSO), ethyl acetate, ethyl benzoate, ethyl lactate, glycerol formal, methyl ethyl ketone, methyl isobutyl ketone, N-ethyl-2-pyrrolidone, Nmethyl-2-pyrrolidinone (NMP ), pyrrolidone-2, tetraglycol, triacetin, tributyrin, tripropionine, glycofurol and mixtures thereof.

In one embodiment, the pharmaceutically active ingredient is hydrophobic. By this we mean that the pharmaceutical active ingredient has positive logP or logD values and a water solubility at physiological pH (pH 7.0 to 7.4) below 1 mg/ml.

In a preferred embodiment, the pharmaceutically active ingredient is meloxicam, bupivacaine, tamsulosin, octreotide, tadalafil, empagliflozin, tenofovir, liothyronine, or combinations thereof.

According to one embodiment of the present invention, the content of at least one pharmaceutically active ingredient is from 0.05 to 60% w/w, optionally from 0.05 to 40%, optionally from 0.05 to 30%, optionally from 0 .05 to 10%, optionally from 0.05 to 7%, optionally from 0.05 to 2% by weight of the total composition.

In a preferred embodiment, the composition is an injectable liquid.

According to one embodiment of the present invention, the multi-branched copolymer content is from 2 to 80%, optionally from 2 to 70%, optionally from 2 to 60%, optionally from 10 to 60%, optionally from 10 to 50%, optionally from 20 to 40 %, optionally from 20 to 35%, optionally from 30 to 50% wt./wt. from the total mass of the composition.

According to one embodiment of the present invention, the composition is as defined in table. 1 or table 4.

Typically, the release of at least one pharmaceutically active ingredient

- 6 045374 can be modulated by composition.

In one embodiment of the present invention, the composition is suitable for delivering a pharmaceutically active ingredient to a subject for at least 1 day, optionally for at least 3 days, optionally for at least 7 days, optionally for at least 30 days, optionally for at least 90 days, optionally for at least 1 year.

In a further aspect, the present invention provides the use of a pharmaceutical composition as defined above to modulate the release kinetics of at least one active ingredient.

In a further aspect, the present invention provides a method for preparing a pharmaceutical composition as defined above, comprising dissolving a multi-branched copolymer as defined above in a pharmaceutically acceptable carrier, such as a diluent. According to one embodiment of the present invention, the method further includes adding a pharmaceutically active ingredient to the composition.

In a further aspect, the present invention provides a bioabsorbable depot that is prepared ex vivo or in situ by contacting the composition as defined above with an aqueous medium, water or biological fluid. The depot is bioabsorbable in the sense that the PLA fragment is degraded in vivo and that the PEG is assimilated by the body and excreted.

In a final aspect, a method of controlled release of a pharmaceutically active ingredient is provided, comprising administering a composition as defined above and allowing depot formation in vivo and in situ by solvent exchange.

Detailed Description of the Invention

As used herein, the term biodegradable or bioabsorbable means that the block copolymers undergo hydrolysis in vivo to form their constituent (m)PEGs and oligomers, or monomers, or repeat units derived from the polyester block. For example, PLA undergoes hydrolysis to form lactic acid. The result of the hydrolysis process leads to a progressive loss of depot mass and, ultimately, to its disappearance.

The term multi-branched copolymer means a polymer with at least three polyester arms attached to a central core that contains polyester. Polyester arms may be called branches, shoulders or chains. The term multi-branched copolymer has the same meaning as the terms star polymer, or star polymer, or multi-arm copolymer, and these terms are used interchangeably throughout the application.

Typically, the molecular weight of the polyether is 10 kDa or less, preferably 5 kDa or less, 4 kDa or less, 3 kDa or less, or 2 kDa or less, or 1 kDa or less, or 0.5 kDa or less. Preferably the polyester has a molecular weight of at least 0.2 kDa or at least 0.5 kDa.

Typically, the multi-branched copolymer is represented by the formula A(B) _n , wherein A is the central core and B is the polyester arms, and n is an integer equal to at least 3. In embodiments of the present invention, n is at least at least 4, or at least 6, or at least 8. Preferably, n is 4. Below is an example structure of a multi-branched PEG-PLA block copolymer with n = 4.

The letters m and u correspond to the number of repeating ethylene oxide units and lactic acid units composing each block, respectively.

A polyol is an organic compound containing many hydroxyl groups. Typically, a polyol contains at least three hydroxyl groups. Typically, the polyol is a hydrocarbon functionalized with at least three hydroxyl groups, for example, 3, 4, 5, 6 or 8 hydroxyl groups. The polyol may also contain one or more ether groups. Typically, the polyol is pentaerythritol (PE), dipentaerythritol (DPE), trimethylolpropane (TMP), glycerol, hexaglycerol, erythritol, xylitol, di(trimethylolpropane) (diTMP), sorbitol, or inositol.

- 7 045374

Polyether is an organic compound containing many ether groups.

In a preferred embodiment, the central core is a multi-branched polyether poly(ethylene glycol) (PEG) polyol. For example, a multi-branched polyether can be formed by reacting ethylene oxide with a polyol. A multi-branched polyether may be called a star-shaped PEG. Ethylene oxide reacts with the hydroxyl group of the polyol to form a PEG-based arm. For example, pentaerythritol can react with ethylene oxide to form the four- or three-arm or four- or three-arm polyester shown in Formula 1 below.

Formula 1 where m is an integer from 2 to 150. R is H or alkyl or PEG.

In an alternative embodiment, the multi-branched polyether is an eight-armed or eight-armed polyester (p = 6) given in Formula 2 below.

Formula 2 where m is an integer from 2 to 150 and p is 6.

In an alternative embodiment, the multi-branched polyether is a six-arm or six-arm polyester, as set forth in Formula 3 and Formula 4 below.

Formula 3 where m is an integer from 2 to 150.

- 8 045374

Formula 4 where m is an integer from 2 to 150.

Typically, each branch of a multi-branched polyester contains a terminal hydroxyl group, but other reactive terminal groups capable of reacting with the polyesters or their monomers or precursors may also be provided. Typically, the polyester is formed from at least one polymer or copolymer selected from the group consisting of poly(lactic acid) (PLA), poly(glycolic acid) (PGA), poly(epsilon-caprolactone) (PCL), poly( ethylene adipate) (PEA), copolymer of lactic acid and glycolic acid (LAGC) and poly(hydroxyalkanoate) (PGA) or mixtures thereof. Typically, the terminal hydroxyl group of each branch of a multi-branched polyester reacts with a polyester monomer or precursor to form a polyester arm. For example, D-L-lactide can react with a multi-branched polyester to form a PLA-based arm.

In one embodiment, the polyester is a homopolymer.

In one embodiment, the polyester is derived from more than one monomer. When a polyester is derived from more than one monomer, the polyester may be a random copolymer or a block copolymer.

In a preferred embodiment, the polyester is or contains PLA.

In one embodiment, the number of repeating ester units in each arm independently ranges from 4 to 200.

The term depot injection means the injection of a flowable pharmaceutical composition, typically subcutaneously, intradermally or intramuscularly, in which the drug is deposited as a localized, eg solid, mass called a depot. Depot, as defined in the present invention, is obtained in situ by injection. Thus, the compositions can be prepared in the form of solutions or suspensions and can be administered to the body by injection.

An in-situ depot is a solid, localized mass formed by precipitation of a pharmaceutical composition following injection of the composition into a subject. The pharmaceutical composition contains a multi-branched copolymer that is substantially insoluble in aqueous solution. Thus, when a pharmaceutical composition is brought into contact with the aqueous environment of a human or animal body, a phase inversion occurs, resulting in a change in the state of the composition from a liquid to a solid, i.e. precipitation of the composition occurs, which leads to the formation of an in-situ depot.

In-situ depot can be clearly distinguished from hydrogel pharmaceutical formulations described in the prior art.

Hydrogels can be formed from star-shaped polymers containing a polyether core and PLA-based arms. Some star polymers containing a polyether core and PLA-based arms can form micelles in aqueous solution. The hydrophobic outer blocks of PLA associate with neighboring micelles to form a network of interconnected micelles or large aggregates, resulting in the formation of gels in certain temperature and concentration ranges. Hydrogels have three-dimensional networks that are capable of absorbing large amounts of water. The polymers that form hydrogels are soluble in aqueous solution. In contrast, the multi-branched polymers used in the present invention are substantially insoluble in aqueous solution. The pharmaceutical compositions of the present invention contain no or substantially no water. For example, the pharmaceutical compositions according to the present invention contain less than 0.5% wt./wt. water.

Typically, the multi-branched copolymer is substantially insoluble in aqueous solution. Typically, this means that the multi-branched copolymer has a solubility in aqueous solution of less than 15 mg/ml, optionally less than 10 mg/ml, less than 5 mg/ml, less than 2 mg/ml, optional

- 9 045374 specifically less than 1 mg/ml. Typically, solubility is measured at 37°C.

In a preferred embodiment, the solubility of the multi-branched copolymer in water is determined as follows: 500 mg of the copolymer is placed in an empty 20 ml vial. 5 ml of ultrapure water was added, the vial was placed at 37°C on a continuous vortex mixer for 2 hours. The vial was then centrifuged for 10 min at 3000 rpm. The supernatant was transferred to another vial of known weight, which was placed at -80°C overnight before lyophilization for 24 hours. The amount of solubilized copolymer was determined as the difference in weight of the empty vial and the lyophilized vial.

Temperature-sensitive hydrogels or thermogels described in the prior art are typically solid within a certain narrow temperature range, for example, 3035°C, and this curing is reversible. In contrast, the in-situ depot formed according to the present invention is solid when introduced over a much wider temperature range, for example from 20 to 37°C.

Additionally, PEG- and PLA-based hydrogels have been shown to function as sustained-release formulations for about 6 months. In contrast, the depot composition of the present invention allows the release of the API over a longer period.

In a further embodiment, the composition of the present invention may comprise one or more additional biodegradable multi-branched copolymers as defined above. Providing a composition containing two or more multi-branched copolymers provides an additional means of modulating the release of a pharmaceutically active agent. Various combinations of multi-branched copolymers may be provided, and the relative amounts of two or more multi-branched copolymers may be varied.

In one embodiment, the composition comprises a first biodegradable multi-branched copolymer as defined above and a second different biodegradable multi-branched copolymer as defined above. The first biodegradable multi-branched copolymer may be present in an amount from 15 to 25% w/w, and the second biodegradable multi-branched copolymer may be present in an amount from 15 to 25% w/w. from the total mass of the composition.

In one embodiment, each of the first and second biodegradable multi-branched copolymers has a structure of formula 5

where u is an integer from 4 to 200 and m is an integer from 2 to 150;

and wherein the first biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa and a molar ratio of the repeating ester unit to ethylene oxide is 2; and wherein the second biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 6; or the first biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 5 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 2; and wherein the second biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 6.

According to an alternative embodiment of the present invention, the composition further comprises a biodegradable triblock copolymer characterized by the formula

Av-Bw-Ax, where A is a polyester, B is a polyethylene glycol, v and x are the number of repeat units ranging from 1 to 3000, and w is the number of repeat units ranging from 3 to 300, wherein v= x or v^x.

The combination of a multi-branched copolymer with a linear triblock copolymer results in

- 10 045374 for an additional option for modulating the release of a pharmaceutically active ingredient.

Typically, the weight of the polyethylene glycol chain in a triblock copolymer ranges from

180 Da to 12 kDa, or 194 Da to 12 kDa, or 200 Da to 12 kDa, or 100 Da to 4 kDa, preferably 1 to 2 kDa.

Typically, the molar ratio of ester repeating unit to ethylene oxide repeating unit in the triblock copolymer is from 0.5 to 22.3, optionally from 0.5 to 10, preferably from 0.5 to 3.5.

In a preferred embodiment, the triblock copolymer contains a polyethylene glycol chain having a mass of 1 kDa, and the molar ratio of ester repeat unit to ethylene oxide repeat unit is 4, and the multi-branched copolymers have the structure of formula 5

where u is an integer from 4 to 200 and m is an integer from 2 to 150;

containing a polyether core having a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 3; or wherein the triblock copolymer contains a polyethylene glycol chain having a mass of 2 kDa, and the molar ratio of the ester repeat unit and the ethylene oxide repeat unit is 2, wherein the multi-branched copolymers have a structure of formula 5 containing a polyether core having a molecular weight of 2 kDa, wherein the molar ratio of the ester repeating unit and the ethylene oxide repeating unit is 2; or wherein the triblock copolymer contains a polyethylene glycol chain having a mass of 2 kDa, and the molar ratio of the ester repeat unit and the ethylene oxide repeat unit is 2, wherein the multi-branched copolymers have a structure of formula 5 containing a polyether core having a molecular weight of 2 kDa, wherein the molar ratio of the ester repeating unit and the ethylene oxide repeating unit is 6;

optionally the content of multi-branched copolymer is from 15 to 25% wt./wt. of the total weight of the composition, and the triblock copolymer content remains from 15 to 25% wt./wt. from the total mass of the composition.

According to a further alternative embodiment, the composition also contains a biodegradable diblock copolymer characterized by the formula

wherein A is a polyester and C is a capped polyethylene glycol, and y and z are the number of repeating units, with y ranging from 2 to 250 and z ranging from 1 to 3000.

The term capped polyethylene glycol refers to cPEG that has a single hydroxyl end group to react, and includes alkoxy-terminated PEG, urethane-terminated PEG, ester-terminated PEG, and the like. The terminal group is a chemical group that does not contain chemical functional groups susceptible to interaction with cyclic esters such as lactide, glycolactide, caprolactone and the like, or other esters and mixtures thereof. The reaction of the capped PEG-based polymer with a lactide produces a PEG-PLA diblock copolymer.

The combination of a multi-branched copolymer with a linear diblock copolymer provides an additional option for modulating the release of a pharmaceutically active ingredient.

In one embodiment, the molecular weight of the diblock copolymer-terminated polyethylene glycol chain is 100 Da to 10 kDa, or 164 Da to 2 kDa, preferably 1 to 2 kDa.

In one embodiment, the molar ratio of ester repeat unit to ethylene oxide repeat unit in the diblock copolymer is from 0.8 to 15, optionally from 1 to 10.

Typically, the polyester A in the triblock or diblock copolymer is selected from the group consisting of polylactic acid (PLA), polyglycolic acid, polycaprolactone, polyethylene adipate,

- 11 045374 polyhydroxyalkanoate, ε-caprolactone-lactide copolymer (PCLA), lactic acid-glycolic acid copolymer (LACA) and mixtures thereof, and optionally the polyethylene glycol with the end capping group is methoxy polyethylene glycol. Preferably, polyester A is polylactic acid.

Linear triblock and diblock copolymers suitable for use in the compositions of the present invention are described in WO 2012/090070 A1, WO 2019016233 A1, WO 2019016234 A1 and WO 2019016236 A1, incorporated herein by reference.

According to one embodiment of the present invention, the composition further comprises a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is optionally an organic solvent. The solvent is typically a biocompatible solvent. Preferably, the pharmaceutically acceptable carrier is selected from the group: benzyl alcohol, benzyl benzoate, dimethyl isosorbide (DMI), dimethyl sulfoxide (DMSO), ethyl acetate, ethyl benzoate, ethyl lactate, glycerol formal, methyl ethyl ketone, methyl isobutyl ketone, N-ethyl-2-pyrrolidone, N-methyl-2-pyrrolidinone (NMP) , pyrrolidone-2, tetraglycol, triacetin, tributyrin, tripropionine, glycofurol and mixtures thereof. The amount of said carrier is typically at least 25% or at least 35% w/w. from the total mass of the composition.

The composition contains at least one pharmaceutically active ingredient.

In one embodiment, the pharmaceutically active ingredient is hydrophobic.

In a preferred embodiment, the pharmaceutically active ingredient is meloxicam, bupivacaine, tamsulosin, octreotide, tadalafil, empagliflozin, tenofovir, liothyronine, or combinations thereof.

According to one embodiment of the present invention, the content of the at least one pharmaceutically active ingredient is from 0.05 to 60% w/w, optionally from 0.05 to 40%, optionally from 0.05 to 30%, optionally from 0. 05 to 10%, optionally from 0.05 to 7%, optionally from 0.05 to 2% of the total weight of the composition.

In a preferred embodiment, the composition is an injectable liquid.

According to one embodiment of the present invention, the multi-branched copolymer content is from 2 to 80%, optionally from 2 to 70%, optionally from 2 to 60%, optionally from 10 to 60%, optionally from 10 to 50%, optionally from 20 to 40 %, optionally from 20 to 35%, optionally from 30 to 50% wt./wt. from the total mass of the composition. This may correspond to the amount of one multiple branched copolymer if only one multiple branched copolymer is present, or the total amount of multiple branched copolymers if two or more multiple branched copolymers are present. If a diblock or triblock copolymer is present, the amount of multi-branched copolymer is preferably varied such that the total amount of copolymer remains substantially constant.

Typically, the molar ratio of ester repeating units and ethylene oxide in the composition is from 1 to 10, preferably from 2 to 6.

Typically, the release of at least one pharmaceutically active ingredient can be modulated by the composition.

In one embodiment of the present invention, the composition is suitable for delivering a pharmaceutically active ingredient to a subject for at least 1 day, optionally for at least 3 days, optionally for at least 7 days, optionally for at least 30 days, optionally for at least 90 days, optionally for at least 1 year.

In a further aspect, the present invention provides the use of a pharmaceutical composition as defined above to modulate the release kinetics of at least one pharmaceutically active ingredient.

In a further aspect, the present invention provides a method for preparing a pharmaceutical composition as defined above, comprising dissolving a multi-branched copolymer as defined above in a pharmaceutically acceptable carrier and then adding a pharmaceutically active ingredient to the composition.

In a further aspect, the present invention provides a bioabsorbable depot that is prepared ex vivo or in situ by contacting the composition as defined above with an aqueous medium, water or biological fluid.

In a final aspect, a method of controlled release of a pharmaceutically active ingredient is provided, comprising administering to a subject a composition as defined above and allowing the formation of an in-situ depot in vivo.

The pharmaceutical composition is preferably suitable for parenteral administration. The term parenteral administration includes intramuscular, intraperitoneal, intraperitoneal, subcutaneous, intravenous and intra-arterial. The term also includes intradermal, intracavernosal, intravitreal, intracerebral, intrathecal, epidural, intra-articular and intraosseous administration.

The subject may be an animal or a plant. The term animals covers all representatives of the animal kingdom. The animal may be a human or a non-human animal.

As used herein, the term plant covers all members of the plant kingdom.

Pharmaceutically active ingredient means a drug or drug for the treatment or prevention of various medical diseases. For the purposes of this patent application, the term active principle has the same meaning as active ingredient. Thus, the terms active ingredient, active principle, drug or drug product are used interchangeably. The term active pharmaceutical ingredient or API is also used. As used herein, the terms drug or active ingredient include, without any limitation, physiologically or pharmacologically active substances that act locally or systemically in the animal or plant body.

As used herein, the term disease means any disorder in a person, animal or plant caused by infection, diet or abnormal process.

The term bulk composition covers any composition that can be applied to or introduced into the body of an animal or plant body and does not necessarily need to be administered by syringe.

As used herein, the term repeat units refers to the underlying repeating units of the polymer.

As used herein, the term polyethylene glycol, abbreviated as PEG throughout the present invention, is sometimes referred to as poly(ethylene oxide) or poly(oxyethylene), and these terms are used interchangeably in the present invention.

The abbreviation PLA stands for poly(lactic acid).

The abbreviation PMCGK corresponds to a copolymer of lactic acid and glycolic acid.

The abbreviation PKMK corresponds to a copolymer of ε-caprolactone and lactide.

The abbreviation PE stands for polyester.

The copolymers were named as follows:

The linear triblock copolymers described herein are referred to as PxRy, where x is the molecular weight of the PEG chain expressed in kDa and y is the molar ratio of the ester repeat unit and ethylene oxide, for example the molar ratio of lactic acid and ethylene oxide (LA/EO), which makes it possible to calculate the length of the PLA chain inside the copolymer.

The linear diblock copolymers described herein are referred to as dPxRy, where x is the molecular weight of the PEG chain expressed in kDa and y is the molar ratio of ester monomer to ethylene oxide, for example the molar ratio of lactic acid to ethylene oxide (LA/EO).

sz-PxRy denotes the PEG-PLA star copolymer with z arms, x and y provide the same information as in linear copolymers, namely x represents the molecular weight of the polyether core formed by the reaction of the polyol with PEG (often called star PEG) , and y represents the molar ratio of ester monomer and ethylene oxide.

As an example, s4-P2R6 is a 4-arm star copolymer with a 2 kDa star PEG block with a total UA/EO molar ratio of 6.

As used herein, the term needle penetration capacity for a composition is defined as the force, expressed in Newtons (N), required to inject the composition using predetermined parameters. These parameters include: injection speed, injection volume, injection duration, syringe type or needle type, and the like. These parameters may vary depending on the at least one pharmaceutically active ingredient used or the desired route of administration, such as subcutaneous, intraocular, intra-articular, and so on. They can be adjusted depending on the at least one pharmaceutically active ingredient present in the formulations so that differences and variations between formulations can be observed. The ability to pass through the needle upon administration should be so low that the composition can be easily administered by a qualified healthcare professional in an acceptable period of time. Acceptable needle passability may be 0.1 to 20 N using the measurement method described below, with needle passability of 0.1 to 10 N being most preferred. Needle passability when inserted in suboptimal conditions may be greater than 20 to 30 N. Formulations are difficult to inject at 30 to 40 N and are not injectable at greater than 40 N. The ability to pass through the needle during injection can be measured using a texture meter, preferably a Lloyd Instruments texture meter.

FT plus using the following assay conditions: 500 µl of the formulation is injected through a 1 ml syringe with a 1'' 23G Terumo needle at a flow rate of 1 ml/min as described in Example 6.

Viscosity, by definition and as used herein, is a measure of a fluid's resistance to flow and step deformation under shear stress or tensile strength. The term describes the internal friction of a moving fluid. For liquids, the term corresponds to the informal concept of density. Dynamic viscosity is a measure of the resistance to fluid flow under the influence of applied force. The dynamic speed can range from 1 mPa-s to 3000 mPa-s, or from 5 mPa-s to 2500 mPa-s, or from 10 mPa-s to 2000 mPa-s, or from 20 mPa-s to 1000 mPa -With. Dynamic viscosity is determined using an Anton Paar viscometer equipped with a cone-plane system. As a rule, 250 μl of the test composition is placed on the measuring flat disk. The temperature is controlled at +25°C. The measuring system used is a cone with a flat disk with a diameter of 25 mm and a cone angle of 1° (CP25-1). The operating range is 10-1000 s ^-1 . After shaking for 10 s, the formulations are placed in the center of a temperature-controlled measuring flat disk using a spatula. The measuring system is lowered down, leaving a clearance of 0.051 mm between the measuring system and the measuring flat disk. 21 viscosity measurement points are determined in the shear rate range of 10-1000 s ^-1 . The given values correspond to those obtained at 100 s ^-1 .

Exemplary drugs and biologically active agents for use in accordance with the present invention include, but are not limited to, peptides, proteins, antibodies, antibody fragments, desensitizers, antigens, vaccines, vaccine antigens, anti-infectives, antidepressants, stimulants, opiates, antipsychotics, atypical antipsychotics, drugs from glaucoma, anxiolytics, antiarrhythmics, antibacterials, anticoagulants, anticonvulsants, antidepressants, antiemetics, antifungals, antitumor agents, antivirals, antibiotics, antimicrobials, antiallergic agents, antidiabetic agents, steroidal anti-inflammatory agents, decongestants, miotic agents , anticholinergics, sympathomimetic agents, sedatives, hypnotics, thymoleptics, tranquilizers, hormones, androgenic steroids, estrogens, progestogens, humoral agents, prostaglandins, analgesics, corticosteroids, antispasmodics, antimalarials, antihistamines, cardiotonic agents, non-steroidal anti-inflammatory agents , antiparkinsonian agents, antihypertensive agents, beta-adrenergic receptor blockers, nutritional agents, gonadotropin-releasing hormone agonists, insecticides, anthelmintic agents, or combinations thereof.

The pharmaceutically active ingredient may be meloxicam, bupivacaine, tamsulosin, octreotide, tadalafil, empagliflozin, tenofovir, liothyronine, or combinations thereof.

Combinations of drugs can be used in a biodegradable drug delivery composition according to the present invention. For example, if treatment of lupus erythematosus (Lupus erythematosus) is necessary, then according to the present invention, non-steroidal anti-inflammatory agents and corticosteroids can be administered together.

Veterinary drugs, such as drugs for the treatment of helminthic infestations or vaccines for animals, are also part of the present invention.

Antiviral agents for plants against viruses such as viruses of the families Potyviridae, Geminiviridae, virus of the genus Tospovirus of the family Bunyaviridiae and banana streak virus are also covered by the present invention. Also, agents against tobacco mosaic virus, turnip crinkle virus, barley yellow dwarf virus, watermelon ringspot virus and cucumber mosaic virus can be used in the biodegradable drug delivery composition of the present invention.

Those skilled in the art may utilize other drugs or biologically active agents that may be released into the aqueous environment in the described delivery system. Various forms of drugs or biologically active agents can also be used. These forms include, but are not limited to, forms such as uncharged molecules, molecular complexes, salts, esters, esters, amides, etc., which are biologically activated when administered to an animal or plant or are used in bulk form so that they can be applied to the surface of the body of an animal or plant organism or inserted into them in the form of a rod-type implant.

The pharmaceutically effective amount of the active ingredient may vary depending on the active ingredient, the state of health of the animal or plant, and the time required to deliver the active ingredient. There is no critical upper limit for the amount of active ingredient included in the polymer solution because the solution or suspension has a viscosity that is acceptable for administration through a syringe needle and because it

- 14 045374 can provide effective treatment for a medical condition without exposing the animal or plant to overdose. The lower limit of the active ingredient included in the delivery system depends only on the potency of the active ingredient and the length of time required for treatment.

In the biodegradable drug delivery composition of the present invention, a pharmaceutically effective amount can be released gradually over an extended period of time. This sustained release may be continuous or intermittent, linear or non-linear, and may vary depending on the composition of the multi-branched copolymer.

The active ingredient may be released over a period of time ranging from 1 day to 1 year or longer depending on the type of treatment required and the biodegradable drug delivery composition used. In one embodiment, the biodegradable drug delivery composition may provide delivery of the active ingredient within at least 1 day, optionally at least 3 days, optionally at least 7 days. In another embodiment, the biodegradable drug delivery composition may provide delivery of the active ingredient over at least 30 days. In one embodiment, the biodegradable drug delivery composition may provide delivery of the active ingredient over at least 90 days. In yet another embodiment, the biodegradable drug delivery composition may provide delivery of the active ingredient for 1 year or longer.

The biodegradable drug delivery composition may be an injectable liquid, preferably at room temperature, and can be administered through a syringe without much effort. These biodegradable drug delivery compositions are also in-situ depot-forming and biodegradable, and convert to solid depots when administered to an animal or plant.

The composition may further comprise a pharmaceutically acceptable carrier, adjuvant or vehicle.

The adjuvant can be formulated simultaneously by mixing the drug. Regarding this, adjuvants that can be used are alum, aluminum phosphate, calcium phosphate, MPL™, CpG motifs, modified toxins, saponins, endogenous stimulatory adjuvants such as cytokines, complete and incomplete Freund's adjuvants, ISCOM type adjuvants, muramyl peptides and things like that.

The carrier may be any diluent, additional solvent, excipient or binder that may, if desired, affect the delivery of the active ingredient in the biodegradable drug delivery composition. Examples include small amounts of triglycerides such as triacetin or tripropionine.

In one embodiment, the composition may contain an organic solvent. The organic solvent may be selected from the following group: benzyl alcohol, benzyl benzoate, dimethyl isosorbide (DMI), dimethyl sulfoxide (DMSO), ethyl acetate, ethyl benzoate, ethyl lactate, glycerol formal, methyl ethyl ketone, methyl isobutyl ketone, N-ethyl-2-pyrrolidone, N-methyl-2- pyrrolidinone (NMP), 2-pyrrolidone, tetraglycol, triacetin, tributyrin, tripropionine and mixtures thereof. In one embodiment, the solvents may be DMSO, NMP, tripropionine, or mixtures thereof.

List of abbreviations

DBS - Diblock copolymer (mPEG-PLA)

DMSO - Dimethyl sulfoxide

EO - Ethylene Oxide

GPC - Gel permeation chromatography

UPLC - Ultraperformance Liquid Chromatography

KRT - Krebs-Ringer-Tris buffer solution

MK - Lactic acid mPEG - Methoxypoly(ethylene glycol) mPEG-PLA Block copolymer methoxypoly(ethylene glycol)-poly(lactic acid)

PBS - Phosphate buffer solution

IP - Polydispersity Index

PEG - Poly(ethylene glycol)

PLA-PEG-PLA - Block copolymer poly(lactic acid)-poly(ethylene glycol)-poly(lactic acid)

PLA - Poly(lactic acid)

TBS - Triblock copolymer (PMK-PEG-PMK)

Brief description of graphic materials

In fig. 1 shows the cumulative release of meloxicam in vitro as a percentage over time

- 15 045374 time from two different formulations: formulation F396 (□) containing 40.00% P2R6 triblock copolymer with

2.00% active ingredient (API) and 58.00% DMSO, and formulation F397 (Δ) containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation exhibits slower release kinetics compared to the linear copolymer formulation with comparable molecular weight and identical total copolymer content. Indeed, formulation F397 exhibits slower release kinetics than formulation F396.

In fig. Table 2 shows the values of the ability to pass through a needle during administration for compositions F396 and F397. The data demonstrates that, at the same copolymer loading and comparable molecular weight, the star copolymer formulation has a lower needle pass rate than the linear copolymer formulation. Thus, the values of the ability to pass through the needle upon administration for composition F397 are lower than those for F396. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 3 shows the viscosity values for compositions F396 and F397. The data shows that at the same copolymer loading and comparable molecular weight, the star copolymer formulation has a lower viscosity than the linear copolymer formulation. Thus, the viscosity values for F397 are lower than the viscosity values for F396. In table 4 provides detailed viscosity data.

In fig. 4 shows the cumulative percentage release of meloxicam in vitro over time from two different formulations: formulation F511 (O) containing 38.00% dP2R6 diblock copolymer with 2.00% active ingredient (API) and 60.00% DMSO, and formulation F397 (Δ) containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation results in slower release kinetics compared to a linear copolymer formulation of comparable molecular weight and similar total copolymer content. Indeed, formulation F397 exhibits slower release kinetics than formulation F511.

In fig. Figure 5 shows the values of the ability to pass through a needle upon administration for composition F397 and composition F511. The data demonstrates that with similar copolymer loading and comparable molecular weight, both formulations have comparable needle pass rates upon administration. Thus, while having a similar ability to pass through a needle upon administration, the star copolymer formulation exhibits slower release kinetics. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 6 shows the viscosity values for compositions F397 and F511. The data shows that at similar copolymer loading and comparable molecular weight, both the star copolymer formulation and the linear copolymer formulation have similar viscosity values. Thus, for similar viscosities, the star copolymer formulation exhibits slower release kinetics. In table 4 provides detailed viscosity data.

In fig. Figure 7 shows the cumulative percentage release of meloxicam in vitro over time from two different formulations: formulation F510(®) containing 61.00% dP2R1.5 diblock copolymer with 2.00% active ingredient (API) and 37.00% DMSO , and formulation F397 (Δ) containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

Data indicate that a star copolymer formulation results in slower release kinetics compared to a linear copolymer formulation with comparable polyester chain length per branch. Indeed, formulation F397 exhibits slower release kinetics compared to formulation F510.

In fig. Figure 8 shows the values of the ability to pass through a needle when administered for compositions F397 and F510. The data demonstrate that, with similar needle permeability upon administration and similar polyester chain length per branch, the star copolymer formulation exhibits slower release kinetics. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 9 shows the viscosity values for compositions F397 and F510. The data demonstrates that at similar viscosity and similar polyester chain length per branch, a formulation based on

- 16 045374 star copolymer results in slower release kinetics. In table 4 provides detailed viscosity data.

In fig. 10 shows the cumulative percentage release of meloxicam in vitro over time from two different formulations: Formulation F391 (O) containing 40.00% P2R3.5 triblock copolymer with 2.00% active ingredient (API) and 58.00% DMSO , and formulation F451 (V), containing 47.00% s4-P2R3 star copolymer with 2.00% active ingredient (API) and 51.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation results in slower release kinetics than the linear copolymer formulation of comparable molecular weight. Indeed, formulation F451 exhibits slower release kinetics compared to formulation F391.

In fig. Figure 11 shows the values of the ability to pass through a needle upon administration for compositions F391 and F451. The data demonstrate that, with similar needle permeability upon administration and similar molecular weight, the star copolymer formulation exhibits slower release kinetics. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 12 shows the viscosity values for F391 and F451. The data demonstrates that, at similar viscosity and molecular weight, the star copolymer formulation exhibits slower release kinetics than the linear copolymer formulation. In table 4 provides detailed viscosity data.

In fig. 13 shows the total cumulative percentage release of meloxicam in vitro over time from two different formulations: formulation F449 (O) containing 45.00% dP2R3 diblock copolymer with 2.00% active ingredient (API) and 53.00% DMSO, and formulation F451 (V) containing 47.00% s4-P2R3 star copolymer with 2.00% active ingredient (API) and 51.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation results in slower release kinetics than the linear copolymer formulation of comparable molecular weight. Indeed, formulation F451 exhibits slower release kinetics compared to F449.

In fig. 14 shows the values of the ability to pass through a needle upon administration for compositions F449 and F451. The data demonstrate that, with a lower needle penetration rate and similar molecular weight, the star copolymer formulation exhibits slower release kinetics than the linear copolymer formulation. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 15 shows the viscosity values for compositions F449 and F451. The data demonstrates that at lower viscosities and similar molecular weights, the star copolymer formulation results in slower release kinetics than the linear copolymer formulation. In table 4 provides detailed viscosity data.

In fig. 16 shows the cumulative percentage release of meloxicam in vitro over time from two different formulations: Formulation F509 (O) containing 68.00% dP2R0.8 diblock copolymer with 2.00% active ingredient (API) and 30.00% DMSO , and formulation F451 (V), containing 47.00% s4-P2R3 star copolymer with 2.00% active ingredient (API) and 51.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

Data indicate that the star copolymer formulation results in slower release kinetics compared to linear copolymer formulations with comparable polyester chain length per branch. Indeed, formulation F451 exhibits slower release kinetics compared to F509.

In fig. Figure 17 shows the values of the ability to pass through a needle upon administration for compositions F451 and F509. The data demonstrate that, with comparable needle penetration values and similar polyester chain length per arm, the star copolymer formulation exhibits slower release kinetics compared to the linear copolymer formulation. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 18 shows the viscosity values for compositions F451 and F509. The data demonstrates that at comparable viscosity and similar polyester chain length per arm, the star copolymer formulation exhibits slower release kinetics compared to the linear copolymer formulation. In table 4 provides detailed viscosity data.

In fig. 19 shows the cumulative release of meloxicam in vitro as a percentage over time

- 17 045374 time from two different compositions: composition F388 (O) containing 40.00% triblock copolymer P2R2 with

2.00% active ingredient (API) and 58.00% DMSO, and formulation F389 (Δ) containing 40.00% s4-P2R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation exhibits slower release kinetics compared to the linear copolymer formulation of comparable molecular weight. Formulation F389 exhibits slower release kinetics than formulation F388.

In fig. 20 shows the values of the ability to pass through a needle upon administration for compositions F388 and F389. The data demonstrate that, at identical copolymer content and comparable molecular weight, the star copolymer formulation exhibits lower needle penetration and slower release kinetics compared to the linear copolymer formulation. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 21 shows the viscosity values for compositions F388 and F389. The data demonstrate that, at the same copolymer content and comparable molecular weight, the star copolymer formulation exhibits lower viscosity and slower release kinetics than the linear copolymer formulation. In table 4 provides detailed viscosity data.

In fig. 22 shows the cumulative percentage release of meloxicam in vitro over time from three different formulations: Formulation F483 (O) containing 44.00% P2R3.5 triblock copolymer with 2.00% active ingredient (API) and 54.00% NMP ; formulation F484 containing 47.00% dP2R3 diblock copolymer with 2.00% active ingredient (API) and 51.00% NMP, and formulation F489 (V) containing 51.00% s4-P2R3 star copolymer with 2.00% active ingredient (API) and 47.00% NMP. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation results in slower release kinetics than linear copolymer formulations of comparable molecular weight. Indeed, formulation F489 exhibits slower release kinetics compared to F483 and F484.

In fig. Figure 23 shows the values of the ability to pass through a needle upon administration for compositions F483, F484 and F489. The data demonstrate that, with identical needle penetration and comparable molecular weight, the star copolymer formulation exhibits slower release kinetics compared to linear copolymer formulations. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 24 shows the viscosity values for F483, F484 and F489. The data demonstrate that, at similar viscosity and comparable molecular weight, the star copolymer formulation exhibits slower release kinetics compared to linear copolymer formulations. In table 4 provides detailed viscosity data.

Fig. 25 is a graph showing the cumulative percentage release of meloxicam in vitro over time from three different formulations: Formulation F485 (O) containing 22.00% P2R3.5 triblock copolymer with 2.00% active ingredient (API) and 76.00% triacetin; formulation F486 containing 24.00% dP2R3 diblock copolymer with 2.00% active ingredient (API) and 74.00% triacetin, and formulation F488 (V) containing 26.00% s4-P2R3 star copolymer with 2.00 % active ingredient (API) and 72.00% triacetin. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation results in slower release kinetics than linear copolymer formulations of comparable molecular weight. Formulation F488 exhibits slower release kinetics than F485 and F486.

In fig. 26 shows the total cumulative percentage release of meloxicam in vitro over time from three different formulations: Formulation F389 (O) containing 40.00% s4-P2R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO ; formulation F405 (Δ) containing 40.00% s4-P2R3 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F397 (V) containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

Data show that increasing PLA chain length within a star copolymer

- 18 045374 leads to modulation of release kinetics in formulations with the same copolymer content. Formulation F397 exhibits slower release kinetics than F405 and F389. Similarly, formulation F405 exhibits slower release kinetics than F389.

In fig. 27 shows the cumulative percentage release of meloxicam in vitro over time from two different formulations: formulation F389 (V) containing 40.00% s4-P2R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F402 (Δ), containing 40.00% s4-P5R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

Data show that increasing PEG chain length within a star copolymer with a fixed UA/EO ratio results in increased release rates for formulations with the same copolymer content. Formulation F389 exhibits slower release kinetics compared to F402.

In fig. 28 shows the cumulative percentage release of meloxicam in vitro over time from two different formulations: formulation F405 (Δ) containing 40.00% s4-P2R3 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F451 (V), containing 47.00% s4-P2R3 star copolymer with 2.00% active ingredient (API) and 51.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

The data show that increasing the star copolymer content results in a decreased release rate. Indeed, formulation F451 exhibits slower release kinetics compared to F405.

In fig. 29 shows the cumulative in vitro release of bupivacaine as a percentage over time from two different formulations: formulation F413 (O) containing 40.00% P2R6 triblock copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F414 (Δ) containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

The results indicate that the star copolymer formulation exhibits slower release kinetics compared to linear copolymer formulations of comparable molecular weight. Indeed, formulation F414 exhibits slower release kinetics compared to formulation F413.

In fig. 30 shows the cumulative percentage in vitro release of meloxicam, bupivacaine and tamsulosin over time from three different formulations: formulation F414 (Δ) containing 40.00% s4-P2R6 star copolymer with 2.00% bupivacaine and 58.00% DMSO; formulation F397 (O) containing 40.00% s4-P2R6 star copolymer with 2.00% meloxicam and 58.00% DMSO, and formulation F460 (V) containing 40.00% s4-P2R6 star copolymer with 2.00 % tamsulosin and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

Data show that it is possible to achieve long-term sustained release of various APIs using the star copolymer formulations described in the present invention.

Fig. 31 is a graph showing the cumulative percentage release of meloxicam in vitro over time from F397. Formulation F397 (O) contains 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

The results demonstrate that the star copolymer formulation results in sustained drug release over a period of no more than at least 6 months.

In fig. 32 shows the cumulative percentage release of tamsulosin in vitro over time from two different formulations: formulation F463 (Δ) containing 40.00% s4-P2R6 star copolymer, 14.40% active ingredient (API) and 45.60% DMSO, and formulation F460 (V), containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

The results show that star copolymer formulations provide sustained release with two different API loadings.

In fig. 33 shows the release rate of meloxicam in micrograms per day over time from four different formulations: Formulation F391 (O) containing 40.00% P2R3.5 triblock copolymer with 2.00% active ingredient (API) and 58.00% DMSO ; composition F449 (□), containing 45.00% dP2R3 diblock copolymer with 2.00% active ingredient (API) and 53.00% DMSO; composition F451 (Δ), containing 19 045374 47.00% star copolymer s4-P2R3 with 2.00% active ingredient (API) and 51.00%

DMSO, and formulation F397 (V), containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results indicate that star copolymer formulations result in slower release kinetics than linear copolymer formulations of comparable molecular weight. Indeed, formulations F451 and F397 exhibit slower release kinetics compared to F391 and F449.

It can be seen that the star copolymer formulation of F397 results in the lowest release rate.

Fig. 34 is a graph depicting the total plasma concentration of meloxicam active moiety, expressed in nanograms per milliliter, over time from the four different formulations shown in FIG. 33. In vivo release tests were carried out in accordance with scheme 1 in table. 5, example 7.

The results indicate that, based on in vitro observations, star copolymer formulations exhibit slower in vivo release kinetics compared to linear copolymer formulations of comparable molecular weight. Indeed, formulations F451 and F397 exhibit slower release kinetics compared to F391 and F449.

Similar to the in vitro results, it was observed that the star copolymer formulation of F397 resulted in the lowest release rate.

In fig. Figure 35 shows the values of the ability to pass through a needle upon administration for compositions F391, F449, F451 and F397. The data demonstrate that, with similar needle penetration capacity (about 20 N), star copolymer formulations exhibit slower release kinetics compared to linear copolymer formulations. In table 3 provides detailed data on the ability to pass through the needle during injection.

In fig. 36 presents the cumulative percentage in vitro release of lactic acid over time from two different formulations as a measure of the degree of degradation of the PEGPLLA copolymer. Formulation F496 (O) contains 40.00% triblock copolymer P2R2 with 2.00% tamsulosin and 58.00% DMSO, and formulation F497 (□) contains 40.00% star copolymer s4-P2R2 with 2.00% tamsulosin and 58.00% DMSO. Quantitative determination of lactic acid was carried out in accordance with example 4. In table. 1 below shows specific compositions based on block copolymers.

Data show that the linear copolymer formulation of F496 results in a continuous release of lactic acid over a period of time. The star copolymer formulation F497 results in slower lactic acid release kinetics than F496 for the first 50 days, with release then accelerating for the remainder of the study period.

Thus, unexpectedly, the star copolymer formulation results in accelerated decomposition of the depot compared to the linear copolymer formulation.

Fig. 37 is a graph showing the cumulative percentage in vitro release of lactic acid over time from two different formulations: formulation F498 (O) containing 40.00% dP2R3 diblock copolymer with 2.00% tamsulosin and 58.00% DMSO, and formulation F499 (□), containing 40.00% s4-P2R3 star copolymer with 2.00% tamsulosin and 58.00% DMSO. Quantitative determination of lactic acid was carried out in accordance with example 4. In table. 1 below shows specific compositions based on block copolymers.

Data show that the linear copolymer formulation of F498 results in a slow, quasi-constant release of lactic acid during the first 90 days, followed by an accelerated release for the remainder of the study period. The star copolymer formulation of F499 results in slower lactic acid release kinetics than F498 for the first 50 days, with release then accelerating for the remainder of the study period. Thus, depots made from star copolymer formulations undergo accelerated degradation compared to depots made from linear copolymer formulations.

In fig. 38 shows the cumulative percentage release of meloxicam in vitro over time from three different formulations: formulation F389(®) containing 40.00% s4-P2R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, composition F401 (Δ), containing 20.00% star copolymer s4-P2R6 and 20.00% star copolymer s4-P2R2 with 2.00% active ingredient (API) and 58.00% DMSO, and composition F397 (O), containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

The results demonstrate that the mixture of two star copolymers in the formulation composition results in a modulated or improved release profile.

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In fig. 39 shows the cumulative percentage release of meloxicam in vitro over time from three different formulations: formulation F397 (O) containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, formulation F402 (V), containing 40.00% s4-P5R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F404 (□), containing 20.00% s4-P2R6 star copolymer and 20.00% s4-P5R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

The results demonstrate that the mixture of two star copolymers in the formulation results in a modulated release profile.

In fig. 40 shows the cumulative percentage release of meloxicam in vitro over time from three different formulations: Formulation F388 (O) containing 40.00% linear copolymer P2R2 with 2.00% active ingredient (API) and 58.00% DMSO, Formulation F389 (O), containing 40.00% s4-P2R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F390 (□), containing 20.00% s4-P2R2 star copolymer and 20 .00% linear copolymer P2R2 with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

Data indicate that a mixture of linear and star copolymers in the formulation composition can result in a modulated release profile. Surprisingly, the mixed star and linear copolymer formulation results in an optimized release profile compared to the star or linear copolymer formulation.

In fig. 41 shows the cumulative percentage release of bupivacaine in vitro over time from three different formulations: formulation F407 (Δ) containing 40.00% linear copolymer P2R2 with 2.00% active ingredient (API) and 58.00% DMSO, formulation F408 (V) containing 40.00% s4-P2R2 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F409 (®) containing 20.00% s4-P2R2 star copolymer, 20 .00% linear copolymer P2R2 with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

Data indicate that a mixture of linear and star copolymers in the formulation composition can result in a modulated release profile. Thus, surprisingly, a mixed star and linear copolymer formulation results in an optimized release profile compared to a star or linear copolymer formulation.

In fig. 42 shows the cumulative percentage release of meloxicam in vitro over time from three different formulations: formulation F388 (O) containing 40.00% linear copolymer P2R2 with 2.00% active ingredient (API) and 58.00% DMSO, formulation F397 (O) containing 40.00% s4-P2R6 star copolymer with 2.00% active ingredient (API) and 58.00% DMSO, and formulation F399 (□) containing 20.00% s4-P2R6 star copolymer, 20 .00% linear copolymer P2R2 with 2.00% active ingredient (API) and 58.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3. In table. 1 shows specific compositions based on block copolymers.

Data indicate that a mixture of linear and star copolymers in the formulation composition can result in a modulated release profile.

In fig. 43 shows the cumulative percentage release of empagliflozin in vitro over time from three different formulations: formulation F135 (Δ) containing 20.00% s4-P2R3 star copolymer with 20.00% active ingredient (API) and 60.00% DMSO, formulation F136 ('=) containing 20.00% s4-P2R6 star copolymer with 20.00% active ingredient (API) and 60.00% DMSO, and formulation F137 (V) containing 30.00% s4- star copolymer P2R6 with 20.00% active ingredient (API) and 50.00% DMSO. In vitro release tests were carried out in accordance with scheme 2 in table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

The results indicate that star copolymer formulations exhibit sustained release kinetics of empagliflozin over time. The data demonstrate that changing the R ratio causes a modulation of the resulting release profiles. Indeed, formulation F136 exhibits slower release kinetics than formulation F135. The data show that increasing the total copolymer content leads to a modulation of the release profile. F137 exhibits slower release kinetics than formulation F136.

Fig. 44 is a graph showing the cumulative percentage release of tadalafil in vitro over time from F47 and F48. Formulation F47 (Δ) contains 25.00% s4-P2R6 star copolymer with 20.00% active ingredient (API) and 55.00% DMSO. Composition F48 (V)

- 21 045374 contains 25.00% star copolymer s4-P2R3 with 20.00% active ingredient (API) and 55.00%

DMSO. In vitro release tests were carried out in accordance with scheme 3 in table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

Data show that star copolymer formulations provide sustained release of tadalafil over time. The data demonstrate that changing the R ratio causes a modulation of the resulting release profiles. Indeed, formulation F47 exhibits slower release kinetics than formulation F48.

Fig. 45 is a graph showing the cumulative percentage release of streptavidin in vitro over time from F149. Formulation F149 (Δ) contains 28.00% s4-P2R3 star copolymer with 1.50% active ingredient (API) and 70.50% tripropionine. In vitro release tests were carried out in accordance with scheme 4 in table. 2, example 3. In table. 1 below shows specific compositions based on block copolymers.

Data show that star copolymer formulations provide sustained release of streptavidin over time.

Fig. 46 is a graph depicting the total plasma concentration of tadalafil active moiety, expressed in nanograms per milliliter, over time for formulation F48, wherein F48(V) contains 25.00% s4-P2R3 star copolymer with 20.00% active ingredient (API) and 55.00% DMSO. In vivo release tests were carried out in accordance with scheme 2 in table. 5, example 7.

The results indicate that, consistent with in vitro observations, the formulation provides sustained release of tadalafil over time in vivo.

Fig. 47 is a graph depicting the total plasma concentration of meloxicam active moiety, expressed in nanograms per milliliter, over time for formulation F2. F2(O) contains 20.00% s4-P2R3 star copolymer, 20.00% P2R2 triblock copolymer with 2.00% active ingredient (API) and 58.00% DMSO. In vivo release tests were carried out in accordance with scheme 1 in table. 5, example 7.

The results indicate that the combination of star and linear copolymers in F2 results in a sustained release profile of meloxicam over time in vivo.

In fig. 48 shows the cumulative percentage release of tenofovir in vitro over time from two different formulations: formulation F19 (Δ) containing 40.00% s4-P2R3 star copolymer with 19.00% active ingredient (API) and 41.00% DmSO, and formulation F20 ('=) containing 40.00% s4-P2R6 star copolymer with 19.00% active ingredient (API) and 41.00% DMSO. In vitro release tests were carried out in accordance with scheme 5 in table. 2, example 3.

The results demonstrate that star copolymer formulations provide sustained release of tenofovir over time. The data demonstrate that changing the R ratio causes a modulation of the resulting release profiles. Indeed, formulation F20 exhibits slower release kinetics than formulation F19.

Fig. 49 is a graph depicting the total plasma concentration of octreotide active moiety, expressed in nanograms per milliliter, over time for formulation F145. F145(O) contains 28.00% s4-P2R3 star copolymer, 18.00% P1R4 triblock copolymer with 0.70% active ingredient (API) and 53.30% DMSO. In vivo release tests were carried out in accordance with scheme 3 in table. 5, example 7.

The results indicate that the combination of star and linear copolymers in F145 results in a sustained release profile of octreotide over time in vivo.

In fig. 50 shows the cumulative percentage release of tenofovir in vitro over time from two different formulations: formulation F29 (V) containing 40.00% s4-P2R3 star copolymer with 0.10% active ingredient (API) and 59.90% DMSO, and formulation F30 (O), containing 40.00% s4-P2R6 star copolymer with 0.10% active ingredient (API) and 59.90% DMSO. In vitro release tests were carried out in accordance with scheme 6 in table. 2, example 3.

The results demonstrate that star copolymer formulations provide sustained release of liothyronine over time. The data demonstrate that changing the R ratio causes a modulation of the resulting release profiles. Indeed, formulation F29 exhibits slower release kinetics than formulation F30.

In fig. 51 shows the cumulative percentage release of meloxicam in vitro over time from two different formulations: formulation F521 (V) containing 55.00% S3-P0.45R3 star copolymer with 2.00% active ingredient (API) and 43.00% DMSO, and formulation F522 (□), containing 55.00% S3-P1R3 star copolymer with 2.00% active ingredient (API) and 43.00% DMSO. In vitro release tests were carried out according to Scheme 1 in Table. 2, example 3.

The results demonstrate that increasing the PEG chain length within a star copolymer with a fixed UA/EO ratio results in a modulated release rate for formulations with the same copolymer content. Indeed, formulation F521 exhibits slower release kinetics than formulation F522.

Examples

Example 1. Materials

Star block copolymers

Below is a general reaction scheme for preparing multi-branched PEG-PLA used in the pharmaceutical composition according to the present invention. The letters m and and describe the number of repeat units in each PEG and PLA block, respectively. Considering the synthesis route and experimental conditions, it is assumed that polymers with multiple arms are symmetrical and each arm is characterized by the same structure and composition. It should be understood that although Scheme 1 below uses a 4-arm PEG derivative; a similar reaction scheme can be used with multi-branched PEG having a different number of PEG-based arms.

Multi-branched block copolymers were synthesized by ring-opening polymerization of D,L-lactide initiated by multi-branched polyethers called multi-branched PEGs or star-shaped PEGs (S. J. Buwalda et al., Influence of amide versus ester linkages on the properties of eight-armed PEGs -PLA star block copolymer hydrogels, Biomacromolecules 11 (2010) 224-232). Star-shaped PEGs are commercially available, for example, from PEG development efforts, such as the 4-arm PEG-OH shown in Scheme 1. Alternatively, multi-branched PEG can be formed by reacting the lenoxide with a polyol.

The catalyst (e.g., at a monomer/catalyst mol/mol ratio of 10,000) was then added to the star-shaped PEG at 80°C in the reaction vessel and allowed to mix with the macroinitiator before adding the appropriate amount of D,L-lactide (the amount of monomer is determined depending on the target ratio R). To remove any water in the system, the mixture was further dried using several successive cycles of vacuum and nitrogen. The reaction mixture was then heated and allowed to react for several hours (until complete conversion of the monomer). At the end of the polymerization reaction, the polymer is removed from the reaction vessel and allowed to cool. The polymer is then further purified to remove any unreacted monomer, catalyst, and oligomers. To do this, the polymer mixture is dissolved in an appropriate solvent, namely acetone, and left to dissolve overnight. The polymer solution is then precipitated in a block copolymer precipitant (ie ethanol). This step is repeated several times to ensure that only the target polymer is recovered. The collected polymer is left to dry under vacuum until all solvents have been removed from the sample.

Star block copolymers were analyzed and characterized after synthesis and purification to ensure that the resulting polymers had the desired structure and composition. For this purpose, analysis was carried out by 1H NMR in CDCl ₃ and by GPC in chloroform.

1H NMR analysis was performed by a third party company according to their standard procedure on a Brucker advance 300 MHz spectrometer. For all 1H NMR spectrograms, MestReNova software was used for peak integration and analysis. Chemical shifts were presented relative to the δ=7.26 ppm value for the CDCl ₃ solvent.

Gel permeation chromatography (GPC) measurements were performed on a Gel Permeation Chromatography Triple Diode Array Detector (GPC-TDA) instrument supplied by Malvern. About 150-200 mg of polymers were solubilized in 10 mL HPLC grade chloroform overnight before being placed in a 1.5 mL capped assay vial. After determining the dn/dc value for each polymer, 100 μl of the polymer solution was introduced into the GPC system in 3 replicates. Each replicate was then separately analyzed and integrated. The values of _Mn and polydispersity index (PI) given above correspond to the average value determined taking into account all injections.

Linear block copolymers

Linear triblock copolymers, compared to star block copolymers, are usually characterized by the formula

Av-Bw-Ax, where A is a polyester, B is a polyethylene glycol, v and x are the number of repeat units ranging from 1 to 3000, and w is the number of repeat units ranging from 3 to 300, wherein v= x or vax.

Linear diblock copolymers, compared to star block copolymers, are usually characterized by the formula

Cy-Az, wherein A is a polyester, C is a capped polyethylene glycol, and y and z are the number of repeat units, with y ranging from 2 to 250 and z ranging from 1 to 3000.

Typically, linear triblock copolymers, compared to star block copolymers, are characterized by the formula

LAv-EOw-LAx, where v and x represent the number of repeat units in the range from 1 to 3000 and w represents the number of repeat units in the range from 3 to 300, with v = x or vax.

In general, linear diblock copolymers, as compared to star block copolymers, are characterized by the formula m(EO)y-LAz, where y and z represent the number of repeating units, with y ranging from 2 to 250 and z ranging from 1 to 3000.

The block copolymers were synthesized in accordance with the method described in US patent No. 6350812, incorporated herein by reference with minor modifications. Typically, the required amount of PEG (in the triblock copolymer) or methoxy-PEG (in the diblock copolymer) was heated at 80°C and dried under vacuum for 30 min in the reaction vessel. D,L-lactide (according to the target LA/EO molar ratio) and catalyst (1/1000 of the amount of lactide) were added. To avoid any side reaction due to the presence of water, the reaction mixture was subjected to two short vacuum/No. cycles. The reaction mixture was then heated at 130°C under a constant flow of nitrogen (0.2 bar). After the reaction stopped, the block copolymer was removed from the vessel and left to cool. The resulting polymer sample was then solubilized in acetone before precipitation in a large volume of precipitant (precipitant/solvent ratio = 6-10) to remove any unreacted monomer, catalyst or oligomers present in the sample. This purification process was repeated twice. The collected polymer was then left to dry under vacuum to remove any traces of solvent and extract only the target polymer.

The resulting product was characterized by 1H NMR to determine its residual lactide content and to determine the R ratio.

1H NMR spectroscopy was performed using a Brucker advance spectrometer at 300 MHz. For all 1H NMR spectrograms, MestReNova software was used for peak integration and analysis. Chemical shifts were presented relative to the δ=7.26 ppm value for the CDCl ₃ solvent.

To determine the R ratio, which describes the ratio of lactic acid-based units to ethylene oxide-based units (LA/EO), integration of all peaks was performed separately. The signal intensity (the result of integration) is directly proportional to the number of hydrogen atoms that cause the signal. Therefore, in order to determine the ratio R (MK/EO ratio), the integration results must be homogeneous and correspond to the same number of protons (for example, all signal values are determined for 1H). The characteristic peak of PLA and one peak of PEG are then used to determine the LA/EO ratio. This technique is applicable for calculating the molecular weight of PEG above 1000 g/mol, when the signal obtained for the end groups in the polymer can be neglected.

Example 2. Analysis of the soluble fraction of star copolymers in water

Water solubility tests were performed to determine the soluble fraction of the star copolymer in water.

The water solubility analysis consisted of the following steps:

Empty 20 ml bottles were weighed (1). A sample of 500 mg of copolymer was taken and added to the appropriate vial. 5 ml of ultrapure water was added to each bottle. The vials were kept for 2 hours at 37°C with vortex mixing. Conducted visual observations and

- 24 045374 received images. Then the vials (1) were centrifuged for 10 minutes at 3000 rpm. A glass bottle (2) with a capacity of 10 ml was weighed. The supernatant from (1) was transferred to (2) and the masses were recorded. The wet copolymer in (1) was weighed. (1) and (2) were placed at -80°C overnight. (1) and (2) were placed in a lyophilizer for 22 hours. Vials (1) and (2) were weighed. Water solubility was determined after drying and weighing the remaining dried copolymer. The amount of dissolved copolymer was determined as the difference in weight of the empty vial and the lyophilized vial. The water solubility assay was performed as a single replicate assay.

The results show water solubility values of 2.7 mg/mL, 1.7 mg/mL, and 1.7 mg/mL for s4P2R2, s4-P2R6, and s4-P5R4, respectively.

Example 3: In Vitro Release Tests

Detailed description of the procedure according to the scheme 1 mg of the meloxicam-containing composition was added to 20 ml of buffer solution in an Erlenmeyer flask. The buffer solution used was phosphate buffered saline (PBS) pH 7.4 which contained 137 mM sodium chloride, 2.7 mM potassium chloride, 10 mM disodium phosphate, 1.8 mM monopotassium phosphate and 0.1% azide sodium Upon injection, the solvent diffuses out of the formulation and the remaining polymer forms an in situ depot in the aqueous environment.

Closed Erlenmeyer flasks were kept with constant shaking at 180 rpm (Unimax 1010 device, Heidolph) at 37°C. At predetermined time intervals, 2 ml of medium was withdrawn and analyzed by UHPLC; the remaining medium was discarded and 20 ml of fresh buffer solution was added to the Erlenmeyer flask. Sufficient dilution conditions were maintained throughout the study. The amount of meloxicam released from the formulation was calculated from a calibration curve where the meloxicam concentration ranged from 0 to 160 μg/ml.

Meloxicam included in the polymer solution was encapsulated into the polymer matrix as the polymer cured.

An in vitro release (IVR) assay was performed after performing the sequential steps described below:

Preparation of the composition:

The required portion of the copolymer was taken into an empty and tared glass bottle with a capacity of 3 ml. The glass bottle was tared again. An exact mass of DMSO was added using a Pasteur pipette. Then the carrier media (copolymer + solvent) were placed on a roller mixer at room temperature (RT) for 6-7 hours until the copolymer was completely dissolved. Then the glass vials were tared and the required portion of the API was taken. The compositions were then placed overnight at room temperature on a roller mixer.

Start IVR assessment:

50 μl of the composition, pre-mixed on a vortex mixer, was taken from the appropriate glass vial into a 0.5 ml Codan syringe. The syringe was cleaned, tared and the formulation was directly injected from the syringe without a needle into a prefilled 50 ml glass vial containing 20 ml of release assay buffer (PBS 1X). After sedimentation and depot formation, the depot was cut out from the syringe using scissors. The syringe was weighed to determine the exact mass of the depot.

After all depots had been formed, the glass vials were placed on a stirrer at 37°C.

IVR Sample Selection and IVR Sample Preparation for API Quantitation

At each required time point, a sufficient amount of buffer solution was withdrawn from a 50 mL glass vial for analysis before the buffer solution was completely refreshed. 1 mL of each sample was filtered through a 0.2 μm hydrophilic filter into a 1 mL HPLC glass vial. The content of the API in the buffer solution after release was determined using UHPLC.

Some parameters, such as the weight of the formulation, the type of buffer solution, or the volume of the buffer solution, can be tailored depending on the API being studied, its solubility in different buffer solutions, and its target dose and duration of release. A diagram with various parameters is presented in table. 2 below.

All studied compositions are presented in table. 1 below.

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Table 1

Compound API Copolymer Solvent Number Name % (w/w) Code Structure PEG-based chain weight (kDa) Ratio (MK/EO) % (w/w) Name % (w/w) 2 Meloxicam 2.00 S4-P2R3 Branched 2000 3 20.00 DMSO 58.00 P2R2 Linear 2000 2 20.00 19 Tenofovir 19.00 S4-P2R3 Branched 2000 3 40.00 DMSO 41.00 20 Tenofovir 19.00 S4-P2R6 Branched 2000 6 40.00 DMSO 41.00 29 Liothyronine 0.10 S4-P2R3 Branched 2000 3 40.00 DMSO 59.90 thirty Liothyronine 0.10 S4-P2R6 Branched 2000 6 40.00 DMSO 59.90 47 Tadalafil 20.00 S4-P2R6 Branched 2000 6 25.00 DMSO 55.00 48 Tadalafil 20.00 S4-P2R3 Branched 2000 3 25.00 DMSO 55.00 135 Empagliflozin 20.00 S4-P2R3 Branched 2000 3 20.00 DMSO 60.00 136 Empagliflozin 20.00 S4-P2R6 Branched 2000 6 20.00 DMSO 60.00 137 Empagliflozin 20.00 S4-P2R6 Branched 2000 6 30.00 DMSO 50.00 145 Octreotide 0.70 S4-P2R3 Branched 2000 3 28.00 DMSO 53.30 P1R4 Linear 1000 4 18.00 149 Streptavidin 1.50 S4-P2R3 Branched 2000 3 28 Tripropionine 70.50 388 Meloxicam 2.00 P2R2 Linear 2000 2 40.00 DMSO 58.00 389 Meloxicam 2.00 S4-P2R2 Branched 2000 2 40.00 DMSO 58.00 390 Meloxicam 2.00 P2R2 Linear 2000 2 20.00 DMSO 58.00 S4-P2R2 Branched 2000 2 20.00 391 Meloxicam 2.00 P2R3.5 Linear 2000 3.5 40.00 DMSO 58.00 396 Meloxicam 2.00 P2R6 Linear 2000 6 40.00 DMSO 58.00 397 Meloxicam 2.00 S4-P2R6 Branched 2000 6 40.00 DMSO 58.00 399 Meloxicam 2.00 P2R2 Linear 2000 2 20.00 DMSO DMSO 58.00 58.00 S4-P2R6 Branched 2000 6 20.00 401 Meloxicam 2.00 S4-P2R2 Branched 2000 2 20.00 DMSO 58.00 S4-P2R6 Branched 2000 6 20.00 402 Meloxicam 2.00 S4-P5R2 Branched 5000 2 40.00 DMSO 58.00 404 Meloxicam 2.00 S4-P5R2 Branched 5000 2 20.00 DMSO 58.00 S4-P2R6 Branched 2000 6 20.00 405 Meloxicam 2.00 S4-P2R3 Branched 2000 3 40.00 DMSO 58.00 407 Bupivacaine 2.00 P2R2 Linear 2000 2 40.00 DMSO 58.00 408 Bupivacaine 2.00 S4-P2R2 Branched 2000 2 40.00 DMSO 58.00 409 Bupivacaine 2.00 S4-P2R2 Branched 2000 2 20.00 DMSO 58.00 P2R2 Linear 2000 2 20.00 413 Bupivacaine 2.00 P2R6 Linear 2000 6 40.00 DMSO 58.00 414 Bupivacaine 2.00 S4-P2R6 Branched 2000 6 40.00 DMSO 58.00 449 Meloxicam 2.00 dP2R3 Linear 2000 3 45.00 DMSO 53.00 451 Meloxicam 2.00 S4-P2R3 Branched 2000 3 47.00 DMSO 51.00 460 Tamsulosin 2.00 S4-P2R6 Branched 2000 6 40.00 DMSO 58.00 463 Tamsulosin 14.40 S4-P2R6 Branched 2000 6 40.00 DMSO 45.60 483 Meloxicam 2.00 P2R3.5 Linear 2000 3.5 44.00 NMP 54.00 484 Meloxicam 2.00 dP2R3 Linear 2000 3 47.00 NMP 51.00 485 Meloxicam 2.00 P2R3.5 Linear 2000 3.5 22.00 Triacetin 76.00 486 Meloxicam 2.00 dP2R3 Linear 2000 3 24.00 Triacetin 74.00 488 Meloxicam 2.00 S4-P2R3 Branched 2000 3 26.00 Triacetin 72.00 489 Meloxicam 2.00 S4-P2R3 Branched 2000 3 51.00 NMP 47.00 496 Tamsulosin 2.00 P2R2 Linear 2000 2 40.00 DMSO 58.00 497 Tamsulosin 2.00 S4-P2R2 Branched 2000 2 40.00 DMSO 58.00 498 Tamsulosin 2.00 dP2R3 Linear 2000 3 40.00 DMSO 58.00 499 Tamsulosin 2.00 S4-P2R3 Branched 2000 3 40.00 DMSO 58.00 509 Meloxicam 2.00 dP2R0.8 Linear 2000 0.8 68.00 DMSO 30.00 510 Meloxicam 2.00 dP2R1.5 Linear 2000 1.5 61.00 DMSO 37.00 511 Meloxicam 2.00 dP2R6 Linear 2000 6 38.00 DMSO 60.00

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table 2

IVR scheme Education depot Buffer solution Number Procedure Weight of substance administered (mg) Using a syringe Using a needle Type Volume (ml) 1 The medium was injected from a syringe without a needle 60 Codan 0.5 ml syringe No PBS-1X 20 2 The carrier medium was injected from a syringe with a needle 250 1ml capacity with softJect luer lock 19G*1.1x50mm PBS-1X 100 3 The carrier medium was injected from a syringe with a needle 150 1ml capacity with softJect luer lock 21G * 0.8 x 16 mm PBS-1X + 1% Tween 80 150 4 The medium was injected from a syringe without a needle 100 Codan 0.5 ml syringe No SEC-1X 20 5 The carrier medium was injected from a syringe with a needle 250 1ml capacity with softJect luer lock 21G*1.1x50mm KRT-1X + 2% Twin 80 33 6 The carrier medium was injected from a syringe with a needle 1000 1ml capacity with softJect luer lock 21 G 1.1X50 mm PBS-1X + 1% Triton X100 20

Example 4. Evaluation of depot decomposition

Depot degradation was assessed by quantifying the lactic acid content of the buffer solution used for the IVR assessment tests at each sampling time point. The amount of lactic acid in the medium is associated with the decomposition of PLA-based chains.

First, the samples were subjected to hydrolysis. After release, 500 μl of the medium was transferred into a 1.5 ml Eppendorf tube. 250 μl of 5 M NaOH was added. The Eppendorf tube was placed for 1 hour at 40°C. The reaction was stopped by adding 250 μl of 5 M HC1.

The material used was a commercial kit called Megazyme L-lactic acid kit®, commercially available from Libios, Pontchard-sur-Turdin, France. Lactic acid quantification was performed using a standard protocol without any modifications.

Example 5 Ability to Pass Through a Needle upon Insertion

The purpose of this experiment was to evaluate the potential impact of the use of star copolymers on the ability of formulations to pass through a needle during injection by comparing the values for these formulations with those of formulations based on similar linear copolymers.

Needle penetration tests were performed using a Lloyd Instruments FT plus texturometer according to the procedure described below:

The compositions (copolymer dissolved in an organic solvent) were mixed on a vortex mixer for 15 s. 500 μl of the composition was withdrawn using a 1 ml Codan syringe without a needle. Air bubbles were removed to avoid any interference during the measurement of the ability to pass through the needle during insertion. Then a Terumo 1 gauge 23 G needle was installed on the syringe for medium carriers or formulations, respectively. The syringe was placed on the texturemeter. The flow rate was fixed at 1 ml/min. The speed was recorded at 56.3 mm/min. The introduction of the composition began at a fixed rate. The injection device (ie, syringe + needle) was changed for each replicate.

The average force in newtons (N) required to insert each repetition was calculated using the software supplied with the texturemeter. Using the scheme described above, the present inventors have determined 20H as the maximum value for a formulation that can be easily added manually.

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Table 3

Compound API Copolymer Solvent Ability to pass through a needle when inserted Number Name % (mass/mass) Code Structure % (mass/mass) Name % (mass/mass) Number of repetitions Strength (N) Standard deviation 388 Meloxicam 2.00 P2R2 Linear 40.00 DMSO 58.00 6 6.3 1.1 389 Meloxicam 2.00 S4-P2R2 Another branch 40.00 DMSO 58.00 6 4.2 0.6 391 Meloxicam 2.00 P2R3.5 Linear 40.00 DMSO 58.00 6 15.7 1.0 396 Meloxicam 2.00 P2R6 Linear 40.00 DMSO 58.00 6 57.1 3.9 397 Meloxicam 2.00 S4-P2R6 Another branch 40.00 DMSO 58.00 6 21.6 1.6 449 Meloxicam 2.00 dP2R3 Linear 45.00 DMSO 53.00 6 20.4 0.5 451 Meloxicam 2.00 S4-P2R3 Another branch 47.00 DMSO 51.00 6 15.9 1.4 483 Meloxicam 2.00 P2R3.5 Linear 44.00 NMP 54.00 6 19.2 0.9 484 Meloxicam 2.00 dP2R3 Linear 47.00 NMP 51.00 6 19.2 1.1 489 Meloxicam 2.00 S4-P2R3 Another branch 51.00 NMP 47.00 6 17.2 1.3 509 Meloxicam 2.00 dP2R0.8 Linear 68.00 DMSO 30.00 6 19.0 1.1 510 Meloxicam 2.00 dP2R1.5 Linear 61.00 DMSO 37.00 6 21.3 2.7 511 Meloxicam 2.00 dP2R6 Linear 38.00 DMSO 60.00 6 20.6 0.5

Example 6 Dynamic Viscosity Analysis

Dynamic viscosity analysis was carried out using an Anton Paar viscometer equipped with a cone-plane system, with the following analysis conditions:

Temperature controlled at 25°C.

Amount of carrier medium: 0.25 ml

Measuring system: cone with a flat disk with a diameter of 25 mm and a cone angle of 1° (CP25-1).

Operating range: from 10 to 1000 mPa-s.

Before analysis, the composition was mixed on a vortex mixer for 10 s. 250 μl of the composition was placed in the center of a temperature-controlled measuring flat disk using a spatula. The measuring system was lowered down, leaving a clearance of 0.051 mm between the measuring system and the measuring flat disk. Twenty-one viscosity measurement points were determined at shear rates from 10 s ^-1 to 1000 s ^-1 (10 points per ten replicates). The viscosity data corresponds to the data calculated at a shear rate of 100 s ^-1 , which corresponds to the average value at the plateau of the curve. Analysis of the dynamic viscosity of the composition was carried out in triplicate.

Table 4

Compound API Copolymer Solvent Viscosity Number Name % w/ma ss. Code Structure wt.%) Name % (mass/mass) Number of repetitions Dynamic viscosity (mPa-s) Standard deviation 388 Meloxicam 2.00 P2R2 Linear 40.00 DMSO 58.00 6 341.2 5.9 389 Meloxicam 2.00 S4-P2R2 Branched 40.00 DMSO 58.00 6 213.5 1.8 391 Meloxicam 2.00 P2R3.5 Linear 40.00 DMSO 58.00 6 1001.7 3.3 396 Meloxicam 2.00 P2R6 Linear 40.00 DMSO 58.00 6 4602.9 123.1 397 Meloxicam 2.00 S4-P2R6 Branched 40.00 DMSO 58.00 6 1790.7 26.1 449 Meloxicam 2.00 dP2R3 Linear 45.00 DMSO 53.00 6 1427.0 28.2 451 Meloxicam 2.00 S4-P2R3 Branched 47.00 DMSO 51.00 6 1083.7 5.7 483 Meloxicam 2.00 P2R3.5 Linear 44.00 NMP 54.00 6 1286.4 20.7 484 Meloxicam 2.00 dP2R3 Linear 47.00 NMP 51.00 6 1376.3 6.6 489 Meloxicam 2.00 S4-P2R3 Branched 51.00 NMP 47.00 6 1182.0 7.1 509 Meloxicam 2.00 dP2R0.8 Linear 68.00 DMSO 30.00 6 1232.3 17.2 510 Meloxicam 2.00 dP2R1.5 Linear 61.00 DMSO 37.00 6 1579.9 15.1 511 Meloxicam 2.00 dP2R6 Linear 38.00 DMSO 60.00 6 1465.3 23.4

Example 7 Pharmacokinetic Study

Detailed description of in vivo regimen 1

Several meloxicam formulations were tested in a pharmacokinetic study on adult male rats weighing between 300 and 350 g. Drugs containing 3.6 mg meloxicam were administered subcutaneously into the interscapular region of rats using 1 ml Soft Ject® syringes and needles caliber 23G (1'' 0.6x25 mm) Terumo®. The volumes of the injected composition were fixed at 160 μl. Blood samples were collected in EDTA tubes at different time points: T0.5 h, T1 h, T3 h, T8 h, T24 h (day 1), T48 h (day 2), T96 h (day 4), T168 h ( day 7), T240 h (day 10), T336 h (day 14). Blood samples were centrifuged and plasma was retained from each time point. Plasma samples were analyzed by LC/MS/MS to quantify meloxicam.

Certain parameters, such as formulation weight, animal model, or needle size, may be tailored depending on the API being studied, its intended medical use, and its target dose and duration of release. In table 5 below are diagrams with different

- 28 045374 new parameters.

Table 5

In vivo scheme Procedure Animal Number Syringe Needle Injection Volume (µL) Model Injection site Route of administration 1 1ml capacity with soft ject® luer lock 23G (1 0.6 x 25 mm) Terumo® 160 Rat Between the shoulder blades Subcutaneously 2 1.5ml capacity with Terumo luer lock 21G (1 0.80 x 25 mm) Terumo® 1700 Pygmy pig Armpit Subcutaneously 3 1ml capacity with soft ject® luer lock 23G (1 0.6 x 30 mm) Terumo® 180 Rat Between the shoulder blades Subcutaneously

Embodiments of the present invention are set forth below with reference to the following numbered paragraphs:

1. A pharmaceutical composition suitable for in-situ depot formation comprising a biodegradable multi-branched copolymer comprising at least three polyester arms attached to a central polyether-containing core, and wherein said multi-branched copolymer is substantially insoluble in an aqueous solution.

2. The composition according to claim 1, characterized in that the molecular weight of said polyether is 10 kDa or less, preferably 5 kDa or less, 4 kDa or less, 3 kDa or less, or 2 kDa or less.

3. A pharmaceutical composition suitable for in-situ depot formation comprising a biodegradable multi-branched polyester copolymer comprising at least three polyester arms attached to a central polyester core, wherein the molecular weight of said polyester is 10 kDa or less, preferably 5 kDa or less, 4 kDa or less, 3 kDa or less, or 2 kDa or less

4. The composition according to claim 3, characterized in that said multi-branched copolymer is substantially insoluble in an aqueous solution.

5. A composition according to any one of claims 1 to 4, characterized in that said multi-branched copolymer is characterized by a solubility in an aqueous solution of less than 15 mg/ml, optionally less than 10 mg/ml, less than 5 mg/ml, less than 5 g/ml, less than 2 mg/ml or less than 1 mg/ml.

6. Composition according to claim 5, characterized in that the solubility is measured at 37°C.

7. The composition according to any one of claims 1 to 6, characterized in that said multi-branched copolymer is represented by the formula A(B) _n , where A represents the central core, and B represents polyester arms and n represents an integer equal to at least 3.

8. Composition according to claim 7, characterized in that n is at least 4, or at least 6, or at least 8.

9. Composition according to any one of claims 1 to 8, characterized in that the central core is a multi-branched polyether based on poly(ethylene glycol) (PEG) and a polyol.

10. The composition according to claim 9, characterized in that said polyol contains at least three hydroxyl groups.

11. The composition according to claim 10, characterized in that the polyol is a hydrocarbon substituted with at least three hydroxyl groups, optionally 3, 4, 5, 6 or 8 hydroxyl groups.

12. Composition according to any one of claims 9 to 11, characterized in that said polyol additionally contains one or more ether groups.

13. Composition according to any one of claims 9-12, characterized in that said polyol is pentaerythritol (PE), dipentaerythritol (DPE), trimethylolpropane (TMP), glycerol, hexaglycerol, erythritol, xylitol, di(trimethylolpropane) (diTMP) , sorbitol or inositol.

14. The composition according to any one of claims 9 to 13, characterized in that each branch of said multi-branched polyester contains a terminal reactive group capable of reacting with the polyester or its monomer or precursor.

15. The composition according to claim 14, characterized in that said terminal reactive group is a hydroxyl group.

16. Composition according to any one of claims 9-15, characterized in that said multi-branched polyether is characterized by formula 1a or formula 2a:

- 29 045374

Formula 1a where m is an integer from 5 to 150;

Formula 2a where m is an integer from 5 to 150;

17. Composition according to any one of claims 1 to 16, characterized in that said polyester is or is formed by at least one polymer or copolymer selected from the group consisting of poly(lactic acid) (PLA), poly(glycolic acid) , poly(epsiloncaprolactone), poly(ethylene adipate), copolymer of lactic acid and glycolic acid (LAGC) and poly(hydroxyalkanoate) or mixtures thereof.

18. Composition according to any one of claims 1 to 17, characterized in that said polyester is a homopolymer.

19. Composition according to any one of claims 1 to 17, characterized in that said polyester is obtained from more than one monomer.

20. The composition according to claim 19, characterized in that said polyester is a random copolymer or a block copolymer.

21. Composition according to any one of paragraphs. 1 -20, characterized in that said polyester is or contains PLA.

22. The composition according to claim 21, characterized in that said multi-branched copolymer is obtained by reacting a multi-branched polyether as defined in any one of claims 9 to 13 with D,L-lactide.

23. The composition according to claim 22, characterized in that said multi-branched copolymer is obtained by ring-opening polymerization of D,L-lactide initiated by said multi-branched polyether.

24. The composition according to claim 23, characterized in that said multi-branched copolymer is characterized by formula 5:

Formula 5 where n is an integer from 10 to 200, and m is an integer from 5 to 150.

25. The composition according to any one of claims 1 to 24, characterized in that the number of repeating polyester units in each arm is independently in the range from 10 to 200.

26. Composition according to any one of claims 1 to 25, characterized in that the mass of polyester is in the range from 500 to 40 kg/mol, optionally from 500 to 20 kg/mol, optionally from 10 to 40 kg/mol, preferably from 2 up to 10 kg/ml.

27. The composition according to any one of claims 1 to 26, characterized in that said composition further contains a pharmaceutically acceptable carrier, optionally said pharmaceutically acceptable carrier being an organic solvent.

28. The composition according to claim 27, characterized in that said pharmaceutically acceptable carrier is selected from the group consisting of benzyl alcohol, benzyl benzoate, dimethyl isosorbide (DMI), dimethyl sulfoxide (DMSO), ethyl acetate, ethyl benzoate, ethyl lactate, glycerol formal, methyl ethyl ketone, methyl isobutyl ketone, N- ethyl-2-pyrrolidone, N-methyl-2-pyrrolidinone (NMP), pyrrolidone-2, tetraglycol, triacetin, tributyrin, tripropionine, glycofurol and mixtures thereof.

-

Claims (28)

29. The composition according to any one of claims 1 to 28, characterized in that said composition additionally contains at least one pharmaceutically active ingredient. 30. The composition according to claim 29, characterized in that said pharmaceutically active ingredient is hydrophobic. 31. The composition according to claim 29, characterized in that said pharmaceutically active ingredient is meloxicam, bupivacaine, tamsulosin or combinations thereof. 32. The composition according to any one of claims 29-31, where the content of at least one pharmaceutically active ingredient is from 0.05 to 60%, optionally from 0.05 to 40%, optionally from 0.05 to 30%, optionally from 0.05 to 10%, optionally from 0.05% to 7%, optionally from 0.05 to 2% w/w. from the total mass of the composition. 33. The composition according to any one of claims 1 to 32, characterized in that said composition is a liquid for injection. 34. Composition according to any one of claims 1 to 33, characterized in that the content of multi-branched copolymer is from 2 to 80%, optionally from 2 to 70%, optionally from 2 to 60%, optionally from 10 to 60%, optionally from 10 to 50%, optionally from 20 to 40%, optionally from 20 to 35%, optionally from 30 to 50% w/w. from the total mass of the composition. 35. The composition according to any one of claims 1 to 34, characterized in that the molar ratio of the polyester repeating unit and ethylene oxide in the composition is from 1 to 10. 36. Composition according to any one of claims 1 to 35, characterized in that the release of at least one active ingredient can be modulated. 37. A composition according to any one of claims 1 to 34, suitable for delivering a pharmaceutically active ingredient to a subject for at least 1 day, optionally for at least 3 days, optionally for at least 7 days, optionally for at least at least 30 days, optionally for at least 90 days, optionally for at least 1 year. 38. Use of a pharmaceutical composition according to any one of claims 1 to 37 for modulating the release kinetics of at least one active ingredient. 39. A method for preparing a pharmaceutical composition according to any one of claims 1 to 37, comprising dissolving the multi-branched copolymer defined in any of claims 1 to 35 in a pharmaceutically acceptable carrier. 40. The method of claim 39, further comprising adding a pharmaceutically active ingredient to said composition. 41. Biodegradable depot obtained ex vivo or in situ by bringing the composition according to any one of claims 1 to 37 into contact with an aqueous environment, water or biological fluid. 42. A method for the controlled release of a pharmaceutically active ingredient, comprising administering a composition according to any one of claims 1 to 37 and allowing the formation of an in-situ depot in vivo. CLAIM 1. A pharmaceutical composition containing at least one pharmaceutically active ingredient and a biodegradable multi-branched copolymer containing at least three polyester arms attached to a central core containing a polyether, where said multi-branched copolymer has a solubility in an aqueous solution of less than 15 mg/ ml at 37°C and is obtained by reacting a multi-branched polyether with D,L-lactide, where the multi-branched polyether is derived from poly(ethylene glycol) (PEG) and a polyol. 2. The composition according to claim 1, characterized in that the molecular weight of said multi-branched polyether is 10 kDa or less, preferably 5 kDa or less, 4 kDa or less, 3 kDa or less, or 2 kDa or less, or 1 kDa or less, or 0.5 kDa or less. 3. A pharmaceutical composition containing at least one pharmaceutically active ingredient and a biodegradable multi-branched copolymer containing at least three polyester arms attached to a central core containing polyether, wherein said multi-branched copolymer is obtained by reacting the multi-branched polyester with D ,Llactide, and wherein said multi-branched polyether is derived from poly(ethylene glycol) (PEG) and a polyol, wherein the molecular weight of said multi-branched polyether is 10 kDa or less. 4. A composition according to any one of claims 1 to 3, characterized in that said multi-branched copolymer is characterized by a solubility in an aqueous solution of less than 15 mg/ml, optionally less than 10 mg/ml, less than 5 mg/ml, less than 2 mg/ml or less than 1 mg/ml, optionally solubility in aqueous solution measured at 37°C. 5. Composition according to any one of claims 1 to 4, suitable for in-situ formation of depots. - 31 045374 6. Composition according to any one of claims 1 to 5, characterized in that the multi-branched copolymer is represented by the formula A(B)n, where A represents the central core, and B represents polyester arms and n represents an integer equal to at least at least 3, wherein optionally n is at least 4, or at least 6, or at least 8, preferably wherein n is 4. 7. A composition according to any one of claims 1 to 6, characterized in that said polyol contains at least three hydroxyl groups, wherein said polyol is optionally a hydrocarbon substituted with at least three hydroxyl groups, optionally 3, 4, 5, 6 or 8 hydroxyl groups, wherein said polyol optionally further contains one or more ether groups. 8. The composition according to any one of claims 1 to 7, wherein said polyol is pentaerythritol (PE), dipentaerythritol (DPE), trimethylolpropane (TMP), glycerol, hexaglycerol, erythritol, xylitol, di(trimethylolpropane) (diTMP), sorbitol or inositol 9. The composition according to any one of claims 1 to 8, wherein each branch of said multi-branched polyether contains a reactive end group capable of reacting with the polyester or a monomer or precursor thereof, wherein said end reactive group is optionally a hydroxyl group. 10. Composition according to any one of claims 1 to 9, wherein said multi-branched polyether is characterized by formula 1, or formula 2, or formula 3, or formula 4 Formula 1 where m is an integer from 2 to 150, R is H or alkyl or PEG Formula 2 where m is an integer from 2 to 150 and p is 6. Formula 3 where m is an integer from 2 to 150 Formula 4 - 32 045374 where m is an integer from 2 to 150. 11. The composition according to any one of claims 1 to 10, wherein said multi-branched copolymer is obtained by ring-opening polymerization of D,L-lactide initiated by said multi-branched polyether. 12. Composition according to any one of claims 1 to 11, where said multi-branched copolymer is characterized by formula 5 or formula 6, formula 7 or formula 8 Formula 5 where u is an integer from 4 to 200 and m is an integer from 2 to 150 Formula 6 where u is an integer from 4 to 200, m is an integer from 2 to 150 and p is 6. Formula 7 where u is an integer from 4 to 200 and m is an integer from 2 to 150 - 33 045374 Formula 8 where u is an integer from 4 to 200 and m is an integer from 2 to 150. 13. The composition according to claim 12, wherein said multi-branched copolymer is characterized by formula 5, wherein the polyether core has a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 2, 3 or 6. 14. The composition according to any one of claims 1 to 13, wherein the number of repeating polyester units in each arm is independently in the range from 4 to 200, and/or where the molecular weight of the polyether is from 0.5 to 10 kDa, optionally from 1 to 10 kDa, preferably 2 to 10 kDa, or preferably 0.5 to 2 kDa, or most preferably 2 to 5 kDa, and/or wherein the molar ratio of the repeating ester unit to ethylene oxide in the multi-branched copolymer in said composition is from 1 up to 10, preferably from 2 to 6. 15. The composition according to any one of claims 1 to 14, containing one or more additional biodegradable multi-branched copolymers as defined in any of claims 1 to 14, optionally containing a first biodegradable multi-branched copolymer as defined in any of claims 114, and a second , different from the first, biodegradable multi-branched copolymer as defined in any one of claims 1 to 14, where optionally the content of the first biodegradable multi-branched copolymer is from 15 to 25% wt./wt. and the content of the second biodegradable multi-branched copolymer is from 15 to 25% w/w. from the total mass of the composition. 16. The composition of claim 15, wherein each of the first and second biodegradable multi-branched copolymers has a structure of formula 5 where u is an integer from 4 to 200 and m is an integer from 2 to 150; and wherein the first biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 2; and wherein the second biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 6; or the first biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 5 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 2; and wherein the second biodegradable multi-branched copolymer contains a polyether core with a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 6. 17. The composition according to any one of claims 1 to 16, additionally containing a biodegradable triblock copolymer characterized by the formula Av-Bw-Ax, where A is a polyester and B is polyethylene glycol, v and x - 34 045374 represent the number of repeat units in the range from 1 to 3000 and w represents the number of repeat units in the range from 3 to 300, with v=x or v^x. 18. The composition according to claim 17, where the mass of the polyethylene glycol chain is in the range from 180 Da to 12 kDa, or 194 Da to 12 kDa, or 200 Da to 12 kDa, or 100 Da to 4 kDa, preferably 1 to 2 kDa. 19. The composition according to claim 17 or 18, wherein the molar ratio of the ester repeating unit to the ethylene oxide repeating unit in the triblock copolymer is from 0.5 to 22.3, optionally from 0.5 to 10, preferably from 0.5 to 3, 5. 20. The composition according to claim 19, wherein said triblock copolymer contains a polyethylene glycol chain having a mass of 1 kDa, and the molar ratio of ester repeating unit to ethylene oxide repeating unit is 4, and the multi-branched copolymer has the structure of formula 5 where u is an integer from 4 to 200 and m is an integer from 2 to 150; containing a polyether core having a molecular weight of 2 kDa, and the molar ratio of the repeating ester unit to ethylene oxide is 3; or wherein said triblock copolymer contains a polyethylene glycol chain having a mass of 2 kDa and the molar ratio of ester repeat unit to ethylene oxide repeat unit is 2, wherein said multi-branched copolymers have a structure of formula 5 containing a polyether core having a molecular weight of 2 kDa, wherein the molar ratio of the ester repeating unit to ethylene oxide is 2; or wherein said triblock copolymer contains a polyethylene glycol chain having a mass of 2 kDa and the molar ratio of ester repeat unit to ethylene oxide repeat unit is 2, wherein said multi-branched copolymers have a structure of formula 5 containing a polyether core having a molecular weight of 2 kDa, wherein the molar ratio of the ester repeating unit to ethylene oxide is 6; optionally the content of multi-branched copolymer is from 15 to 25% wt./wt. of the total weight of the composition, and the triblock copolymer content is from 15 to 25% wt./wt. from the total mass of the composition. 21. The composition according to any one of claims 1-20, additionally containing a biodegradable diblock copolymer characterized by the formula Cy-Az, wherein A is a polyester, C is a capped polyethylene glycol, and y and z are the number of repeat units, with y ranging from 2 to 250 and z ranging from 1 to 3000. 22. The composition according to claim 21, wherein the molecular weight of the polyethylene glycol chain with a terminal capping group is in the range from 100 Da to 10 kDa or from 164 Da to 2 kDa, preferably from 1 to 2 kDa. 23. The composition according to claim 21 or 22, wherein the molar ratio of ester repeating unit to ethylene oxide repeating unit in the diblock copolymer is from 0.8 to 15, optionally from 1 to 10. 24. The composition according to any one of claims 17 to 23, wherein said polyester A in the triblock or diblock copolymer is selected from the group consisting of polylactic acid (PLA), polyglycolic acid, polycaprolactone, polyethylene adipate, polyhydroxyalkanoate, ε-caprolactone-lactide copolymer (PCLA), copolymer of lactic acid and glycolic acid (PLCA) and mixtures thereof, and optionally the polyethylene glycol terminated with a capping group is methoxy polyethylene glycol. 25. The composition according to any one of claims 1 to 24, further comprising a pharmaceutically acceptable carrier, wherein optionally said pharmaceutically acceptable carrier is an organic solvent, wherein optionally said organic solvent is a biocompatible organic solvent, wherein optionally the amount of said carrier is at least 25% or at least 35% w/w. based on the total weight of the composition, wherein said pharmaceutically acceptable carrier is optionally selected from the group: benzyl alcohol, - 35 045374 benzyl benzoate, dimethyl isosorbide (DMI), dimethyl sulfoxide (DMSO), ethyl acetate, ethyl benzoate, ethyl lactate, glycerol formal, methyl ethyl ketone, methyl isobutyl ketone, N-ethyl-2-pyrrolidone, N-methyl-2-pyrrolidinone (NMP), pyrrolidone-2, tetraglycol, triacetin, tributyrin, tripropionin, glycofurol and mixtures thereof. 26. The composition according to any one of claims 1 to 25, wherein said pharmaceutically active ingredient is hydrophobic, wherein said pharmaceutically active ingredient is optionally meloxicam, bupivacaine, tamsulosin, octreotide, tadalafil, empagliflozin, tenofovir, liothyronine, or combinations thereof. 27. The composition according to any one of claims 1 to 26, where the content of at least one pharmaceutically active ingredient is from 0.05 to 60%, optionally from 0.05 to 40%, optionally from 0.05 to 30%, optionally from 0.05 to 10%, optionally from 0.05 to 7%, optionally from 0.05 to 2% w/w. based on the total weight of the composition and/or wherein said composition is a liquid for injection, and/or wherein the content of said multi-branched copolymer is from 2 to 80%, optionally from 2 to 70%, optionally from 2 to 60%, optionally from 10 to 60%, optionally from 10 to 50%, optionally from 20 to 40%, optionally from 20 to 35%, optionally from 30 to 50% w/w. from the total mass of the composition. 28. The composition according to any one of claims 1 to 27, which is a composition defined in the table Table Compound API Copolymer Solvent Weight Number Name % (mass/mass) Code Structure PEG-based chains Ratio (MK/EO) % (mass/mass) Name % (w/w) (kDa) s4P2R3 Branched 2000 3 20.00 2 Meloxicam 2.00 DMSO 58.00 P2R2 Linear 2000 2 20.00 19 Tenofovir 19.00 s4P2R3 Branched 2000 3 40.00 DMSO 41.00 20 Tenofovir 19.00 s4P2R6 Branched 2000 6 40.00 DMSO 41.00 29 Liothyronine 0.10 s4P2R3 Branched 2000 3 40.00 DMSO 59.90 thirty Liothyronine 0.10 s4P2R6 Branched 2000 6 40.00 DMSO 59.90 47 Tadalafil 20.00 s4P2R6 Branched 2000 6 25.00 DMSO 55.00 48 Tadalafil 20.00 s4P2R3 Branched 2000 3 25.00 DMSO 55.00 135 Empagliflozin 20.00 s4P2R3 Branched 2000 3 20.00 DMSO 60.00 136 Empagliflozin 20.00 s4P2R6 Branched 2000 6 20.00 DMSO 60.00 137 Empagliflozin 20.00 s4P2R6 Branched 2000 6 30.00 DMSO 50.00 s4P2R3 Branched 2000 3 28.00 145 Octreotide 0.70 DMSO 53.30 P1R4 Linear 1000 4 18.00 149 Streptavidin 1.50 s4P2R3 Branched 2000 3 28 Tripropionine 70.50 389 Meloxicam 2.00 s4P2R2 Branched 2000 2 40.00 DMSO 58.00 P2R2 Linear 2000 2 20.00 390 Meloxicam 2.00 s4P2R2 Branched 2000 2 20.00 DMSO 58.00 397 Meloxicam 2.00 s4P2R6 Branched 2000 6 40.00 DMSO 58.00 399 Meloxicam 2.00 P2R2 s4P2R6 Linear Branched 2000 2000 2 6 20.00 20.00 DMSO DMSO 58.00 58.00 s4P2R? Branched 2000 2 20.00 401 Meloxicam 2.00 s4P2R6 Branched 2000 6 20.00 DMSO 58.00 402 Meloxicam 2.00 s4P5R2 Branched 5000 2 40.00 DMSO 58.00 s4P5R2 Branched 5000 2 20.00 404 Meloxicam 2.00 DMSO 58.00 s4P2R6 Branched 2000 6 20.00 405 Meloxicam 2.00 s4P2R3 Branched 2000 3 40.00 DMSO 58.00 408 Bupivacaine 2.00 s4P2R2 Branched 2000 2 40.00 DMSO 58.00 409 Bupivacaine 2.00 s4P2R2 Branched 2000 2 20.00 DMSO 58.00 P2R2 Linear 2000 2 20.00 414 Bupivacaine 2.00 s4P2R6 Branched 2000 6 40.00 DMSO 58.00 -