Polymer compositions comprising a thermal gel polymer and a tyrosine kinase inhibitor and methods related thereto

Abstract

Provided are polymer compositions comprising a multi-block thermogel polymer comprising a hydrophilic poly(alkylene glycol), a hydrophobic polymer, and a polyether or polyester chemically coupled together by at least one of a urethane / carbamate linkage, a carbonate linkage, an ester linkage, a urea linkage, an amide linkage, an ether linkage, an amine linkage, a triazole linkage, or a combination thereof, and a tyrosine kinase inhibitor (TKI) mixed with the multi-block thermogel polymer.

Description

Technical Field

[0001] This disclosure generally relates to polymer compositions comprising a thermogellin polymer and a tyrosine kinase inhibitor (TKI) and related methods thereof. Background Technology

[0002] Currently, delivering sustained release of biologics at specific sites to treat a variety of diseases remains a challenging area. This includes the treatment of diseases such as retinal neovascularization and cancer, where unmet clinical needs persist to provide effective sustained delivery solutions as alternatives to current treatments.

[0003] For example, the current gold standard treatment for ocular neovascular diseases or posterior segment proliferative vascular diseases (such as age-related macular degeneration (AMD) and diabetic retinopathy (DR)) involves expensive, invasive, and inconvenient monthly or bi-monthly injections of anti-vascular endothelial growth factor (anti-VEGF), leading to poor patient compliance. This frequent injection regimen places a significant burden on both patients and caregivers.

[0004] For diseases currently reliant on oral medication, prolonged oral administration can lead to the development of overall drug resistance and / or overall drug toxicity. For example, in diseases such as cancer that require long-term tyrosine kinase inhibitor (TKI) therapy, TKIs can be administered orally for periods exceeding two years. Consequently, these patients have reported developing resistance after eight months of treatment and experiencing toxic effects caused by prolonged oral supplementation.

[0005] In view of the above, it is necessary to address or at least improve the aforementioned problems. Specifically, it is necessary to provide injectable drug reservoirs capable of providing local drug release over a sustained period of time. Summary of the Invention

[0006] According to one aspect, a polymer composition is provided comprising:

[0007] Multiblock thermogel polymers comprising hydrophilic poly(alkylene glycols), hydrophobic polymers, and polyethers or polyesters chemically coupled together via at least one of urethane / carbamate, carbonate, ester, urea, amide, ether, amine, triazole, or combinations thereof; and

[0008] Tyrosine kinase inhibitors (TKIs) mixed with this multi-block thermogel polymer.

[0009] In one embodiment, TKI interacts with the multiblock thermogel polymer to promote the gelation of the multiblock thermogel polymer.

[0010] In one embodiment, TKI increases the viscosity of the polymer composition by at least 10 times compared to the absence of TKI.

[0011] In one implementation, the TKI comprises a VEGFR inhibitor (e.g., a VEGFR-2 inhibitor) and / or an FGFR inhibitor.

[0012] In one implementation, the TKI includes one or more of the following:

[0013] i. CP-547632, its analogues, or pharmaceutically acceptable salts thereof; or

[0014] ii. Compounds of formula (I),

[0015]

[0016] Formula (I).

[0017] In one embodiment, the hydrophilic poly(alkylene glycol) comprises poly(ethylene glycol)(PEG).

[0018] In one embodiment, the hydrophobic polymer is selected from poly(propylene glycol) (PPG), poly(lactic acid-co-glycolic acid) (PLGA), polylactic acid (PLA), poly(N-isopropylacrylamide) (PNIPAAM), polypeptides, or combinations thereof.

[0019] In one embodiment, the polyether or polyester comprises one or more of polycaprolactone (PCL), polytetrahydrofuran (PTHF), polyhydroxybutyrate (PHB), polylactic acid-co-glycolic acid (PLGA), and polylactic acid (PLA).

[0020] In one embodiment, the concentration of TKI in the polymer composition is not less than about 10 mg / L.

[0021] In one embodiment, the molar ratio of the hydrophilic poly(alkylene glycol) to the hydrophobic polymer is 1-10:1.

[0022] In one embodiment, the polyether or polyester is present in the multiblock polymer in an amount of 1% to 10% by weight.

[0023] In one embodiment, the molar ratio of the hydrophilic poly(alkylene glycol) to the hydrophobic polymer to the polyether or polyester in the multiblock polymer is in the range of about 1-10:1:0.01-1.5.

[0024] In one embodiment, the multiblock polymer is present in an aqueous medium at a level of up to 30% w / v.

[0025] In one embodiment, the pH of the composition is in the range of 7.1 to 7.4.

[0026] In one embodiment, the polymer composition has a critical gelation temperature of not less than 4°C.

[0027] In one embodiment, the polymer composition further comprises one or more pharmaceutically active ingredients that are different from TKIs.

[0028] In one implementation, the pharmaceutically active ingredient, which is different from a TKI, is selected from aflibercept, sunitinib malate, and combinations thereof.

[0029] In one embodiment, the polymer composition has a drug release profile of not less than 2 months.

[0030] In one embodiment, the polymer composition has a drug release profile of not less than 12 months.

[0031] According to another aspect, the polymer compositions disclosed herein are provided for use in medicine.

[0032] According to another aspect, the polymer compositions disclosed herein are provided for the treatment or prevention of eye diseases, for the treatment or prevention of tumors, for the treatment or reduction of angiogenesis, and / or for the treatment of cancer.

[0033] According to another aspect, the use of the polymer compositions disclosed herein in the manufacture of medicaments for treating or preventing eye diseases, for treating or preventing tumors, for treating or reducing angiogenesis, and / or for treating cancer is provided.

[0034] According to another aspect, methods for treating or preventing eye diseases, treating or preventing tumors, treating or reducing angiogenesis, and / or treating cancer are provided, the methods comprising administering the polymer composition disclosed herein to a subject in need of such treatment.

[0035] In one implementation, the eye disease is selected from neovascular retinal disease, age-related macular degeneration (AMD), diabetic macular edema (DME), and retinal vein occlusion (RV).

[0036] In one implementation, the cancer is selected from breast cancer, leukemia, gastrointestinal cancer, kidney cancer, lung cancer (e.g., non-small cell lung cancer), non-Hodgkin lymphoma, melanoma, ovarian cancer, fallopian tube cancer, and eye cancer.

[0037] According to another aspect, a method for preparing the compositions disclosed herein is provided, the method comprising:

[0038] Provided are multi-block thermogel polymers comprising a hydrophilic poly(alkylene glycol), a hydrophobic polymer, and a polyether or polyester chemically coupled together via at least one of urethane / methionine ester bonds, carbonate bonds, ester bonds, urea bonds, amide bonds, ether bonds, amine bonds, triazole bonds, or combinations thereof; and

[0039] Tyrosine kinase inhibitors (TKIs) are mixed with multi-block thermogel polymers.

[0040] definition

[0041] The term "substantially transparent," when used herein to describe an object, should be interpreted broadly to mean that 50% or more of incident light perpendicular to the object's surface can be transmitted through it. In some instances, objects that are substantially transparent to light allow 60% or more, 65% or more, 70% or more, 80% or more, 85% or more, 90% or more, or 95% or more of incident light perpendicular to the object's surface to be transmitted. In one instance, an object that is substantially transparent to light allows more than 70% of incident light perpendicular to the object's surface to be transmitted.

[0042] As used in this specification, the terms “coupled” or “connected” are intended to cover a direct connection or a connection between the two through one or more intermediate means, unless otherwise stated.

[0043] The term "alkyl" as a group or part of a group refers to a straight-chain or branched aliphatic hydrocarbon group having 1 to 20 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. Examples of suitable straight-chain and branched alkyl substituents include methyl, ethyl, n-propyl, 2-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, hexyl, pentyl, 1,2-dimethylpropyl, 1,1-dimethylpropyl, pentyl, isopentyl, hexyl, 4-methylpentyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 1,2,2 -Trimethylpropyl, 1,1,2-trimethylpropyl, 2-ethylpentyl, 3-ethylpentyl, heptyl, 1-methylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 4,4-dimethylpentyl, 1,2-dimethylpentyl, 1,3-dimethylpentyl, 1,4-dimethylpentyl, 1,2,3-trimethylbutyl, 1,1,2-trimethylbutyl, 1,1,3-trimethylbutyl, 5-methylheptyl, 1-methylheptyl, octyl, nonyl, decyl, etc. This group can be a terminal group or a bridging group.

[0044] The term “and / or”, such as “X and / or Y”, is understood to mean “X and Y” or “X or Y”, and should be regarded as providing explicit support for both meanings or either meaning.

[0045] Furthermore, throughout this description, the term “substantially” is understood to mean, whenever used, including but not limited to, “all” or “completely.” Additionally, terms such as “comprising” are intended to be non-limiting descriptive language, broadly encompassing elements / components described after these terms, as well as other components not explicitly described. For example, when “comprising” is used, a reference to “one” feature is also intended to refer to “at least one” of the features. Terms such as “consisting of” may be considered, where appropriate, a subset of terms such as “comprising.” Therefore, in embodiments disclosed herein using terms such as “comprising,” it will be appreciated that these embodiments provide guidance for corresponding embodiments using terms such as “consisting of.” Furthermore, terms such as “about” or “approximately”, whenever used, generally refer to reasonable variations, such as a variation of + / -5% of the disclosed value, or a variation of 4% of the disclosed value, or a variation of 3% of the disclosed value, or a variation of 2% of the disclosed value, or a variation of 1% of the disclosed value.

[0046] Furthermore, in the description herein, certain values ​​may be disclosed within a range. The values ​​at the endpoints of a range are intended to illustrate a preferred range. Whenever a range is described, it is intended to indicate that the range encompasses and teaches all possible subranges within that range, as well as individual numerical values. That is, the endpoints of a range should not be interpreted as rigid limitations. For example, the description of a range of 1% to 5% is intended to specifically disclose subranges such as 1% to 2%, 1% to 3%, 1% to 4%, 2% to 3%, etc., and individual values ​​within that range, such as 1%, 2%, 3%, 4%, and 5%. The aforementioned intention to specifically disclose applies to any depth / breadth of range.

[0047] Furthermore, when describing some embodiments, this disclosure may have already disclosed methods and / or processes in a specific order of steps. However, unless otherwise required, it will be appreciated that the methods or processes should not be limited to the specific order of steps disclosed. Other orders of steps may be possible. The specific order of steps disclosed herein should not be construed as an excessive limitation. Unless otherwise required, the methods and / or processes disclosed herein should not be limited to performing the steps in the order they are written. The order of steps may be changed and will still remain within the scope of this disclosure.

[0048] Furthermore, it will be appreciated that although this disclosure provides embodiments having one or more of the features / characteristics discussed herein, one or more of these features / characteristics may be omitted in other alternative embodiments, and this disclosure provides support for such omissions and these related alternative embodiments.

[0049] It will also be recognized that, when claiming priority of an earlier application, the entire contents of that earlier application are also considered to form part of this disclosure and may serve as support for the embodiments disclosed herein. Detailed Implementation

[0050] The following discloses exemplary non-limiting embodiments of polymer compositions, methods for preparing polymer compositions, and related uses of polymer compositions.

[0051] polymer composition

[0052] In various embodiments, polymer compositions are provided comprising a multi-block thermogel polymer and a tyrosine kinase inhibitor (TKI) mixed with the multi-block thermogel polymer. Advantageously, this polymer composition can act as a drug reservoir for the slow release of a drug or pharmaceutically active ingredient (e.g., when placed in a human or animal). The inventors have found that TKI encapsulation delays the dissociation or dissolution of the hydrogel. Without being bound by theory, it is believed that the TKI and the multi-block thermogel polymer interact synergistically to produce a further cross-linked structure, thereby allowing for an extended drug release period. Advantageously, the long-term release profile of the TKI-gel composition can provide a longer therapeutic effect, preventing the need for repeated treatments.

[0053] In various embodiments, TKI interacts with the multiblock thermogel polymer to promote its gelation. In various embodiments, the interaction between TKI and the multiblock thermogel polymer may involve hydrogen bonding and hydrophobic interactions, which can induce secondary gelation. In various embodiments, the TKI-induced gelation does not alter the thermosensitive properties of the multiblock thermogel polymer. For example, in various embodiments, the TKI-gel composition is still injectable at 4°C and gels at body temperature. The polymer composition may be thermosensitive, and its physical state changes based on temperature.

[0054] In various embodiments, the presence of TKI increases the viscosity of the composition by at least about 10 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 30 times, at least about 35 times, at least about 40 times, at least about 45 times, at least about 50 times, at least about 55 times, at least about 60 times, at least about 65 times, at least about 70 times, at least about 75 times, at least about 80 times, at least about 85 times, at least about 90 times, at least about 95 times, at least about 100 times, at least about 150 times, at least about 200 times, at least about 250 times, at least about 300 times, at least about 350 times, at least about 360 times, at least about 370 times, at least about 380 times, at least about 390 times, and at least about 400 times, compared to the absence of TKI.

[0055] In various implementations, the TKI comprises a VEGFR inhibitor (e.g., a VEGFR-2 inhibitor) and / or an FGFR inhibitor.

[0056] In various embodiments, the TKI comprises small molecules, such as those with a molecular weight of less than / not more than about 1000 Da, less than / not more than about 900 Da, less than / not more than about 850 Da, less than / not more than about 800 Da, less than / not more than about 750 Da, less than / not more than about 700 Da, or less than / not more than about 650 Da.

[0057] In various implementations, TKI includes one or more of the following:

[0058] i. CP-547632, its analogues, or pharmaceutically acceptable salts thereof (e.g., HCl, trifluoroacetate, etc.); or

[0059] ii. Compounds of formula (I),

[0060]

[0061] Formula (I).

[0062] In various implementations, the use of small molecules like CP-547632 hydrochloride (CP-HCl) is advantageous compared to biologics / proteins such as aflibercept or bevacizumab. This is because those biologics cannot self-assemble to form a gel and therefore cannot reinforce the thermogel polymer matrix in the same way as CP-HCl. Consequently, when aflibercept or bevacizumab is used instead of CP-HCl, the release of aflibercept or bevacizumab from the hydrogel occurs much faster compared to when CP-HCl is used.

[0063] In various embodiments, CP-HCl exhibits increased water solubility. It allows more soluble drug to interact with the polymer and form a secondary gel. In various embodiments, the TKI is water-soluble CP-547632 HCl, in contrast to water-insoluble drugs such as highly water-insoluble paclitaxel. When CP-HCl is encapsulated in a multi-block thermogel polymer (such as poly(PEG / PPG / PCL urethane)), it is partially encapsulated within the micelle core, and it also simultaneously forms a secondary gel matrix to enhance the polymer matrix of the thermogel. This secondary gel matrix significantly reduces the gel erosion rate, and this allows the CP-HCl multi-block thermogel assemblies to achieve extended release exceeding one year. Conversely, when paclitaxel is used instead of CP-HCl, the release of paclitaxel only lasts for about 30 days because paclitaxel does not contribute to gel matrix enhancement.

[0064] In various embodiments, the concentration of the TKI (e.g., CP-HCl) is not less than about 10 mg / L, not less than about 20 mg / L, not less than about 30 mg / L, and not less than about 40 mg / L. Advantageously, it has been found that when a TKI (e.g., CP-HCl) at a concentration of about 40 mg / mL is used in the polymer composition, sustained drug release for one year can be achieved.

[0065] In various implementations, the TKI (e.g., CP-HCl) is mixed with the polymer in a substantially homogeneous manner. Advantageously, long-term sustained drug release can be achieved from homogeneous thermosensitive hydrogels compared to hydrogels loaded with drug powder.

[0066] In some implementations, TKI (e.g., CP-HCl) is the only pharmaceutically active ingredient in the polymer composition.

[0067] In various implementation schemes, the TKI (e.g., CP-HCl) has release profiles for not less than about 2 months, not less than about 3 months, not less than about 4 months, not less than about 5 months, not less than about 6 months, not less than about 7 months, not less than about 8 months, not less than about 9 months, not less than about 10 months, not less than about 11 months, or not less than about 12 months.

[0068] In various embodiments, the polymer composition may also include one or more pharmaceutically active ingredients that are different from TKIs (e.g., CP-HCl). In various embodiments, the pharmaceutically active ingredient that is different from a TKI (e.g., CP-HCl) is selected from aflibercept and sunitinib malate.

[0069] In various embodiments, the multiblock thermogel polymer comprises a hydrophilic poly(alkylene glycol), hydrophobic polymer, and a polyether or polyester chemically coupled together by at least one of urethane / methionine ester bonds, carbonate bonds, ester bonds, urea bonds, amide bonds, ether bonds, amine bonds, triazole bonds, or combinations thereof. In various embodiments, the selection of different types of polymer chains in the multiblock thermogel polymer is based on achieving desired thermogel properties. The combination of hydrophilic and hydrophobic polymers can advantageously promote self-assembly to form a gel at desired temperatures and environments, thereby providing a structure that can act as a favorable drug reservoir under suitable conditions.

[0070] In various embodiments, the thermogel is made of one or more multiblock copolymers, which are three-component multiblock polymers. For example, the multiblock thermogel polymer comprises three different types of polymer segments. In various embodiments, the multiblock thermogel polymer is essentially composed of A: a hydrophilic poly(alkylene glycol), B: a hydrophobic polymer, and C: a polyether or polyester. In various embodiments, the multiblock polymer may have at least one unit with the following structural sequence ABC. It will be appreciated that in some embodiments, the positions of A, B, and C may be interchanged among them. In various embodiments, the multiblock polymer may comprise a plurality of hydrophilic poly(alkylene glycol) blocks, a plurality of hydrophobic polymer blocks, and / or a plurality of polyether or polyester blocks. In various embodiments, the multiblock copolymer comprises more than three polymer blocks. The blocks may be randomly distributed / arranged within the polymer. In various embodiments, the polymer blocks in the copolymer are linked to each other by bonds selected from: methionine esters, carbonates, ureas, esters, amides, ethers, amines, triazoles, and any combination thereof.

[0071] In some embodiments, the hydrophilic poly(alkylene glycol) comprises hydrophilic poly(ethylene glycol) (PEG), and the hydrophobic polymer comprises a second poly(alkylene glycol), such as poly(propylene glycol) (PPG), poly(butanediol), etc., or combinations thereof. In various embodiments, the hydrophobic polymer comprises a temperature-responsive / thermosensitive polymer.

[0072] In various embodiments, the polyether or polyester comprises one or more of polycaprolactone (PCL), polytetrahydrofuran (PTHF), polyhydroxybutyrate (PHB), polylactic acid-co-glycolic acid (PLGA), and polylactic acid (PLA).

[0073] In various embodiments, the molar ratio of the hydrophilic poly(alkylene glycol) to the hydrophobic polymer is 1-10:1. For example, the molar ratio of the hydrophilic poly(alkylene glycol) to the hydrophobic polymer may be about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1 or about 10:1, preferably about 4:1.

[0074] In various embodiments, the polyether or polyester is hydrophobic. In various embodiments, the polyether or polyester is present in an amount of 1% to 10% by weight of the multiblock polymer. For example, the polyether or polyester may be present in an amount / concentration of about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% by weight of the multiblock polymer, preferably about 1% to about 3% by weight.

[0075] In various embodiments, the molar ratio of the hydrophilic poly(alkylene glycol) to the hydrophobic polymer to the polyether or polyester in the multiblock polymer is in the range of about 1–10:1:0.01–1.5.

[0076] In various embodiments, the multiblock polymer is present in an aqueous medium at an amount of up to 30% w / v. In various embodiments, the polymer composition contains about 1% to about 30% w / v of the multiblock thermogel polymer in the aqueous medium. In various embodiments, the composition contains up to about 30% w / v of the polymer in the aqueous / aqueous solution / buffered solution, or contains about 1% w / v, about 2% w / v, about 3% w / v, about 4% w / v, about 5% w / v, about 6% w / v, about 7% w / v, about 8% w / v, about 9% w / v, about 10% w / v, about 11% w / v, about 12% w / v, or about 1% w / v in the aqueous / aqueous solution / buffered solution. The amount of polymer at 3% w / v, about 14% w / v, about 15% w / v, about 16% w / v, about 17% w / v, about 18% w / v, about 19% w / v, about 20% w / v, about 21% w / v, about 22% w / v, about 23% w / v, about 24% w / v, about 25% w / v, about 26% w / v, about 27% w / v, about 28% w / v, about 29% w / v, or about 30% w / v.

[0077] In various embodiments, the aqueous medium may be a balanced salt solution. In various embodiments, the balanced salt solution is a solution having a physiological pH and isotonic salt concentration. In various embodiments, the balanced salt solution comprises at least one of sodium, potassium, calcium, and magnesium salts, such as calcium chloride, potassium chloride, magnesium chloride, sodium acetate, sodium citrate, and sodium chloride.

[0078] In various embodiments, the polymer composition may have a high water content of more than about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% by weight. Therefore, the material may be a water-based polymer.

[0079] In various embodiments, the pH of the composition is in the range of 7.1 to 7.4. In various embodiments, the pH of the composition has a pH value that is substantially similar to the physiological pH values ​​of about pH 7.1 to about pH 7.7, about pH 7.2 to about pH 7.6, about pH 7.3 to about pH 7.5, or about pH 7.4.

[0080] In various embodiments, the polymer composition has the following critical gelation temperature / thermally reversible sol-gel transition temperature / temperature at which it transitions from a liquid / flowable state to a non-flowable / gel state: not less than about 4°C, not less than about 5°C, not less than about 6°C, not less than about 7°C, not less than about 8°C, not less than about 9°C, not less than about 10°C, not less than about 15°C, not less than about 20°C, not less than about 25°C, not less than about 30°C, not less than about 31°C, not less than about 32°C, not less than about 33°C, not less than about 34°C, not less than about 35°C, not less than about 36°C, not less than about 36.5°C, up to about 37°C, or a temperature substantially similar to the human body temperature at about 36.5°C or about 37°C. In some embodiments, the transition from the liquid / flowable state to the gel state is reversible.

[0081] In various embodiments, because the polymer composition is a sol at low temperatures, it can be injected using a needle (e.g., 18G and above). Advantageously, less tissue damage is expected because larger-diameter instruments are not required. In various embodiments, the polymer composition can be injected intravitreally (IVT) for posterior segment diseases or intratumorally for local drug delivery in cancer treatment. Advantageously, the injectability of the polymer composition allows for treatment in a less invasive manner compared to implants.

[0082] In various embodiments, the polymer composition is substantially free of solvent contaminants. For example, the composition or multiblock polymer may be substantially free of benzene and / or carbon tetrachloride and / or 1,2-dichloroethane and / or 1,1-dichloroethylene and / or 1,1,1-trichloroethane and / or acetonitrile and / or chlorobenzene and / or chloroform and / or cyclohexane and / or 1,2-dichloroethylene and / or dichloromethane and / or 1,2-dimethoxyethane and / or N,N-dimethylacetamide and / or N,N-dimethylformamide and / or 1,4-dioxane and / or 2-ethoxyethanol and / or ethylene glycol and / or formamide and / or hexane and / or methanol and / or 2-methoxyethanol and / or methyl butyl ketone and / or methylcyclohexane and / or N-methylpyrrolidone and / or nitromethane and / or pyridine and / or sulfolane. And / or tetrahydrofuran and / or naphthalene and / or toluene and / or 1,1,2-trichloroethylene and / or xylene (m, para, or ortho-isomers) and / or acetic acid and / or acetone and / or anisole and / or 1-butanol and / or 2-butanol and / or butyl acetate and / or tert-butyl methyl ether and / or cumene and / or dimethyl sulfoxide and / or ethanol and / or ethyl acetate and / or diethyl ether and / or ethyl formate and / or formic acid and / or heptane and / or isobutyl acetate and / or isopropyl acetate and / or methyl acetate and / or 3-methyl-1-butanol and / or methyl ethyl ketone and / or methyl isobutyl ketone and / or 2-methyl-1-propanol and / or pentane and / or 1-pentanol and / or 1-propanol and / or 2-propanol and / or propyl acetate.

[0083] In various embodiments, the polymer or composition is biocompatible and / or non-toxic and / or does not cause inflammation or adverse immune responses in animals or humans, particularly in the eyes of animals or humans.

[0084] In various embodiments, the multiblock thermogel polymer is substantially transparent and / or exhibits high optical transparency and / or a refractive index substantially similar to that of naturally occurring vitreous fluids, such as about 1.20 to about 1.48, about 1.21 to about 1.47, about 1.22 to about 1.46, etc. About 1.23 to about 1.45, about 1.24 to about 1.44, about 1.25 to about 1.43, about 1.26 to about 1.42, about 1.27 to about 1.41, about 1.28 to about 1.40, about 1.29 to about 1.39, about 1.30 to about 1.38, about 1.31 to about 1.37, about 1.32 to about 1.36, about 1.33 to about 1.35, about 1.339 to about 1.349, about 1.338 to about 1.348, about 1.337 to about 1.347, about 1.336 to about 1.346, about 1.335 to about 1.345, or about 1.334 to about 1.344.

[0085] In various embodiments, at least one or more blocks throughout the polymer or the polymer or composition are biodegradable and / or biodegradable (in some instances, all polymer blocks are biodegradable).

[0086] In various embodiments, the polymer composition is capable of naturally degrading / dissolving in humans / animals within about 16 months, about 15 months, about 14 months, about 13 months, or about 12 months.

[0087] In various embodiments, the polymer composition is permissible in humans / animals for at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 110 days, at least 120 days, at least 130 days, at least 140 days, at least 150 days, at least 160 days, at least 170 days, at least 180 days, at least 190 days, and at least 200 days. Complete degradation / dissolution within approximately 210 days, approximately 220 days, approximately 230 days, approximately 240 days, approximately 250 days, approximately 260 days, approximately 270 days, approximately 280 days, approximately 290 days, approximately 300 days, approximately 310 days, approximately 320 days, approximately 330 days, approximately 340 days, approximately 350 days, approximately 360 days, or approximately 365 days.

[0088] In various embodiments, the polymer composition comprises / consistently consists of / consistent with: the multiblock polymer disclosed herein; the TKI disclosed herein; water / aqueous medium / aqueous buffer; a pharmaceutically active ingredient optionally different from the TKI; and optionally a pharmaceutically acceptable excipient.

[0089] In various embodiments, the polymer composition may exist as a drug reservoir or a drug formulation.

[0090] In various embodiments, the polymer compositions disclosed herein are also provided for use in medicine. In various embodiments, a drug reservoir is provided comprising: a multiblock thermogel polymer comprising poly(ethylene glycol) (PEG), poly(propylene glycol) (PPG), and poly(caprolactone) (PCL) chemically coupled together by at least urethane bonds; and a drug composition mixed with the multiblock thermogel polymer, the drug composition comprising CP-547632 or a pharmaceutically acceptable salt thereof (e.g., HCl).

[0091] In various embodiments, the polymer compositions disclosed herein are also provided for ocular drug delivery.

[0092] In various embodiments, the polymer compositions disclosed herein are also provided for treating or preventing eye diseases and / or eye disorders, for treating or preventing tumors, for treating or reducing angiogenesis, for treating or reducing tumor metastasis and recurrence, for treating cancer, and / or for preventing or reducing the frequency of IVT injections.

[0093] In various embodiments, the polymer compositions disclosed herein are also provided for use in the manufacture of medicaments for the treatment or prevention of eye diseases and / or ocular disorders, for the treatment or prevention of tumors, for the treatment or reduction of angiogenesis, for the treatment or reduction of tumor metastasis and recurrence, for the treatment of cancer, and / or for the prevention or reduction of IVT injection frequency.

[0094] In various embodiments, methods are also provided for treating or preventing eye diseases and / or ocular disorders, treating or preventing tumors, treating or reducing angiogenesis, treating or reducing tumor metastasis and recurrence, treating cancer, and / or preventing or reducing the frequency of IVT injections using the polymer compositions disclosed herein. In various embodiments, the method includes administering the polymer compositions disclosed herein to a subject in need of this treatment. In various embodiments, the step of administering the polymer compositions includes injecting the polymer compositions into the subject in need of this treatment.

[0095] In various implementation schemes, the eye disease is selected from neovascular retinal disease, age-related macular degeneration (AMD), diabetic macular edema (DME), and retinal vein occlusion (RV).

[0096] In various implementation schemes, the cancer is selected from breast cancer, leukemia, gastrointestinal cancer, kidney cancer, lung cancer (e.g., non-small cell lung cancer), non-Hodgkin lymphoma, melanoma, ovarian cancer, fallopian tube cancer, and eye cancer.

[0097] Methods for preparing polymer compositions

[0098] In various embodiments, a method for preparing the polymer compositions disclosed herein is also provided, the method comprising: providing a multiblock thermogel polymer comprising a hydrophilic poly(alkylene glycol), hydrophobic polymer, and polyether or polyester chemically coupled together by at least one of urethane / methionine ester bonds, carbonate bonds, ester bonds, urea bonds, amide bonds, ether bonds, amine bonds, triazole bonds, or combinations thereof; and mixing a tyrosine kinase inhibitor (TKI) with the multiblock thermogel polymer.

[0099] In various implementation schemes, the TKI and polymer are mixed at a temperature not exceeding about 30°C, not exceeding about 25°C, not exceeding about 20°C, not exceeding about 15°C, not exceeding about 10°C, not exceeding about 5°C, or not exceeding about 4°C for a period of not less than about 20 hours, not less than about 24 hours, not less than about 25 hours, not less than about 30 hours, not less than about 35 hours, not less than about 40 hours, not less than about 45 hours, not less than about 48 hours, not less than about 50 hours, not less than about 55 hours, not less than about 60 hours, not less than about 65 hours, not less than about 70 hours, not less than about 72 hours, or not less than about 75 hours.

[0100] In various embodiments, the TKI is provided in a buffer solution (e.g., AMO buffer, phosphate-buffered saline (PBS), etc.). In various embodiments, the buffer solution is water-based. The presence of additives can be adjusted in the water-based buffer solution as needed.

[0101] In various embodiments, the step of providing a multi-block thermogel polymer includes the following steps: mixing a hydrophilic poly(alkylene glycol), a hydrophobic polymer, and a polyether or polyester with a coupling agent in the presence of a metal catalyst and a suitable solvent to form the polymer. The metal catalyst may comprise a tin catalyst selected from alkyltin compounds, aryltin compounds, and dialkyltin diesters (such as dibutyltin dilaurate, dibutyltin diacetate, dibutyltin dioctanoate, and dibutyltin distearate). The solvent may comprise an anhydrous solvent selected from toluene, benzene, and xylene. The hydrophilic poly(alkylene glycol), hydrophobic polymer, and polyether or polyester may be mixed in a molar ratio of about 1-10:1:0.01-1.5. In some embodiments, the amount of coupling agent added is equal to the number of reactive groups in the composition.

[0102] The mixing of hydrophilic poly(alkylene glycol), hydrophobic polymers and polyethers or polyesters with coupling agents can be carried out at the following elevated temperatures: about 70°C to about 150°C, about 72°C to about 148°C, 74°C to about 146°C, about 76°C to about 144°C, about 78°C to about 142°C, about 80°C to about 140°C, about 82°C to about 138°C, about 84°C to about 136°C, about 86°C to about 134°C, about 88°C to about 132°C, about 90°C to about 130°C, about 92°C to about 128°C, about 94°C to about 126°C, about 96°C to about 124°C, about 98°C to about 122°C, about 100°C to about 120°C, about 102°C to about 118°C, about 104°C to about 116°C, about 106°C to about 114°C, about 108°C to about 112°C, or about 110°C.

[0103] The mixing of hydrophilic poly(alkylene glycol), hydrophobic polymer and polyether or polyester with coupling agent is carried out for at least about 12 hours, at least about 14 hours, at least about 16 hours, at least about 18 hours, at least about 20 hours, at least about 22 hours, at least about 24 hours, at least about 26 hours, at least about 28 hours, at least about 30 hours, at least about 32 hours, at least about 34 hours, at least about 36 hours, at least about 38 hours, at least about 40 hours, at least about 42 hours, at least about 44 hours, at least about 46 hours, or at least about 48 hours.

[0104] In various embodiments, the mixture of hydrophilic poly(alkylene glycol), hydrophobic polymer and polyether or polyester is dissolved at a temperature of about 110°C for no more than about 10 minutes.

[0105] The mixing of hydrophilic poly(alkylene glycols), hydrophobic polymers, and polyethers or polyesters with coupling agents can be carried out in the absence of air and / or water / moisture and / or in the presence of an inert gas such as nitrogen. The coupling agent may comprise an isocyanate monomer containing at least two (or more) isocyanate functional groups. In various embodiments, the coupling agent is a diisocyanate selected from the following: hexamethylene diisocyanate, tetramethylene diisocyanate, cyclohexane diisocyanate, tetramethylxylene diisocyanate, dodecylene diisocyanate, toluene 2,4-diisocyanate, and toluene 2,6-diisocyanate.

[0106] In various embodiments, the method further includes removing contaminants from the multiblock polymer and dissolving the multiblock polymer in an aqueous medium to form a multiblock thermogel polymer. The step of removing contaminants from the multiblock polymer may include purifying and / or washing the multiblock polymer. The step of dissolving the multiblock polymer in an aqueous medium may include redissolving the polymer (e.g., the final polymer powder) in a balanced salt solution (BSS). In various embodiments, the BSS is water-based.

[0107] In various embodiments, the method further includes a step of removing contaminants from the multiblock polymer, which includes dialysis to remove unreacted reactants, solvents, and catalysts (e.g., extensive dialysis to remove unreacted PEG, solvents, and metal catalysts, etc.).

[0108] It will be appreciated that multiblock thermogel polymers can be obtained or prepared by one or more methods and steps disclosed in Singapore Patent Application No. 10202001798R, the entire contents of which are incorporated herein by reference. Attached Figure Description

[0109] Figure 1 The figures show drug release profiles of aflibercept (A), sunitinib malate (STB), and CP547682-HCl (CP) from different 20 wt% hydrogel reservoirs: (1) Pluronic F127 (F127) with a 20 wt% hydrogel concentration, (2) poly(ethylene glycol (PEG) / polypropylene glycol (PPG) / polycaprolactone (PCL) carbamate (20 wt% EPC1) with a 1 wt% PCL content and a 20 wt% hydrogel concentration, and (3) poly(PEG / PPG / PCL carbamate (20 wt% EPC3) with a 3 wt% PCL content and a 20 wt% hydrogel concentration, according to the various embodiments disclosed herein. Figure 1 A-1C shows tunable in vitro drug release profiles for three different drugs using three different hydrogel reservoirs. The numbers in parentheses represent the drug concentration (mg / mL) encapsulated within the hydrogel. For example, A(10)-EPC1 indicates that the EPC1 hydrogel contains 10 mg / mL of drug A. Figure 1 D shows the in vitro release profiles of three different drugs, A, STB, and CP, from the same hydrogel reservoir (20 wt% EPC1), indicating that the properties of the drugs also play an important role in drug release. Figure 1 E shows the in vitro release curves of 10 mg / mL CP from three different hydrogel reservoirs. When the concentration of CP was increased from 40 mg / mL (… Figure 1 C) Reduce to 10 mg / mL ( Figure 1 At time E), the complete drug release was significantly shortened, indicating that the CP concentration in the hydrogel reservoir plays an important role in sustained drug release.

[0110] Figure 2This is a graph showing the drug release profiles of CP from three different hydrogel reservoirs, F127, EPC1, and EPC3 (each containing CP at a concentration of 40 mg / mL or 10 mg / mL), according to the various embodiments disclosed herein. The numbers in parentheses represent the drug concentration (mg / mL) encapsulated within the hydrogel. For example, CP(10)-EPC1 indicates that the EPC1 hydrogel contains 10 mg / mL of the drug CP.

[0111] Figure 3 The graph shows the mass loss (%) of three types of hydrogel reservoirs F127, EPC1 and EPC3 after incubation in PBS at pH 7.4 and 37°C on a shaker at 50 rpm, according to the various embodiments disclosed herein. Figure 3 A shows the mass loss (%) of the hydrogel reservoir prepared in AMO at a hydrogel concentration of 10 wt%. Figure 3 B shows the mass loss (%) of the hydrogel reservoir prepared in AMO at a hydrogel concentration of 20 wt%. Figure 3 C shows the mass loss (%) of the hydrogel reservoir prepared in AMO with a hydrogel concentration of 20 wt% and a CP solution of 40 mg / mL.

[0112] Figure 4 It is a graph showing the flow behavior of different hydrogels according to various embodiments disclosed herein, represented by viscosity as a function of oscillating strain rate. Figure 4 A shows the flow behavior of EPC1 and its corresponding STB (10 or 20 mg / mL) and CP (10 or 40 mg / mL) complexes. Figure 4 B shows the flow behavior of EPC3 and its corresponding STB (10 or 20 mg / mL) and CP (10 or 40 mg / mL) complexes. Figure 4 C shows the flow behavior of F127 and its corresponding STB (10 or 20 mg / mL) and CP (10 or 40 mg / mL) complexes.

[0113] Figure 5 The Fourier transform infrared (FT-IR) spectra of the copolymers (i.e., EPC1, EPC3, and F127) and their corresponding CP-gel reservoirs (i.e., EPC1+CP, EPC3+CP, and F127+CP) according to the various embodiments disclosed herein are shown, from 1800 to 4000 cm⁻¹ for clarity. -1 and 1200-1800 cm -1 Plotting. The FT-IR spectrum of CP547682-HCl(CP) was used as a reference.

[0114] Figure 6This figure presents scanning electron microscopy (SEM) images of hydrogels EPC1 and EPC3, according to the various embodiments disclosed herein, before and after incorporation with CP at a concentration of 40 mg / mL or STB at a concentration of 20 mg / mL. The figure includes overview and cross-sectional images of the hydrogels and their respective drug-incorporated gel reservoirs. All gels were prepared in deionized (DI)-water at 20 wt%, with and without drug incorporation. SEM images were taken on lyophilized hydrogel residues. SEM images of the individual drugs (CP at 40 mg / mL and STB at 20 mg / mL) are used for reference.

[0115] Figure 7 These are graphs and diagrams relating to the in vitro bioactivity characterization of CP and STB released from the gel, based on the various embodiments disclosed herein. Figure 7 A shows that human umbilical vein endothelial cell (HUVEC) death depends on the concentration of the drug STB or CP. Figure 7 B is a schematic diagram of the experimental design, which includes two aspects: 1) quantifying the drug concentration after obtaining the released drug from the hydrogel; and 2) diluting the in vitro release concentration of the drug by 10 times to expose the cells to the drug. Figure 7 C shows the detected CP concentration of CP40, which was initially encapsulated at 40 mg / mL and released from F127, EPC1, and EPC3 on a predetermined date. Figure 7 D shows the detected STB concentration of STB10, which was initially encapsulated at 10 mg / mL and released from F127, EPC1, and EPC3 on a predetermined date. Figure 7 E-7F demonstrates the cytotoxicity of CP and STB released in vitro from EPC1, EPC3, and F127 against HUVECs.

[0116] Figure 8 The images and graphs show that, according to the various embodiments disclosed herein, intravitreal (IVT) administration of CP released in vitro from EPC1 hydrogel resulted in the regression of choroidal neovascularization (CNV) in a laser-induced mouse model. Figure 8 A consists of fundus fluorescein angiography (FFA) images taken from a representative eye on days 7 and 14 after modeling. PBS: phosphate-buffered saline used as a control; A: 10 mg / mL in PBS; CP: 10 mg / mL CP-HCl in PBS; DX: "X" represents the day when the images were taken. Figure 8B is a graph showing the degree of fluorescence leakage in choroidal lesion areas (n=6) quantified by ImageJ based on FFA images. The reduction in leakage area was calculated using the following formula: (Leakage area on the day of IVT injection: Day 0 – Leakage area on day 7 after IVT injection) / 7 days p<0.05 P<0.01 P<0.001 compared with PBS control. Figure 8 C is a graph quantified by isolectin B4 staining of choroidal lesions using ImageJ, showing an overall reduction in CNV lesion size after treatment.

[0117] Figure 9 The images and graphs show that, according to the various embodiments disclosed herein, intratumoral injection of a CP-EPC1 reservoir causes regression of in situ breast tumors and slows further growth. Figure 9 Image A is an in vivo imaging system (IVIS) image used to monitor tumor volume in nude mice carrying tumors. These mice were injected with PBS, 40 mg / mL CP (CP40), EPC3, and 40 mg / mL CP encapsulated in EPC3 (CP-EPC3). Tumor injection was performed on day 0, gel injection on day 8, and mice were euthanized on day 22. Images were taken on days 7, 15, and 21. PBS solution was used as a control. Figure 9 B is a graph showing changes in tumor volume quantified as luciferase reporter gene signaling (expressed as radioactivity) over time. The larger the tumor, the stronger the radioactivity emitted relative to its volume.

[0118] Figure 10 Images are shown of ulcers in mouse models treated with different hydrogels or pharmaceutical compositions (i.e., 40 mg / mL CP (CP40), EPC3, and 40 mg / mL CP encapsulated in EPC3 (CP-EPC3)) according to various embodiments disclosed herein. In experiments where CP was present, it was administered intratumorally. PBS solution was used as a control.

[0119] Figure 11 These are immunofluorescence (IF) staining images of cells treated with different thermogels or pharmaceutical compositions (i.e., 40 mg / mL CP (CP40) and 40 mg / mL CP encapsulated in EPC3 (CP-EPC3)) according to various embodiments disclosed herein. The staining includes DAPI for nuclear visualization, Ki67 for assessing cell proliferation, and CD31 for assessing endothelial cells and angiogenesis. These images demonstrate inhibition of angiogenesis. Vessels appear white in the images. PBS solution was used as a control.

[0120] Figure 12 This is a schematic diagram illustrating the preparation of a biodegradable 3-block copolymer thermosensitive hydrogel made of poly(ether urethane)-poly(ethylene glycol)-poly(propylene glycol) (EPC: PEG-PPG-PCL) as an injectable drug reservoir according to the various embodiments disclosed herein. The hydrogel dissolves in an aqueous environment and facilitates easy drug encapsulation. When injected at 4°C, it remains liquid and forms a gel upon reaching body temperature (approximately 37°C). The gel then undergoes bio-erosion, thereby allowing controlled and sustained drug release.

[0121] Example

[0122] The exemplary embodiments of this disclosure will be better understood and readily apparent to those skilled in the art through the following examples, tables, and, where applicable, the accompanying drawings. It should be recognized that other modifications relating to structural, biological, and / or chemical changes may be made without departing from the scope of the invention. The exemplary embodiments are not necessarily mutually exclusive, as some may be combined with one or more embodiments to form new exemplary embodiments. The exemplary embodiments should not be construed as limiting the scope of this disclosure.

[0123] Example 1. List of drugs and hydrogel types used in this study.

[0124] This study includes three types of thermosensitive hydrogels (Table 1): commercially available Pluronic F127, EPC1, and EPC3. EPC1 and EPC3 are internally prepared polyurethanes composed of random multiblock copolymers with a PEG:PPG ratio of 4:1, and containing an additional 1% or 3% PCL, respectively. F127 is a commercially available thermosensitive hydrogel. It is included due to its chemical similarity; it is composed of a triblock copolymer (PEG-PPG-PEG) with a PEG:PPG molar ratio of 40:13 and exhibits thermogel properties.

[0125] Table 1 summarizes the types of polymers used in drug release studies and the corresponding conditions for gel preparation.

[0126]

[0127] A range of drugs, including aflibercept (A), tyrosine kinase inhibitors (TKIs) such as sunitinib malate (STB), and CP547682-HCl, were included in this work (Table 2). These drugs are commonly used to treat cancer and some eye conditions, such as age-related macular degeneration (AMD), due to their angiogenesis-inhibiting mechanisms.

[0128] Table 2. Types and concentrations of drugs used in this study.

[0129]

[0130] In short, all drug solutions were directly dissolved or dispersed in AMO buffer and added to the polymer powder for gel incorporation. To promote dissolution, the solutions were incubated at 4°C for 3 nights and then vortexed and centrifuged.

[0131] Example 2. Hydrogel preparation and characterization and drug-gel interactions.

[0132] 2.1. Synthesis of random multiblock poly(PEG / PPG / PCL urethane) copolymer.

[0133] Table 3. Molecular composition of EPC1 and EPC3.

[0134]

[0135] Poly(PEG / PPG / PCL) carbamates, namely EPC1 and EPC3, are synthesized by addition polymerization of a macromonomer diol with hexamethylene diisocyanate (HMDI) in the presence of a dibutyltin dilaurate (DBTL) catalyst. The amounts of PEG (M...) required for the synthesis of EPC1 and EPC3 are listed below. n 2,050 g·mol -1 ), PPG (M n 2,000 g·mol -1 ) and PCL-diol (M n 2,000 g·mol -1 Weigh the contents into separate, clean 250 mL round-bottom flasks. Dissolve the macromonomer diol in 20 mL of anhydrous toluene at 60 °C and dry by double azeotropic distillation. Introduce inert dry argon gas into the reaction flask and add 60 mL of anhydrous toluene, then heat to 110 °C. Add 10 μL of DBTL catalyst, followed by the desired amount of HMDI. The reaction time for EPC1 is 1 hour, while the reaction time for EPC3 is 30 minutes, because if the copolymer is too long, EPC3 may become insoluble after synthesis. Quench the reaction by adding 5 mL of anhydrous ethanol. Obtain the crude copolymer by precipitation into anhydrous diethyl ether, and then further purify the crude copolymer by dialysis. For dialysis, dissolve 10 g of the crude copolymer in 100 mL of CMOS grade isopropanol at 60 °C and fill into a regenerated cellulose dialysis tube with a molecular weight cutoff of 3500 Da. Dialyze with 2 L of deionized water for 3 days, changing the water twice daily. The dialyzed copolymer solution is frozen and lyophilized to obtain purified copolymer (typical yield ≈90%).

[0136] 2.2. Molecular characterization.

[0137] The apparent molecular weight of the copolymers was determined using gel permeation chromatography (GPC) with an Agilent 1260 Infinity II system. The GPC system was equipped with a 1260 vial injector, a 1260 isocratic pump, a 1260 refractive index detector (RID), and an Agilent PLgel 5 μm MIXED-D column with a molecular weight range of 200 Da to 400,000 Da. The system was pumped with HPLC-grade THF at 1 mL / min. -1 Cycling was performed at 40°C. A calibration curve was obtained using monodisperse polystyrene standards. Copolymer samples were collected at 5 mg / mL. -1 20 μL of sample was injected for GPC measurements. 1H nuclear magnetic resonance (NMR) spectra were recorded at room temperature using a JEOL 500 MHz NMR spectrometer (Tokyo, Japan), with chemical shifts referenced to the solvent peak of deuterated chloroform (CDCl3, δ = 7.26 ppm). The acquisition time was 4.37 seconds, the pulse repetition time was 9.37 seconds, the pulse width was 90°, and 64 scans were performed per sample.

[0138] 2.3. Drug encapsulation and release.

[0139] Aflibercept stock solution was diluted to 10 mg / mL in AMO buffer, STB was prepared as a suspension in 20 mg / mL AMO, and CP was prepared as suspensions in 10 mg / mL and 40 mg / mL AMO buffer. The copolymer, Pluronic F127, EPC1, and EPC3 (20 mg) were placed in 1.5 mL centrifuge tubes, followed by the addition of 100 μL of drug solution. For control tubes, 100 μL of AMO was used instead of the drug solution. The solutions were thoroughly mixed and incubated at 4°C for 3 nights to form a drug-encapsulated hydrogel reservoir. The prepared drug gel reservoir was transferred to 37°C, and 1 mL of pre-warmed fresh PBS was added over half an hour to initiate drug release. The tubes were kept at 37°C and shaken at 50 rpm. At regular intervals, 500 μL of supernatant was collected, followed by the addition of an equal volume of pre-warmed fresh PBS. The collected supernatant was kept at -20°C for drug concentration quantification. The released aflibercept was quantified using the Pierce microBCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA). Quantification was performed using quartz cuvettes based on the typical absorbance of STB and CP at 426 nm and 261 nm, respectively. Drug quantification was based on calibration curves obtained using appropriate stock solutions prepared at gradient concentration ranges.

[0140] Aflibercept (A) is a hydrophilic anti-vascular endothelial growth factor (anti-VEGF) protein with a molecular weight of 119 kDa. It is used as a standard intravitreal anti-VEGF treatment for age-related macular degeneration (AMD). Sunitinib malate (STB) and C547682-HCl (CP) are both tyrosine kinase inhibitors; they are synthetic small molecule drugs (<1000 Da). STB is FDA-approved for the treatment of gastrointestinal stromal tumors. It is also formulated into a microparticle reservoir for sustained release of STB for the treatment of wet AMD, currently in Phase 2b clinical trials. PAN-90806 is a CP-HCl eye drop formulation for wet AMD, currently in Phase 1 / 2 clinical trials.

[0141] Hydrogel matrices play an important role in drug release via diffusion or matrix degradation. For example, A( Figure 1 A) and STB ( Figure 1 B) exhibited the same drug release pattern, with the gel matrix arranged from fastest to slowest as F127>>EPC3>EPC1, despite significant drug differences. This trend was found to be consistent with... Figure 3 The gel dissociation rates shown in A and 3B are consistent, i.e., complete dissolution / degradation of 20% gel: F127, EPC3, and EPC1 takes approximately 10 days, 30 days, and 30-40 days, respectively. Interestingly, this drug release trend changes when CP is incorporated into the gel at 40 mg / mL. First, the drug release trend becomes F127 >> EPC1 > EPC3. Second, the drug release period is significantly prolonged from 2 months to 1 year. This drug release trend was found to be related to the gel dissociation rate (…). Figure 3 C) Completely consistent, i.e., when CP is incorporated into the gel at 40 mg / mL, gel dissociation decreases and the trend becomes F127 >> EPC1 > EPC3. A, the release profiles of STB and CP40 from EPC1 (20 wt%) are shown in... Figure 1 In D, the effect of drug type on release rate was further disclosed. The low initial burst release of A was attributed to the electrostatic interaction between A and the EPC gel and its high protein molecular weight. The high burst release from STB was attributed to its small molecule properties and improved water solubility through the use of the malate form. However, contrary to expectations, CP40 exhibited a low initial burst release and a long sustained release despite the use of the CP·HCl form and its small molecular weight to enhance water solubility in this study. When the CP concentration was reduced from 40 mg / mL to 10 mg / mL ( Figure 1 At point E), the release trend becomes the same as at points A and STB, and the release period is significantly shortened.

[0142] 2.4. Hydrolytic degradation of hydrogels and their corresponding drug reservoirs.

[0143] Using the same design as the drug release assay, 20 wt% copolymer ± drug (100 μL) was prepared in PBS in 1.5 mL centrifuge tubes, mixed, and equilibrated at 4°C for 3 nights. When the temperature equilibrated to 37°C, PBS (1 mL) was added to each hydrogel sample. The samples were then incubated at 37°C with shaking at 50 rpm. Fresh PBS buffer was replaced at predetermined time intervals. This process was allowed to continue for up to one year. The experiment was repeated three times. At different time points, the tubes were lyophilized and weighed. The mass loss of the copolymer gel after dissolution and degradation was defined as...

[0144] Mass loss (%) - [1 - (W) t / W0)]×100%,

[0145] Among them, W0 and W t These are the initial weight and the weight of the copolymer remaining in the tube after dissolution and / or degradation at time t, respectively.

[0146] Table 4. Mass loss (in mg or %) of hydrogel reservoirs EPC1, CP40+EPC1, EPC3, CP40+EPC3, F127 and CP40+F127 over 200 days.

[0147]

[0148] like Figure 3 As shown, the hydrolytic erosion process is accompanied by mass loss of the hydrogel. It was found that the F127 hydrogel degraded / dissolved the fastest in all cases, regardless of gel concentration (…). Figure 1 A and 1B) and drug incorporation ( Figure 3 How about B and 3C? Increasing the hydrogel concentration reduced the rate of weight loss, especially at earlier time points (<20 days). Figure 3 B). This contributes to an increase in the degree of cross-linking within the hydrogel as the polymer concentration increases. Figure 3 C indicates that when CP is incorporated into 20% by weight of polyurethane gel, weight loss only begins to occur 20 days after the onset of drug release. For pure 20% by weight polyurethane gel, EPC1 and EPC3, 100% gel dissolution occurs at approximately 50 days. However, when CP is incorporated at 40 mg / mL, less than 60% weight loss is observed at 100 days.

[0149] 2.5. Drug-gel interactions quantified by rheological measurements, Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM).

[0150] 2.5.1. Rheological Measurement

[0151] Steady-state and dynamic rheological experiments were conducted on an AR-G2 stress-controlled rheometer (TA Instruments, Newark, DE). The steady-state shear flow behavior of the samples was evaluated under strain-controlled mode at 60 °C, from 0.001 to 100 s⁻¹. -1 Perform a shear rate scan.

[0152] The flow curve (steady-state shear flow) describes the viscosity of the hydrogel as a function of the applied shear rate, which determines the tendency of the material flow. Figure 4 When applying 0.0006 to 0.06 S -1 During periods of low shear rate, the viscosity of the hydrogel remained relatively stable, primarily due to the influence of Brownian forces. Compared to the hydrogel and its corresponding drug complex, the CP40-composite EPC exhibited the highest viscosity increment, being 370 times that of EPC3 and 20 times that of EPC1. However, no such increment was observed in F127, which, after being composited with CP40, was only about 1.5 times that of F127.

[0153] 2.5.2. Infrared Spectroscopy (FT-IR)

[0154] FT-IR spectroscopic analysis of hydrogels and drug-gel complexes was performed on a Perkin Elmer Spectrum 2000 FT-IR spectrometer. All hydrogels and drug-gel complexes were prepared in aqueous solution and lyophilized. Samples were run by pressing with spectral-grade KBr pellets or by attenuated total reflectance (ATR) using a zinc selenide optical crystal.

[0155] Correlation superposition FTIR spectra of CP40, EPC1, EPC3 and F127 ( Figure 5 This shows evidence of enhanced supramolecular interactions between CP40 and the EPC polymer compared to F127:

[0156] Through the OH stretching of CP40 (center approximately 3470 cm) -1 The significant broadening of the OH group of CP40 in the presence of FTIR identifies the extensive hydrogen bonding between CP40 and EPC. In contrast, the OH broadening of CP40 in the presence of FTIR is much smaller, indicating a weaker degree of hydrogen bonding. This is further supported by the following: (F127 (1680→1688 cm⁻¹)) -1 Compared to the case where EPC is present, the perturbation of C=O stretching in CP40 is larger (1680→1695 cm). -1 ).

[0157] The perturbation of the aromatic C=C stretching of CP40 (1540→1550 cm) can be observed. -1The hydrophobic interactions between the aromatic groups of CP40 and the hydrophobic portions of EPC and F127 were identified. In the solid state, the degree of hydrophobic interaction appears to be similar, based on similar displacements of C=O stretching.

[0158] 2.5.3. Scanning Electron Microscopy (SEM)

[0159] The surface morphology and pores within the hydrogel were observed using SEM (scanning electron microscopy). The hydrogel composite was cryogenically cut and fixed onto a cylindrical microscope base covered with carbon strips, coated with a 100-200 Å thick gold layer, and then observed using SEM.

[0160] The microstructure of the hydrogel was studied using SEM and as follows: Figure 6 As shown, the surfaces of EPC1 and EPC3 are tightly packed, and their cross-sectional images exhibit a mesh-like network. CP40 alone at 40 mg / mL shows an irregular cross-linked network structure, indicating that it forms a soft gel at this concentration. By incorporating CP40, EPC1 / EPC3+CP40 shows morphological differences, differing in having a regular cage-like cross-linked structure.

[0161] Example 3. Biological applications of TKI gel reservoirs for the treatment of neovascular retinal diseases and cancer.

[0162] 3.1. The in vitro bioactivity of released CP and STB, exhibiting release-dependent drug concentration-dependent cytotoxicity against HUVECs.

[0163] Human umbilical cord endothelial cells (HUVECs, 12,000 cells per well in a 96-well plate) were grown overnight in endothelial growth medium (EGM). Both CP and STB stock solutions were prepared in EGM at 1 mg / mL and serially diluted to EGM to obtain experimental concentrations of 0.1, 1, 5, and 10 μg / mL. The medium was then replaced with 100 μL of serially diluted drug solutions. Cells directly exposed to EGM were used as a negative control. After 24 hours of culture, HUVECs were washed twice with PBS, and 50 μL of a 10-fold diluted lysis buffer (from a lactate dehydrogenase release (LDH) assay kit) was added to each well. One hour after cell lysis, 50 μL of working solution was added. After 30 minutes, stop buffer (50 μL per well) was added, and the absorbance was read at 490 nm (650 nm as a reference) using a microplate reader to determine viable cells. The percentage of viable cells was calculated as follows:

[0164] Live cells % = 100 × (Ab test / Ab control).

[0165] Long-term, sustained delivery of active therapeutic agents is important for the treatment of AMD. Tyrosine kinases are important cell signaling proteins with diverse biological activities, including cell proliferation and migration. Multiple kinases are involved in angiogenesis, including receptor tyrosine kinases. Inhibition of angiogenic tyrosine kinases has been developed as a systemic therapeutic strategy for cancer, and clinical trials are underway for several angiogenic retinal diseases. In this study, concentration-dependent cytotoxicity of TKIs against HUVECs was identified. Figure 7 A), this method was used to test CP released in vitro from 20 wt% F127, EPC1, and EPC3 (within 200 days). Figure 7 E) and STB (within 30 days, Figure 7 F) Bioactivity. The concentrations of released CP and STB obtained on specific dates were quantified and recorded. Figure 7 In C and 7D. Notably, the CP released from F127 on day 3 was significantly higher than the CP released from EPC1 and EPC3. Figure 7 C), which leads to Figure 7 Cell survival was significantly lower in E cells. On the other hand, almost complete release of CP was achieved from F127 cells on day 40. Figure 7 C), therefore, a high corresponding cell viability was observed using samples obtained from F127 on day 40. Figure 7 E). Until day 158, the cytotoxicity of CP released from EPC1 was higher than that of CP released from EPC3. Figure 7 E), but no significant difference was observed. The same higher CP release observed on those days was seen in... Figure 7 In C. Once this release trend reversed on day 200, a corresponding change was observed in the cytotoxicity results of CP released on day 200. The same higher STB release was observed. Figure 7 D) and lower HUVEC survival rate Figure 7 (F) This was particularly evident for STBs released from F127 gels. High STB release from F127 on day 1 resulted in low cell viability; however, cell viability greater than 75% on day 10 and approximately 100% on days 20 and 30 corresponded to low and almost complete STB release on days 10, 20, and 30. These results strongly suggest that CP and STBs released in vitro remain biologically active, regardless of gel type and release day.

[0166] 3.2. In vitro released CP injected intravitreally can reduce vascular leakage in a laser-induced CNV mouse model.

[0167] Male wild-type C57B / 6J mice aged 6 to 8 weeks were obtained from In-vivos (Singapore) for in vivo experiments. Mice were anesthetized with intraperitoneal injections of ketamine (150 mg / kg) and toluidine (10 mg / kg). Photocoagulation was induced in these eyes using an image-guided laser system (Micro IV, Phoenix Research Laboratories, Pleasanton, CA). Seven days after laser treatment, fundus fluorescein angiography (FFA) images were taken as T=0, and mice were divided into four groups: PBS, aflibercept (10 mg / mL) in PBS (A), CP-HCl (10 mg / mL) in PBS (CP), and in vitro-acquired CP released from EPC1 hydrogel (at predetermined time points). Samples from all four groups were injected intravitreally (1 μL) into the vitreous humor of the eye immediately after T=0 imaging. Seven days after IVT, FFA images were taken at T=7. Mice were then euthanized and the eyes were enucleated to prepare choroidal lamina plana. Eyes were fixed in 4% paraformaldehyde in PBS and incubated overnight at 4°C. The eyecups were incubated with isolectin B4 at 4°C for choroidal vascular staining, followed by three PBS washes. After four incisions along the radial direction of the optic nerve, the tissue was mounted flat, and Z-stack images of the CNV lesions were captured using a confocal microscope (LSM700, Zeiss, Thornwood, NY). Angiographic and Z-stack images were imported into ImageJ. The maximum boundary of the CNV lesion on each image was manually drawn at magnification, and the area was quantized to pixels per 100 μm. The fluorescence intensity of the CNV lesions was graded using ImageJ (National Institutes of Health, Bethesda, MD) by two single-blind independent graders. Results are as follows: Figure 8 As shown.

[0168] The biological activity of the released CP was further confirmed using a mouse model of laser-induced CNV. When IVT was administered with PBS buffer, the leakage was reduced to 17.4%, which was minimal.

[0169] 3.3. Intratumoral injection of CP-EPC1 reservoir caused regression in mouse model of orthotopic breast cancer xenograft.

[0170] The application of CP as a local drug release reservoir was further tested in an in vivo orthotopic xenograft breast cancer model in NCr-Foxn1 nude mice. The MCF7-Luc orthotopic breast cancer model was generated by injecting the MCF7-Luc reporter cell line (MCF7-Luc) into the two abdominal mammary pads of nude mice. On day 0 ( Figure 9A) After confirming solid tumor formation via IVIS, CP-EPC1 was injected into a tumor in the contralateral breast. Other sites within the tumor served as negative controls (phosphate-buffered saline, injected intratumorally). Regression at the CP-EPC1 injection site was observed on IVIS imaging on day 7 post-injection. Significant regression or growth retardation was observed even as late as day 14 post-injection.

[0171] Example 4. Summary and Discussion

[0172] In the foregoing embodiments, three types of thermosensitive hydrogels were prepared: commercially available pluronic F127, EPC1, and EPC3, prepared at 20% by weight, and used to contain drugs. The PEG to PPG ratio of these polymers was fixed at 4:1, but the PCL content ranged from 0% to 1% to 3%, respectively. When the hydrogels were drug-free, F127 completely dissolved (20% by weight) at approximately 10 days, EPC3 at approximately 40 days, and EPC1 at approximately 50 days.

[0173] Three types of anti-VEGF (aflibercept (a macromolecule against VEGF); and two small-molecule TKIs, sunitinib malate (STB) and CP) were directly incorporated into hydrogels by dissolving polymer powder in a desired drug solution in phosphate-buffered saline (PBS). At a CP concentration of 20 mg / mL, release was extended to one year, while gel dissolution was significantly delayed in EPC1 and EPC3. Achieving long-term drug release from biodegradable hydrogel reservoirs is typically challenging due to their inherent hydrophilic nature. However, examples demonstrate that sustained in vitro release of a tyrosine kinase inhibitor (TKI), CP-547632 (CP), for more than one year can be achieved using a biodegradable 3-block copolymer poly(ether ester urethane)-poly(ethylene glycol)-poly(propylene glycol) (EPC: PEG-PPG-PCL) thermosensitive hydrogel.

[0174] It will be recognized that the prolonged release of CP from the EPC hydrogel is attributed to a CP-promoted EPC gelation process, which is PCL content-dependent and linearly correlated with CP concentration. No prolonged drug release and gel dissolution were observed with aflibercept and STB, nor when using F127 gel. Scanning electron microscopy (SEM) morphology and cross-sectional images of the gels indicate that the same drug promotes gel crosslinking within CP-incorporated EPC1 and EPC3, as well as increased porosity and decreased pore size. The surprising technical effect of TKIs promoting further gelation of the hydrogel reservoir through physical interactions has not been previously known or suggested.

[0175] FT-IR data reveal that the crosslinking is primarily attributed to the hydrophobic interactions of alkyl groups between the CP and EPC polymers, as well as hydrogen bonding formed by hydroxyl and carbonyl groups.

[0176] The bioactivity of STB and CP released from the hydrogel reservoir was demonstrated by in vitro HUVEC cytotoxicity assays. Cell death increased with increasing drug concentration throughout the drug release process, indicating the stability of the drug within the hydrogel over a one-year drug release period and in an in vivo mouse laser-induced choroidal neovascularization (CNV) model.

[0177] Accelerated vascular leakage following injection of 1 μL of in vitro released CP into the vitreous humor of CNV-treated mice confirmed the potential biological applications of the released CP for neovascular retinal disease. Anticancer activity of the CP-EPC1 reservoir was demonstrated using a nude mouse model carrying breast tumors, through significant reductions in cancer volume at 1 and 2 weeks post-injection into the reservoir.

[0178] To the inventors' knowledge, based on results from clinical trials, the longest treatment duration using the clinical trial product, the intravitreal axitinib implant OTX-TKI, is approximately one year. Therefore, this technology provides an alternative that potentially offers treatment outcomes comparable to or even longer than those achieved with OTX-TKIs.

[0179] In fact, the implementation scheme of this technology is considered to offer several advantages over the prior art, including but not limited to the following:

[0180] Localized sustained release of active tyrosine kinase inhibitors (TKIs) can significantly affect the treatment of retinal diseases (such as age-related macular degeneration (AMD), diabetic macular edema (DME), retinal vein occlusion (RV), etc.) and diseases requiring long-term TKI treatment (such as cancer).

[0181] Easy drug encapsulation in aqueous environments;

[0182] Injectable polymer compositions that minimize damage to surrounding tissues;

[0183] Utilizing the soft tissue properties of materials;

[0184] Local drug delivery helps minimize side effects; for example, local, long-term drug release can help overcome drug resistance problems that are typically reported 8 months after treatment and reduce toxic effects associated with long-term oral supplements.

[0185] Since both the drug delivery system (CP) and the hydrogel reservoir (EPC) have undergone excellent ophthalmic application testing, the need to verify ophthalmic efficacy and address safety concerns can be alleviated.

[0186] A hydrogel reservoir loaded with CP can be obtained through a simple mixing process;

[0187] The gelation process between CP and gel molecules is a physical interaction, thus avoiding the use of toxic reactive chemicals and strict reaction conditions;

[0188] The gelation process is highly repeatable and has a high success rate;

[0189] A hydrogel reservoir loaded with CP can be applied as a transparent implant to treat eye diseases and therefore will not adversely interfere with the patient's vision; and

[0190] Sustained release of TKIs can reduce the frequency of intravitreal injections of anti-VEGF from once every two months to an annual treatment.

[0191] Those skilled in the art will recognize that other changes and / or modifications can be made to the embodiments disclosed herein without departing from the spirit or scope of this disclosure, which is broadly described. For example, features of different exemplary embodiments may be mixed, combined, exchanged, incorporated, adopted, modified, included, or similar among different exemplary embodiments in the description herein. Therefore, the present embodiments should be considered illustrative rather than restrictive in all respects.

Claims

1. A polymer composition comprising: Multiblock thermogel polymers comprising hydrophilic poly(alkylene glycols), hydrophobic polymers, and polyethers or polyesters chemically coupled together via at least one of urethane / methionine ester bonds, carbonate bonds, ester bonds, urea bonds, amide bonds, ether bonds, amine bonds, triazole bonds, or combinations thereof; and Tyrosine kinase inhibitors (TKIs) mixed with the multi-block thermogel polymer.

2. The polymer composition of claim 1, wherein the TKI interacts with the multiblock thermogel polymer to promote the gelation of the multiblock thermogel polymer.

3. The polymer composition according to any one of the preceding claims, wherein the TKI increases the viscosity of the polymer composition by at least 10 times compared to the absence of the TKI.

4. The polymer composition according to any one of the preceding claims, wherein the TKI comprises a VEGFR inhibitor (e.g., a VEGFR-2 inhibitor) and / or an FGFR inhibitor.

5. The polymer composition according to any one of the preceding claims, wherein the TKI comprises one or more of the following: i. CP-547632, its analogues, or pharmaceutically acceptable salts thereof; or ii. Compounds of formula (I), Formula (I).

6. The polymer composition according to any one of the preceding claims, wherein the hydrophilic poly(alkylene glycol) comprises polyethylene glycol (PEG).

7. The polymer composition according to any one of the preceding claims, wherein the hydrophobic polymer is selected from poly(propylene glycol) (PPG), poly(lactic acid-co-glycolic acid) (PLGA), polylactic acid (PLA), poly(N-isopropylacrylamide) (PNIPAAM), polypeptides, or combinations thereof.

8. The polymer composition according to any one of the preceding claims, wherein the polyether or polyester comprises one or more of polycaprolactone (PCL), polytetrahydrofuran (PTHF), polyhydroxybutyrate (PHB), polylactic acid-co-glycolic acid (PLGA), and polylactic acid (PLA).

9. The polymer composition according to any one of the preceding claims, wherein the concentration of TKI in the polymer composition is not less than about 10 mg / L.

10. The polymer composition according to any one of the preceding claims, wherein the molar ratio of the hydrophilic poly(alkylene glycol) to the hydrophobic polymer is 1-10:

1.

11. The polymer composition according to any one of the preceding claims, wherein the polyether or polyester is present in the multiblock polymer in an amount of 1% to 10% by weight.

12. The polymer composition according to any one of the preceding claims, wherein the molar ratio of the hydrophilic poly(alkylene glycol) in the multiblock polymer to the hydrophobic polymer and the polyether or polyester is in the range of about 1-10:1:0.01-1.

5.

13. The polymer composition according to any one of the preceding claims, wherein the multiblock polymer is present in an aqueous medium in an amount of up to 30% w / v.

14. The polymer composition according to any one of the preceding claims, wherein the pH of the composition is in the range of 7.1 to 7.

4.

15. The polymer composition according to any one of the preceding claims, wherein the polymer composition has a critical gelation temperature of not less than 4°C.

16. The polymer composition according to any one of the preceding claims, further comprising one or more pharmaceutically active ingredients different from the TKI.

17. The polymer composition of claim 16, wherein the pharmaceutically active ingredient, which is different from the TKI, is selected from aflibercept, sunitinib malate, and combinations thereof.

18. The polymer composition according to any one of the preceding claims, wherein the polymer composition has a drug release profile of not less than 2 months.

19. The polymer composition of claim 18, wherein the polymer composition has a drug release profile of not less than 12 months.

20. The polymer composition according to any one of the preceding claims, for use in medicine.

21. The polymer composition according to any one of the preceding claims, used for treating or preventing eye diseases, treating or preventing tumors, treating or reducing angiogenesis, and / or treating cancer.

22. Use of the polymer composition according to any one of the preceding claims in the manufacture of medicaments for treating or preventing eye diseases, for treating or preventing tumors, for treating or reducing angiogenesis, and / or for treating cancer.

23. A method for treating or preventing eye diseases, treating or preventing tumors, treating or reducing angiogenesis, and / or treating cancer, said method comprising administering to a subject in need of the polymer composition according to claims 1 to 19.

24. The polymer composition of claim 21, the use of claim 22, or the method of claim 23, wherein the eye disease is selected from neovascular retinal disease, age-related macular degeneration (AMD), diabetic macular edema (DME), and retinal vein occlusion (RV).

25. The polymer composition of claim 21, the use of claim 22, or the method of claim 23, wherein the cancer is selected from breast cancer, leukemia, gastrointestinal cancer, kidney cancer, lung cancer (e.g., non-small cell lung cancer), non-Hodgkin's lymphoma, melanoma, ovarian cancer, fallopian tube cancer, and eye cancer.

26. A method for preparing the composition according to any one of claims 1 to 21, the method comprising: Provided are multi-block thermogel polymers comprising a hydrophilic poly(alkylene glycol), a hydrophobic polymer, and a polyether or polyester chemically coupled together via at least one of urethane / methionine ester bonds, carbonate bonds, ester bonds, urea bonds, amide bonds, ether bonds, amine bonds, triazole bonds, or combinations thereof; and The tyrosine kinase inhibitor (TKI) was mixed with the multiblock thermogel polymer.

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