Responsive elastomeric polymers and methods of making and using same

Abstract

Functionalized hyaluronic acids (HAs), responsive elastomeric polymer systems comprising functionalized HAs, and methods of making and using the same are disclosed. The polymer system can be used for controlled local or systemic drug delivery and release of analgesics, anesthetics, antibiotics and other drugs as well as tissue engineering articles.

Description

[0001] This application is a divisional application of the Chinese invention patent application (filing date is June 13, 2017, application number is 201780044188.1, PCT application number is PCT / US2017 / 037248), and the invention name is "Responsive elastic polymers and preparation and use methods thereof").

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 349,475, filed June 13, 2016, and U.S. Provisional Application No. 62 / 366,160, filed July 25, 2016, the entire contents of which are expressly incorporated herein. Technical Field

[0003] The present invention relates generally to polymer systems, and in particular to functionalized hyaluronic acid (HA), responsive elastic polymer systems comprising functionalized HA, and methods of making and using the same. Background Art

[0004] This section introduces aspects that may help to better understand the present invention. Accordingly, these statements should be read in this light and should not be construed as admissions of what is or is not prior art.

[0005] Controlled drug delivery offers many advantages over conventional dosage forms, including improved efficacy, reduced toxicity, reduced need for specific drug administration (e.g., repeated injections), and improved patient compliance and convenience.[ 1 ]. Several controlled systems based on biomaterials, such as polymersomes, polymer micelles, microspheres, nanospheres, nanoparticles, polymer membranes, and silica nanoparticles, are currently being investigated for the delivery of drugs in a spatiotemporally controlled manner. Hydrogels composed of polymer networks swollen in water offer a promising delivery platform for controlled drug release applications, as depot formulations can be created to allow slow elution of the drug, maintaining high local drug concentrations in the surrounding tissue for extended periods of time.[ 2 ]. In addition, this hydrogel can also be used for controlled systemic drug release. Because responsive hydrogels can simultaneously respond to triggers such as pH, temperature, light, ions, and proteins, they represent attractive candidates for the development of drug delivery vehicles to achieve prolonged effects [ 3 ].

[0006] However, the utility of conventional hydrogels for clinical applications is often hampered by their poor mechanical properties. For example, they are generally very weak and do not have high stretchability [ 4]. Therefore, the use of conventional hydrogels in load-bearing anatomical sites such as synovial joints is a huge challenge. To function effectively in these environments, hydrogels must be stretchable and swellable under compression and tension without rupture. Therefore, there is an unmet need for the development of responsive, highly elastic injectable hydrogels. Summary of the invention

[0007] The present invention provides a composition comprising a polymer matrix comprising a functionalized hyaluronic acid (HA) having at least 100 monomer units cross-linked with at least one telechelic polymer unit.

[0008] In some embodiments, the aforementioned functionalized hyaluronic acid is selected from Formula I-IV:

[0009]

[0010] Wherein R1, R2, R3, R4 and R5 can include any one or combination of the following: haloacetate, dihydrazide, amine, thiol, carboxylic acid, aldehyde, ketone, active hydrogen site on aromatic ring, diene, azidoisothiocyanate, isocyanate, acyl azide, N-hydroxysuccinimide (NHS) ester, sulfo-NHS, sulfonyl chloride, epoxide, carbonate, aryl halide, imidate, carbodiimide (e.g., N,N'-dicyclohexylcarbodiimide (DCC) and 1-ethylhexane)

[0043] The invention also includes but is not limited to 3-(3-dimethylaminopropyl)carbodiimide (EDC), alkyl phosphate compounds, acid anhydrides, fluorophenyl esters, hydroxymethyl phosphines, guanidine groups, iodoacetyl derivatives, maleimides, aziridines, acryloyl derivatives, arylating agents, disulfide derivatives, vinyl sulfones, phenyl thioesters, cisplatin, diazoacetates, carbonyldiimidazoles, oxirane, N,N'-disuccinimidyl carbonate, N-hydroxysuccinimidyl chloroformate, alkyl halogens, hydrazines, alkynes, and phosphorus-bound chlorine.

[0011] In some embodiments, the telechelic polymer is selected from any one or combination of the following: poly(aliphatic ester) (e.g., poly(lactide) (PLA), poly(ε-caprolactone) (PCL), poly(glycolic acid) (PGA), poly(lactic acid-co-glycolic acid) (PLGA), poly(trimethylene carbonate) (PTMC), polydioxanone (PDS), poly(orthoester), polyanhydride, poly(anhydride-co-imide), poly(anhydride-ester), polyurethane (e.g., degrapol), poly(amide), poly(esteramide), poly(orthoester), poly(dioxanone), poly(acetal), poly(ketal), poly(carbonate), poly(orthocarbonate), poly(hydroxybutyrate), poly(hydroxy-valerate), poly(alkylene oxalate), poly(alkylene succinate) (poly(alkylene succinate) succunate)), poly(malic acid), poly(amino acids), poly(vinyl pyrrolidone), poly(hydroxycellulose), poly(glycerol sebacate), poly(ethylene imine), poly(acrylic acid) (PAA), poly(N,N′-diethylaminoethyl methacrylate, polyethylene glycol (PEG), poly(propylene oxide) (PPO), PEO-b-PPO block copolymers (e.g., pluronic (or poloxamer), and tetronic), poly(vinyl alcohol) (PVA), Poly(N-isopropylacrylamide) (PNIPAm), poly(N,N-diethylacrylamide) (PDEAAm), poly(oxazolines) (e.g., poly(2-methyloxazoline and poly(2-ethyl-2-oxazoline), oligo(ethylene glycol) fumarate (OPF), poly(propylene fumarate), poly(alkyl cyanoacrylate), poly(acrylamide), synthetic poly(amino acids) (e.g., poly(L-glutamic acid) (L-PGA) and poly(aspartic acid)), polyphosphazenes, and poly(phosphates).

[0012] In some embodiments, the aforementioned telechelic polymer is selected from any one or combination of the following: fibrin, collagen, matrigel, elastin, elastin-like peptides, albumin, natural poly (amino acids) (e.g., phycocyanin, poly (ε-l-lysine), poly (γ-glutamic acid)), polysaccharides (e.g., chitosan, dextran, chondroitin sulfate, agarose, alginate, methylcellulose and heparin).

[0013] In some embodiments, the aforementioned functionalized hyaluronic acid comprises at least one monomer unit of hyaluronic acid functionalized with an amine group.

[0014] In some embodiments, the aforementioned functionalized hyaluronic acid comprises at least one monomer unit of hyaluronic acid functionalized with an acrylate group.

[0015] In some embodiments, the aforementioned telechelic polymer is thiolated poly(N-isopropylacrylamide) PNIPAm.

[0016] The present invention also provides a method for preparing a polymer matrix based on hyaluronic acid (HA), the polymer matrix comprising a functionalized hyaluronic acid having at least 100 monomer units cross-linked with at least one unit of a telechelic polymer. The method comprises the following steps:

[0017] preparing a functionalized hyaluronic acid having at least 100 monomer units;

[0018] preparing pre-processed functional telechelic polymers;

[0019] Functionalized hyaluronic acid is cross-linked with functional telechelic polymers via carbodiimide-mediated reactions, esterification, amidation, aldehyde and ketone reactions, active hydrogen reactions, photochemical reactions, azide-alkyne cycloadditions (e.g., copper-catalyzed azide-alkyne cycloadditions (CuAAC), copper-free azide-alkyne huisgen cycloadditions), thiol-click reactions, Diels-Alder reactions, nitrile oxide cycloadditions, and enzymatic cross-linking strategies (e.g., horseradish peroxidase and hydrogen peroxide).

[0020] In some embodiments, the aforementioned functional telechelic polymer is thiolated PNIPAm.

[0021] In some embodiments, the aforementioned HA-based polymer matrix further comprises at least one monomer unit of a polymer that can be prepared by polymerization reactions of i) 'grafting to' and (ii) 'grafting from' strategies.

[0022] In some embodiments, the aforementioned cross-linking reaction between the functionalized hyaluronic acid and the thiolated PNIPAm is a thiol-ene reaction.

[0023] In some embodiments, the aforementioned 'grafting from' method involves a functionalized hyaluronic acid comprising at least one polymerizable moiety, an initiator, a RAFT agent, and an infertier.

[0024] In some embodiments, the aforementioned polymerizable moiety is an acrylate.

[0025] In some embodiments, the aforementioned RAFT agent is S-1-dodecyl-S'-(α,α'-dimethyl-α"-acetic acid) trithiocarbonate (DATC).

[0026] In some embodiments, the aforementioned polymerization reaction is a 'graft from' process using surface initiated RAFT polymerization of PNIPAm.

[0027] The present invention also provides a polymer-based drug delivery platform comprising the aforementioned composition, which further encapsulates anesthetics, analgesics or antibiotics with a polymer matrix system through physical or chemical interactions.

[0028] In some embodiments, the aforementioned physical interaction includes any one or a combination of the following: hydrophobic interaction, hydrophilic interaction, hydrogen bonding, and intermolecular electrostatic interaction.

[0029] In some embodiments, the aforementioned drug delivery platform comprises any one of the following: an opioid (eg, morphine) or a nonsteroidal anti-inflammatory drug (NSAID), which includes chloroprocaine, bupivacaine, lidocaine, and procaine.

[0030] The present invention also provides a method for controlled drug delivery. The method comprises:

[0031] preparing a polymer matrix comprising a functionalized hyaluronic acid (HA) having at least 100 monomer units cross-linked with a telechelic polymer having a theoretical degree of substitution of 100% to 30%;

[0032] preparing a polymer matrix comprising a functionalized hyaluronic acid (HA) having at least 100 monomer units cross-linked with a telechelic polymer having a theoretical degree of substitution of amine groups of 20% to 80%;

[0033] preparing a conjugate of the drug in the polymer matrix at a concentration of 0 to 30% (w / v); and

[0034] It was observed that the drug release rate decreased as the molecular weight of the polymer matrix increased.

[0035] In some embodiments, the aforementioned drug delivery polymer matrix is ​​HA-g-PNIPAm, and the drug is morphine.

[0036] These and other features, aspects and advantages of the present invention will become better understood with reference to the following drawings, associated description and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 is a schematic diagram depicting the synthetic pathway of a hyaluronic acid-based polymer matrix.

[0038] Figure 2 Functionalized monomeric hyaluronic acid represented by Formula I, II, III, IV is shown.

[0039] Figure 3 A method for the synthesis of HA-g-PNIPAm via a combination of reversible addition-fragmentation chain transfer (RAFT) polymerization and thiol-olefin click reaction is shown.

[0040] Figure 4 The synthesis of HA-g-PNIPAm by a 'graft-from' approach using reversible addition-fragmentation chain transfer (RAFT) polymerization is shown.

[0041] Figure 5 Schematic diagram showing the advantages of HA-g-PNIPAm hydrogel.

[0042] Figure 6 FTIR spectra of HA (black line), HA-ADH (blue line), HA-NAS (pink line), and HA-g-PNIPAm (red line) are shown, demonstrating the successful synthesis of each step.

[0043] Figure 7 The results show that HA, HA-ADH, HA-NAS and HA-g-PNIPAm 1 H NMR spectra confirmed the successful synthesis of each step.

[0044] Figure 8 The reversibility of HA-g-PNIPAm hydrogel was shown.

[0045] Fig. 9 Confirmation of HA-g-PNIPAm hydrogel formation is shown.

[0046] Fig.10 Shown is a time course graph of HA-g-PNIPAm hydrogel loaded and unloaded at 37 °C, demonstrating the elastic nature of the hydrogel.

[0047] Fig.11 Frequency sweeps of HA-g-PNIPAm hydrogels are shown. The linear modulus plateau versus frequency was determined. The gel stiffness increased with increasing HA-g-PNIPAm concentration.

[0048] Fig.12 The proposed mechanism of HA-g-PNIPAm gel formation is shown: the formation of intra- and intermolecular hydrogen bonds.

[0049] Fig.13 The in vitro release curves of morphine from HA-g-PNIPAm (13k) loaded with morphine are shown at concentrations (w / v) of 5%, 7.5%, 10% and 15%, respectively, and a HA-g-PNIPAm (13k) hydrogel control. The morphine concentration was 0.468% (w / v). As the concentration of HA-g-PNIPAm increased, the release rate of morphine slowed down.

[0050] Fig.14Pharmacokinetic studies of morphine in rats are shown. (A) Carotid catheterization; (B) Subcutaneous (sc) injection (0.25 mL) of morphine-loaded 10% HA-g-PNIPAm hydrogel; (C) Blood samples were collected through a pre-placed carotid catheter using an automated system (Culex); and (D) 3 days after subcutaneous injection (0.25 mL), recovery tissue showing hydrogel pockets can be seen, confirming hydrogel formation after subcutaneous injection.

[0051] Fig.15 In vivo drug release profiles following subcutaneous injection (0.25 mL) in rats are shown. Morphine plasma concentration (ng / mL) as a function of time. The blue line indicates the therapeutic plasma concentration (10 ng / mL).

[0052] Fig.16 Schematic diagram of an injectable hydrogel for intra-articular delivery of analgesics or anesthetics is shown.

[0053] Fig.17 Frequency sweep of HA-g-PNIPAm hydrogels at 10% (w / v) concentration. The linear modulus plateau versus frequency was determined. As the molecular weight of HA-g-PNIPAm increases, the gel stiffness increases.

[0054] Fig.18 (A) FTIR spectra of 10% PNIPAm (w / v) at temperatures of 0°C, 31°C, and 37°C, respectively. (B) FTIR spectra of 10% HA-g-PNIPAm13k (w / v) at temperatures of 0°C, 31°C, and 37°C, respectively. The results indicate that hydrogen bonding plays an important role in the formation of HA-g-PNIPAM hydrogels.

[0055] Fig.19 In vitro release curves of morphine from morphine-loaded HA-g-PNIPAm 13k and 20k. The morphine concentration was 0.468% (w / v). As the molecular weight of HA-g-PNIPAm increased, the release of morphine decreased.

[0056] Fig. 20 .Intra-articular injections in cadaveric dogs. The hydrogel was well distributed in the cranial compartment of the joint. The gel formed sheets most likely in the trochlear groove and on the lateral and medial sides of the condyle.

[0057] Fig.21Effect of grafting density and concentration of HA-g-PNIPAm on (A) gelation temperature and (B) gelation time. Theoretical degree of substitution (DS): at a concentration of 10% (w / v), S5 is 100%, S4 is 80%, S3 is 64%, S2 is 40%, and S1 is 30%. S6: at 100% DS and at a concentration of 15%.

[0058] Fig. 22 . Among the adjustable amine groups in HA-g-PNIPAm 17k, structures No.5, No.2, No.6, and No.4 have PNIPAm DS of 100%, 40%, 50%, and 80%, respectively.

[0059] Fig.23 Injectability and degradability of HA-g-PNIPAm hydrogels using a rat model (at a concentration of 15% (w / v)). Photo images show (A) Intra-articular injection into rats. (B) Hydrogel formation immediately after injection. (C) Hydrogel degradation 21 days after injection.

[0060] Fig.24 In vivo fluorescence imaging of the release of Alexa Fluor 680-conjugated BSA from HA-g-PNIPAm hydrogels. (n=4)

[0061] Fig.25 Total radiation efficiency emitted from the knee joint as a function of time. BSA conjugated with Alexa Fluor 680 (BSA-AF) released from HA-g-PNIPAm hydrogels showed maximum concentrations 6-12 hours after intra-articular injection and displayed a sustained release profile, and BSA-AF was detected in the knee joint for more than 24 hours.

[0062] Fig.26 . Ex vivo fluorescence imaging of different tissues 72 hours after injection. DETAILED DESCRIPTION

[0063] For the purposes of promoting an understanding of the principles of the invention, reference will now be made to the embodiments illustrated in the drawings and specific language will be used to describe these embodiments. It will be understood, however, that no limitation of the scope of the invention is intended thereby.

[0064] Although the concepts of the present invention are illustrated and described in detail in the drawings and specification of the present invention, the results in the drawings and their description are considered to be exemplary rather than restrictive; it should be understood that only exemplary embodiments are shown and described, and all changes and modifications that fall within the spirit of the present invention are desired to be protected.

[0065] Unless defined otherwise, scientific and technical terms have the same meanings as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0066] As used herein, the "degree of substitution" (DS) of a substituent (e.g., PNIPAm or an amine group) is the (average) number of substituents (e.g., PNIPAm or an amine group) attached to the total monomer units of the polymer HA. For example, the theoretical DS of HA is equal to (the number of substituents) / (the number of monomer units of the polymer HA). Since each monomer unit of the polymer HA contains one -COOH, DS = (the number of substituents) / (the number of -COOH).

[0067] In response to the unmet need, the present invention discloses the development of responsive, highly elastic injectable hydrogels comprising functionalized hyaluronic acid (HA).

[0068] HA is an immunoneutral polysaccharide composed of alternating disaccharide units linked by [β(1,4)-D-glucuronic acid-β(1,3)-N-acetyl-D-glucosamine] 5 ]. HA is the only non-sulfated glycosaminoglycan that is widely distributed throughout the body, particularly in the synovium of joints, the vitreous humor of the eye, and the dermis of the skin. HA is primarily located in the extracellular and pericellular matrix. Functionally, HA contributes to the elasticity and viscosity of fluid connective tissues, including synovial and vitreous humor, and regulates hydration and water transport through tissues.[ 6 ]. The enzymatic degradation of HA results from the action of three enzymes: hyaluronidase, β-glucuronidase, and β-n-acetyl-hexosaminidase, which explains the biodegradable nature of HA-derived hydrogels.[ 7 ]. In addition, HA has been used clinically for more than 30 years[ 8 , 9 ]. For example, it has been approved by the U.S. Food and Drug Administration for the treatment of osteoarthritis in humans since 1997.[ 10 ].

[0069] Cross-linked HA-based polymer biomaterials are widely used as drug delivery vehicles due to the major advantages of HA, including: (i) biodegradability and biocompatibility; (ii) ease of chemical modification due to the abundance of carboxylic acid and hydroxyl groups; (iii) high potential drug loading; (iv) its intrinsic targeting properties due to selective interactions with receptors such as CD44, Toll2, Toll4, RAAMM receptors or hyaluronic acid receptors for endocytosis; (vi) HA degraded from HA-based polymer biomaterials can be used as a lubricant and shock absorber in joints. HA can stabilize joint function because of its lubricating properties at low shear and increased friction at high shear. The HA thin layer acts as a shock absorber between the cartilage and cartilage / meniscus surfaces; and (vii) HA coats pain receptors to prevent binding to peptide agonists [ 11 , 12].

[0070] Cross-linked HA-based polymer biomaterials can be used for tissue engineering, in part due to their ability to effectively encapsulate cells. Mechanical and structural properties can be controlled by varying the cross-link density, which controls network pore size, water content, mechanical properties, and cell-material interactions. In some cases, cross-linked polymers or gels can have a high tissue-like water content, which can allow nutrient and waste transport. HA has been used in tissue integration, including wound healing. 13 ], cell adhesion and proliferation[ 14 ], cell motility, angiogenesis, cell signaling, and matrix organization[ 9 ], has many useful biological properties.

[0071] According to at least one embodiment, two main strategies can be followed to prepare HA-based polymer matrices: (i) 'grafting to' and (ii) 'grafting from' strategies, e.g. Figure 1 shown.

[0072] The “grafting onto” strategy involves the attachment of a preformed polymer via physical adsorption or covalent bond formation (chemisorption). 15 ]. HA can be modified in a variety of ways to change the properties of the resulting material, including modifications that result in hydrophobicity and bioactivity

[10] . In part, the present invention provides a composition comprising at least one HA monomer unit functionalized with at least one functional group moiety. Chemical modification of HA can target three functional groups: glucuronic acid carboxylic acid, primary and secondary hydroxyl groups, and N-acetyl (after deamidation). In some embodiments, a HA composition of the present invention is provided, which can be represented by Formulas I, II, III and IV ( Figure 2 ).

[0073] The carboxylates in the HA backbone can be modified by carbodiimide-mediated reactions, esterification, and amidation. The hydroxyl groups in the HA backbone can be modified by etherification cations, divinyl sulfone crosslinking, esterification, and diepoxide crosslinking. Additionally, conversion of diols to aldehydes can be achieved by periodate oxidation of HA. Finally, deacetylation of the N-acetyl groups of HA restores the amino groups, which can then react with acids using the same amidation.

[0074] The functional groups R1, R2, R3, R4 and R5 include any one or combination of the following: haloacetate, dihydrazide, amine, thiol, carboxylic acid, aldehyde, ketone, active hydrogen site on an aromatic ring, diene, azidoisothiocyanate, isocyanate, acyl azide, N-hydroxysuccinimide (NHS) ester, sulfo-NHS, sulfonyl chloride, epoxide, carbonate, aryl halide, imidate, carbodiimide (e.g., N,N'-dicyclohexylcarbodiimide (DCC) and 1-ethyl-3-(3-hydroxy-2-nitropropene) carbodiimide (DCC)). -dimethylaminopropyl)carbodiimide (EDC)), alkyl phosphate compounds, anhydrides, fluorophenyl esters, hydroxymethyl phosphine, guanidine, iodoacetyl derivatives, maleimide, aziridine, acryloyl derivatives, arylating agents, disulfide derivatives, vinyl sulfone, phenyl thioester, cisplatin, diazoacetate, carbonyldiimidazole, oxirane, N,N'-disuccinimidyl carbonate, N-hydroxysuccinimidyl chloroformate, alkyl halides, hydrazine, maleimide, alkyne and phosphorus-bound chlorine.

[0075] Telechelic polymers can be synthetic polymers and natural polymers. Synthetic polymers include any one or combination of the following: poly(aliphatic esters) (e.g., poly(lactide) (PLA), poly(ε-caprolactone) (PCL), poly(glycolic acid) (PGA), poly(lactic acid-co-glycolic acid) (PLGA), poly(trimethylene carbonate) (PTMC), polydioxanone (PDS), poly(orthoesters), polyanhydrides, poly(anhydride-co-imides), poly(anhydride-esters), polyurethanes (e.g., degrapol), poly(amides), poly(esteramides), poly(orthoesters), poly(dioxanone), poly(acetals), poly(ketals), poly(carbonates), poly(orthocarbonates), poly(hydroxybutyrates), poly(hydroxy-valerates), poly(alkylene oxalates), poly(alkylene succinates), poly(malic acid), poly(amino acids), poly(vinyl pyrrolidone), poly(hydroxycellulose), poly(glycerol sebacic acid), esters), poly(ethylene imine), poly(acrylic acid) (PAA), poly(methacrylate N,N′-diethylaminoethyl ester, polyethylene glycol (PEG), poly(propylene oxide) (PPO), PEO-PPO block copolymers (e.g., pluronic (or poloxamer), and tetronic), poly(vinyl alcohol) (PVA), poly(N-isopropylacrylamide) (PNIPAm), poly(N,N-diethylacrylamide) (PDEAAm), poly(oxazolines) (e.g., poly(2-methyloxazoline and poly(2-ethyl-2-oxazoline), oligo(ethylene glycol) fumarate (OPF), poly(propylene fumarate), poly(alkyl cyanoacrylate), poly(acrylamide), synthetic poly(amino acids) (e.g., poly(L-glutamic acid) (L-PGA) and poly(aspartic acid)), polyphosphazenes, poly(phosphates), and blends thereof.

[0076] Synthetic telechelic polymers can be prepared by conventional methods, such as bulk polymerization, solution (or homogeneous) polymerization, suspension polymerization, emulsion polymerization, radiation polymerization (using gamma rays, electron beams, etc.). Synthetic telechelic polymers can be prepared by addition or chain growth polymerization, coordination polymerization, condensation or step growth polymerization. Addition or chain growth polymerization includes free radical polymerization, controlled living radical polymerization (such as atom transfer radical polymerization (ATRP), reversible addition fragmentation transfer (RAFT) polymerization, and nitroxide-mediated radical polymerization (NMP)), cationic polymerization, anionic polymerization, etc.

[0077] Natural polymers include any one or combination of the following: fibrin, collagen, matrigel, elastin, elastin-like peptides, albumin, natural poly(amino acids) (e.g., phycocyanin, poly(ε-l-lysine), poly(γ-glutamic acid)), polysaccharides (e.g., chitosan, dextran, chondroitin sulfate, agarose, alginate, methylcellulose and heparin), and blends thereof.

[0078] Conjugation of functional HA and telechelic polymers can be achieved through carbodiimide-mediated reactions, esterification, amidation, aldehyde and ketone reactions, active hydrogen reactions, photochemical reactions, azide-alkyne cycloadditions (e.g., copper-catalyzed azide-alkyne cycloaddition (CuAAC), copper-free azide-alkyne huisgen cycloaddition), thiol-click reactions, Diels-Alder reactions, nitrile oxide cycloadditions, and enzymatic cross-linking strategies (e.g., horseradish peroxidase and hydrogen peroxide).

[0079] Conjugated linkages include any one or combination of the following: isothiourea, isourea, amide, sulfonamide, Schiff base, secondary amine, carbamate, arylamine, amidine, phosphoramidate, thioether, disulfide, β-thiosulfonyl, ester, carbamate, hydrazone, diazo, triazole, carbohydrate, and amino acid ester bonds.

[0080] Alternatively, cross-linked HA-based polymer matrices can be prepared by a 'grafting from' approach ( Figure 1 ). The 'grafting from' method involves preparing a precursor of the backbone polymer using a monomer unit containing a functional group that is ultimately capable of initiating polymerization of a second monomer. 15 ]. In this embodiment, at least one monomer unit of HA is conjugated to at least one of a polymerizable moiety, an initiator, a RAFT agent, and an iniferter.

[0081] The polymerizable part is included in any part that can be polymerized when exposed to a polymerization initiator. The polymerizable part can include an alkenyl part, such as acrylate, methacrylate, dimethacrylate, oligoacrylate, oligomeric methacrylate, ethyl acrylate, itaconate acrylamide, aldehyde, ethylenically unsaturated monomers. Ethylenically unsaturated monomers can include acrylic or methacrylic alkyl esters, nitrile and amide of the same acid, unsaturated monomers containing carboxylic acid groups and polyethylenically unsaturated monomers. The example of acrylic or methacrylic alkyl esters is methyl methacrylate, ethyl methacrylate, butyl methacrylate, ethyl acrylate, butyl acrylate, hexyl acrylate, n-octyl acrylate, lauryl methacrylate, 2-ethylhexyl methacrylate, nonyl acrylate, benzyl methacrylate, hydroxyalkyl esters of the same acid (such as 2-hydroxyethyl acrylate, 2-hydroxyethyl methacrylate and 2-hydroxypropyl methacrylate). Examples of nitriles and amides of the same acid are acrylonitrile, methacrylonitrile and methacrylamide, vinyl acetate, vinyl propionate, vinylidene chloride, vinyl chloride and vinyl aromatic compounds such as styrene, tert-butyl styrene and vinyl toluene, dialkyl maleates, dialkyl itaconic acid esters, dialkyl methylene malonates, isoprene and butadiene. Examples of unsaturated monomers containing carboxylic acid groups include acrylic acid, methacrylic acid, ethacrylic acid, itaconic acid, maleic acid, fumaric acid, monoalkyl itaconic acid. Examples of polyethylenically unsaturated monomers include butadiene, isoprene, allyl methacrylate, diacrylates of alkyl glycols (e.g., dibutylene glycol diacrylate and dihexanediol diacrylate, and divinylbenzene).

[0082] RAFT agent moieties include any moiety that is capable of capturing a growing polymer radical and releasing polymer fragments as radicals to achieve highly controlled polymerization [ 16 ] The RAFT agent portion may include dithiobenzoates (e.g., cumyl dithiobenzoate, cyanopentanoate), trithiocarbonates (e.g., 4-cyano-4-(dodecylsulfanylthiocarbonyl)sulfanylpentanoic acid, phthalimidomethyl trithiocarbonate, S-1-dodecyl-S'-(α,α')-dimethyl-α"-acetic acid) trithiocarbonate (DATC), 3,5-bis(2-dodecylsulfanylthioformylthio-1-oxopropoxy)benzoic acid, 4-cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl]pentanol, 4-cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl]pentanoic acid) and xanthates.

[0083] The ATRP initiator moiety includes any moiety capable of initiating polymerization during the ATRP process. 17Examples of ATRP initiator moieties include, but are not limited to, 2-bromopropionitrile (BPN), ethyl 2-bromoisobutyrate (BriB), ethyl 2-bromopropionate (EBrP), methyl 2-bromopropionate, 1-phenylethyl bromide (1-PEBr), toluenesulfonyl chloride (TsCl), 1-cyano-1-methylethyldiethyldithiocarbamate (MANDC), ethyl 2-(N,N-diethyldithiocarbamoyl)-isobutyrate (EMADC), and dimethyl 2,6-dibromoheptanedioate (DMDBHD).

[0084] NMP iniferters include, but are not limited to, 2,2,6,6-tetramethylpiperidinyloxy (TEMPO) and TEMPO-based derivatives. TEMPO-based derivatives include, but are not limited to, 4-acetamido-TEMPO, 4-acetamido-2,2,6,6-tetramethylpiperidinyl 1-oxypurine (purum), 4-amino-TEMPO, 2-azaadamantane-N-oxyl, 4-(2-bromoacetamido)-TEMPO, 4-carboxyl-TEMPO, 4-cyano-TEMPO, 4-hydroxy-TEMPOpurine, 4-hydroxy-TEMPOpurine, 4-hydroxy-TEMPObenzoate, 4-(2-iodoacetamido)-TEMPO, 4-isothiocyanate-TEMPO, 4-maleimido-TEMPO, 4-methoxy-TEMPO, 4-oxo-TEMPO, 4-phosphonooxy-TEMPO hydrate, 2,2,6,6-tetramethyl-4-(methylsulfonyloxy)-1-piperidinyloxy.

[0085] The polymerization reaction of the present invention can be carried out by conventional methods, such as bulk polymerization, solution polymerization, suspension polymerization, emulsion polymerization, radiation polymerization (using gamma rays, electron beams, etc.), free radical polymerization, controlled living free radical polymerization (such as ATRP, RAFT polymerization and NMP), photopolymerization, ring-opening polymerization and step-growth polymerization, etc.

[0086] Polymerization initiators include, but are not limited to, electromechanical radiation, thermal initiators, redox initiators, and photoinitiators. Initiation of polymerization can be accomplished by irradiation with light having a wavelength between about 200 and about 700 nm. Redox initiators can be one or a combination of these initiators, tetramethylethylene, ferrous salts, sodium bisulfite, or similar reducing agents. Examples of useful photoinitiators include 2,2-dimethoxy-2-phenylacetophenone or ethyl eosin (10 -4 Up to 10 -2The invention relates to a combination of 4,4-azobis(4-cyanovaleric acid), benzoyl peroxide, azobisisobutyronitrile (AIBN), di-tert-butyl peroxide, etc. Such a system initiates free radical polymerization at physiological temperature and includes, for example, potassium persulfate in the presence or absence of tetramethylethylenediamine; benzoyl peroxide in the presence or absence of triethanolamine; ammonium persulfate in the presence of sodium bisulfite.

[0087] HA-based drug delivery platforms have been developed by encapsulating anesthetics, analgesics, and antibiotics within polymer matrix systems through physical interactions or chemical interactions (e.g., polymer-drug conjugation).

[0088] Physical interactions include any one or a combination of the following: hydrophobic interactions, hydrophilic interactions, hydrogen bonding, and intermolecular electrostatic interactions.

[0089] The polymer-therapeutic conjugation reactions include any one or combination of the following: amine reactions, thiol reactions (e.g., thiol-olefin click reactions, Michael additions), carboxylate reactions, hydroxyl reactions, aldehyde and ketone reactions, active hydrogen reactions, photochemical reactions, and cycloaddition reactions (e.g., Diels-Alder reactions, CuAAC, copper-free azide-alkyne huisgen cycloadditions).

[0090] The conjugated linkers include any one or combination of the following: isothiourea, isourea, amide, sulfonamide, Schiff base, secondary amine, carbamate, arylamine, amidine, phosphoramidate, thioether, disulfide, β-thiosulfonyl, ester, carbamate, hydrazone, diazo, 2+4 cycloaddition, 1,2,3-triazole, carbohydrate and amino acid ester bonds.

[0091] Embodiment:

[0092] Example 1

[0093] Material

[0094] Hyaluronic acid sodium salt (HA) was purchased from Carbosynth Limited (Berkshire, UK). N-isopropylacrylamide (NIPAm) was purchased from Aldrich and purified by recrystallization in hexane (3:1) before use. S-1-Dodecyl-S'-(α,α'-dimethyl-α"-acetic acid) trithiocarbonate (DATC) was prepared according to the literature [ 18 ]synthesis.

[0095] Example 2

[0096] HA-g-PNIPAm( Figure 3 PNIPAm was selected based on its biocompatibility [ 19 ] and thermoresponsive phase transition properties (i.e., the hydrophilic ring-hydrophobic sphere transition occurs at around 32°C)[ 20 , 21 ]. The combination of RAFT polymerization to prepare polymers of well-defined molecular weight and the efficient coupling mechanism of thiol-olefin “click” chemistry allows for the highly controlled formation of HA hydrogels with different physical properties. Advantages of HA-g-PNIPAm such as Figure 4 shown.

[0097] (1) Synthesis of HA-ADH. HA (100 mg) was dissolved in 20 mL of water to prepare a 5 mg / mL HA solution. A 40-fold molar excess of solid adipic acid dihydrazide (ADH) (1.736 g) was added to the solution and completely dissolved by mixing for 10 minutes. The pH of the mixed solution was adjusted to 4.8 by adding 1.0 N HCl. Thereafter, a four-fold molar excess of solid EDC (0.191 g) was added. The pH of the mixed solution was maintained at 4.8 by adding 1.0 N HCl. The reaction was terminated by raising the pH of the reaction solution to 7.0 with 1.0 N NaOH. The reaction solution was dialyzed against a large excess of 100 mM NaCl solution and then dialyzed against 25 vol% ethanol and deionized water using a dialysis membrane (MWCO, 12-14 kDa). The resulting solution was finally freeze-dried for 3 days. 13 ].

[0098] (2) Synthesis of HA-NAS. 100 mg HA-ADH was dissolved in 20 mL distilled water. N-acryloyloxysuccinimide (NAS) (0.5 g, 3 mmol) was then added to the HA-ADH solution. The reaction was continued for 12 hours by stirring at room temperature. Using a dialysis membrane (MWCO, 12-14 kDa), HA-NAS was thoroughly dialyzed against a 100 mM NaCl solution, and then dialyzed against 25% by volume ethanol and deionized water. The product was then freeze-dried for 3 days to obtain solid acrylated HA (HA-NAS).

[0099] (3) Carboxyl-terminated PNIPAm was synthesized by RAFT polymerization. A mixture of NIPAm (3.0 g, 94 mmol), DATC (0.1000 g, 0.35 mmol), AIBN (10.0 mg, 0.0625 mmol) and DMF (5.0 mL) was placed in a 10 mL polymerization tube. After removing oxygen by purging argon, the sealed tube was immersed in a temperature-controlled oil bath maintained at 60 °C and stirred for 24 hours. After stopping the heating, the reaction mixture was dissolved with THF and then precipitated in 10 times diethyl ether. The polymer was collected by filtration and dried in a vacuum oven at 40 °C. 21 ].

[0100] (4) Aminolysis of PNIPAm: A THF solution of PNIPAm and hexylamine was reacted at room temperature overnight, and the reaction mixture was precipitated from hexane three times to obtain an aminolysis product, thiolated PNIPAm (PNIPAm-SH).

[0101] (5) HA-NAS was conjugated to PNIPAm-SH via a thiol-olefin click reaction. PNIPAm-SH and HA-NAS were then dissolved in deionized (DI) water. After stirring overnight, the resulting solution was purified by dialysis against DI water (MWCO 50 kDa). The product (HA-g-PNIPAm) was then recovered by freeze drying as a white powder. The product was analyzed by Fourier transform infrared spectroscopy (FTIR) ( Figure 6 )and 1 H nuclear magnetic resonance (NMR) spectrum ( Figure 7 ) confirmed the chemical structure of the product in each step. NMR spectra were obtained on a Bruker ARX400 MHz. D2O was used as the solvent for all samples and the reported spectra represent an average of 64 scans.

[0102] Example 3

[0103] HA-g-PNIPAm( Figure 4 ).

[0104] (1) Synthesis of a macromolecular RAFT agent based on HA. In order to make hyaluronic acid soluble in DMSO, the sodium ions of HA were exchanged with lipophilic tetrabutylammonium (TBA). An aqueous solution (1 L) of HANa (10 mg / mL) was subjected to ion exchange column chromatography (Dowex 50w×8 [H+], Dow Chemicals, Midland, MI) to obtain an aqueous solution (1.5 L) of HA. 22 ]. Next, tetrabutylammonium bromide was added to 1000 mL of 1% (w / w) HA aqueous solution and mixed at room temperature for 2 hours. The mixture was centrifuged at 5000 rpm for 2 minutes to remove the resin. The obtained HA-TBA solution was lyophilized. 1 H NMR confirmed the successful synthesis of HA-TBA. A 1% (w / v) HA-TBA solution in DMSO (100 mL) was prepared at 50°C under a nitrogen atmosphere. Subsequently, 0.2 g DMAP and calculated amounts of DATC and DCC were added according to the desired degree of substitution (DS). The solution was stirred at 50°C for 48 hours. HA-DATC was obtained by precipitation in ether three times. The successful synthesis of HA-DATC was confirmed by UV-vis.

[0105] (2) RAFT polymerization. The polymerization conditions for synthesizing HA-g-PNIPAm nanocomposites are as follows: HA-DATC:AIBN:PNIPAm=1:0.2:500 and reaction mixture. In detail, 80 mg HA-DATC, 1 g NIPAm, 2 mL anhydrous DMSO and 0.55 mg AIBN were added to a Schlenk flask. The reaction mixture was degassed by four freeze-pump-thaw cycles and then placed in an oscillator at 70°C. After the reaction time, the polymerization reaction was terminated by liquid nitrogen cooling, and the reaction mixture was precipitated in 10 times cold ether and dried in vacuo at 40°C. The conversion rate was determined by gravimetric analysis.

[0106] Example 4

[0107] Hydrogel formation and gelation time. Briefly, HA-g-PNIPAm solutions (1-20 w / v%) at room temperature were quickly placed in a 37°C water bath. The time to form a gel (expressed as gelation time) was defined as the time when the gel had no fluidity in an inverted state for 1 min [ 23 The experiment was performed in triplicate. Table 1 shows the gelation temperature and gelation time of HA-g-PNIPAm of various concentrations and molecular weights.

[0108] Table 1. Gelation temperature and time of HA-g-PNIPAm hydrogels.

[0109]

[0110] Example 5

[0111] Rheological Characterization. Rheological experiments were performed in oscillating mode at 37°C using the new Discovery Series Hybrid Rheometer (DHR)-3 (TA) using a parallel plate (20 mm diameter, 0°C) configuration. Strain sweeps and frequency sweeps were performed at 37°C using 20 mm parallel plate geometry. Time sweeps were performed to determine the gelation time of the hydrogels. Each hydrogel sample was used for only one test. Each test was performed in triplicate and the data represent the average of three tests with the corresponding standard deviation. The test times for determining the gelation time and modulus are very short, so there is no need to use a humidified chamber or trap to perform these experiments.

[0112] HA-g-PNIPAm is soluble in water at room temperature but rapidly forms hydrogels within its concentration range (5-15% w / v) at physiological temperature, as demonstrated by the reverse method ( Figure 8 and Fig. 9 ). Hydrogels are able to maintain their elasticity and shape when compressed ( Fig.10 ). Rheological test( Fig.11) showed that the increase in HA-g-PNIPAm concentration significantly increased the hydrogel stiffness. Thus, the elastic moduli of 15% and 10% HA-g-PNIPAm hydrogels were ~42 kPa and ~32 kPa, respectively, indicating that their high elastic properties were due to the double hydrogen bond network within the hydrogel ( Fig.12 ).

[0113] Example 6

[0114] In vitro drug release

[0115] In vitro morphine release was evaluated in a membrane-based diffusion system. Morphine and HA-g-PNIPAm with different concentrations (5-15% w / v) were prepared and loaded into a 3mL syringe. The hydrogel was balanced in a 37°C incubator for 10 minutes. The hydrogel was then distributed into a cell culture insert (12mm diameter, 3μm pore size (Corning Incorporated, USA)) in a 12-well plate. The hydrogel was then submerged with PBS and the well plate was placed in a 37°C water bath. At specified time intervals, 1mL of solution was taken out from each well and replaced with preheated water. The morphine concentration from the buffer solution was measured at 263nm by UV spectrophotometry.

[0116] A sustained morphine release profile was observed over 48 h due to the diffusion of morphine from the morphine-loaded HA-g-PNIPAm hydrogels ( Fig.13 ). Our hydrogel system also showed the feasibility of tuning the release profile simply by adjusting the HA-g-PNIPAm concentration: after 48 h, 15% HA-g-PNIPAM hydrogel had the slowest distribution with ~70% cumulative release; 10% HA-g-PNIPAM hydrogel showed an intermediate release profile with ~80% cumulative release. This result is consistent with the conclusion drawn from the rheological data. In another in vitro study, the hydrogel supported the cell growth of chondrocytes and bone marrow-derived stem cells with high viability during culture, which means that our hydrogel is cytocompatible.

[0117] Example 7

[0118] In vivo drug release via subcutaneous injection

[0119] To evaluate the effectiveness of our hydrogel delivery system for analgesia, a conscious rat model was used, which allows for quantification of morphine release in plasma as well as evaluation of the biocompatibility of the hydrogel. The specific protocol for the in vivo efficacy evaluation was designed as follows:

[0120] (1) Animal population. Rats (sample size n = 10) were enrolled in the study. The study was randomized and blinded in preclinical trials. Each Sprague Dawley rat was assigned to 1 of 3 treatment groups, including morphine-loaded hydrogel, positive control (morphine, subcutaneous), negative control (saline / hydrogel).

[0121] (2) Surgical procedures. Surgery was performed on each rat under sterile conditions. Isoflurane (3-5%) was used in an anesthesia chamber, and isoflurane (1.5-3%) and mask were used for induction and maintenance. The hair on the back and ventral neck was shaved, scrubbed with Nolvasan, and replaced with alcohol. Both sides of the flank were shaved in a 1x1 square. In our preliminary study, hydrogel pockets were visible after 3 days, confirming that hydrogels were formed by subcutaneous injection of macromolecular solutions by syringe, and allowed sustained release over several days ( Fig.14 ).

[0122] (3) Pharmacokinetic study. 24 hours after carotid cannulation, blood samples were collected from the subcutaneous port at designed time points using a carotid artery placement method in an awake rat model. The decanted plasma was extracted by SPE. In addition, an HPLC-mass spectrometry (HLPC-MS) method was identified for the reliable determination of plasma levels of morphine in rats. Prior to HPLC-MS, analytes and internal standards (morphine-d3) were extracted from plasma samples by a single solid phase extraction (SPE) method. Our standard calibration curve was linear in the range of 10-1000 ng / mL (r=0.999). The in vivo drug release of morphine from HA-g-PNIPAm hydrogel was shown in Fig.15 In vivo studies have shown that HA-g-PNIPAm hydrogels enable sustained release of morphine at concentrations exceeding therapeutic plasma concentrations (10 ng / mL) [ 24 ] Up to 48h( Fig.15 ).

[0123] (4) Statistical analysis. Descriptive statistics and data normality tests were performed using the Shapiro Wilk test. Depending on the normality of the data, ANOVA (if the data were normal) or the Kruskal Wallis ANOVA statistical test (if non-parametric) was used to compare the treatment groups. Significance was set at P < 0.05.

[0124] Example 8

[0125] In vivo drug release via intra-articular injection

[0126] In vivo studies of the novel hydrogel loaded with morphine were conducted in a canine animal model to determine the effectiveness of the system in pain relief studies and the duration of analgesia in dogs and possibly men. A schematic diagram of the injectable hydrogel for intra-articular delivery of analgesics or anesthetics is shown in Fig.16 The specific design of the in vivo efficacy evaluation is as follows [ 25 ]:

[0127] (1) Animals. Client-owned dogs with anterior cruciate ligament rupture will participate in the study with informed consent from the owners.

[0128] (2) Experimental Design. The study design is a prospective, randomized, and blinded clinical trial. A random number table will be used to assign dogs to 1 of 4 treatment groups for intra-articular administration of the following drugs: saline solution (control group), morphine group (control group), hydrogel only (control group), and morphine-loaded hydrogel (treatment group).

[0129] (3) Procedure. Prior to surgery, all dogs will be premedicated with hydromorphone (0.1 mg / kg IM) and acepromazine (0.01 mg / kg IM), and anesthesia will be induced with propofol (4-6 mg / kg, IV) and maintained with isoflurane (1-2.5% to produce effect) in 100% oxygen. Conventional tibial tuberosity advancement (TTA) or tibial plateau corrective osteotomy (TPLO) via a medial skin and arthrotomy approach will be performed by a board-certified veterinarian or surgical resident experienced in these techniques.

[0130] (4) Pain assessment and scoring. All dogs will be assessed for signs of pain using modified criteria adopted from two pain scoring systems, the Dynamic and Interactive Visual Analog Scale for Soft Tissue Surgery (DIVAS) and the Multifactorial Pain Score (MPS) developed for hind knee arthrotomy studies. Pain score assessments will be performed at 2, 4, 6, 8, 10, 12, 16, 20, and 24 hours after intra-articular injection by a trained investigator who is blinded to the intra-articular formulation used.

[0131] Rescue Parameters. Dogs will be given systemic rescue analgesia if the scoring system exceeds: 70 mm (for DIVAS); 6 on the Glascow Scale composite pain scale at the time of assessment.

[0132] Additional disclosure is found in Appendix A, filed herein, the entire contents of which are incorporated herein by reference.

[0133] Those skilled in the art will recognize that various modifications may be made to the above specific embodiments. Implementations should not be limited to the specific limitations described. Other implementations are also possible. In addition, all references cited in the present invention indicate the state of the art and are incorporated herein by reference as a whole.

[0134] In summary, the present invention relates to the following aspects:

[0135] 14. A composition comprising a polymer matrix comprising at least 100 monomer units of functionalized hyaluronic acid (HA) cross-linked with at least one unit of a telechelic polymer.

[0136] 15. A composition according to claim 1, wherein the functionalized hyaluronic acid comprises formula I-IV:

[0137]

[0138] Wherein R1, R2, R3, R4 and R5 can include any one or combination of the following: haloacetate, dihydrazide, amine, thiol, carboxylic acid, aldehyde, ketone, active hydrogen site on aromatic ring, diene, azidoisothiocyanate, isocyanate, acyl azide, N-hydroxysuccinimide (NHS) ester, sulfo-NHS, sulfonyl chloride, epoxide, carbonate, aryl halide, imidate, carbodiimide (for example, N, N'-dicyclohexylcarbodiimide (DCC) and 1-ethyl-3-hydroxy-4-nitrocarbodiimide (3-nitrocarbodiimide (4-nitrocarbodiimide (5-nitrocarbodiimide (6-nitrocarbodiimide (7-nitrocarbodiimide (8-nitrocarbodiimide (9-nitrocarbodiimide ( -(3-dimethylaminopropyl)carbodiimide (EDC)), alkyl phosphate compounds, anhydrides, fluorophenyl esters, hydroxymethylphosphines, guanidines, iodoacetyl derivatives, maleimides, aziridines, acryloyl derivatives, arylating agents, disulfide derivatives, vinyl sulfones, phenyl thioesters, cisplatin, diazoacetates, carbonyldiimidazoles, oxirane, N,N'-disuccinimidyl carbonate, N-hydroxysuccinimidyl chloroformate, alkyl halides, hydrazines, alkynes and chlorine bound to phosphorus.

[0139] 16. A composition according to claim 1, wherein the telechelic polymer is selected from any one or a combination of the following: poly(aliphatic esters) (e.g., poly(lactide) (PLA), poly(ε-caprolactone) (PCL), poly(glycolic acid) (PGA), poly(lactic acid-co-glycolic acid) (PLGA), poly(trimethylene carbonate) (PTMC), polydioxanone (PDS), poly(orthoesters), polyanhydrides, poly(anhydride-co-imides), poly(anhydride-esters), polyurethanes (e.g., degrapol), poly(amides), poly(esteramides), poly(orthoesters), poly(dioxanone), poly(acetals), poly(ketals), poly(carbonates), poly(orthocarbonates), poly(hydroxybutyrates), poly(hydroxy-valerates), poly(alkylene oxalates), poly(alkylene succinates), poly(malic acid), poly(amino acids), poly(vinyl pyrrolidone), poly(hydroxy cellulose), poly(glycerol sebacate), poly(ethyleneimine), poly(acrylic acid) (PAA), poly(N,N′-diethylaminoethyl methacrylate), polyethylene glycol (PEG), poly(propylene oxide) (PPO), PEO-PPO block copolymers (e.g., pluronic (or poloxamer), and tetronic), poly(vinyl alcohol) (PVA), poly(N-isopropylacrylamide) (PNIPAm), poly(N,N-diethylacrylamide) (PDEAAm), poly(oxazolines) (e.g., poly(2-methyloxazoline and poly(2-ethyl-2-oxazoline), oligo(ethylene glycol) fumarate (OPF), poly(propylene fumarate), poly(alkyl cyanoacrylate), poly(acrylamide), synthetic poly(amino acids) (e.g., poly(L-glutamic acid) (L-PGA) and poly(aspartic acid)), polyphosphazenes, and poly(phosphates).

[0140] 17. A composition according to item 1, wherein the telechelic polymer is selected from any one or combination of the following: fibrin, collagen, matrigel, elastin, elastin-like peptides, albumin, natural poly (amino acids) (e.g., phycocyanin, poly (ε-l-lysine) poly (γ-glutamic acid)), polysaccharides (e.g., chitosan, dextran, chondroitin sulfate, agarose, alginate, methylcellulose and heparin).

[0141] 18. The composition of claim 1, wherein the functionalized hyaluronic acid comprises at least one monomer unit of hyaluronic acid functionalized with an amine group.

[0142] 19. The composition of claim 1, wherein the functionalized hyaluronic acid comprises at least one monomer unit of hyaluronic acid functionalized with an acrylate group.

[0143] 20. The composition of claim 1, wherein the telechelic polymer is thiolated poly(N-isopropylacrylamide) PNIPAm.

[0144] 21. A method for preparing a polymer matrix based on hyaluronic acid (HA), said polymer matrix comprising a functionalized hyaluronic acid having at least 100 monomer units cross-linked with at least one unit of a telechelic polymer, comprising the following steps:

[0145] preparing a functionalized hyaluronic acid having at least 100 monomer units;

[0146] preparing pre-processed functional telechelic polymers;

[0147] The functionalized hyaluronic acid is cross-linked to the functional telechelic polymer via carbodiimide-mediated reactions, esterification, amidation, aldehyde and ketone reactions, active hydrogen reactions, photochemical reactions, azide-alkyne cycloadditions (e.g., copper-catalyzed azide-alkyne cycloadditions (CuAAC), copper-free azide-alkyne huisgen cycloadditions, thiol-click reactions, Diels-Alder reactions, nitrile oxide cycloadditions, and enzymatic cross-linking strategies (e.g., horseradish peroxidase and hydrogen peroxide).

[0148] 22. The method of claim 8, wherein the functional telechelic polymer is thiolated PNIPAm.

[0149] 23. The method of claim 8, wherein the HA-based polymer matrix further comprises at least one monomer unit of a polymer that can be prepared by polymerization reactions of i) 'grafting to' and (ii) 'grafting from' strategies.

[0150] 24. The method of item 9, wherein the cross-linking reaction between the functionalized hyaluronic acid and the thiolated PNIPAm is a thiol-olefin reaction.

[0151] 25. The method of item 10, wherein the 'grafting from' process involves a functionalized hyaluronic acid comprising at least one of a polymerizable moiety, an initiator, a RAFT agent, and an iniferter.

[0152] 26. The method of claim 10, wherein the polymerizable moiety is an acrylate.

[0153] 27. The method of claim 10, wherein the RAFT agent is S-1-dodecyl-S'-(α,α'-dimethyl-α"-acetic acid) trithiocarbonate (DATC).

[0154] 28. The method of item 10, wherein the polymerization reaction is a 'graft from' process using RAFT polymerization of PNIPAm.

[0155] 29. A polymer-based drug delivery platform comprising the composition of item 1, wherein the drug delivery platform further encapsulates anesthetics, analgesics or antibiotics with a polymer matrix system through physical interaction or chemical interaction.

[0156] 30. The platform of claim 16, wherein the physical interaction comprises any one or combination of the following: hydrophobic interaction, hydrophilic interaction, hydrogen bonding, and intermolecular electrostatic interaction.

[0157] 31. The platform of claim 16, comprising any one of: an opioid (eg, morphine) or a nonsteroidal anti-inflammatory drug (NSAID), the nonsteroidal anti-inflammatory drugs including chloroprocaine, bupivacaine, lidocaine, and procaine.

[0158] 32. A method for controlled drug delivery, comprising:

[0159] preparing a polymer matrix comprising a functionalized hyaluronic acid (HA) having at least 100 monomer units cross-linked with a telechelic polymer having a theoretical degree of substitution of 100% to 30%;

[0160] preparing a polymer matrix comprising a functionalized hyaluronic acid (HA) having at least 100 monomer units cross-linked with a telechelic polymer having a theoretical degree of substitution of amine groups of 20% to 80%;

[0161] preparing a conjugate of the drug in the polymer matrix at a concentration of 0 to 30% (w / v); and

[0162] It was observed that the release rate of the drug decreased as the molecular weight of the polymer matrix increased.

[0163] 33. The method of claim 19, wherein the polymer matrix is ​​HA-g-PNIPAm and the drug is morphine.

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Claims

1. A composition comprising a polymer matrix comprising: At least 100 monomer units of functionalized hyaluronic acid (HA), each unit comprising a first functional amino group having a theoretical degree of substitution of 20% to 80% per HA monomer unit, thereby producing at least one unreacted amino group, and a second functional group comprising a polymerizable acrylate moiety; and at least one unit of a telechelic polymer comprising a functional group reactive toward said second functional group of HA, wherein The functionalized hyaluronic acid and the telechelic polymer are linked via a thioether bond formed between the second functional group of HA and the telechelic polymer, and The telechelic polymer is thiolated poly(N-isopropylacrylamide) (PNIPAm).

2. The composition according to claim 1, wherein the functionalized hyaluronic acid comprises Formula I-IV: Wherein R1, R2, R3, R4 and R5 can include any one or combination of the following: haloacetate, dihydrazide, amine, thiol, carboxylic acid, aldehyde, ketone, active hydrogen site on aromatic ring, diene, azidoisothiocyanate, isocyanate, acyl azide, N-hydroxysuccinimide (NHS) ester, sulfo-NHS, sulfonyl chloride, epoxide, carbonate, aryl halide, imidate, carbodiimide (for example, N, N'-dicyclohexylcarbodiimide (DCC) and 1-ethyl-3-hydroxy-4-nitrocarbodiimide (3-nitrocarbodiimide (4-nitrocarbodiimide (5-nitrocarbodiimide (6-nitrocarbodiimide (7-nitrocarbodiimide (8-nitrocarbodiimide (9-nitrocarbodiimide ( -(3-dimethylaminopropyl)carbodiimide (EDC)), alkyl phosphate compounds, anhydrides, fluorophenyl esters, hydroxymethylphosphines, guanidines, iodoacetyl derivatives, maleimides, aziridines, acryloyl derivatives, arylating agents, disulfide derivatives, vinyl sulfones, phenyl thioesters, cisplatin, diazoacetates, carbonyldiimidazoles, oxirane, N,N'-disuccinimidyl carbonate, N-hydroxysuccinimidyl chloroformate, alkyl halides, hydrazines, alkynes and chlorine bound to phosphorus.

3. A method for preparing a polymer matrix based on hyaluronic acid (HA), comprising a functionalized hyaluronic acid having at least 100 monomer units cross-linked with at least one unit of a telechelic polymer, comprising the following steps: Preparing a functionalized hyaluronic acid having at least 100 monomer units, each unit comprising a first functional amino group and a second functional group, wherein the first functional amino group has a theoretical degree of substitution of 20% to 80% per HA monomer unit, thereby generating at least one unreacted amino group, and the second functional group comprises a polymerizable acrylate moiety; preparing a pre-processed functional telechelic polymer comprising a functional group reactive toward the second functional group of HA; cross-linking the second functional group of the functionalized hyaluronic acid and the functional telechelic polymer via a thioether bond; The telechelic polymer is thiolated poly(N-isopropylacrylamide) (PNIPAm), and the cross-linking reaction between the functionalized hyaluronic acid and the thiolated PNIPAm is a thiol-olefin reaction.

4. The method of claim 3, wherein the HA-based polymer matrix further comprises at least one monomer unit of a polymer that can be prepared by polymerization reactions of i) 'grafting to' and (ii) 'grafting from' strategies.

5. The method of claim 4, wherein the 'grafting from' process involves a functionalized hyaluronic acid comprising at least one of a polymerizable moiety, an initiator, a RAFT agent, and an iniferter.

6. The method of claim 5, wherein the polymerizable moiety is an acrylate.

7. The method of claim 5, wherein the RAFT agent is S-1-dodecyl-S'-(α,α'-dimethyl-α"-acetic acid) trithiocarbonate (DATC).

8. The method of claim 4, wherein the polymerization reaction is a 'graft from' process using RAFT polymerization of PNIPAm.

9. A polymer-based drug delivery platform comprising the composition of claim 1, wherein the drug delivery platform further encapsulates an anesthetic, analgesic or antibiotic with a polymer matrix system through physical interaction or chemical interaction.

10. The platform of claim 9, wherein the physical interaction comprises any one or a combination of the following: hydrophobic interaction, hydrophilic interaction, hydrogen bonding, and intermolecular electrostatic interaction.

11. The platform of claim 9, comprising any one of the following: an opioid (eg, morphine) or a nonsteroidal anti-inflammatory drug (NSAID), including chloroprocaine, bupivacaine, lidocaine, and procaine.

12. Use of a polymer matrix in the preparation of a drug capable of controlled delivery, comprising: A polymer matrix is ​​prepared, comprising: At least 100 monomer units of functionalized hyaluronic acid (HA), each unit comprising a first functional amino group having a theoretical degree of substitution of 20% to 80% per HA monomer unit, thereby producing at least one unreacted amino group, and a second functional group comprising a polymerizable acrylate moiety; and at least one unit of a telechelic polymer having a theoretical degree of substitution of carboxyl groups of 100% to 30% and comprising a functional group reactive toward the second functional group of HA, wherein The functionalized HA and the telechelic polymer are connected via a thioether bond formed between the second functional group of HA and the telechelic polymer; loading the drug into the polymer matrix at a concentration of 0 to 30% (w / v); and It was observed that the release rate of the drug decreased as the molecular weight of the polymer matrix increased.

13. The use of claim 12, wherein the polymer matrix is ​​HA-g-PNIPAm and the drug is morphine.

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