Functionalized and crosslinked polymers

Abstract

To provide polymers, methods of making the polymers, and methods of using the polymers.SOLUTION: Polyhydric polymers may be converted to derivatives thereof by reaction with divinyl sulfone to provide vinyl sulfone substituted polymers, where the polymers may additionally be further derivatized, e.g., crosslinked, and the crosslinked and non-crosslinked derivatives may be used in biomedical and other applications.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Cross-reference of related applications This application asserts the interests of U.S. Provisional Patent Application No. 62 / 553,371, filed on September 1, 2017, under Section 119 of the U.S. Patent Act, and this application, without regard to its purpose, is incorporated herein by reference in its entirety as an integral part of this specification.

[0002] Area of ​​Disclosure This disclosure generally relates to functionalized polymers, including crosslinked polymers, and methods for producing and using them. The polymers of this disclosure offer useful properties that cannot be obtained from currently available polymers. [Background technology]

[0003] background The use of polymer materials as biomaterials has increased in recent years, forming an ever-expanding polymer toolbox. Both synthetic and natural polymers are used as components of biomaterials, and their unique chemical structures can provide specific functions for desired applications. The use of polymers as biomaterials has expanded due to advances in polymer synthesis with controlled functional design, thereby improving the range of possible materials and their biocompatibility. While these biocompatible polymers are useful because they do not specifically interact with biological systems, this hinders their use in applications where natural physiological interactions are desired to manipulate biological responses such as wound healing, binding of cellular or growth factors, or enzymatic degradation. Therefore, there is interest in the use of polymers found in living organisms, or polymers that have been modified to provide properties improved from or different from those of unmodified polymers. Examples of such polymers include those containing polyhydric alcohols. A not limited example of such polymers is hyaluronic acid (HA) polymer.

[0004] HA is a non-sulfated glycosaminoglycan (GAG) composed of repeating high molecular weight disaccharides of D-glucuronic acid and N-acetyl-D-glucosamine linked by glucuronide β(1→3) bonds. In aqueous solution, HA forms a specific stable three-dimensional structure. Despite its simple composition, unchanging sugar composition, and lack of branching points, HA possesses a variety of physicochemical properties. HA polymers exist in several configurations and shapes depending on their size, salt concentration, pH, and the cations they are bound to. Unlike other GAGs, in living organisms, HA does not covalently bond to protein cores but can form aggregates with proteoglycans. HA can encapsulate large volumes of water, making solutions highly viscous even at low concentrations.

[0005] HA is involved in several physiological processes, such as skin. A key molecule involved in skin hydration is hyaluronan or hyaluronic acid (HA), a glycosaminoglycan (GAG) with the unique ability to bind to and retain water molecules. HA belongs to the extracellular matrix (ECM) molecules. ECM molecules present between cells exert major effects on cellular function in addition to providing a structural framework. Although these ECM molecules appear amorphous under a light microscope, they form a highly organized structure mainly consisting of GAGs, proteoglycans, growth factors, and structural proteins such as collagen, and HA is the major component of the skin ECM. [Overview of the project] [Problems that the invention aims to solve]

[0006] What is needed are derivatives of polyvalent polymers, compositions of such polymers, and the use of such polymers for medical procedures and the manufacture of medical devices and components. [Means for solving the problem]

[0007] overview In short, this disclosure provides polymers, methods for producing polymers, and methods for using polymers. 1) For example, in one embodiment, the present disclosure relates to a derivative of a polysaccharide, such as a polyhydric polymer such as polyhyaluronic acid, wherein one or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, and the derivative of hyaluronic acid or other polyhydric polymer has the structure HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n The compound is such that HA is a polyvalent polymer such as hyaluronic acid, X is S or NH, and R1 is a substituted or unsubstituted C1-C. 20 The present invention provides a derivative which is an aliphatic or aromatic moiety, where n is the number of modified hydroxyl groups, n is an integer and n≧1, and Y is one or more of H, a carboxylic acid group or its salt or ester, a hydroxyl group, a sulfonic acid group or its salt, or an amine group.

[0008] In another embodiment, the present disclosure relates to a derivative of a polysaccharide, such as a polyvalent polymer such as hyaluronic acid, wherein two or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, and the derivative of hyaluronic acid or other polyvalent polymer has the structure (Y-R2-X-CH2CH2SO2CH2CH2O) m -HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n The compound is such that HA is hyaluronic acid or other polyvalent polymer, X is S or NH, and R1 is substituted or unsubstituted C1-C 20 The aliphatic or aromatic moiety is R2, and R2 is a substituted or unsubstituted C1-C 20 The present invention provides a derivative having an aliphatic or aromatic moiety, where R1 and R2 are distinct from each other, n and m are integers, and n≧1 and m≧1, and Y is H; a carboxylic acid group or its salt or ester; a hydroxyl group; a sulfonic acid group or its salt, or an amine group.

[0009] In another aspect, the present disclosure provides a derivative of a polysaccharide, such as a polyvalent polymer like hyaluronic acid, wherein two or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, and the derivative of hyaluronic acid has the structure (CH2=CH-SO2CH2CH2O)m-HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n where HA is hyaluronic acid or another polyvalent polymer, X is S or NH, R1 is a substituted or unsubstituted C1-C 20 aliphatic or aromatic moiety, each of n and m is an integer, and n≧1 and m≧1, and Y is H; a carboxylic acid group or its salt or ester; a hydroxyl group; a sulfonic acid group or its salt; or an amine group.

[0010] In a further aspect, the present disclosure provides a derivative of a polyvalent polymer such as those described above, further characterized in that 0.25 to 50% of the total of the hydroxyl groups and the modified hydroxyl groups are modified hydroxyl groups.

[0011] In another aspect, the present disclosure includes a crosslinked polymer comprising a reaction product of a derivative of a polysaccharide, such as a polyvalent polymer disclosed herein like hyaluronic acid, optionally including a crosslinking agent. As used herein, the crosslinking agent can be a crosslinking compound, such as a known crosslinking agent like an OH crosslinking agent or a vinyl crosslinking agent, FeCl3, or a compound and / or an energy source including, but not limited to, UV and related photoinitiator compounds.

[0012] In one aspect, the present disclosure includes a crosslinked polymer comprising a reaction product of a derivative of a polysaccharide, such as a polyvalent polymer disclosed herein like hyaluronic acid, optionally including a crosslinking agent, where a) the derivative of a polyvalent polymer such as hyaluronic acid has the structure HA-(OCH2CH2SO2CH2CH2-X-R1-Y) nThe compound is such that, where one or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, HA is a polyvalent polymer containing hydroxyl groups, such as hyaluronic acid, X is S or NH, and R 1 is a substitution or non-substitution of C1-C 20 It is an aliphatic or aromatic moiety, and n is the number of modified hydroxyl groups, n ≥ 1, and Y is H; a carboxylic acid group, or a salt or ester thereof; a hydroxyl group; a sulfonic acid group or a salt thereof; or an amine group; and b) The crosslinking agent comprises at least two functional groups that can react with the hydroxyl group of the hyaluronic acid derivative.

[0013] In another aspect, the present disclosure provides a crosslinked polymer comprising a reaction product of a polysaccharide, such as a derivative of a polyvalent polymer disclosed herein, such as hyaluronic acid, and a crosslinking agent, where, a) The polyvalent polymer derivative comprises two or more hydroxyl groups of hyaluronic acid as modified hydroxyl groups, where hyaluronic acid or the other polyvalent polymer derivative has the structure (Y-R2-X-CH2CH2SO2CH2CH2O) m -HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n The compound is such that HA is hyaluronic acid or other polyvalent polymer, X is S or NH, and R1 is substituted or unsubstituted C1-C 20 The aliphatic or aromatic moiety is R2, and R2 is a substituted or unsubstituted C1-C 20 It is an aliphatic or aromatic moiety, R1 and R2 are distinct from each other, n and m are integers, and n≧1 and m≧1, and Y is H; a carboxylic acid group, or a salt or ester thereof; a hydroxyl group; a sulfonic acid group or a salt thereof; or an amine group; and b) The crosslinking agent comprises at least two functional groups that can react with the hydroxyl group of the hyaluronic acid derivative.

[0014] In another aspect, the present disclosure provides a crosslinked polymer comprising a reaction product of a polysaccharide, for example, a derivative of a polyvalent polymer such as hyaluronic acid, and a crosslinking agent, wherein a) Hyaluronic acid derivatives contain a vinyl group and have the structure (CH2=CH-SO2CH2CH2O) m -HA-(OCH2CH2SO2CH2CH2-XR 1 -Y) n The compound is such that two or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, HA is hyaluronic acid containing hydroxyl groups, X is S or NH, and R 1 is a substitution or non-substitution of C1-C 20 It is an aliphatic or aromatic moiety, n≧1 and m≧1, and Y is H; a carboxylic acid group or its salt or ester; a hydroxyl group; a sulfonic acid group or its salt; or an amine group; and b) The crosslinking agent comprises at least two functional groups that can react with the vinyl group of the hyaluronic acid derivative, or c) The crosslinking agent contains at least two functional groups that can react with the hydroxyl group of the hyaluronic acid derivative, or d) The crosslinking agent contains a functional group that can be ionically crosslinked, or e) The crosslinking agent contains a functional group that can be thermally crosslinked, or f) The crosslinking agent generates free radicals that can undergo free radical crosslinking.

[0015] In a further embodiment, the crosslinked polymers disclosed herein may be further characterized in that 0.25 to 50% of the total of hydroxyl groups and modified hydroxyl groups are modified hydroxyl groups.

[0016] In another aspect, this disclosure is: a) To provide a first derivative of a polymer by reacting a hydroxyl group bonded to a polymer, such as a hydroxyl group on hyaluronic acid (HA) or other polyvalent polymers, with divinyl sulfone (DVS); and b) First derivative of the polymer and X'-R 1 -Y and X'-R2 A second derivative of the polymer is provided by reacting it with a nucleophile of a formula selected from -Y. Provide a process that includes; Here, R 1 is a substitution or non-substitution of C1-C 20 It is an aliphatic or aromatic moiety, R 2 is a substitution or non-substitution of C1-C 20 The polymer is an aliphatic or aromatic moiety, where X' is a nucleophile containing a thiol or amine, and Y is one or more of H, a carboxylic acid group or its salt or ester, a hydroxyl group, a sulfonic acid group or its salt, or an amine group. A polymer that has undergone one or more derivatization reactions includes a derivative of a polyvalent polymer, even if it has undergone one or more derivatizations of the original polymer, for example, one, two, three, four times, etc., and is referred to as such herein. A derivative of a polymer is also called the first derivative, second derivative, third derivative, etc. of the polymer.

[0017] In a further embodiment, this process may further feature one or more of the following: 1) A process in which 0.25-50% of the hydroxyl groups present on the initial (non-derivativeized) polymer are converted to oxyethylethenylsulfone groups of the formula -OCH2CH2-SO2CH=CH2. 2) The polymer is hyaluronic acid, and the first derivative of the polymer is an oxyethyl ethenyl sulfone derivative of hyaluronic acid HA-(-OCH2CH2SO2CH=CH2) n A process that is (HA-DVS). 3) The second derivative is HA-(OCH2CH2SO2CH2CH2-XR 1 A process that is )n(HA-DVS-N). 4) 0.25-50% of the hydroxyl groups present on the polymer are -OCH2CH2SO2CH2CH2-XR 1 The process of converting to the base. 5) A process in which X' is a thiol or amine and X is -S- or -NH-. 6) The second derivative is HA-(-OCH2CH2SO2CH2CH2-XR 2 -Y) n A process that is (HA-DVS-NY). 7) When calculated based on the total of hydroxyl groups and modified hydroxyl groups, 0.25-50% of the hydroxyl groups present on the polymer are -OCH2CH2SO2CH2CH2-XR 2 - The process of converting to the Y base. 8) A process where X is -S-. 9) A process in which Y is a hydroxyl molecule. 10) A process in which Y is a carboxylic acid or a salt or ester thereof. 11) A process in which Y is a sulfonic acid or a salt or ester thereof. 12) A process comprising reacting a second derivative of a polymer with a crosslinking agent to provide a third derivative of a polymer, wherein the third derivative is a crosslinked polymer.

[0018] In another aspect, the disclosure provides derivatives of polyvalent polymers, such as hyaluronic acid derivatives, which are produced by any of the processes described herein. In another aspect, the disclosure provides crosslinked polymers, which are produced by any of the processes described herein.

[0019] In another embodiment, the present disclosure provides a composition comprising a high molecular weight polyhydric alcohol, for example, a derivative of hyaluronic acid, wherein the derivative of the high molecular weight polyhydric alcohol may include derivatives such as those disclosed above with respect to polyhydric polymers. For example, HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n ;(Y-R2-X-CH2CH2SO2CH2CH2O) m -HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n (CH2=CH-SO2CH2CH2O) m -HA-(OCH2CH2SO2CH2CH2-XR 1 -Y) n; or (CH2=CH-SO2CH2CH2O) m -HA, where HA is hyaluronic acid or other polyvalent polymer, X is S or NH, and R1 is substituted or unsubstituted C1-C 20 The aliphatic or aromatic moiety is R2, and R2 is a substituted or unsubstituted C1-C 20 The moiety is aliphatic or aromatic, and where applicable, R1 and R2 are distinct from each other, and Y is one or more of H, a carboxylic acid group or its salt or ester, a hydroxyl group, a sulfonic acid group or its salt, or an amine group; n and m are integers from each other, and n≧1 and m≧1. The composition may further contain excipients.

[0020] In another embodiment, the present disclosure provides compositions comprising crosslinked polymers, such as crosslinked derivatives of high molecular weight hyaluronic acid, as described herein. The compositions may further comprise excipients. Each of the compositions disclosed herein may optionally comprise one or more of pharmaceutically acceptable synthetic polymers, thermoreversible polymers, biodegradable polymers, buffers, complexing agents, tonicity modifiers, ionic strength modifiers, solvents, antioxidants, preservatives, viscosity modifiers, pH modifiers, surfactants, emulsifiers, phospholipids, stabilizers, and pologens. Furthermore, compositions such as those disclosed herein may optionally further comprise bioactive agents.

[0021] The derivatized polymers disclosed herein and compositions comprising one or more derivatized polymers exhibit shear reduction. Shear reduction is the non-Newtonian behavior of a fluid in which viscosity decreases under shear strain.

[0022] In further embodiments, the Disclosure provides methods for using polymers and compositions as disclosed herein. For example, the Disclosure provides the following embodiments: 1) A wound healing device comprising a composition as described herein. 2) A method for wound healing comprising administering an effective amount of a composition as described herein to a subject requiring attention. 3) A filler comprising a composition such as those described herein. 4) Skin fillers comprising compositions such as those described herein. 5) A method for filling a void in a subject as needed, comprising administering a skin filler as described herein to the subject. 6) An intra-articular replacement agent comprising a composition such as those described herein. 7) A method for reducing joint pain in subjects in need, comprising administering an intra-articular replacement agent as described herein to the subject. 8) A method for preventing postoperative adhesions in a subject in need, comprising administering an effective amount of a composition as described herein to the subject. 9) A tissue sealant comprising a composition such as those described herein. 10) A method for sealing tissue in a subject as needed, comprising administering an effective amount of a tissue sealant as described herein to the subject. 11) A method for treating bacterial vaginosis in a subject requiring treatment, comprising administering to the subject an effective amount of a composition as described herein. 12) A nasal treatment device comprising a composition such as those described herein. 13) A method for treating a nasal condition in a subject requiring treatment, comprising administering an effective amount of a composition as described herein to the subject. 14) Eye drops comprising a composition as described herein. 15) A method for treating an ocular condition in a subject requiring treatment, comprising administering an effective amount of a composition as described herein to the subject. 16) A punctal plug comprising a composition such as those described herein. 17) A method for treating mucositis in a subject requiring treatment, comprising administering to the subject an effective amount of a composition as described herein. 18) Antimicrobial formulations comprising compositions such as those described herein. 19) An ear treatment device comprising a composition as described herein. 20) A method for treating an ear condition, comprising administering an effective amount of a composition as described herein to a subject requiring treatment. 21) A method for delivering a drug to a target, comprising administering to the target an effective amount of a composition containing a drug, such as those described herein. 22) A biopsy plug comprising a composition such as those described herein. 23) A female contraceptive plug comprising a composition such as those described herein. 24) A method of female contraception for a subject in need, comprising administering to the subject an effective amount of a composition as described herein. 25) A tissue scaffold comprising a composition such as those described herein. 26) A method for assisting tissue growth in a subject in need, comprising transplanting a tissue scaffold as described herein to the subject. 27) A piercing hole plug comprising a composition as described herein. 28) A nerve guide comprising a composition such as that described herein. 29) A vaginal lubricant comprising a composition such as those described herein. 30) A coating agent for apparatus comprising a composition such as those described herein. 31) A method for coating an apparatus, comprising applying a coating to the surface of the apparatus as described herein. 32) A method for administering an injectable preparation comprising a composition as described herein. 33) A polymer as described herein, for example, a derivative of hyaluronic acid produced by a process as described herein or as described herein for providing such a polymer, for example, a derivative of hyaluronic acid, and a method for addition manufacturing, comprising attaching the derivative to a substrate to obtain an article formed by addition manufacturing. 34) A method for addition manufacturing by attaching a polymer as described herein, for example, a hyaluronic acid derivative as described herein, to a surface or within a polymer substrate. 35) A method of coating an article formed by addition with a polymer as described herein, for example, a hyaluronic acid derivative as described herein, or impregnating an article formed by addition with a polymer as described herein, for example, a hyaluronic acid derivative as described herein. 36) Electrospun materials or electrospun articles comprising compositions such as those described herein. 37) A method for producing an electrospun material or electrospun article, comprising producing a material or article containing a hyaluronic acid derivative described herein using an electrospinning apparatus. 38) A method of coating an article formed by electrospinning with a polymer as described herein, for example, a hyaluronic acid derivative as described herein, or impregnating an article formed by electrospinning with a polymer as described herein, for example, a hyaluronic acid derivative as described herein. 39) Textile materials or textile articles comprising compositions such as those described herein. 40) A method for producing a textile material or textile article, comprising using an electrospinning apparatus to produce a material or article containing a hyaluronic acid derivative described herein. 41) A method of coating an article formed on a textile substrate by electrospinning with a polymer as described herein, for example, a hyaluronic acid derivative as described herein, or impregnating an article formed on a textile substrate by electrospinning with a polymer as described herein, for example, a hyaluronic acid derivative as described herein.

[0023] The above and additional features of this disclosure, as well as how to obtain them, will become clearer upon further detailed explanation below, and this disclosure will be best understood. All references disclosed herein are incorporated in their entirety as part of this specification, just as each of them is incorporated individually as part of this specification.

[0024] This brief overview is provided to introduce certain concepts in a simplified form, which will be further elaborated upon in the detailed explanations below. Unless otherwise explicitly stated, this brief overview is not intended to identify any essential or important features of the claimed subject matter, nor is it intended to limit the scope of the claimed subject matter.

[0025] Details of one or more embodiments are shown in the following description. Features illustrated or described in relation to one exemplary embodiment may be combined with features of other embodiments. Thus, any combination of the various embodiments described herein may provide further embodiments. Embodiments of embodiments may, if necessary, be modified to adopt concepts from various patents, patent applications and publications shown herein to provide even further embodiments. Other features, purposes and advantages will become apparent from this specification, the drawings and the claims.

[0026] The exemplary features, nature, and various advantages of this disclosure will become apparent from the attached drawings and the detailed descriptions of the various embodiments below. Non-limiting and non-exclusive embodiments are described with reference to the attached drawings, and unless otherwise noted, similar designations or reference numbers refer to similar parts from different perspectives. The size and relative positions of elements in the drawings are not necessarily depicted in exact proportions. For example, the shapes of various elements are selected, enlarged, and arranged to make the drawings easy to view. The specific shapes of elements depicted are selected for their visibility in the drawings. One or more embodiments are described below with reference to the attached drawings. [Brief explanation of the drawing]

[0027] [Figure 1] Figure 1 shows the 1H NMR spectrum of divinyl sulfone-modified hyaluronic acid according to this disclosure. [Figure 2] Figure 2 shows an exemplary response of the present disclosure. [Figure 3] Figure 3 shows an exemplary response of the present disclosure. [Figure 4]Figure 4 shows the 1H-NMR spectrum of 2-mercaptobenzoic acid (MBA) modified hyaluronic acid according to this disclosure. [Figure 5] Figure 5 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 6] Figure 6 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 7] Figure 7 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 8] Figure 8 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 9] Figure 9 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 10] Figure 10 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 11] Figure 11 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 12] Figure 12 is a graph illustrating the characteristics of exemplary derivatized polymers disclosed herein. [Figure 13] Figure 13 is a graph showing cell proliferation on exemplary derivatized polymers disclosed herein. [Figure 14] Figure 14 is a graph showing cell proliferation on electrospun products containing exemplary derivatized polymers disclosed herein. [Modes for carrying out the invention]

[0028] Detailed explanation of disclosure This disclosure may be more readily understood by referring to the detailed description of preferred embodiments of this disclosure below and the examples included herein.

[0029] In one embodiment, the disclosure provides functionalized polymers, including crosslinked polymers. A functionalized polymer refers to an organic polymer that contains a hydroxyl group and optionally a second functional group such as a carboxylic acid group, an amine group, or a sulfonic acid group.

[0030] This disclosure relates to a derivative of a polysaccharide, such as a polyvalent polymer such as hyaluronic acid, wherein one or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, and the derivative of hyaluronic acid or other polyvalent polymer has the structure HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n The compound is such that HA is a polyvalent polymer such as hyaluronic acid, X is S or NH, and R1 is a substituted or unsubstituted C1-C. 20 The present invention provides a derivative which is an aliphatic or aromatic moiety, where n is the number of modified hydroxyl groups, n is an integer and n≧1, and Y is one or more of H, a carboxylic acid group or its salt or ester, a hydroxyl group, a sulfonic acid group or its salt, a phosphonic acid group or its salt, or an amine group.

[0031] In another embodiment, the present disclosure relates to a derivative of a polysaccharide, such as a polyvalent polymer such as hyaluronic acid, wherein two or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, and the derivative of hyaluronic acid or other polyvalent polymer has the structure (Y-R2-X-CH2CH2SO2CH2CH2O) m -HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n The compound is such that HA is hyaluronic acid or other polyvalent polymer, X is S or NH, and R1 is substituted or unsubstituted C1-C 20 The aliphatic or aromatic moiety is R2, and R2 is a substituted or unsubstituted C1-C 20 The present invention provides derivatives in which the aliphatic or aromatic moiety is a carboxylic acid group or a salt or ester thereof, n and m are integers, n≧1 and m≧1, and Y is H; a carboxylic acid group or a salt or ester thereof; a hydroxyl group; a sulfonic acid group or a salt thereof; a phosphonic acid group or a salt thereof; or an amine group.

[0032] In another embodiment, the present disclosure relates to a derivative of a polysaccharide, such as a polyvalent polymer such as hyaluronic acid, wherein two or more hydroxyl groups of hyaluronic acid are modified hydroxyl groups, and the derivative of hyaluronic acid has the structure (CH2=CH-SO2CH2CH2O)m-HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n The compound is such that HA is hyaluronic acid or other polyvalent polymer, X is S or NH, and R1 is substituted or unsubstituted C1-C 20 The present invention provides derivatives in which the aliphatic or aromatic moiety is an aliphatic or aromatic moiety, where n and m are integers, and n≧1 and m≧1, and Y is H; a carboxylic acid group or a salt or ester thereof; a hydroxyl group; a sulfonic acid group or a salt thereof; a phosphonic acid group or a salt thereof; or an amine group.

[0033] In a further embodiment, the disclosure provides derivatives of polyvalent polymers such as the above, further characterized in that the derivatives consist of 0.25 to 50% of the total of hydroxyl groups and modified hydroxyl groups being modified hydroxyl groups.

[0034] In another embodiment, the disclosure includes reaction products of polysaccharides, such as derivatives of polyvalent polymers disclosed herein, such as hyaluronic acid, and includes crosslinked polymers which may optionally include a crosslinking agent, where the crosslinking agent may include crosslinking compounds, such as known crosslinking agents such as OH crosslinking agents or vinyl crosslinking agents, FeCl3, or energy sources including compounds and / or, but not limited to, UV and related photoinitiator compounds.

[0035] In one embodiment, the disclosure provides a polysaccharide having one or more available hydroxyl groups, wherein one or more of these hydroxyl groups are reacted with divinyl sulfone under specific conditions as disclosed herein by an addition reaction such that only one vinyl group of the divinyl sulfone forms an ether bond between the polysaccharide and the divinyl sulfone residue. The degree of reaction ranges from about 0.5% to about 50% of the available hydroxyl groups. At higher substitutions, i.e., around 50%, some degree of crosslinking generally occurs. Thus, the disclosure provides vinyl sulfone-substituted polysaccharide polymers with minimal or no crosslinking, or polysaccharide polymers having a certain level of vinyl sulfone-substituted crosslinking for the double reaction of divinyl sulfone (i.e., the reaction of both ethenyl and hydroxyl groups in DVS).

[0036] Next, the vinyl residue of vinyl sulfone can be reacted with a compound having a reactive thiol group. This reaction occurs by Michael addition between the vinyl residue of divinyl sulfone and the free thiol group such that a thioether bond is formed. Many variations exist in the degree of substitution, the thiol derivative used, the order of the reaction, and the repetition of the reaction, which provide a variety of derivatives and compositions of high molecular weight polyhydric alcohols as intended and disclosed herein. Derivatives of high molecular weight polyhydric alcohols can be crosslinked in many different ways, and compositions containing such crosslinked derivatives of high molecular weight polyhydric alcohols are intended and disclosed herein. Derivatives of high molecular weight polyhydric alcohols and their compositions have many medical and non-medical applications. Methods of use or treatment disclosed herein may include derivatives of high molecular weight polyhydric alcohols and their compositions. Derivatives of high molecular weight polyhydric alcohols may also be referred to herein as polyhydric polymer derivatives.

[0037] Derivatives of polymeric polyhydric alcohols and their compositions of the present disclosure are prepared as described herein. Generally, polymers having hydroxyl groups are combined with divinyl sulfone (DVS) under suitable reaction conditions. Such reaction conditions include a suitable pH of the solution, and the reaction generally occurs under basic conditions, for example, pH 11-14 or 12-13, for example, about 12.3. The reaction conditions include a suitable solvent, water or DMSO is a suitable solvent, for example, the reaction can be carried out in water. The description of the reaction conditions may further include stirring of the reaction mixture, for example, stirring at a stirring speed >200 rpm (revolutions per minute), for example, 250-800 rpm. Furthermore, the description of the reaction conditions may also include specifying the relative amounts of DVS and polymer (e.g., polysaccharide) to be combined, and these relative amounts can be expressed as moles of DVS relative to moles of repeating units in the polymer. For example, a method for producing a functionalized polymer may be described as a DVS:polymer repeating unit ratio, which can be at least 0.5:1, for example, up to about 5:1, or up to about 7.5:1, or up to about 10:1, or up to about 15:1, or up to about 20:1.

[0038] The exemplary functionalized polymer is a polysaccharide, and the exemplary polysaccharide is hyaluronic acid (HA). HA is a polysaccharide exemplified by the structure shown below.

[0039] [ka]

[0040] HA contains two distinct functional groups, namely a hydroxyl group and a carboxylic acid group. HA also contains ether and acetamide groups, which are essentially chemically inert. In commercially available preparations of HA, some or all of the carboxylic acids may exist as corresponding salts, for example, sodium salts, potassium salts, or ammonium salts. In this disclosure, unless the context indicates otherwise, HA exclusively refers to the polymers of the structures shown above and the corresponding carboxylic acid salts of those polymers. Another exemplary polysaccharide polymer is dextran. Other exemplary polysaccharide polymers useful in this disclosure include, but are not limited to, sodium alginate, calcium alginate, dextran, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, hyaluronic acid, hyaluronic acid derivatives, dextran, heparin, chitosan, xanthan gum, xylan, guar gum, pullulan, or locust bean gum. The term “polyvalent polymer” includes, but is not limited to, such high molecular weight polysaccharides. The term "polyvalent polymer" is not limited to polymers having two or more hydroxyl groups, and in this specification, it may also be referred to as a high molecular weight polyhydric alcohol.

[0041] In one embodiment, the disclosure provides a process for reacting a high molecular weight polysaccharide having available hydroxyl groups, i.e., hydroxyl groups that can react with divinylsulfone, with DVS under basic conditions. By appropriately selecting the conditions, the reaction can be controlled so that one of the vinyl groups of divinylsulfone reacts with a free hydroxyl group of the polysaccharide, without crosslinking to the extent that the polysaccharide forms a hydrogel. This results in a polysaccharide in which one vinyl group reacts with the hydroxyl group of the polysaccharide, while the other vinyl group is functionalized with divinylsulfone to retain its functionality. The vinyl group of divinylsulfone reacts with the hydroxyl group by an addition reaction that produces an ether bond.

[0042] This reaction can be carried out under basic conditions with a pH greater than 11. Preferably, the pH is in the range of 12.0 to 13.5. Preferably, the pH is in the range of 12.0 to 12.5. Preferably, the pH range is in the range of 12.2 to 12.7.

[0043] To ensure that the main reaction is a single reaction of one vinyl group of divinylsulfone and not a crosslinking reaction in which both vinyl groups react with the hydroxyl groups of the polysaccharide to form a crosslinked gel, the molar ratio of divinylsulfone to polysaccharide repeating units is greater than 1. In one embodiment, the molar ratio of divinylsulfone to polysaccharide repeating units is greater than 5. In one embodiment, the molar ratio of divinylsulfone to polysaccharide repeating units is greater than 7. In one embodiment, the molar ratio of divinylsulfone to polysaccharide repeating units is greater than 10. In one embodiment, the molar ratio of divinylsulfone to polysaccharide repeating units is greater than 15. In one embodiment, the molar ratio of divinyl sulfone to polysaccharide repeating units is approximately 1 to approximately 20, or approximately 1 to approximately 15, or approximately 1 to approximately 10, or approximately 1 to approximately 5, or approximately 5 to approximately 20, or approximately 5 to approximately 15, or approximately 5 to approximately 10, approximately 10 to approximately 20, or approximately 10 to 15, or approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

[0044] To provide close contact with the reactants, the reaction mixture can be stirred. Methods and apparatus for thorough mixing are known to those skilled in the art. For example, to ensure sufficient stirring of the reaction solution during the reaction, the rotation speed of the mixing impeller should be controlled. In one embodiment, the revolutions per minute (rpm) of the mixing impeller should be in the range of 200 to 400 rpm. In another embodiment, the revolutions per minute (rpm) of the mixing impeller should be in the range of 400 to 600 rpm. In yet another embodiment, the revolutions per minute (rpm) of the mixing impeller should be in the range of 600 to 800 rpm.

[0045] The amount of substitution achieved can be controlled, in part, by the duration of exposure of the polysaccharide to divinyl sulfone at a pH above 11 (reaction time). In one embodiment, the reaction time may be in the range of 10 seconds to 60 minutes. In another embodiment, the reaction time may be in the range of 2 minutes to 35 minutes. In yet another embodiment, the reaction time may be in the range of 4 minutes to 30 minutes, or 20 minutes to 60 minutes, 15 minutes to 20 minutes, 5 minutes to 10 minutes, 10 seconds to 30 seconds, 30 seconds to 1.5 minutes, and in between.

[0046] The solvents that can be used in the reaction may be water, water containing an ion modifier, such as NaCl, or a combination of water and a water-miscible solvent. Water-miscible solvents, though not limited to these, may include methanol, ethanol, isopropanol, dimethylformamide (DMF), acetone, 1,4-dioxane, pyridine, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), and acetonitrile.

[0047] Furthermore, the temperature of the reaction mixture can also be used to influence the amount of polysaccharide substitution by divinyl sulfone. In one embodiment, the reaction mixture can be maintained at a temperature below 25°C to slow the reaction rate. This allows for a lower level of substitution in the same period compared to room temperature, or a longer reaction time at room temperature to obtain the same amount of substitution. In one embodiment, the temperature may be in the range of 15°C to 20°C. In another embodiment, the reaction mixture may be in the range of 10°C to 15°C. In yet another embodiment, the temperature may be in the range of 2°C to 10°C. In yet another embodiment, the temperature can be higher than 25°C to allow for a shorter reaction time compared to 25°C to obtain the same amount of substitution, or to obtain a greater amount of substitution in the same reaction time compared to 25°C. In one embodiment, the reaction mixture may be in the range of 28°C to 35°C. In another embodiment, the reaction mixture may be in the range of 36°C to 50°C. In yet another embodiment, the reaction mixture may be in the range of 51°C to 75°C.

[0048] The amount of substitution, as measured by the molar ratio of the attached vinyl group derived from divinyl sulfone to the polysaccharide repeating unit, may exceed 5%. In one embodiment, for a polysaccharide having at least one hydroxyl group, the amount of substitution is in the range of 5% to 35%. In another embodiment, for a polysaccharide having at least one hydroxyl group, the amount of substitution is in the range of 36% to 70%. In yet another embodiment, for a polysaccharide having at least one hydroxyl group, the amount of substitution is in the range of 71% to 100%. In yet another embodiment, for a polysaccharide having at least two hydroxyl groups, the amount of substitution is in the range of 101% to 200%.

[0049] In one embodiment, a polysaccharide polymer containing at least one hydroxyl group available for reaction with divinyl sulfone under conditions of pH greater than 11 is preferred for use in the present disclosure. Such polysaccharides include, but are not limited to, hyaluronic acid and its sodium or potassium salts, hyaluronic acid derivatives, dextran and dextran derivatives, dextran sulfate, heparin, chitosan and its derivatives, xylan, guar gum, locust bean gum, chondroitin 6-sulfate, chondroitin 4-sulfate, heparan sulfate, keratin sulfate, dermatan sulfate, and chitin. In one embodiment, the polysaccharide polymer is hyaluronic acid or sodium hyaluronate. In another embodiment, the polysaccharide is dextran. As used herein, polysaccharide means a high molecular weight polysaccharide molecule.

[0050] The molecular weight of the polysaccharide can be selected. Molecular weights from 10,000 to 5,000,000 are usable. In one embodiment, the polysaccharide has a molecular weight greater than 10,000. In another embodiment, the polysaccharide has a molecular weight in the range of 10,000 to 50,000. In yet another embodiment, the polysaccharide has a molecular weight in the range of 50,000 to 200,000. In yet another embodiment, the polysaccharide has a molecular weight in the range of 200,000 to 600,000. In one embodiment, the polysaccharide has a molecular weight in the range of 600,000 to 1,000,000. In one embodiment, the polysaccharide has a molecular weight in the range of 1,000,000 to 2,500,000. In yet another embodiment, the polysaccharide has a molecular weight in the range of 2,500,000 to 5,000,000. The molecular weight can be measured by known methods, including gel permeation chromatography or intrinsic viscosity, although this is not limited to these methods.

[0051] After reacting the functionalized polymer with DVS to produce a first derivative of the polymer, this first derivative is then subjected to XR 1 and XR 2 -A nucleophile of a formula selected from Y, for example, a thiol derivative, is reacted to provide a second derivative of the polymer. In these formulas, R 1 is a substitution or non-substitution of C1-C 20 Aliphatic or aromatic, R 2 is a substitution or non-substitution of C1-C 20 The compound is aliphatic or aromatic, X is a nucleophile, and Y is selected from carboxylic acids, sulfonic acids, and hydroxyls. The nucleophile contains a thiol group as X of the thiol derivative. Generally, thiol derivatives can be a single compound or a mixture of thiol compounds. Examples include alkylthiols, which can be linear, branched, or cyclic, such as methanethiols and ethanethiols. Alternatively, thiols can be arylthiols, charged thiols, high molecular weight thiols, peptides containing thiol groups, proteins containing thiol groups, heterocycles containing thiol groups, drugs containing thiol groups, such as active pharmacological components, growth factors containing thiol groups, and bioactive agents containing thiol groups.

[0052] For example, the thiol compounds available in this disclosure may be compounds containing at least one free thiol group that can react with a vinyl sulfone group by a Michael addition reaction.

[0053] Thiol compounds can be represented by the formula R1SH or R2SH, where R1 and R2 may be aliphatic or aromatic moieties, and either of them may have one or more substituents, for example, a substituted aliphatic moiety or a substituted aromatic moiety. The aliphatic moiety refers to an alkyl or cycloalkyl moiety having 1 to 20 carbon atoms.

[0054] "Alkyl" refers to a linear or branched hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms that are unsaturated, have 1 to the indicated number of carbon atoms, and are bonded to the rest of the molecule by single bonds, such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, etc. In one embodiment, the alkyl group has 1 carbon. In one embodiment, the alkyl group has 2 carbons. In one embodiment, the alkyl group has 3 carbons. In one embodiment, the alkyl group has 4 carbons. In one embodiment, the alkyl group has 4 carbons. In one embodiment, the alkyl group has 5 carbons. In one embodiment, the alkyl group has 6 carbons. Two or more of these embodiments may be combined to describe the derivatives of the present disclosure.

[0055] "Cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms, having 3 to 15 carbon atoms, preferably including a condensed or crosslinked ring system having 3 to 10 carbon atoms, and being saturated or unsaturated, bonded to the rest of the molecule by a single bond. Examples of monocyclic radicals include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic radicals include adamantyl, norbornyl, dekalinyl, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl. Unless otherwise specified, cycloalkyl groups may be optionally substituted with one or more substituents independently selected in each presence.

[0056] The aromatic moiety refers to a carbocyclic aromatic moiety having 1 to 20 carbon atoms, also known as an aryl moiety, or a heteroaromatic moiety, also known as a heteroaryl moiety, or a heteroaromatic moiety having at least one heteroatom selected from sulfur, oxygen, and nitrogen.

[0057] "Aryl" refers to a hydrocarbon ring radical comprising hydrogen, 6 to 18 carbon atoms, and at least one aromatic ring. In one embodiment, the aryl ring system has 6 to 12 carbon atoms. In one embodiment, the aryl ring system has 6 to 10 carbon atoms. For the purposes of this disclosure, the aryl radical may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include condensed or crosslinked ring systems. Examples of aryl radicals include, but are not limited to, aryl radicals derived from acetantrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluorantene, fluorene, as-indacene, s-indacene, indan, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless otherwise stated herein, the aryl group may be optionally substituted with one or more substituents independently selected in each presence.

[0058] "Heteroaryl" refers to an "aryl" as defined herein, wherein the aromatic ring contains one or more heteroatoms, preferably selected from N, O, and S. Thus, a heteroaryl radical refers to an aromatic ring system radical in which the ring atoms are selected from carbon, nitrogen, oxygen, and sulfur, and which contain at least one of nitrogen, oxygen, and sulfur. For the purposes of this disclosure, heteroaryl radicals may be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, which may include condensed or bridging ring systems. Optionally, heteroaryl radicals are 5-membered, 6-membered, or 7-membered heteroaryl groups. If multiple O and S atoms are present in the heteroaryl ring system, the O atoms and / or S atoms are preferably not directly linked to each other. Examples of heteroaryl groups include five-membered rings such as pyrrole, pyrazole, imidazole, 1,2,3-triazole, 1,2,4-triazole, tetrazole, furan, thiophene, selenophene, oxazole, isoxazole, 1,2-thiazole, 1,3-thiazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole, 1,2,5-thiadiazole, and 1,3,4-thiadiazole.Heteroaryl groups include six-membered rings such as pyridine, pyridazine, pyrimidine, pyrazine, 1,3,5-triazine, 1,2,4-triazine, 1,2,3-triazine, 1,2,4,5-tetrazine, 1,2,3,4-tetrazine, and 1,2,3,5-tetrazine, or indole, isoindole, indidine, indazole, benzimidazole, benzotriazole, purine, naphthoimidazole, phenanthrimidazole, pyridoimidazole, pyrazinimidazole, quinoxalineimidazole, benzoxazole, naphthoxazole, anthroxazole, and phenanthroxazole. The heteroaryl group may be a fused ring containing a six-membered ring, such as throxazole, isoxazole, benzothiazole, benzofuran, isobenzofuran, dibenzofuran, quinoline, isoquinoline, pteridine, benzo-5,6-quinoline, benzo-6,7-quinoline, benzo-7,8-quinoline, benzoisoquinoline, acridine, phenothiazine, phenoxazine, benzopyridazine, benzopyrimidine, quinoxaline, phenazine, naphthyridine, azacarbazole, benzocarbolin, phenanthridine, phenanthroline, thieno[2,3b]thiophene, thieno[3,2b]thiophene, dithienothiophene, isobenzothiophene, dibenzothiophene, and benzothiadiazothiophene. Unless otherwise specified, the ring atoms of the heteroaryl group may be optionally substituted with one or more substituents independently selected at each ring atom.

[0059] Substitution C1-C 20 The aliphatic or aromatic moiety is a C1-C molecule having one or more substituents. 20Aliphatic or aromatic moiety is used, where “substituent” refers to a monovalent group that can be bonded to the moiety described. For example, “substituted phenyl” refers to a phenyl ring having 1, 2, 3, or 4 substituents bonded to the phenyl ring. Substituents include halogens, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, -OH, -O(C1-C6 alkyl), -O(C1-C6 haloalkyl), -O(C1-C6 hydroxyalkyl), -S(C1-C6 alkyl), -S(C1-C6 haloalkyl), -S(C1-C6 hydroxyalkyl), cyano, amino (-NH2), formyl (-CHO), carboxylic acid (-COOH), carboxylic acid ester (-COOR, where R is C1-C 10 It can be selected from (which are alkyl groups).

[0060] These thiol compounds include alkylthiols, which may be linear, branched, or cyclic arylthiols, charged thiol compounds, polymers containing free thiols, peptides containing free thiols, heterocyclic compounds containing free thiols, drugs or bioactive compounds having free thiols, growth factors having free thiols, antibodies or antibody fragments having free thiols, and proteins having free thiols. Examples of such thiol compounds include, but are not limited to, thiophenol, 2-phenylethanethiol, triphenylmethanethiol, 4-methylbenzenethiol, 4-aminothiophenol, 2-aminothiophenol, 4-methoxy-α-toluenethiol, 4-nitrothiophenol, 4-tert-butylbenzenethiol, 2-mercapto-2-phenylacetic acid, 4-mercaptobenzoic acid, 2-mercaptobenzoic acid (thiosalicylic acid), 3-mercapto-1-propanol, 1-mercapto-2-propanol, 4-mercapto-1-butanol, 3-mercapto-1-hexanol, 6-mercapto-1-hexanol, 8-mercapto-1-octanol, 9-mercapto-1-nonanol, 11-mercapto-1-undecanol, 4-mercapto-4-methylpentan-2-ol, ethanethiol, 1-propanthiol, and 2-propanthiol. L, 1-butanethiol, 1-pentanethiol, 1-hexanethiol, 2-ethylhexanethiol, 1-heptanethiol, 1-octanthiol, 1-nonanthiol, 1-decanethiol, 1-undecanethiol, 1-dodecanethiol, 1-tetradecanethiol, 1-hexadecanethiol, cis-9-octadecene-1-thiol, 1-octadecanethiol, 2-methyl-1-butanethiol, 3-methyl-1-butanethiol Thiols, cycloalkyls, cyclohexanethiol, cyclopentanethiol, sodium 3-mercapto-1-propanesulfonate, sodium mercaptopyruvate, 6-mercaptohexanoic acid, 8-mercaptooctanoic acid, 11-mercaptoundecanoic acid, 16-mercaptohexadecanoic acid, sodium 2-mercaptoethanesulfonate, 3-mercaptopropionic acid, 2-amino-4-mercaptobutyric acid (DL-homocysteine),L-cysteine, 11-mercaptoundecyl phosphate, 2-mercapto-1-methylimidazole, 1-benzyl-2-mercaptoimidazole, 2-mercapto-6-methylpyridine, 3-mercapto-2-butanone, 3-mercapto-3-methyl-1-butyl-1-formate, 3-mercapto-3-methylbutan-1-ol, 7-mercapto-4-methylcoumarin, 2-mercapto-4-methyl-5-thiazoleacetic acid, 2- It contains mercapto-5-nitrobenzimidazole, 2-mercapto-5-benzimidazole sulfonate sodium dihydrate, 3-mercapto-N-nonylpropionamide, 2-mercapto-4-methylpyrimidine hydrochloride, 2-mercapto-2-phenylacetic acid, 2-mercapto-3-(trifluoromethyl)pyridine, 2-mercapto-Nm-tolylacetamide, and 4-mercapto-4-methylpentan-2-ol.

[0061] Polymers containing free thiols include, but are not limited to, thiol-PEG3-phosphonic acid, poly(L-lactide), thiol-terminated 5000, poly(L-lactide), thiol-terminated 2500, PEG-SH 3000, PEG-SH 5000, thiol-functionalized hyaluronic acid, thiol-functionalized chitosan, thiol-functionalized arunate, thiol-functionalized dextran, thiol-functionalized chondroitin sulfate, and thiol-functionalized carboxymethylcellulose.

[0062] Examples of thiol-functionalized hyaluronic acid include, but are not limited to, hyaluronic acid with a thiol group linked to it, hydrazide compounds as described in U.S. Patent No. 7,981,871, carbodiimide groups as described in U.S. Patent No. 6,884,788, and those described in U.S. Patent No. 8,124,757.

[0063] Examples of thiol-functionalized chitosans include, but are not limited to, chitosan-cysteine ​​conjugates, chitosan-thioglycolic acid conjugates, and chitosan-4-thio-butylamidine conjugates.

[0064] Non-degradable thiol-functionalized polymers include, but are not limited to, polycarbophil-cysteamine conjugates, polycarbophil-cysteine ​​conjugates, and poly(acrylic acid)-homocysteine ​​conjugates.

[0065] Examples of thiolated peptides or peptides containing at least free thiols include, but are not limited to, cysteine-terminal peptides containing residues 73-92 of the BMP-2 knuckle epitope (N→C:KIPKASSVPTELSAISTLYLSGGC), thiolated gelatin (see, for example, U.S. Patent Nos. 7,928,069 and 7,981,871), and cysteine-terminal intercellular adhesion epitopes, such as Arg-Gly-Asp. This includes (RGD), Arg-Gly-Asp-Ser (RGDS) and Ile-Lys-Val-Ala-Val (IKVAV), cysteine-terminal TAT peptide (GRKKRRQRRRPQ), laminin peptide sequence Cys-Ser-Arg-Ala-Arg-Lys-Gln-Ala-Ala-Ser-Ile-Lys-Val-Ala-Val-Ser-Ala-Asp-Arg (CSRARKQAASIKVAVSADR;lam-IKVAV), and cysteine-terminal elastin-like polypeptides, such as sequence (VPGXG) (where X = any amino acid other than proline).

[0066] Thiol-containing drugs include, but are not limited to, captopril, thiophan, thiopronine, and penicillamine.

[0067] Suitable proteins containing a cysteine ​​group include, but are not limited to, IL-3 variants (e.g., see U.S. Patent No. 5,166,322), IL-2 variants (e.g., see U.S. Patent No. 5,206,344), protease nexin-1 variants (e.g., see U.S. Patent No. 5,766,897), cysteine ​​variants of granulocyte-macrophage colony-stimulating factor (see, e.g., U.S. Patent No. 7,148,333; and Bioconjugate Chem., 2005, 16(5), pp1291-1298; DOI:10.1021 / bc050172r), and cysteine-modified maize ribosome inactivating proteins (maize RIP) [e.g., Toxins]. These include [see 2016,8,298;doi:10.3390 / toxins8100298], cysteine ​​analogs of erythropoietin [e.g., see Int J Nanomedicine.2011;6:1217-1227;doi:10.2147 / IJN.S19081], truncated antibody fragments [e.g., see Protein Eng Des Sel(2007)20 (5):227-234.DOI:https: / / doi.org / 10.1093 / protein / gzm015], and cysteine ​​analogs of osteogenic factor-2 (e.g., see Bioconjugate Chem.,2010,21(10),pp1762-1772;DOI:10.1021 / bc9005706).

[0068] Suitable growth factors containing free thiol groups include, but are not limited to, cysteine ​​analogs of human basic fibroblast growth factor (hbFGF) [see, for example, Tropical Journal of Pharmaceutical Research October 2014;13(10):1601-1607;http: / / dx.doi.org / 10.4314 / tjpr.v13i10.5; and Protein Expr.Purif.2006 Jul;48(1):24-7https: / / doi.org / 10.1016 / j.pep.2006.02.002].

[0069] In one embodiment, the disclosure relates to reacting a hydroxyl group bonded to a polymer, for example, a hydroxyl group on hyaluronic acid (HA), with divinyl sulfone (DVS) to obtain a first derivative of the polymer; and the first derivative of the polymer with XR 1 and XR 2 The present invention provides a process comprising reacting a nucleophile of a formula selected from -Y to obtain a second derivative of a polymer. The first derivative has several ethenyl (vinyl) groups bonded to a sulfone group, which are then bonded to the polymer via an oxyethylene group. Some or all of these vinyl groups react with a nucleophile such as a thiol derivative. The extent to which these vinyl groups are reacted can be made explicit in this disclosure. In one embodiment, all or almost all, e.g., 100%, or 99-100%, or 98-100%, or 97-100%, or 96-100%, or 95-100%, are substituted with a thiol derivative. In another embodiment, some substitution is achieved with a thiol derivative, e.g., 1-95% of the available free vinylsulfone groups are derivatized.

[0070] For example, in one embodiment, the number of vinyl sulfone residues that can bind to a polysaccharide and react with a free thiol-containing compound is variable. The percentage of vinyl sulfone residues that react with the free thiol-containing compound is variable from 1% to 100%. 1 NMR, such as 1H-NMR, can be used to determine the substitution percentage. If 100% substitution of vinyl sulfone groups occurs, essentially all available vinyl sulfone residues bound to the polysaccharide react with the free thiol-containing compound to form thioether bonds. If less than 100% of available vinyl sulfone groups react with the free thiol-containing compound, the polysaccharide will contain both vinyl sulfone groups and compounds bound via thioether bonds. The percentage of repeating units of a polysaccharide substituted via thioether bonds can be determined by NMR, usually... 1 This can be determined by 1H-NMR. The substitution percentage (often calculated in moles) is in the range of 1% to 100%, preferably exceeding 10%, and more preferably exceeding 25%.

[0071] In one embodiment, the Michael addition reaction of a free thiol compound with a vinyl sulfone residue on a polysaccharide may occur using a single free thiol-containing compound. In another embodiment, this addition reaction may occur using two or more different free thiol-containing compounds.

[0072] Figure 2 illustrates options for carrying out the polymer derivatization reaction according to this disclosure. In Figure 2, "A" represents a hydroxyl-substituted polymer such as hyaluronic acid or polyvinyl alcohol. Polymer A can be characterized with respect to its molecular weight. In one embodiment, the intrinsic viscosity of polymer A is used as an indicator of the polymer's molecular weight. Optionally, the intrinsic viscosity of polymer A is 0.3 to 3 m 3 The range is / kg. In another embodiment, chromatography is used to characterize the molecular weight of polymer A. Optionally, the weight-average molecular weight of polymer A is approximately 75,000 Da to 3,000,000 Da.

[0073] In Figure 2, "B" represents the product obtained by reacting polymer A with divinyl sulfone (DVS) under basic conditions (NaOH in aqueous solvent). Polymer B is the compound of the present disclosure. Polymer B is shown as two polymers. When linked to each other via X bonds, X represents the diethyl sulfone group of the formula -CH2-CH2-SO2-CH2-CH2-, which was originally part of the hydroxyl group of polymer A and linked at each of its ends. These X groups are created by the reaction of two hydroxyl groups reacting with two vinyl groups of divinyl sulfone. Although these X groups have been shown to link two different A polymers to each other, a single X group may also link two hydroxyl groups of a single A polymer to form polymer B according to the present disclosure.

[0074] In Figure 2, "B" contains three VS groups in addition to three X bonds between two A polymers. The VS groups are the result of a divinyl sulfone substitution reaction by the hydroxyl groups of polymer A. To create the VS groups, only one of the two vinyl groups of the divinyl sulfone molecule reacts with only one of the hydroxyl groups of polymer A. In one embodiment of the present disclosure, a hydroxyl-substituted polymer ("A") reacts with divinyl sulfone (DVS) to create bonds between two or more hydroxyl groups in a mixture of hydroxyl-substituted polymer chains (shown as polymer B in Figure 2), and in addition, to create vinyl sulfone substituents on one or more hydroxyl-substituted polymer chains.

[0075] In one embodiment, polymer B still contains unreacted hydroxyl groups. For example, if a desired amount of polymer A containing a specified number of hydroxyl groups is loaded into a flask, the addition of DVS will consume at least 5%, or at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80% of the initial hydroxyl groups in the formation of X and VS groups present in polymer B. The number of hydroxyl groups present after the reaction of DVS can be described with respect to hydroxyl residues such that, in addition or in place, at least 15%, or at least 20%, or at least 25%, or at least 30%, or at least 35%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80% of the initial hydroxyl groups are still present in polymer B. The number of hydroxyl groups present in polymer B may also be expressed as the range of the initial number of hydroxyl groups present in polymer A, for example, the conversion from polymer A to polymer B consumes 5-10% of the available hydroxyl groups, or in other embodiments, 5-15%, or 5-20%, or 5-25%, or 5-30%, or 5-35%, or 10-15%, or 10-20%, or 10-25%, or 10-30%, or 10-35%, or 10-40% of the initially available hydroxyl groups.

[0076] In one embodiment, polymer B contains both X substituents and VS substituents. In one embodiment, polymer B contains both X substituents and VS substituents, and within its molar ratio, the number of VS groups exceeds the number of X groups. However, in another embodiment, the number of X groups exceeds the number of VS groups. In yet another embodiment, the X groups provide at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or at least 90% of the total number of X and VS groups.

[0077] As shown in Figure 2, polymer B can act as a reactant to produce either polymer C or polymer D, which are polymers according to this disclosure. To produce polymer C, a mixture of nucleophiles represented as R1SH and R2SH in Figure 2 is reacted with polymer B. To produce polymer D, a single nucleophile represented as R1SH in Figure 2 is reacted with polymer B. This disclosure provides polymers B, polymer C, polymer D, and reactions for producing polymer B from polymer A, for producing polymer C from polymer B, and for producing polymer D from polymer B. In one embodiment, polymers A, B, and C are each derivatized hyaluronic acid.

[0078] Polymer D contains an X moiety that interconnects two polymer A chains. In addition, polymer D contains a ZS-R1 moiety produced by the reaction of a vinyl sulfone (VS) group of polymer B with a thiol compound R1SH to provide an -O-CH2-CH2-SO2-CH2-CH2-S-R1 moiety (abbreviated as the ZS-R1 moiety in Figure 2). In one embodiment, the disclosure provides polymer D having a mixture of X groups and ZS-R1 groups. In one embodiment, the X groups make up at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total of the X groups and ZS-R1 groups. In one embodiment, the ZS-R1 group provides at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total of the X group and the ZS-R1 group.

[0079] In one embodiment, the disclosure provides a polymer E having the structure shown in Figure 2. In another embodiment, the disclosure provides a polymer F having the structure shown in Figure 2. In yet another embodiment, the disclosure provides a polymer G having the structure shown in Figure 2. In yet another embodiment, the disclosure provides a polymer H having the structure shown in Figure 2.

[0080] As shown in Figure 2, polymer E can be obtained by reacting polymer A with divinyl sulfone under basic conditions. As shown in Figure 2, polymer E can be formed from polymer A by the conversion of the hydroxyl groups of polymer A to vinyl sulfone (VS) groups through the reaction of divinyl sulfone (DVS). In polymer E, X groups that interconnect the two hydroxyl groups of polymer A are present in small numbers, if any. In various embodiments, VS groups account for at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.9% of the total X and VS groups present in polymer E.

[0081] Polymer E may optionally react with one or more additional nucleophiles, such as R2SH, in combination with R1SH to provide polymers of structures F, G, or H, as shown in Figure 2. Polymer F has a mixture of VS residues and ZS-R1 groups formed by the reaction of VS groups with R1SH. The charge of R1SH formed on a molar basis is less than 100% of the total number of VS groups present on polymer E. Based on this stoichiometry, not all VS groups react with R1SH molecules, and therefore polymer F has a mixture of VS groups and ZS-R1 groups. In one embodiment, VS groups account for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total of VS groups and ZS-R1 groups. In one embodiment, the ZS-R1 group accounts for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total of the VS group and the ZS-R1 group.

[0082] Polymer G is predominantly composed of ZS-R1 groups, with few or no X and VS groups present. In various embodiments, ZS-R1 groups account for at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.9% of the total X, VS, and ZS-R1 groups present in polymer G. Polymer G can be formed by the reaction of polymer E with R1SH molecules in equimolar or molar excess relative to the moles of available VS groups.

[0083] Polymer H is predominantly composed of ZS-R1 and ZS-R2 groups, with few or no X and VS groups present. In various embodiments, the combined ZS-R1 and ZS-R2 groups account for at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.9% of the combined X and VS groups and ZS-R1 and ZS-R2 groups present in polymer H. Polymer H can be formed by the reaction of polymer E with a mixture of nucleophiles, for example, a mixture of R1SH and R2SH, as shown in Figure 2.

[0084] In one embodiment, the disclosure provides a polymer I having the structure shown in Figure 2. In one embodiment, the disclosure provides a polymer J which is a gel manufactured as shown in Figure 2. In another embodiment, the disclosure provides a polymer K which is a gel manufactured as shown in Figure 2.

[0085] Polymer I has a mixture of ZS-R1 substituents and VS substituents. In one embodiment, the disclosure provides polymer I having a mixture of VS groups and ZS-R1 groups. In one embodiment, VS groups account for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total of VS groups and ZS-R1 groups. In one embodiment, ZS-R1 groups account for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total of VS groups and ZS-R1 groups. Polymer I can be formed by reacting polymer G with divinyl sulfone under basic conditions. This reaction converts hydroxyl groups (not shown in Figure 2) present on polymer G into vinyl sulfone (VS) groups.

[0086] Polymers J and K are gels that can be manufactured as shown in Figure 2. Polymer J can be formed by crosslinking polymer G. Polymer K can be formed by crosslinking polymer H.

[0087] In one embodiment, the disclosure provides a polymer L having the structure shown in Figure 2. In another embodiment, the disclosure provides a polymer M having the structure shown in Figure 2. In yet another embodiment, the disclosure provides a polymer N having the structure shown in Figure 2.

[0088] Polymer L, as shown in Figure 2, contains a mixture of ZS-R1 and ZS-R2 substituents. Polymer L may also contain hydroxyl substituents (not shown). In various embodiments, the sum of ZS-R1 and ZS-R2 groups represents at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.9% of the sum of X groups, VS groups, ZS-R1 groups, and ZS-R2 groups present in polymer L. Polymer L can be formed by the conversion of VS substituents to ZS-R2 substituents by the reaction of polymer F containing ZS-R1 and VS substituents with R2SH.

[0089] Polymer M, as shown in Figure 2, contains a mixture of X groups, ZS-R1 groups, and ZS-R2 groups. Polymer M may also contain hydroxyl substituents (not shown). Polymer M can be formed by adding a crosslinking agent such as divinyl sulfone to polymer L, which contains hydroxyl residues. The crosslinking agent creates X groups between the hydroxyl groups present on polymer L.

[0090] Polymer N, as shown in Figure 2, contains a mixture of ZS-R1 and -R- groups, where the R groups form bonds between different polymer A chains. The R groups can be introduced by reacting a precursor polymer, such as polymer F or another polymer containing VS groups, with a polyfunctional nucleophile such as R(SH)n (where n is 2 or greater). In R(SH)n, R represents an aliphatic or aromatic group, which may be optionally substituted.

[0091] In one embodiment, the disclosure provides a polymer O which is a gel that can be formed as shown in Figure 2. In another embodiment, the disclosure provides a polymer P which is a gel that can be formed as shown in Figure 2.

[0092] Polymer O can be formed from polymer I by a two-step reaction. In the first step, polymer I is reacted with a nucleophile such as R1SH to convert the VS groups present on polymer I into corresponding ZS-R1 groups. In the second step, a crosslinking agent X is added to this intermediate polymer to obtain polymer gel O.

[0093] Polymer P can be formed from polymer I by a two-step reaction. In the first step, polymer I is reacted with a nucleophile such as R2SH to convert the VS groups present on polymer I into corresponding ZS-R2 groups. In the second step, a crosslinking agent X is added to this intermediate polymer to obtain polymer gel P.

[0094] Polymer I can also act as a precursor to crosslinked polymers having -R- groups as bonds between polymer chains. The R groups can be introduced by reacting polymer I, or another polymer containing VS groups, with a polyfunctional nucleophile such as R(SH)n (where n is 2 or greater). In R(SH)n, R represents an aliphatic or aromatic group which may be optionally substituted.

[0095] Figure 3 illustrates options for carrying out a polymer derivatization reaction in accordance with this disclosure. In Figure 3, "A" represents a hydroxyl-substituted polymer such as hyaluronic acid or polyvinyl alcohol, which is also shown as polymer A in Figure 2. However, in contrast to Figure 2, the reaction scheme in Figure 3 begins with a crosslinking reaction on polymer A, with little to no conversion of the hydroxyl groups on polymer A to other monofunctional reactive groups.

[0096] As shown in Figure 3, polymer A can also be reacted with a crosslinking agent to obtain a crosslinked polymer A, which is represented as polymer B in Figure 3. Suitable crosslinking reactions for hydroxyl-containing polymers are described elsewhere in this specification.

[0097] In one embodiment, the disclosure provides a polymer C having the structure shown in Figure 3. In another embodiment, the disclosure provides a polymer D having the structure shown in Figure 3. In yet another embodiment, the disclosure provides a polymer E having the structure shown in Figure 3.

[0098] Polymer C can be formed by reacting polymer B with divinyl sulfone (DVS) under basic conditions. Under these reaction conditions, hydroxyl groups (not shown) present on polymer B react with DVS to convert the hydroxyl groups into VS groups. In one embodiment, VS groups account for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total VS and X groups present in polymer C. In one embodiment, X groups account for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the total VS and X groups present in polymer C.

[0099] Polymer D in Figure 3 is a crosslinked polymer having both ZS-R1 and ZS-R2 substituents. Polymer D can be formed by reacting polymer C with a mixture of nucleophiles such as R1SH and R2SH as shown in Figure 3. In one embodiment, the sum of ZS-R1 and ZS-R2 groups accounts for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the sum of ZS-R1, ZS-R2, and X groups present in polymer D. In one embodiment, the X groups account for at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the sum of ZS-R1, ZS-R2, and X groups present in polymer D.

[0100] Polymer E in Figure 3 is a crosslinked polymer having a ZS-R1 substituent (but not a ZS-R2 substituent). Polymer E can be formed by reacting polymer C in Figure 3 with a nucleophile such as R1SH as shown in Figure 3. In one embodiment, ZS-R1 groups account for at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the total ZS-R1 and X groups present in polymer E. In one embodiment, X groups account for at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the total ZS-R1 and X groups present in polymer E.

[0101] Furthermore, as shown in Figure 3, the disclosure provides polymers of structure F and structure G, as well as their crosslinked gels. Polymer F in Figure 3 contains the ZS-R1 substituent, and polymer G contains a mixture of the ZS-R1 substituent and the ZS-R2 substituent. Neither polymer F nor G is a crosslinked polymer. However, when polymers F and G are treated with a crosslinking agent or exposed to crosslinking conditions, the corresponding crosslinked polymer having a gel form is obtained (shown as polymer H in Figure 3).

[0102] While we do not wish to be bound by any particular theory, when vinyl groups are present in a solution of derivatized polyvalent polymer molecules, crosslinking occurs at all or at a low level. This "accidental" crosslinking may be present but is not measurable, and the solution does not exhibit the characteristics of crosslinking; in this specification, it is referred to as a solution or composition containing a derivative of a non-crosslinked polymeric polyhydric alcohol.

[0103] Therefore, in one embodiment, the disclosure provides vinyl sulfone functionalization (i.e., derivatization) of a polysaccharide ("first derivative") and subsequent reaction of the vinyl sulfone substituent with one or more free thiol-containing compounds ("second derivative"), thereby obtaining a polysaccharide functionalized by the compound via a thioether bond and simultaneously with a vinyl sulfone functional group ("third derivative"). In another embodiment, the above compound can be further reacted with a free thiol-containing compound ("fourth derivative"). The molar ratio of the free thiol compound used in this reaction can be changed so that 1% to 100% of the second added vinyl sulfone functional group is reacted. The free thiol-containing compound used in the second Michael addition reaction (derivatization reaction) may be the same as or different from the one used in the first Michael addition reaction. In the second Michael addition reaction, a single free thiol-containing compound may be used, or a mixture of two or more different free thiol-containing compounds may be used. In another embodiment, at least one additional vinyl sulfone / free thiol-containing compound reaction cycle can be performed using the same free thiol-containing compound or one or more different free thiol-containing compounds.

[0104] In one embodiment, the process of the present disclosure further includes crosslinking a second derivative of the polymer, for example, by reacting the second derivative of the polymer with a crosslinking agent. Upon crosslinking, the second derivative is converted into a third derivative of the polymer, the third derivative being a crosslinked polymer.

[0105] For example, in one embodiment, a polysaccharide derivatized with one or more free thiol-containing compounds and containing available vinyl sulfone residues may be crosslinked by exposing a solution of the derivatized polyvalent polymer to conditions sufficiently basic to allow the available vinyl sulfone residues to react with the hydroxyl groups of the polysaccharide. In one embodiment, the reaction pH is greater than 12, for example, in the range of pH 12.5 to 13.0. 1The amount of vinyl sulfone residue, which is often measured as a substitution percentage by 1H-NMR, the reaction time, and the reaction temperature can be selected to achieve the desired degree of crosslinking.

[0106] In another embodiment, a polysaccharide derivatized with one or more free thiol-containing compounds and containing available vinyl sulfone residues can be mixed with a polysaccharide derivatized with one or more free thiol-containing compounds and containing available vinyl sulfone residues, and these free thiol-containing compounds may be the same, different, or a combination thereof. The resulting mixture can be crosslinked by exposing a solution of the derivatized polyvalent polymer to conditions that are sufficiently basic to allow the available vinyl sulfone residues to react with the hydroxyl groups of the polysaccharide. In one embodiment, the reaction pH is greater than 12, for example, in the range of pH 12.2 to 13. 1 The amount of vinyl sulfone residue, which is often measured as a substitution percentage by 1H-NMR, the reaction time, and the reaction temperature can be selected to achieve the desired degree of crosslinking.

[0107] In another embodiment, a non-derivative polysaccharide can be added to the crosslinking reaction mixture, and the resulting mixture can be crosslinked by exposing the solution of the derivatized polymeric polyhydric alcohol and the non-derivative polysaccharide to conditions sufficiently basic to allow the available vinyl sulfone residues of the derivatized polymeric polyhydric alcohol to react with the hydroxyl groups of the polysaccharide. In one embodiment, the reaction pH is greater than 12, for example, in the range of pH 12.2 to 13. The amount of vinyl sulfone residues, which is often measured as substitution percentage by 1H-NMR, the reaction time and reaction temperature can be selected to achieve a desired degree of crosslinking.

[0108] Crosslinking can be achieved by using an external crosslinking agent. In one embodiment, the crosslinking agent is added to a second derivative of the polymer. Exemplary crosslinking agents that can be used include carbodiimides, bis-epoxides, divinyl sulfone derivatives, and combinations thereof. Another preferred crosslinking agent is a polythioether derivative. In one embodiment, at least two (possibly two, three, four, etc.) different thioether derivatives are combined with the crosslinking agent to prepare the derivative of the polyvalent polymer to be fully or partially crosslinked. In this case, exemplary crosslinking agents include, but are not limited to, carbodiimides, bis-epoxides, divinyl sulfone derivatives, and combinations thereof.

[0109] For example, in one embodiment, a polysaccharide derivatized with one or more free thiol-containing compounds can be crosslinked by adding a crosslinking agent and adjusting the pH of the reaction mixture so that the derivatized polysaccharide forms a crosslinked derivatized polysaccharide or composition. Usable crosslinking agents include, but are not limited to, biscarbodiimides, bisepoxides, divinyl sulfone derivatives, diisocyanates, dihalidochlorides, disuccinimidyl derivatives, and combinations thereof.

[0110] Biscarbodiimide compounds may include, but are not limited to, para-phenylenebis-(ethyl)-carbodiimide, 1,6-hexamethylenebis(ethylcarbodiimide), 1,8-octamethylenebis(ethylcarbodiimide), 1,10-decamethylenebis(ethylcarbodiimide), 1,12-dodecamethylenebis(ethylcarbodiimide), PEG-bis(propyl(ethylcarbodiimide)), 2,2'-dithioethylbis(ethylcarbodiimide) (bis(ethylcarbodiimide)), 1,1'-dithio-p-phenylenebis(ethylcarbodiimide); para-phenylene-bis(ethylcarbodiimide), and 1,1'-dithio-m-phenylenebis(ethylcarbodiimide).

[0111] When using a biscarbodiimide crosslinking agent, the biscarbodiimide is mixed with a buffer solution containing a derivatized carboxylic acid-containing polysaccharide. The target pH of the buffer solution is pH 5 to pH 6.5.

[0112] The bis-epoxide compounds may include, but are not limited to, 1,4-butanediol diglycidyl ether (BDDE), 1,2,7,8-diepoxyoctane (DEO), and poly(ethylene glycol) diepoxide. When using a bis-epoxide crosslinking agent, the bis-epoxide is mixed with an aqueous solution of the derivatized polysaccharide, and the pH is raised to pH > 9. To produce a crosslinked derivatized polyvalent polymer, this reaction can be carried out at 40°C for more than 4 hours.

[0113] The divinyl sulfone crosslinking agent may include, but is not limited to, divinyl sulfone and poly(ethylene glycol)bisvinyl sulfone.

[0114] When using divinyl sulfone crosslinking agents, the reaction pH of the aqueous solution can be raised to above 12 in order to achieve crosslinking. The degree of crosslinking can be changed by altering the amount of crosslinking agent added, the reaction time, the reaction pH, and the reaction temperature.

[0115] In another embodiment, a mixture of at least two different thioether-derivative polysaccharides can be mixed with each other, a crosslinking agent can be added, and the reaction conditions can be adjusted so that the derivatized polyvalent polymer is crosslinked. The relative ratio of these different derivatized polysaccharides can be modified to obtain crosslinked derivatized polyvalent polymers with different properties. These properties include, but are not limited to, equilibrium swelling, swelling rate, drug release characteristics, elastic modulus, storage modulus, loss modulus, decomposition, tensile strength, tissue adhesion, and lubricity. As used herein, “derivative polyvalent polymer” may also include a composition comprising one or more derivatized polyvalent polymers.

[0116] In another embodiment, at least two different crosslinking agents can be used to crosslink a derivatized polysaccharide. In one embodiment, two different crosslinking agents derived from the same group can be used to crosslink a derivatized polyvalent polymer. For example, divinyl sulfone and poly(ethylene glycol)bisvinyl sulfone or 1,4-butanediol diglycidyl ether (BDDE) and poly(ethylene glycol) diepoxide can be used.

[0117] In another embodiment, two different crosslinking agents from different groups can be used. For example, divinyl sulfone and 1,4-butanediol diglycidyl ether (BDDE) can be used to crosslink the derivatized polysaccharide. In another embodiment, the crosslinking agents can be added sequentially so that the first crosslinking occurs in the presence of the first crosslinking agent (crosslinked), and then the second crosslinking agent is added so that the second crosslinking occurs. The reaction conditions can be changed after the first crosslinking reaction and before the second crosslinking reaction. Reaction conditions such as temperature, pH, buffer, ionic strength, and solvent composition can be modified.

[0118] In one embodiment, crosslinked derivatized polyvalent polymers can be produced by ionic crosslinking. This can be achieved by mixing a derivatized polyvalent polymer of the disclosure having one negative charge with a compound having two or more positive charges. In one embodiment, a solution of a derivatized polyvalent polymer of the disclosure having one negative charge can be prepared and then mixed with a solution of a compound having two or more positive charges. Usable inorganic compounds include, but are not limited to, ferric chloride, aluminum chloride, chromium sulfate, and aluminum sulfate. Usable positively charged polymers include polymers containing two or more lysine, arginine, or histidine amino acids, chitosan and chitosan derivatives, deacetylated hyaluronic acid, polyethyleneimine (PEI), poly(N,N-dimethylaminoethyl methacrylate), poly(4-vinylpyridine), polyethylene glycol-polylysine block copolymer (PEG-PLL), dextran grafted polylysine copolymer, or combinations thereof.

[0119] In one embodiment, a polymer having a positive or negative charge can be applied first. Subsequently, a polymer with the opposite charge can be applied such that ionic interactions occur at the interface between the two layers, causing the polymers to crosslink with each other. In another embodiment, this process can be repeated at least one more time.

[0120] Polymer (e.g., HA-(OCH2CH2SO2CH2CH2-X-R1-Y) n The second derivative of ) can be crosslinked via internal and external crosslinking. For example, in one embodiment, a polysaccharide that is derivatized with one or more free thiol-containing compounds and also contains available vinyl sulfone residues can be crosslinked in the presence of an external crosslinking agent. In one embodiment, the reaction conditions can be adjusted so that the available vinyl sulfone residues and the added external crosslinking agent react simultaneously. For example, divinyl sulfone can be added as an external crosslinking agent, and then the pH can be raised to pH > 12, resulting in a crosslinked product.

[0121] Alternatively, crosslinking can be performed first via available vinyl sulfone residues, followed by the addition of an external crosslinking agent. Reaction conditions, such as pH, can be altered to facilitate the crosslinking reaction of the added external crosslinking agent. For example, the pH of the derivatized polysaccharide containing available vinyl sulfone residues can be raised to pH > 12. Once the reaction reaches the desired level, the pH can be changed to pH 5-6.5 using a buffer, and then a biscarbodiimide crosslinking agent, such as para-phenylenebis-(ethyl)-carbodiimide, can be added to the reaction mixture and the reaction can be continued until the desired level of crosslinking is achieved. In another embodiment, biscarbodiimide crosslinking can be performed by first adjusting the pH of the derivatized polysaccharide to 5-6.5, adding biscarbodiimide and allowing crosslinking to proceed to the desired level, and then raising the pH to pH > 12 to enable crosslinking of vinyl sulfone residues.

[0122] In one embodiment, a polysaccharide derivatized with one or more free thiol-containing compounds and containing available vinyl sulfone residues can be crosslinked in the presence of an external crosslinking agent having at least two free thiol functional groups. These free thiol groups may be located on the crosslinking center molecule "C". The center molecule may be a linear or cyclic alkane, a polyethylene glycol (PEG) oligomer or polymer, or any other suitable center molecule. In the case of a PEG-based crosslinking agent, the PEG may be linear, branched (having two polymer arms), or multi-armed (e.g., having 3, 4, 5, 6, 7, 8 or more polymer arms). Thus, in such cases, the center molecule is generally linear PEG, branched PEG with two arms, or multi-armed PEG with PEG arms radiating from a central core.

[0123] Exemplary cores of such multi-armed polymers include erythritol, pentaerythritol, trimethylolpropane, glycerol, glycerol dimer (3,3'-oxydipropane-1,2-diol), glycerol oligomer, sorbitol, and hexaglycerol.

[0124] Exemplary thiol crosslinking agents include PEG-dithiol (HS-PEG-SH), 3-arm PEG-tri-thiol (glycerin core), 4-arm PEG-tetrathiol (pentaerythritol core), or 8-arm PEG-octa-thiol (hexaglycerin core). These multi-arm PEG reagents may also have fewer arms functionalized with thiols than the total number of arms. Further preferred thiol reagents having PEG as the central molecule are aromatic dithiols, such as those available from Laysan Bio (Arabia, Alabama) and NanoScience. Other suitable thiol crosslinking agents include dimercaptosuccinic acid, 2,3-dimercapto-1-propanesulfonic acid, trimethylolpropanetris(3-mercaptopropionate), dithiol-functionalized Pluronic F127, dithiol-functionalized F68, dihydrolipoic acid, peptides containing at least two cysteine ​​amino acids, thiol-functionalized dextran, and thiol-functionalized hyaluronic acid.

[0125] The polymers of this disclosure, such as polysaccharides, first, second, and third derivatives of such polymers can be processed into many forms. In the case of non-crosslinked derivatized polyvalent polymers, exemplary compositions of the derivatized polyvalent polymers include, but are not limited to, solutions, suspensions, emulsions, films, gels, coatings for the surface of articles, electrospun matrices, fine particles, fibers, lyophilized solids, cylinders, discs, gels, powders, or particle forms. Particle forms can be prepared by grinding (e.g., jet mill grinding, roller mill grinding, freeze-grinding, mechanical grinding), fragmentation, spray-drying, precipitation, or grinding. In the case of crosslinked derivatized polyvalent polymers, compositions of the derivatized polyvalent polymers may be in the form of suspensions, films, electrospun matrices, fibers, lyophilized solids, cylinders, discs, gels, powders, or particle forms. Particle forms can be prepared by grinding (e.g., jet mill grinding, roller mill grinding, freeze-grinding, mechanical grinding), fragmentation, spray-drying, precipitation, or grinding.

[0126] Solutions of derivatized polyvalent polymers can be prepared by dissolving the derivatized polyvalent polymer in a suitable solvent or combination of solvents. For example, water or a combination of water and a water-miscible solvent can be used. Examples of water-miscible solvents include, but are not limited to, methanol, ethanol, isopropanol, dimethylformamide (DMF), acetone, 1,4-dioxane, pyridine, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), and acetonitrile. The prepared solution can be sterilized by filtering through a 0.2 μm sterilization filter. In one embodiment, the solution can be prepared using a single derivatized polysaccharide. The concentration of the prepared solution may be, for example, in the range of 0.01% (w / v) to about 50% (w / v). In one embodiment, the concentration is in the range of 0.1% (w / v) to 10% (w / v).

[0127] Films of the non-crosslinked derivatized polyvalent polymers of this disclosure can be prepared by preparing a solution of the derivatized polyvalent polymer. This solution can then be placed in a mold and stretched onto a surface using a Gardner knife. The surface used may be glass, metal foil, stainless steel, Teflon®, nylon, polyethylene, polypropylene, or a release liner. The solvent can then be removed to form a film. The solvent removal rate can be modified using at least one of the following parameters: temperature, airflow or inert gas flow, and pressure. To increase the solvent evaporation rate, the temperature can be increased, the airflow velocity or inert gas flow velocity can be increased, or the pressure can be decreased. Combinations of these processes are also available. To decrease the solvent evaporation rate, the temperature can be decreased, the airflow velocity or inert gas flow velocity can be decreased, or the pressure can be increased. Combinations of these processes are also available. The film may contain one of the derivatized polyvalent polymers of this disclosure. The film may also contain two or more different derivatized polyvalent polymers of this disclosure. A composite material film can be produced by casting a second film onto a first film after the first film has been prepared. A composite material film can also be produced by sequentially casting additional layers onto a preceding layer. The layers of the composite material film may include the same derivatized polyvalent polymer of the disclosure, different derivatized polyvalent polymers of the disclosure, or a combination thereof.

[0128] The lyophilized form of the non-crosslinked derivatized polyvalent polymer of this disclosure can be prepared by preparing a solution of the derivatized polyvalent polymer, freezing the solution, and then placing the frozen derivatized polyvalent polymer solution under vacuum so that the solvent sublimes and the derivatized polyvalent polymer composition remains in a solid form. The lyophilized form of the derivatized polyvalent polymer composition of this disclosure may comprise one of the derivatized polyvalent polymers of this disclosure. In another embodiment, the lyophilized form of the derivatized polyvalent polymer and / or composition of this disclosure may comprise two or more different derivatized polyvalent polymers of this disclosure. In another embodiment, the lyophilized composition of one or more derivatized polyvalent polymers may comprise one or more derivatized polyvalent polymers in addition to additives such as chitosan or chitosan derivatives (e.g., chitosan HCl, chitosan acetate, or chitosan lactate). The form of the lyophilized derivatized polyvalent polymer composition may depend on the form of the container in which the solution is poured. In another embodiment, the form of the lyophilized derivatized polyvalent polymer may depend on the form of the container in which the solution is poured and frozen. This form may be a square, rectangular, disc, triangle, trapezoid, cylinder, or any other form from which a mold can be produced.

[0129] The derivatized polyvalent polymer compositions of this disclosure may be in the form of powder or particles. Powder or particles can be obtained directly by precipitation. Powder or particle forms can also be obtained by grinding, abrasion, spray drying or fragmentation. Compositions comprising, but not limited to, films, precipitated derivatized polyvalent polymers, dried derivatized polyvalent polymers and / or compositions, freeze-dried derivatized polyvalent polymers and / or compositions, or derivatized polyvalent polymers and / or compositions dried in some form, may be further processed by grinding (jet mill grinding, roller mill grinding, freeze-milling, mechanical grinding), abrasion or fragmentation. Combinations of these methods may also be used. Derivatized polyvalent polymer compositions having particle sizes in the range of 100 nm to 5 mm can be prepared. Powder or particle derivatized polyvalent polymer compositions of this disclosure with a specific particle size range can be prepared by separating the derivatized polyvalent polymer composition particles according to particle size using a sieve. The particle size distribution may be broad, such as with a standard deviation of the mean particle size exceeding 40%. The particle size distribution may also be narrow, such as with a standard deviation of the mean particle size less than 30%. The final powder or particle form of the derivatized polyvalent polymer composition of this disclosure may include a single average particle size distribution, or it may include two or more particle distributions prepared by mixing particles of different average particle sizes.

[0130] The derivatized polyvalent polymer compositions of this disclosure can be molded into a solid form by preparing a solution of the derivatized polyvalent polymer in a removable solvent, pouring this solution into a mold of a specific shape, and then removing the solvent to obtain a derivatized polyvalent polymer composition in solid form. The molds used may be of various shapes, but are not limited to, and may include cubes, squares, cylinders, semi-circular rodes, and tubes. The solid derivatized polyvalent polymer composition of this disclosure can then be removed from the mold.

[0131] The derivatized polyvalent polymer compositions of this disclosure can be processed as electrospinning matrices. In this process, a solution of the derivatized polyvalent polymer of this disclosure is prepared. The solvent used may be an organic solvent, water, or a combination thereof. For example, in the case of hyaluronic acid-based derivatized polyvalent polymers, a water / ethanol or water / dimethylformamide (DMF) solvent mixture can be used. A solution with a concentration of 0.5 to 5% (w / v) can be prepared. The solution to be electrospinned can be placed in a syringe with a needle. The syringe is then placed in a syringe pump. The needle may have a blunt end and an inner diameter in the range of 0.25 to 1 mm. The needle and collection plate are attached to a high voltage source. A voltage is then applied to this system. The applied voltage may be 10 kV to 45 kV. The syringe pump can then pump out this solution. The flow rate of the syringe pump may be in the range of 10 uL / min to 1000 uL / min. The collector plate may be stationary, rotating, or moving in a specific linear direction to give the fibers some directional orientation. The shape of the collector plate is not limited to but can vary, and may include the following shapes: flat surface, woven surface, curved surface, regular prism, prismatic, round core, elliptical core, semicircular core, or a combination of these shapes. The temperature of the solution, as well as the recovery plate and the surrounding environment, are controllable. The distance between the needle tip and the collector plate is adjustable. The distance between the needle tip and the collector plate may range from 2 to 20 cm. The recovery plate may also be immersed in a solvent to aid in the precipitation of the newly spun fibers, or the solvent may be sprayed onto it. For example, an ethanol bath may be used during the electrospinning of the hyaluronic acid derivatized polyvalent polymer and / or composition of the present disclosure.

[0132] The derivatized polyvalent polymers of this disclosure can be incorporated by solution coating or immersion of an electrospinned matrix produced in the manner described below. In this process, one or more polymer solutions are prepared. The polymers used may be, but are not limited to, polyesters, polyanhydrides, polyorthoesters, polycarbonates, polyester-co-carbonates, polyhydroxybutyrates, or combinations thereof. The biodegradable polymers may include polylactice-co-glycolide copolymers, polydioxanones, polylactice-trimethylene carbonate copolymers, and copolymers containing repeating units derived from at least one of the following monomers: l-lactide, dl-lactide, glycolide, trimethylene carbonate, ε-caprolactone, p-dioxanone, and morpholindione.

[0133] The solvent used may be an organic solvent, water, or a combination thereof. For example, HFIP, DMSO, NMP, chloroform, acetic acid, ethanol, dimethylformamide (DMF) solvent, or a mixture of solvents can be used. Solutions with concentrations of 0.5–25% (w / v) can be prepared. The solution to be electrospun can be placed in a syringe with a needle. The syringe is then placed in a syringe pump. This needle may have a blunt end and an inner diameter in the range of 0.25–2.5 mm. The needle and collection plate are attached to a high voltage source. Voltage is then applied to this system. The applied voltage may be 10kV–45kV. The syringe pump can then pump out this solution. The flow rate of the syringe pump may be in the range of 0.0001uL / min–423mL / min. The collector plate may be stationary, rotating, or moving in a specific linear direction to give the fibers some directional orientation. The shape of the collector plate is not limited to but can be diverse, including the following shapes: flat surface, woven surface, curved surface, regular prism, prismatic prism, round core, elliptical core, semicircular core, or a combination of these shapes. The distance between the needle tip and the collector plate is variable. The distance between the needle tip and the collector plate can range from 2 to 50 cm. The collection plate may also be immersed in a solvent that assists in the precipitation of the newly spun fibers, or the solvent may be sprayed onto it. For example, an ethanol bath may be used when electrospinning the hyaluronic acid-based derivatized polyvalent polymer of this disclosure.

[0134] The derivatized polyvalent polymers and / or compositions of this disclosure can be processed in the form of fibers. A solution of the derivatized polyvalent polymer of this disclosure is prepared. This solution is then extruded through an orifice to produce solvent-containing fibers. These fibers can be extruded into one or more solvent baths that assist in fiber formation. These fibers are then dried to produce solid fibers. The fibers can be prepared as monofilament fibers or multifilament fibers. In one embodiment, these fibers can then be further processed by an annealing step. US9228027, US5520916, US5824335, US8389498, US20130309494, and US20150119783 describe exemplary methods for producing fibers from polysaccharides. These are incorporated herein by reference as means for producing fibers of the derivatized polyvalent polymers and / or compositions of this disclosure.

[0135] The fibers may be further processed by knitting or weaving to form knitted or woven compositions. The knitted or woven compositions may be in the form of meshes. The mesh may comprise a single derivatized polyvalent polymer and / or composition of the present disclosure. In another embodiment, the mesh may comprise two or more different derivatized polyvalent polymers and / or compositions of the present disclosure. In another embodiment, the fibers may be further processed into braids. The braids may comprise a single derivatized polyvalent polymer and / or composition of the present disclosure. In another embodiment, the braids may comprise two or more different derivatized polyvalent polymers and / or compositions of the present disclosure. In meshes and braids using different derivatized polyvalent polymers and / or compositions of the present disclosure, the derivatized polyvalent polymers and / or compositions used may result in meshes or braids having properties that change as a function of time. These include degradation rate, water absorption, elongation, modulus of elasticity, tensile strength, physical shape, lubricity, cell adhesion, or a combination of these properties.

[0136] Braided, woven, or assembled derivatized polyvalent polymers and / or compositions can be prepared in the presence of biodegradable or non-biodegradable non-polysaccharide materials. These materials include polyethylene, polypropylene, polyethylene terephthalate (PET), polytetrafluoroethylene (PTFE), nylon, polyurethane, polyester, polyanhydride, polyorthoester, polycarbonate, polyester-co-carbonate, polyhydroxybutyrate, or combinations thereof.

[0137] The crosslinked polymers of this disclosure may take various physical forms, including particles, films, freeze-dried sponges, powders, and particles (e.g., crushed, fragmented, precipitated, and ground particles), and may be formed in situ (e.g., spray or liquid).

[0138] Films of the crosslinked derivatized polyvalent polymers and / or compositions of this disclosure can be prepared by preparing a solution of the derivatized polyvalent polymer and / or composition to be crosslinked. The derivatized polyvalent polymer can be crosslinked by known methods and / or by methods described herein. Before the final crosslinking process, a crosslinking agent may be added if necessary, and the solution pH may be adjusted to initiate the crosslinking process. The solution can then be placed in a mold or spread onto a surface using, for example, a Gardner knife. The surface used may be glass, metal foil, stainless steel, Teflon, nylon, polyethylene, polypropylene, or a release liner. The solution can then be crosslinked to allow for gel formation. Heating may be used to increase the crosslinking rate. The solvent can then be removed to form a film.

[0139] Films of the crosslinked derivatized polyvalent polymers of this disclosure can be prepared by preparing a solution of the derivatized polyvalent polymer to be crosslinked. The derivatized polyvalent polymer can be crosslinked by one of the methods described above. The solution can then be placed in a mold or spread onto a surface using a Gardner knife. The surface used may be glass, metal foil, stainless steel, Teflon, nylon, polyethylene, polypropylene, polystyrene, or release liner. A crosslinking agent may be added before or after drying the derivatized polyvalent polymer to form a crosslinked film or gel. The rate of removal of residual solvent can be modified by using at least one of the following parameters: temperature, airflow or inert gas flow, and pressure. To increase the solvent evaporation rate, the temperature can be increased, the airflow velocity or inert gas flow velocity can be increased, or the pressure can be decreased. Combinations of these processes are also available. To decrease the solvent evaporation rate, the temperature can be decreased, the airflow velocity or inert gas flow velocity can be decreased, or the pressure can be increased. Combinations of these processes are also available. The film may contain one of the derivatized polyvalent polymers of this disclosure.

[0140] The film may also contain two or more different derivatized polyvalent polymers of the present disclosure. A composite film can be prepared by preparing a first film and then casting a second film on top of the first film. A composite film can also be prepared by sequentially casting additional layers on top of a preceding layer. The layers of the composite film may contain the same derivatized polyvalent polymer of the present disclosure, different derivatized polyvalent polymers of the present disclosure, or a combination thereof. The film may contain both crosslinked and uncrosslinked derivatized polyvalent polymers of the present disclosure.

[0141] The lyophilized form of the crosslinked derivatized polyvalent polymer of this disclosure can be prepared by preparing a solution of the derivatized polyvalent polymer, crosslinking the derivatized polyvalent polymer, freezing the crosslinked derivatized polyvalent polymer composition, and then placing the frozen derivatized polyvalent polymer solution under vacuum so that the solvent sublimes and the resulting derivatized polyvalent polymer composition remains in a solid form. The lyophilized form of the derivatized polyvalent polymer of this disclosure may comprise one of the derivatized polyvalent polymers of this disclosure. In another embodiment, the lyophilized form of the derivatized polyvalent polymer of this disclosure may comprise two or more different derivatized polyvalent polymers of this disclosure. The form of the lyophilized derivatized polyvalent polymer composition depends on the form of the container into which the solution is poured and frozen. This form may be square, rectangular, disc-shaped, triangular, trapezoidal, cylindrical, or any other form from which a mold can be produced. The lyophilized derivatized polyvalent polymer composition of this disclosure may comprise both crosslinked and uncrosslinked derivatized polyvalent polymers of this disclosure. In another embodiment, the crosslinked or non-crosslinked lyophilized polyvalent polymer composition may be rehydrated in the presence of other materials disclosed herein. In another embodiment, the rehydrated lyophilized polyvalent polymer composition may undergo a second lyophilization step.

[0142] In another embodiment, the solution used to rehydrate the first freeze-dried derivatized polyvalent polymer can be crosslinked. In another embodiment, the derivatized polyvalent polymer composition produced from the second crosslinking step can be freeze-dried to produce a dry porous derivatized polyvalent polymer composition. In another embodiment, the derivatized polyvalent polymer solution may be combined with the bioactive agent as is before freeze-drying. In another embodiment, the freeze-dried polyvalent polymer composition may be combined with the bioactive agent via a rehydration process, followed by a first freeze-drying, and further drying may or may not be performed thereafter.

[0143] The crosslinked derivatized polyvalent polymer compositions of this disclosure may be in the form of powder or particles. The powder or particle form may also be obtained by grinding, grinding, spray drying or fragmentation. Films, precipitated derivatized polyvalent polymers and / or compositions, dried derivatized polyvalent polymers and / or compositions, freeze-dried derivatized polyvalent polymers and / or compositions, or derivatized polyvalent polymers and / or compositions dried in some form may be further processed by grinding (jet mill grinding, roller mill grinding, freeze-grinding, mechanical grinding), grinding or fragmentation. Combinations of these methods may also be used. Derivatized polyvalent polymer compositions having particle sizes in the range of 100 nm to 5 mm can be prepared. Powder or particle derivatized polyvalent polymer compositions of this disclosure with a specific particle size range can be prepared by separating the derivatized polyvalent polymer composition according to particle size using a sieve. The particle size distribution may be broad, such as with a standard deviation of the mean particle size exceeding 40%. The particle size distribution may also be narrow, such as with a standard deviation of the mean particle size less than 30%. The final powder or particle form of the derivatized polyvalent polymer and / or composition of the present disclosure may include a single average particle size distribution, or it may include two or more particle distributions prepared by mixing particles of different average particle sizes.

[0144] The crosslinked derivatized polyvalent polymer compositions of this disclosure can be molded into a solid form by preparing a solution of the derivatized polyvalent polymer in a removable solvent, pouring this solution into a mold of a specific shape, crosslinking the derivatized polyvalent polymer within the mold, and then removing the solvent to obtain a crosslinked derivatized polyvalent polymer composition in solid form. The molds used may be of various shapes, but are not limited to, cubes, squares, cylinders, semi-circular rodes, and tubes. The solid derivatized polyvalent polymer composition of this disclosure can then be removed from the mold.

[0145] In one embodiment, the derivatized polyvalent polymers of the present disclosure can be used to prepare in-situ gel-forming compositions. The derivatized polyvalent polymers of the present disclosure containing available vinyl sulfone groups can be reacted with compounds containing at least two available free thiol groups or compounds containing at least two available amine groups, preferably primary or secondary amines. Exemplary thiol-containing compounds include PEG-dithiol (HS-PEG-SH), 3-arm PEG-trithiol (glycerin core), 4-arm PEG-tetrathiol (pentaerythritol core), or 8-arm PEG-octathiol (hexalicerin core). The above multi-arm PEG reagents may also have fewer arms functionalized with thiols than the total number of arms. Further preferred thiol reagents having PEG as the central molecule are aromatic dithiols, such as those available from Laysan Bio (Arabia, Alabama) and NanoScience. Other suitable thiol crosslinking agents include dimercaptosuccinic acid, 2,3-dimercapto-1-propanesulfonic acid, dihydrolipoic acid, peptides containing at least two cysteine ​​amino acids, thiol-functionalized polysaccharides, thiol-functionalized dextran, and thiol-functionalized hyaluronic acid. In one embodiment, the derivatized polyvalent polymer of the present invention can be prepared as a solution. This solution can be mixed with either a solution of a thiol-containing compound or a thiol-containing compound in solid form to produce a gel composition.

[0146] The derivatized polyvalent polymers of this disclosure, for example, the first, second, and third derivatives of the starting polymer, may be combined with one or more other derivatized polyvalent polymers or other components such as pharmaceutically acceptable excipients, or other known or common components of the composition. Accordingly, this disclosure provides compositions comprising the derivatized polyvalent polymers of this disclosure.

[0147] The derivatized polyvalent polymers and their compositions of this disclosure are available for use in treating living organisms. These living organisms include humans, animals, birds, fish, insects, and plants. The derivatized polyvalent polymers and their compositions used in the following applications may include non-crosslinked derivatized polyvalent polymers, crosslinked derivatized polyvalent polymers, or combinations thereof. In another embodiment, the derivatized polyvalent polymer composition used may contain only one of the derivatized polyvalent polymers of this disclosure. In another embodiment, the derivatized polyvalent polymer composition used may contain two or more of the derivatized polyvalent polymers of this disclosure. The derivatized polyvalent polymers and their compositions may further contain one or more excipients. The derivatized polyvalent polymers and their compositions may further contain one or more bioactive agents. The derivatized polyvalent polymers and their compositions used in the following applications are available in sterile form. Sterilization can be achieved by sterilization filtration, sterile manufacturing, gamma irradiation, electron beam irradiation, ethylene oxide, dry heat, autoclaving, or a combination thereof.

[0148] For example, the derivatized polyvalent polymer compositions of this disclosure may also include excipients. Excipients may be pharmaceutically acceptable excipients. Available excipients include, but are not limited to, natural polymers, synthetic polymers, thermoreversible polymers, biodegradable polymers, buffers, complexing agents, tonic modifiers, ionic strength modifiers, solvents, antioxidants, preservatives, viscosity modifiers, pH modifiers, surfactants, emulsifiers, phospholipids, stabilizers, and pologens.

[0149] The excipient polymers that can be used include, but are not limited to, sodium alginate, calcium alginate, dextran, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, hyaluronic acid, hyaluronic acid derivatives, dextran, heparin, chitosan, chitosan acetate, chitosan lactate, chitin, xantham gum, xylan, guar gum, pullulan, locust bean gum, starch, gelatin, collagen, derivatized collagen, and acacia gum (gum arabic).

[0150] Suitable excipient-degradable polymers include, but are not limited to, polyesters, polyether esters, polyorthoesters, polyester carbonates, polycarbonates, polyanhydrides, polyhydroxyalkonates (e.g., polyhydroxybutyrate, polyhydroxyvalerate), polyurethanes, and polyester urethanes. These polymers may be linear, branched, or star-shaped. These polymers may start from compounds with a single, two, three, four, six, or eight origins. The polymers may include, but are not limited to, polymers comprising repeating units derived from at least one of the following monomers: l-lactide, dl-lactide, glycolide, trimethylene carbonate, ε-caprolactone, p-dioxanone, and morpholinedione.

[0151] Suitable excipient synthetic polymers include, but are not limited to, polyacrylic acid and its salts, polyvinylpyrrolidone, Pluronic 127, Pluronic F68, polyethylene glycol, polyethylene oxide, and polyvinyl alcohol.

[0152] The complexing agents are not limited to but may include α-cyclodextrin, β-cyclodextrin (2-hydroxypropyl)-β-cyclodextrin, sulfobutyl ether β-cyclodextrin sodium, and ethylenediaminetetraacetic acid (EDTA).

[0153] The usable phospholipids are not limited to hydrogenated soy phosphatidylcholine, distearoyl phosphatidylglycerol, L-α-dimiristoyl phosphatidylcholine, and L-α-dimiristoyl phosphatidylglycerol.

[0154] Usable surfactants include ionic and nonionic surfactants. Ionic surfactants may include cationic, anionic, and amphoteric surfactants. Nonionic surfactants are not limited to these, but include (Cremofol EL, Cremofol RH40, Cremofol RH60, d--Tocopherol Polyethylene Glycol 1000 Succinate, Brij, Myrj, Polysorbate 20, Polysorbate 80, Polysorbate 40, Polysorbate 60, Polysorbate 65, Polysorbate 85, Soltol HS15, Sorbitan Monooleate (Span 80), Sorbitan Monopalmitate (Span 40), Sorbitan Monostearate (Span 60), Sorbitan Trioleate (Span 8), Poloxamer 407, Labrafil M-1944CS, Labrafil M-2125CS, Labrasol, Gelcia 44 / 14, Nonoxynol-9, Softigen 767, Octyl β-D-Glycopyranoside (OGP), Hexyl β-D-Glucopyranoside This may include (HGP), octyl β-D-1-thioglucopyranoside (TGP), decyl-β-D-glucopyranoside (DGP), dodecyl-β-D-glucopyranoside (DdGP), N-octyl β-D-maltoside (ODM), decyl β-D-maltopyranoside (DMP), cyclohexyl-ethanolyl-maltoside, n-decyl- and n-dodecyl-sucrose, and mono- and di-fatty acid esters of PEG300, 400, or 1750. Anionic surfactants may include, but are not limited to, sodium lauryl sulfate, fatty acid salts, sodium laureth sulfate, and sodium dioctyl sulfosuccinate. Cationic surfactants may include, but are not limited to, phosphatidylcholine (lecithin), cetrimide, cetrimonium bromide, and benzethonium chloride. It may contain dimethyldioctadecylammonium chloride, tetradecyltrimethylammonium bromide, cetylpyridinium chloride, ester quat, and benzalkonium chloride.The amphoteric surfactants may include, but are not limited to, cocamidopropyl betaine, (3-[(3-coramidopropyl)dimethylammonio]-1-propanesulfonate) and cocamidopropyl hydroxysultaine, phosphatidylserine, phosphatidylethanolamine, phosphatidylcholine, and sphingomyelin.

[0155] Suitable solvents include water-soluble organic solvents. These include, but are not limited to, polyethylene glycol 200, polyethylene glycol 300, polyethylene glycol 400, ethanol, propylene glycol, glycerin, N-methyl-2-pyrrolidone, dimethylacetamide, and dimethyl sulfoxide.

[0156] Suitable tension modifiers include, but are not limited to, dextrose, sucrose, mannitol, glycerin, sodium chloride, and potassium chloride.

[0157] Suitable pH adjusters include, but are not limited to, citric acid and its salts, phosphoric acid, tartatic acid, lactic acid, glycolic acid, sodium hydroxide, phosphoric acid, sulfuric acid, oxalic acid, and hydrochloric acid salts.

[0158] Usable antioxidants include, but are not limited to, ascorbic acid, butylated hydroxyanisole, butylated hydroxytoluene, vitamin A, vitamin E, alpha-tocopherol, thioglycerol, cysteine, acetylcysteine, cystine, dithioerythritol, dithiothritol, glutathione, sodium bisulfite, sodium metabisulfite, thiourea, uric acid, melatonin, propyl gallate, tertiary butylhydroquinone, and combinations thereof.

[0159] Suitable emulsifiers include, but are not limited to, glyceryl monostearate, isopropyl palmitate, polyethylene glycol monostearate 400, and the compounds listed as surfactants and combinations thereof.

[0160] Suitable preservatives include, but are not limited to, benzoic acid, sorbic acid, boric acid, methylparaben, ethylparaben, propylparaben, butylparaben, sodium benzoate, sodium propionate, phenylethyl alcohol, chlorobutanol, benzyl alcohol, potassium sorbate, phenol, chlorocresol, o-phenylphenol, thiomersal, nitromersal, phenylmercury nitrate, phenylmercury acetate, benzalkonium, and combinations thereof.

[0161] The excipient may contain at least one solvent. The solvents used may include, but are not limited to, water, ethanol, dimethyl sulfoxide, ethyl lactate, ethyl acetate, benzyl alcohol, benzyl benzoate, triacetin, N-methylpyrrolidone, 2-pyrrolidone, propylene carbonate, polyethylene glycol (PEG200), polyethylene glycol (PEG400), glycoflore, and combinations thereof.

[0162] Suitable buffers include aqueous solutions prepared using one or more of the following: potassium hydrogen phthalate, sodium hydrogen phthalate, potassium or sodium dihydrogen phosphate, dipotassium or disodium hydrogen phosphate, phosphoric acid, boric acid, sodium acetate, acetic acid, ammonium chloride, ammonium acetate, (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), citric acid, and sodium citrate.

[0163] In another embodiment, the derivatized polyvalent polymer composition of the present disclosure may further comprise an inorganic compound. Available inorganic compounds include, but are not limited to, barium sulfate, calcium hydroxyapatite or hydroxyapatite, tricalcium phosphate (TCP) [including various forms, e.g., α-TCP, β-TCP, and biphasic tricalcium phosphate (BCP)], calcium phosphate, and calcium sulfate.

[0164] In one embodiment, the derivatized polyvalent polymer of the Disclosure may be prepared as a composition comprising one or more excipients. In another embodiment, the derivatized polyvalent polymer of the Disclosure may be suspended in a composition comprising one or more excipients. In another embodiment, the derivatized polyvalent polymer of the Disclosure may be rehydrated in a composition comprising one or more excipients. In another embodiment, the derivatized polyvalent polymer of the Disclosure may be prepared with another solution to be mixed before use, as another composition which may comprise one or more excipients. In another embodiment, the derivatized polyvalent polymer of the Disclosure may be prepared in the presence of one or more excipients and then converted into a solid form by one or more of the methods of the Disclosure.

[0165] The compositions of this disclosure may comprise bioactive agents and, optionally, other components, in addition to derivatized polyvalent polymers as described herein. Exemplary bioactive agents include, but are not limited to, small molecule drugs, peptides, proteins, growth factors, hormones, antibodies, agonists, antagonists, antimicrobial and / or antifungal agents.

[0166] Bioactive agents that can be incorporated into formulations together with the described compositions include antiandrogenic agents, antibacterial agents, antiestrogen agents, androgens and anabolic agents, antibiotics, antimigraine agents, antihistamines, anxiolytics, antidiuretics, antihistamines, antirheumatic agents, antigens, analgesics, antidepressants, anti-inflammatory agents, anesthetics, aminoglycosides, antibodies, antivirals, adrenergic stimulants, anticonvulsants, antianginic agents, antiarrhythmics, antimalarial agents, antimitotics, anthelmintics, and appetite suppressants. agents), antitussives, antipruritics, antipyretics, anti-Alzheimer's drugs, anti-Parkinson's drugs, antiemetics and anti-vomiting drugs, antihypertensive drugs, anticoagulants, antifungal drugs, antimicrobial drugs, allergens, antidiarrheals, antihyperuricemia drugs, adrenergic stimulants, antiparasitic drugs, antiproliferative drugs, antipsychotics, antithyroid drugs, β-adrenergic blockers, bronchodilators; bronchospasmodics, blood clotting factors, blood coagulation factors Factors), cytotoxic drugs, cell proliferation inhibitors, chemotherapy drugs, clot inhibitors, clot-dissolving agents, cells, CNS stimulants, corticosteroids, calcium channel blockers, cofactors, ceramides, cardiac glycosides, cytokines (e.g., lymphokines, monokines, chemokines); colony-stimulating factors (e.g., GCSF, GM-CSF, MCSF); topical drugs, decongestants, diuretics, expectorants, endoectosides, growth factors, hemostatic agents, hypoglycemic agents, hormones and hormone analogs, hypercalcemia, hypnotics, interleukins (IL-2, IL-3, IL-4, IL-6); interferons (β-IFN, α-IFN) and γ-IFN); immunosuppressants, muscle relaxants, microorganisms, nonsteroidal anti-inflammatory drugs, nucleic acids, nutritional supplements, neuromuscular blockers, nerve depressants, neurotoxins, nutritional supplements, oligonucleotides, estrogens, obstetric drugs, ovulation inducers, opioids, progestogens, pituitary hormones, pituitary inhibitors, proteins, peptides, polysaccharides, protease inhibitors, prostaglandins, quinolones, reductase inhibitors, sulfonamides, emollients, sedatives, sodium channel blockers, steroids, steroidal anti-inflammatory drugs, smoking cessation drugs, toxins, platelet lysis agents, thyroid hormones, tumor necrosis factor; vesicles, vitamins, viruses, vasodilators, vaccines.

[0167] Further representative examples of bioactive agents that may be suitable for use in the compositions of this disclosure include, but are not limited to, antidiarrheal agents such as diphenoxylate, loperamide, and hyoscyamine; antihypertensive agents such as hydralazine, minoxidil, captopril, enalapril, clonidine, prazosin, debrisoquin, diazoxide, guanethidine, methyldopa, reserpine, and trimaphan; calcium channel blockers such as diltiazem, felodipine, amlodipine, nitrendipine, nifedipine, and verapamil; antiarrhythmics such as amiodarone, flecainidine, disopyramide, procainamide, mexileten, and quinidine; nitroglycerin and erythrityl tetranitrate. Antianginal drugs such as pentaerythritol tetranitrate, mannitol hexanitrate, perhexilene, isosorbide dinitrate, and nicorandil; β-adrenergic blockers such as alprenolol, atenolol, bupranolol, carteolol, labetalol, metoprolol, nadolol, nadoxolol, oxprenolol, pindolol, propranolol, sotalol, timolol, and timolol maleate; digoxin and other cardiac glycosides and theophylline derivatives Cardiac glycosides such as; adrenergic stimulants such as adrenaline, ephedrine, fenoterol, isoprenaline, orciprenaline, rimeterol, salbutamol, salmeterol, terbutaline, dobutamine, phenylephrine, phenylpropanolamine, pseudoephedrine, and dopamine; vasodilators such as cyclandelate, isoxuprine, papaverine, dipyrimadole, isosorbide dinitrate, phentolamine, nicotinyl alcohol, chodergocrine, nicotinic acid, glyceryl trinitrate, pentaerythritol tetranitrate, and xanthinol; antiproliferative agents such as paclitaxel, estradiol, actinomycin D, sirolimus, tacrolimus, everolimus, 5-fluorouracil, and dexamethasone;Antimigraine preparations such as ergotamine, dihydroergotamine, methyserzide, pizotifen, and sumatriptan; anticoagulants and platelet lysis agents such as warfarin, dicumarol, and low molecular weight heparin (e.g., enoxaparin, streptokinase, and its active derivatives); hemostatic agents such as aprotinin, tranexamic acid, and protamine; buprenorphine, dextromoramide, dextropropoxifen, fentanyl, alfentanil, Analgesics and antipyretics including opioid analgesics such as sufentanil, hydromorphone, methadone, morphine, oxycodone, papaveretam, pentazocine, pethidine, fenopefidin, codeine, and dihydrocodeine; acetylsalicylic acid (aspirin), paracetamol, synthetic α2-adrenergic receptor agonists, dexmedetomidine hydrochloride, flunixin meglumine, meperidine, phenylbutazone, and phenazone; rapamycin (sirolimus) and its analogues (everolimus) Immunosuppressants, antiproliferative agents and cell proliferation inhibitors such as tacrolimus; neurotoxins such as capsaicin and botulinum toxin (Botox); hypnotics and sedatives such as barbiturates, amirobarbitone, butobarbitone and pentobarbitone, as well as other hypnotics and sedatives such as chloral hydrate, chlormethiazol, hydroxyzine and meprobamate; benzodiazepines such as alprazolam, bromazepam, chlordiazepoxide, clobazam, and clorazepate. Anxiolytics such as diazepam, flunitrazepam, flurazepam, lorazepam, nitrazepam, oxazepam, temazepam, and triazolam; neurodepressants and antipsychotics such as phenothiazine, chlorpromazine, fluphenazine, periciazine, perphenazine, promazine, thiopropazate, thioridazine, and trifluoperazine; as well as butyrophenone, droperidol, and haloperidol; and other antipsychotics such as pimozide, thiothixen, and lithium;Antidepressants include tricyclic antidepressants such as amitriptyline, clomipramine, desipramine, dotiepine, doxepin, imipramine, nortriptyline, opipramol, protriptyline, and trimipramine, as well as tetracyclic antidepressants such as mianserin, monoamine oxidase inhibitors such as isocarboxazide, pheneridine, tranylcipramine, and moclobemide, and selective serotonin reuptake inhibitors such as fluoxetine, paroxetine, citalopram, fluvoxamine, and sertraline; CNS stimulants such as caffeine and 3-(2-aminobutyl)indole; and antipruritics may include compounds such as synthetic Janus kinase (JAK) inhibitors, NK-1 receptor antagonists, and antibodies that neutralize interleukin-31 (IL-31). These include anti-Alzheimer's drugs such as oclacitinib maleate, cerulopitant, and lokibetomab and tacrine; amantadine, benserazide, carbidopa, levodopa, benztropine, biperiden, benzhexol, procyclidine and dopamine-2 agonists, such as anti-Parkinson's drugs like S(-)-2-(N-propyl-N-2-thienylethylamino)-5-hydroxytetraline (N-0923); phenytoin, valproic acid, primidone, phenobarbiton, methylphenobarbiton and carbamazepine. Anticonvulsants such as ethosuximide, metosuximide, fenxuximide, sultiame, and clonazepam; antiemetics and anti-vomiting agents such as phenothiazines prochlorperazine, thiethylperazine, neurokinin (NK1) receptor antagonists, and maropitant citrate; as well as 5HT-3 receptor antagonists such as ondansetron and granisetron, and dimenhydrinate, diphenhydramine, metoclopramide, domperidone, hyostine, hyostine hydrobromide, hyostine hydrochloride, clevopride, and bromprid;Where applicable, these include racemic mixtures or individual enantiomers, preferably ibuprofen, flurbiprofen, ketoprofen, acrofenac, diclofenac, alloxyprine, aproxen, aspirin, diflunisal, fenoprofen, indomethacin, mefenamic acid, naproxen, phenylbutazone, piroxicam, salicylamide, salicylic acid, slindac, deoxyslindac, tenoxicam, tramadol, ketoralac, fluphenisal, salsalate, and Ethanolamine salicylic acid, aminopyrine, antipyrine, oxyfenbutazone, apazon, synthazon, flufenamic acid, clonixerl, clonixin, meclofenamic acid, 6-chloro-α-methyl-9H-carbazole-2-acetic acid (carprofen), flunixin, colchicine, demecorcin, allopurinol, oxypurinol, benzydamine hydrochloride, dimefadan, indoxol, intrazol, mimban hydrochloride, paranilen hydrochloride, tetridamine, benzydamine hydrochloride Nonsteroidal anti-inflammatory drugs that may be formulated in combination with skin and / or mucous membrane penetration enhancers such as ndopyrin, fluprofen, ibufenac, naproxol, fenbufen, syncofen, diflumidone sodium, phenamol, fluthiazidine, metazamide, retimidone hydrochloride, nexeridine hydrochloride, octazamide, molinazole, neosyncofen, nimazole, proxazole citrate, tesicam, tesimide, tolmetin, and triflumidate; penicillamine, golden thioglucose, golden thiomalate sodium, methotrexate Antirheumatic drugs such as baclofen and auranofin; muscle relaxants such as baclofen, diazepam, cyclobenzaprine hydrochloride, dantrolene, methocarbamol, orphenadrine and quinine; drugs used for gout and hyperuricemia such as allopurinol, colchicine, probenecid and sulfinpyrazone; estrogens such as estradiol, estriol, estrone, ethinylestradiol, mestranol, stilbesterol, dienestrol, epiestriol, estropipate and zeranol;Progesterone and other progestins such as allylestrenol, dydrogesterone, linestrenol, norgestrel, norethinodrel, norethisterone, norethisterone acetate, gestodene, levonorgestrel, medroxyprogesterone and megestrol; antiandrogenic agents such as cyproterone acetate and danazol; antiestrogenic agents such as tamoxifen and epithiostanol; as well as aromatase inhibitors, exemestane and 4-hydroxyandrostenedone and their derivatives; testosterone, methyltestosterone, clostebol acetate, drostanolone, flazabol, nandrolone, oxandrolone, stanozolol, trenbolone acetate, dihydro-testosterone, 17-(α-methyl-19-nortestosterone) ) and androgens and anabolic agents such as fluoxymesterone; 5-α-reductase inhibitors such as finasteride, tulosteride, LY-191704 and MK-306; betamethasone, betamethasone valerate, cortisone, dexamethasone, dexamethasone 21-phosphate, fludrocortisone, flumetasone, fluocinonide, desonide, fluocinolone, fluocinolone acetonide, fluocortolone Corticosteroids such as thorone, halcionide, halopredone, hydrocortisone, hydrocortisone 17-valerate, hydrocortisone 17-butyrate, hydrocortisone 21-acetic acid, methylprednisolone, prednisolone, prednisolone 21-phosphate, prednisone, triamcinolone, and triamcinolone acetonide; glycosylated proteins, proteoglycans, glycosaminoglycans (e.g., chondroitin sulfate); ;Chitin, acetyl-glucosamine, hyaluronic acid;complex polysaccharides such as glucans;cortodoxone, fludrolacetonide, fludrocortisone, difluoronone diacetate, flurandrenolonone acetonide, medrisone, amsinafel, amsinafid, betamethasone and other esters, chloroprednisone, chlorcorterone, descinolone, desonide, dichlorizone, difluprednate, fluchloronide, flumetasone, flunisolide, flucortolone, fluoromethalone, fluperolon, fluprednisolone, meprednisone, methylmeprednisolone, paramethasone, cortisone acetate, hydrocortisone cyclopentylpropionate, cortodoxone, flucetonide, Further examples of steroidal anti-inflammatory drugs include fludrocortisone acetate, flullandrenolone, ainsinaphal, amsinafid, betamethasone, betamethasone benzoate, chloroprednisone acetate, crocortolone acetate, descinolone acetonide, desoxymethasone, dichlorizone acetate, difluprednate, fluchloronide, flumetasone pivalate, flunisolide acetate, fluperolon acetate, fluprednisolone valerate, paramethasone acetate, prednizolamate, prednival, triamcinolone hexaacetonide, cortivazole, formocortal and nivazole; corticotrophin, thyrotropin, follicle-stimulating hormone (FSH), gonadotropin-releasing hormone (GnRH) analogs, deslorerin acetate (u Pituitary hormones such as deslorelin acetate, cetrorelix acetate, gonadrelin acetate, clomiphene, human chorionic gonadotropin (HCG), luteinizing hormone (LH), and gonadotropin-releasing hormone (GnRH), and their active derivatives or analogues; hypoglycemic agents such as insulin, chlorpropamide, glibenclamide, gliclazide, glipizide, trazamide, tolbutamide, and metformin; thyroid hormones such as calcitonin, thyroxine, and liothyronine, as well as antithyroid drugs such as carbimazole and propylthiouracil; other miscellaneous hormonal drugs such as octreotide; pituitary inhibitors such as bromocriptine; ovulation inducers such as clomiphene; thiazides, related diuretics, and loop diuretics such as bendrofluazide and chlorothiazide.Diuretics such as chlorthalidone, dopamine, cyclopentiazide, hydrochlorothiazide, indapamide, mefluside, methicorthiazide, metrazone, quinetazone, bumetanide, ethacrine and flusemide, as well as potassium-sparing diuretics such as spironolactone, amiloride and triamterene; antidiuretics such as desmopressin, repressin and vasopressin (including their active derivatives or analogs); obstetric drugs including uterine-acting drugs such as ergometrine, oxytocin and gemeprost; Prostaglandins such as prostadil (PGE1), prostacyclin (PGI2), dinopresto (prostaglandin F2-α), and misoprostol; cephalosporins such as cephalexin, cefoxitin, and cephalothin; amoxilin, amoxilin and clavulanic acid, ampicillin, bacampicillin, benzathine penicillin, benzyl penicillin, carbenicillin, cloxacillin, methicillin, pheneticillin, phenoxymethylpenicillin, flucloxacillin, meziocillin (mezio Penicillins such as piperacillin, ticarcillin, and azurocillin; tetracyclines such as minocycline, chlortetracycline, tetracycline, demeclocycline, doxycycline, metacycline, and oxytetracycline, and other tetracycline antibiotics; antimicrobial agents including amnioglycosides such as amikacin, amikacin sulfate, gentamicin, kanamycin, neomycin, netylmycin, and tobramycin; Amorol Fin, isoconazole, clotrimazole, econazole, miconazole, nystatin, terbinafine, bifonazole, amphotericin, griseofulvin, ketoconazole, fluconazole and flucytosine, salicylic acid, fezathion, ticlaton, tolnaftate, triacetin, zinc, pyrithione and sodium pyrithione; quinolones such as nalidixic acid, cinoxacin, ciprofloxacin, enoxacin and norfloxacin; phthalisulfthiazole, sulfadoxine, sulfadiazine,Antifungal drugs such as sulfonamides including sulfamethizol and sulfamethoxazole; sulfones including dapsone; chloramphenicol, clindamycin, erythromycin, ethyl erythromycin carbonate, erythromycin estolate, erythromycin glucepate, erythromycin ethyl succinate, erythromycin lactobionate, roxithromycin, lincomycin, natamycin, nitrofurantoin, spectinomycin, vancomycin, Aztreonarn, colistin IV, metronidazole, tinidazole, sekunidazole, ornidazole, fusidic acid, trimethoprim, and other miscellaneous antibiotics such as 2-thiopyridine N-oxide; halogen compounds, especially iodine and iodine compounds, e.g., iodine-PVP complex and diiodohydroxyquine, hexachlorophene; chlorhexidine; chloroamine compounds, silver sulfadiazine, silver, silver nanoparticles, silver nitrate, silver zeolites, silver cations, AgPO3, Ag3PO4, Ag4P2O7, exsalt(registered trademark)SD7 (Exciton Technologies), exsalt(registered trademark)(Exciton Technologies); Lincomycin hydrochloride, tricyclic tetrahydroquinoline antibacterial agent, 8-pyrazinyl-S-spiropyrimidinetrione-oxinoquinoline derivative, 3-spiropyrimidinetrione-quinoline derivative, thiadiazole-spiropyrimidinetrione-quinoline derivative, (2R,4S,4aS)-10-fluoro-2,4-dimethyl-8-(4-methyloxazol-2-yl)-2,4,4a,6-tetra-hydro-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4',6'(-3'H)-trione, (2R,4S,4aS)-9,10-difluoro-2,4-dimethyl-8-(3-methylisoxazole-5-yl)-2,4,4a,6- -Tetrahydro-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',-4',6'(3'H)-trione,(2R,4S,4aS)-10-fluoro-2,4-dimethyl-8-(oxazol-2-yl)-2,4,4a,6-tetrahydro-1H-,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4',6'(3'H)-tri-one, (2R,4S,4aS)-9,10-difluoro-2,4-dimethyl-8-(2-methyloxazol-5-yl)-2,4,4a,6-tetrahydro(t- (tetrahydro)-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4'-,6'(3'H)-trione, (2R,4S,4aS)-9,10-difluoro-2,4-dimethyl-8-(oxazol-4-yl)-2,4,4a,6-tetrahydro(tetrahydr-o)-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4',6'(3'H)- -Trion, (2R,4S,4aS)-9-fluoro-2,4-dimethyl-8-(4-methyloxazol-2-yl)-2,4,4a,6-tetrahydro(tetrah-ydro)-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4',6'(3-'H)-Trion, (2R,4S,4aS)-9,10-difluoro-8-(4-(4-fluorophenyl)oxazol-5-yl)-2,4-dimethyl- -2,4,4a,6-tetrahydro-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine(pyrimid- (ine)-2',4',6'(3'H)-trione, (2S,4R,4aR)-2,4-dimethyl-8-(oxazol-5-yl)-2,4,4a,6-tetrahydro-1H,1'H-spiro-[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4',6'(3'H)-trione, (2S,4R,4aR)-8-(4-ethyloxazol-2-yl)-9,10-difluoro-2,4-dimethyl-2,4,4a,6-tetrahydro(te-trahydro)-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4',-6'(3'H)-trione,(2R,4S,4aS)-9,10-difluoro-2,4-dimethyl-8-(oxazol-2-yl)-2,4,4a,6-tetrahydro(tetrahydr-o)-1H,1'H-spiro[[1,4]oxazino[4,3-a]quinoline-5,5'-pyrimidine]-2',4',6'(3'H)--trione, benzoyl peroxide; antituberculosis drugs such as ethambutol, isoniazid, pyrazinamide, rifampicin and clofazimine; antimalarial drugs such as primaquine, pyrimethamine, chloroquine, hydroxychloroquine, quinine, mefloquine and halofantrine; azithromycin, aztreonam, cefacrol, cefadroxil, cefazolin, cefdinir, cefepime hydrochloride, (cefoperazone sodium, cephthaloline fosamil, avibactam, cefta) Didime sodium, ceftibutene, ceftiofur, tazobactam, cefobecine sodium [(6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-8-oxo-3-[(2S)-tetrahydro-2-furanyl]-5-thia-1-azabicyclo[4.2.0]octa-2-ene-2-carboxylic acid, monosodium salt], cefuroxime acetyl, cefuroxime, cephalexin, chloramphenicol sodium, ciproflo Xacin HCl, clarithromycin, clindamycin hydrochloride, clindamycin palmitate hydrochloride, clindamycin phosphate, dalbavancin hydrochloride, daptomycin, demeclocycline hydrochloride, dicloxacillin, doripenem, doxycycline, doxycycline calcium, doxycycline helicrate, doxycycline monohydrate, ertapenem sodium, erythromycin, erythromycin ethylsuccinate, erythromycin lactobionic acid, erythromycin stearate Romycin, erythromycin, fosfomycin tromethamine, gemifloxacin mesylate, gentamicin sulfate, imipenem, kanamycin, levofloxacin, lincomycin hydrochloride, linezolid, meropenem, methenamine hippurate, metronidazole, metronidazole, micafungin sodium, minocycline hydrochloride, minocycline, moxifloxacin hydrochloride, nafcillin, nalidixic acid, neomycin sulfate, nitrofurantoin, norfloxacin, ofloxacin,This includes compounds such as diphosphate oritabancin, oxacillin, penicillin G, penicillin G benzathine, penicillin G sodium, penicillin V potassium, piperacillin sodium, polymyxin B sulfate, quinupristin, dalfopristin, spectinomycin hydrochloride, streptomycin, sulfamethoxazole, tedizolide phosphate, teravancin, telithromycin, tetracycline hydrochloride, ticarcillin disodium, tigecycline, tobramycin sulfate, tobramycin, trimethoprim hydrochloride, tulathromycin, and vancomycin hydrochloride.

[0168] The compositions of this disclosure may also include antiviral agents, exemplary of which include acyclovir and acyclovir prodrugs, famciclovir, zidovudine, didanosine, stabudine, lamivudine, zalcitabine, saquinavir, indinavir, ritonavir, n-docosanol, tromantadine, and idoxuridine. Other suitable bioactive agents include anthelmintics such as mebendazole, thiabendazole, niclosamide, praziquantel, pyrantel emvonate, and diethylcarbamazine; and plicamycin, cyclophosphamide, dacarbazine, fluorouracil and its prodrugs (e.g., International Journal of This includes cytotoxic agents such as methotrexate, procarbazine, 6-mercaptopurine, and mucophenolic acid (as described in Pharmaceutics, 111, 223-233 (1994)); appetite suppressants and weight-loss agents including dexfenfluramine, fenfluramine, diethylpropion, mazindol, and phentermine; drugs used for hypercalcemia such as calcitriol, dihydrotachisterol, and their active derivatives or analogs; cough suppressants such as ethylmorphine, dextromethorphan, and forcozin; and antiparasitic and endoparasite agents such as moxidectin, ivermectin, niclosamide, praziquantel, pyrantel, pyrvinium, albendazole, flubendazole, mebendazole, and thiabendazole.

[0169] The compositions of this disclosure may include: expectorants such as carbolcysteine, bromhexine, emetine, quanifecin, ipecacuanha, and saponins; decongestants such as phenylephrine, phenylpropanolamine, and pseudoephedrine; and ephedrine, fenoterol, orciprenaline, limiterol, salbutamol, sodium cromoglycate, cromoglycic acid, and its prodrugs (e.g., International Journal of Pharmaceutics). 7,63-75 (1980), bronchospasmodics such as terbutaline, ipratropium bromide, salmeterol, and theophylline and theophylline derivatives; meclozine, cyclidine, chlorcyclidine, hydroxyzine, brompheniramine, chloropheniramine, clemastine, cyproheptadine, dexchlorphenylamine, diphenhydramine, diphenylamine, doxylamine, mebhydroline, pheniramine, tripolidine, azatadine, dif Antihistamines such as phenylpyraline, methidilazine, terfenadine, astemizole, loratidine, and cetirizine; local anesthetics such as benzocaine, bupivacaine, ametocaine, lignocaine, lidocaine, cocaine, cincocaine, dibucaine, mepivacaine, prilocaine, etidocaine, veratridine (specific c-fiber blocker), and procaine; stratum corneum lipids such as ceramides, cholesterol, and free fatty acids for improving skin barrier repair [Man, et al. J. Invest. Dermatol., 106(5), 1096, (1996)]; neuromuscular blocking agents such as succinylcholine, alcuronium, pancuronium, atracurium, galamine, tubocurarine, and vecuronium; sclerocing agents or sclerosants may be surfactants, or they may be selected from the group consisting of ethanol, dimethyl sulfoxide, sucrose, sodium chloride, dextrose, glycerin, minocycline, tetracycline, doxycycline, polidocanol, sodium tetradecyl sulfate, sodium molinate, and sotradecol; angiogenesis inhibitors; 5-lipoxygenase inhibitors or antagonists;Chemokine receptor antagonists; cell cycle inhibitors; taxanes; microtubule inhibitors; paclitaxel; paclitaxel analogs or derivatives; vinca alkaloids; camptothecin or its analogs or derivatives; podophyllotoxin, which may be etoposide or its analogs or derivatives; anthracyclines, which may be doxorubicin or its analogs or derivatives, or which may be mitoxantrone or its analogs or derivatives; platinum compounds; nitroso Urea; Nitroimidazole; Folic acid antagonists; Cytidine analogs; Pyrimidine analogs; Fluoropyrimidine analogs; Purine analogs; Nitrogen mustard or its analogs or derivatives; Hydroxyurea; Mitomycin or its analogs or derivatives; Alkyl sulfonates; Benzamide or its analogs or derivatives; Nicotinamide or its analogs or derivatives; Halogenated sugars or their analogs or derivatives; DNA alkylating agents; Microtubule inhibitors; Topoisomerase inhibitors; DNA cleavage agents; Antimetabolites; Nucleotide interconversion inhibitors; Hydroorotate dehydrogenase inhibitors; DNA intercalation agents; RNA synthesis inhibitors; pyrimidine synthesis inhibitors; cyclin-dependent protein kinase inhibitors; epidermal growth factor kinase inhibitors; elastase inhibitors; factor Xa inhibitors; farnesyltransferase inhibitors; fibrinogen antagonists; guanylate cyclase stimulants; heat shock protein 90 antagonists; which may be geldanamycin or its analogues or derivatives; guanylate cyclase stimulants; HMGCoA reductase inhibitors, which may be simvastatin or so Possibly analogs or derivatives of; IKK2 inhibitors; IL-1 antagonists; ICE antagonists; IRAK antagonists; IL-4 agonists; immunomodulators; sirolimus or its analogs or derivatives; everolimus or its analogs or derivatives; tacrolimus or its analogs or derivatives; biolimus or its analogs or derivatives; tresperimus or its analogs or derivatives; auranofin or its analogs or derivatives; 27-0-demethylrapamycin or its analogs or derivatives; gusperimus or its analogs or derivatives;Pimecrolimus or its analogs or derivatives; ABT-578 or its analogs or derivatives; Inosine monophosphate dehydrogenase (IMPDH) inhibitors, which may be mycophenolic acid or its analogs or derivatives or 1-α-25-dihydroxyvitamin D3 or its analogs or derivatives; Leukotriene inhibitors; MCP-1 antagonists; MMP inhibitors; NFκB inhibitors, which may be Bay 11-7082; NO antagonists; p38 MAP kinase inhibitors, which may be SB Possible causes: phosphodiesterase inhibitors; TGF-β inhibitors; thromboxane A2 antagonists; TNF-α antagonists; TACE inhibitors; tyrosine kinase inhibitors; vitronectin inhibitors; fibroblast growth factor inhibitors; protein kinase inhibitors; PDGF receptor kinase inhibitors; endothelial growth factor receptor kinase inhibitors; retinoic acid receptor antagonists; platelet-derived growth factor receptor kinase inhibitors; fibrinogen antagonists; antifungal agents; sucronizole; bisphosphonates; phospholipase A1 inhibitors; histamine H1 / H2 / H3 receptor antagonists; macrolide antibiotics; GPIIb / IIIa receptor antagonists; endothelin receptor antagonists; peroxisome proliferator-activated receptor agonists; estrone Gen receptor drugs; somatostatin analogs; neurokinin 1 antagonists; neurokinin 3 antagonists; VLA-4 antagonists; osteoclast inhibitors; DNA topoisomerase ATP hydrolysis inhibitors; angiotensin I-converting enzyme inhibitors; angiotensin II antagonists; enkephalinase inhibitors; peroxisome proliferator-activated receptor gamma agonists; insulin sensitizers; protein kinase C inhibitors; ROCK (rho-binding kinase) inhibitors; CXCR3 inhibitors; Itk inhibitors; cytosolic phospholipase A2α inhibitors; PPAR agonists; immunosuppressants; Erb inhibitors; apoptosis agonists; lipocortin agonists; VCAM-1 antagonists; collagen antagonists; α-2 integrin antagonists; TNF-α inhibitors; nitric oxide inhibitors; and cathepsin inhibitors. Antifibrin and fibrinolytic agents including plasmin, streptokinase, single-chain urokinase, urokinase, t-PA (tissue plasminogen activator), and aminocaproic acid; antiplatelet agents including aspirin, prostacyclin (and analogues);Monoclonal antibodies, peptides (e.g., ReoPro, Cilastagel, Eptifibatide, Tirofiban, Ticlopidine, Bapiprost, Dipyridamole, Forskolin, Angiopeptin, Argatroban), glycoprotein IIb / IIIa drugs including thromboxane inhibitors; antithrombin and anticoagulant drugs including Dextan, heparin, LMW heparin (enoxaparin, dalteparin), Hildin, recombinant Hildin, antithrombin, synthetic antithrombin, thrombin inhibitors, warfarin (and other coumarins); Antimitotics, antiproliferatives and cell proliferation inhibitors, including vincristine, vinblastine, paclitaxel, methotrexate, cisplatin, fluorouracil, rapamycin, azathioprine, cyclophosphamide, mycophenolic acid, corticosteroids, colchicine, and nitroprusside; anti-angiogenics and anti-angiogenics, including paclitaxel, angiostatin, and endostatin; genetic material, DNA, DNA sequences, polynucleotides, and oligonucleotides; ACE inhibitors (e.g., cilazapril, lisinopril, captopuriol) (e.g., VEGF, FGF) antagonists; antioxidants and vitamins (e.g., probucol, tocopherol); calcium channel blockers (e.g., nifedipine); fish oil (ω3-fatty acids); phosphodiesterase inhibitors (e.g., dipyridamole); nitrate donors (e.g., morcidomin); somatostatin analogs (e.g., angiopeptin); immunosuppressants and anti-inflammatory drugs (e.g., prednisolone, glucocorticoids and dexamethasone); antibacterial agents (e.g., rifamycin) and α, β and γ radioisotopes ( For example, radionuclides including Re-188, Re-186, I-125, Y-90; COX-2 inhibitors such as celecoxib and Biox; kinase inhibitors such as epidermal growth factor kinase inhibitors, tyrosine kinase inhibitors, MAP kinase inhibitors, and protein transferase inhibitors; Resten-NG; nicotine, bupropion, and ibogaine smoking cessation drugs; insecticides and other parasitic agents suitable for topical application; dermatological drugs such as vitamins A, C, B1, B2, B6, B12, B12α, and E, vitamin E acetate, and vitamin E sorbate;Allergens for desensitization, such as house dust or dust mite allergens; nutritional supplements and dietary products such as vitamins, essential amino acids and fats; high molecular weight pharmacological agents such as proteins, enzymes, peptides, polysaccharides (e.g., cellulose, amylose, dextran, chitin), nucleic acids, cells, and tissues; bone repair biochemical products such as calcium carbonate, calcium phosphate, tricalcium phosphate, hydroxyapatite, or bone morphogenetic proteins (BMPs); angiogenic growth factors such as vascular endothelial growth factor (VEGF) and epidermal growth factor (EFG), cytokines, interleukins, fibroblast and cell migration chemicals (cytotaxic chemicals); and keratolytic agents such as alpha-hydroxy acids, glycolic acid, and salicylic acid;Also, DNA, RNA or other oligonucleotides. Vaccines containing Hendra virus (HeV) G glycoprotein and / or Nipah virus G glycoprotein, luteinizing hormone-releasing hormone (LHRH) peptide, LHRH-diphtheria toxoid conjugate, porcine circovirus type 2 (PCV2) antigen, porcine genital and respiratory syndrome virus antigen, Mycoplasma hyopneumoniae protein antigen. Proteins or protein fragments, e.g., ORFI tolkutenovirus protein or other TTV proteins or fragments, antigens against Aeromonas salmonicida, antigens against Vibrio anguillarum, and antigens against V. salmonicida. Growth factors include, but are not limited to, vascular endothelial growth factor (VEGF) and epidermal growth factor (EFG), growth and differentiation factors (GDF), fibroblast growth factors (FGF-1 to FGF-23), osteoprotegerin, cartilage-derived morphogenic proteins (CDMPs, which can form the basis of soft or hard tissue), Lim mineralized protein (LMP), interleukins (IL-1 to IL-13), insulin-like growth factor-1, connective tissue growth factor (CTGF), platelet-derived growth factor (PDGF), nerve growth factor, neurotrophin brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), transforming growth factors (TGF-α, TGF-β), tumor necrosis factor (TNF), growth factor agonists or antagonists, and antibodies against these growth factors. Bioactive agents available for the treatment of macular degeneration include, but are not limited to, bevacizumab and ranibizumab.

[0170] In one embodiment, the compositions of the present disclosure are formulated for wound healing and are useful for wound healing. The compositions may be formulated for suitable administration, for example, intranasal administration or topical administration. The compositions may comprise one or more bioactive agents suitable for wound healing. Wounds to be treated may include, but are not limited to, diabetic ulcers, burns, pressure wounds, abrasions, cuts, corneal abrasions, incisions after eye surgery, herpes, nasal surgery, abdominal surgery, and damaged tissue after tendon or joint repair.

[0171] For example, in one embodiment, the derivatized polyvalent polymer composition of the Disclosure may be in the form of dried particles. In another embodiment, the derivatized polyvalent polymer composition of the Disclosure may be in the form of a freeze-dried derivatized polyvalent polymer composition. In yet another embodiment, the derivatized polyvalent polymer composition of the Disclosure may be in the form of a nonwoven derivatized polyvalent polymer composition. In one embodiment, the nonwoven derivatized polyvalent polymer composition can be produced by electrospinning. In yet another embodiment, the derivatized polyvalent polymer of the Disclosure may be in the form of a film. The composition may be packaged directly or indirectly in a foil pouch to minimize moisture absorption during storage.

[0172] The derivatized polyvalent polymer compositions of this disclosure can be applied directly to wound sites. The derivatized polyvalent polymer compositions can absorb exudate from the wound. Once sufficient exudate has been absorbed, the dried derivatized polyvalent polymer composition swells to form a gel. In another embodiment, the derivatized polyvalent polymer composition of this disclosure further comprises water or saline solution to obtain a gel. In one embodiment, the gel can be applied directly to the wound.

[0173] In one embodiment, the derivatized polyvalent polymer composition of the present disclosure may, when applied to a wound, cover it with a moisture-retaining semipermeable film. This film may further include an adhesive that holds the film in place at the application site. The moisture-retaining semipermeable adhesive film may be made from a polyurethane or silicone material having an adhesive coating at least on the edges or ends of the film. In one embodiment, the adhesive may be an acrylic adhesive. The semipermeable film allows oxygen, carbon dioxide, and water vapor to pass through, but prevents the transmission of bacteria.

[0174] In another embodiment, the derivatized polyvalent polymers and their compositions of the Disclosure are ready-to-use products in that the product consists of the derivatized polyvalent polymer composition of the Disclosure and a semipermeable film as a single unit, and can be coated onto a semipermeable film. The compositions can be packaged directly or indirectly in a foil pouch. In one embodiment, the derivatized polyvalent polymer of the Disclosure comprises hyaluronic acid derivatized with a sulfonic acid group, and such derivatized polyvalent polymers can be used, for example, in compositions intended for wound healing.

[0175] In another embodiment, the derivatized polyvalent polymers and compositions of the present disclosure can be used as fillers. These fillers can be used to treat stress urinary incontinence, fecal incontinence, gastroesophageal reflux disease (GERD), and as prostate-rectal spacers to reduce rectal damage as a result of radiotherapy for prostate cancer. In one embodiment, the derivatized polyvalent polymer composition to be injected may be in the form of a derivatized polyvalent polymer that may or may not include crosslinking.

[0176] In one embodiment, the derivatized polyvalent polymers and compositions of the present disclosure can be used as skin fillers for filling voids and defects, and for treating moderate to severe wrinkles and folds. The derivatized polyvalent polymer compositions are injectable as solutions or suspensions. In one embodiment, at least one derivatized polyvalent polymer in the derivatized polyvalent polymer composition is crosslinked. In one embodiment, the crosslinked derivatized polyvalent polymers of the present disclosure used in skin filler compositions have a hyaluronidase (or corresponding polysaccharide-degrading enzyme for other polysaccharides) degradation rate equivalent to or lower than that of underivatized polyvalent polymers (e.g., hyaluronic acid). The derivatized polyvalent polymers and compositions can be used to treat areas where skin depressions, wrinkles, or scars are present, including, but are not limited to, nasolabial folds, forehead wrinkles, frown lines, and vertical lip wrinkles. In another embodiment, the derivatized polyvalent polymers and compositions can be used in lip augmentation and breast augmentation.

[0177] In another embodiment, the derivatized polyvalent polymers and compositions used as skin fillers may contain drugs (e.g., bioactive agents) to reduce pain associated with the procedure. As used herein, bioactive agents include compounds or molecules that can be designated as drugs. Such compounds include benzocaine, bupivacaine, ametokine, lignocaine, lidocaine, cocaine, cincocaine, dibucaine, mepivacaine, prilocaine, etidocaine, veratridine (a specific c-fiber blocker), and procaine. In another embodiment, the derivatized polyvalent polymers and compositions used as skin fillers may include degradable water-insoluble polymers (e.g., polyesters; e.g., PLGA, PLLA, etc.), water-insoluble non-degradable polymers (e.g., polymethyl methacrylate [PMMA]), or inorganic materials (e.g., calcium hydroxyapatite). In another embodiment, the derivatized polyvalent polymers and compositions used as skin fillers are in the form of cross-linked hydrogel particles. In one embodiment, the median particle size (Dv50) is in the range of 100 μm to 800 μm. In another embodiment, the median particle size (Dv50) is in the range of 200 μm to 600 μm. In one embodiment, the cross-linked hydrogel particles are suspended in a saline solution. In another embodiment, the hydrogel particles are suspended in a solution of hyaluronic acid or (oa) hyaluronic acid derivative of the Disclosure. In one embodiment, the cross-linked hydrogel suspension is contained in a pre-filled syringe, the contents of which are sterile. In another embodiment, the hydrogel particle suspension is injectable with a needle of at least 27 G.

[0178] In one embodiment, derivatized polyvalent polymers and compositions as disclosed herein are formulated for intra-articular replacement and are useful for intra-articular replacement. The derivatized polyvalent polymers may be crosslinked or uncrosslinked, and these compositions may optionally contain bioactive agents.

[0179] Intra-articular replacement is a process of injecting a derivatized polyvalent polymer composition into a joint to relieve pain. In a preferred embodiment, the polyvalent polymer is hyaluronic acid or a derivative thereof. The derivatized polyvalent polymer composition can be injected into one or more joint cavities of the body. Suitable joints include, but are not limited to, the knee, shoulder, ankle, elbow, hip, rhombometacarpal joint, finger joints, wrist, temporomandibular joint, back, and neck. In another embodiment, the derivatized polyvalent polymer used may include a crosslinked derivatized polyvalent polymer. The derivatized polyvalent polymer composition may contain one or more excipients or diluents. The derivatized polyvalent polymer compositions of this disclosure that are usable for the treatment of osteoarthritis can be injected with an 18-gauge to 21-gauge needle. The derivatized polyvalent polymer compositions of this disclosure may contain a bioactive agent. In one embodiment, the bioactive agent may be, but is not limited to, a corticosteroid, a local anesthetic, an antibody, a peptide, or an anti-inflammatory compound or molecule. The volume of the solution containing the derivatized polyvalent polymer composition of this disclosure may be in the range of 0.5 ml to 10 mL, and a preferred embodiment is 2 mL to 6 mL for injection into the knee. In one embodiment, the crosslinked derivatized polyvalent polymer hydrogel particles are suspended in a saline solution. In another embodiment, the derivatized polyvalent polymer particles are suspended in a solution of hyaluronic acid or a hyaluronic acid derivative of this disclosure. In one embodiment, the crosslinked derivatized polyvalent polymer suspension is contained in a pre-filled syringe, the contents of which are sterile.

[0180] In one embodiment, derivatized polyvalent polymers and compositions as disclosed herein are formulated for and useful in preventing adhesions. Derivatized polyvalent polymers may be crosslinked or uncrosslinked, and these compositions may optionally contain bioactive agents. Areas of the body where treatment methods for preventing adhesions are desired include the spinal and abdominal regions, particularly when used postoperatively, as a coating for dura substitutes, during or on nasal sphincter surgery, and in conjunction with medical procedures on the ears, elbows, and tendons. Exemplary bioactive agents include, but are not limited to, anti-inflammatory and analgesic agents.

[0181] In one embodiment, the derivatized polyvalent polymers of the Disclosure may be used to reduce the incidence and severity of adhesions and scar tissues that may occur after trauma or surgery. These adhesions may include abdominal adhesions, pelvic adhesions, cardiac adhesions, joint adhesions, tendon adhesions (e.g., flexor tendons, Achilles tendons, patellar tendons), spinal adhesions, lumbar adhesions, nerve adhesions, dural adhesions, and sinus adhesions. The derivatized polyvalent polymer composition may further comprise one or more excipients. The derivatized polyvalent polymer composition of the Disclosure may further comprise a bioactive agent. In one embodiment, the bioactive agent may be, but is not limited to, a corticosteroid, a local anesthetic, an antibody, a peptide, or an anti-inflammatory agent. In one embodiment, the derivatized polyvalent polymers of the Disclosure are derived from hyaluronic acid or hyaluronic acid derivatives. In one embodiment, the derivatized polyvalent polymers may be in the form of crosslinked hydrogels. In another embodiment, the derivatized polyvalent polymer of the present disclosure may be in a lyophilized, crosslinked form to form a porous foam, or it may be a solid film or a perforated film.

[0182] In one embodiment, derivatized polyvalent polymers and compositions as disclosed herein are formulated for and useful for tissue encapsulation. The derivatized polyvalent polymers may be crosslinked or uncrosslinked, and these compositions may optionally contain bioactive agents.

[0183] In one embodiment, a derivatized polyvalent polymer of the present disclosure containing a vinyl sulfone residue can be reacted with a compound having two or more free thiol functional groups to produce a crosslinked derivatized polyvalent polymer. In one embodiment, a derivatized polyvalent polymer of the present disclosure containing free vinyl sulfone groups can be prepared as a solution. In one embodiment, this solution can be prepared using physiological saline. In one embodiment, a derivatized polyvalent polymer of the present disclosure containing a vinyl sulfone residue can be prepared as a first solution, and a derivatized polyvalent polymer having two or more free thiol functional groups can be prepared as a second solution. The pH of either the first or second solution can be adjusted so that the pH of the solution is greater than pH 8. This can be achieved by using a solution with a pH greater than 8 to dissolve the derivatized polyvalent polymer of the present disclosure containing a vinyl sulfone residue or a compound having two or more free thiol functional groups, and by adding a buffer component to either the derivatized polyvalent polymer of the present disclosure containing a vinyl sulfone residue or a compound having two or more free thiol functional groups.

[0184] In one embodiment, the first solution and the second solution can be combined and applied to a tissue surface to obtain a mixture. In one embodiment, this mixture can be applied with a needle or cannula. In another embodiment, this mixture can be applied using a spray applicator. Examples of spray applicators, but not limited to, include Fibrijet SA-3674 and SA-3675 (Nordson Medical, 261 Cedar Hill Street, Marlborough, MA 01752, United States). In another embodiment, this mixture can be applied using a gas-assisted spray applicator. Examples of gas-assisted spray applicators, but not limited to, include Fibrijet SA-3651 and SA-3652 (Nordson Medical, 261 Cedar Hill Street, Marlborough, MA 01752, United States).

[0185] In one embodiment, the derivatized polyvalent polymer composition can be applied to a tissue in liquid form, and after 3 minutes, the derivatized polyvalent polymer composition becomes a gel. The time required for conversion from liquid to gel depends on the specific application. In one embodiment, the conversion from liquid to gel may take less than 2 minutes. In another embodiment, the conversion from liquid to gel may take less than 30 seconds. In yet another embodiment, the conversion from liquid to gel may take less than 15 seconds.

[0186] The derivatized polyvalent polymer composition for tissue encapsulation may further contain excipients. The derivatized polyvalent polymer composition may further contain bioactive agents.

[0187] In one embodiment, the derivatized polyvalent polymer composition of the present disclosure is combined with a bioactive agent to treat bacterial vaginosis. The derivatized polyvalent polymer composition of the present disclosure can be formulated so that it acts as a tissue adhesive and adheres to vaginal tissue for a period of time greater than two hours. The derivatized polyvalent polymer composition may further contain one or more excipients. The derivatized polyvalent polymer composition of the present disclosure may further contain a bioactive agent. In one embodiment, the bioactive agent may be an antimicrobial agent, but is not limited to clindamycin, tinidazole, metronidazole, secnidazole, and ornidazole. The formulation containing the derivatized polyvalent polymer of the present disclosure can be applied intravaginally.

[0188] In one embodiment, the derivatized polyvalent polymers and compositions of the present invention are selected to provide applications to the eye, for example, eye drops for dry eyes / lubricating eye drops for contact lenses.

[0189] In one embodiment, the derivatized polyvalent polymer compositions of the present disclosure are usable as eye drops. These eye drops can be used as lubricants for the surface of the eye, as lubricants for use with contact lenses, and to aid in the healing of the eye after trauma or surgery to the eye or surrounding tissues, in order to treat dry eye, eye diseases, infected eye tissue, or inflamed eye tissue. Eye surgeries include, but are not limited to, cataract surgery, intraocular lens replacement, fixation of detached retina, tumor removal, glaucoma surgery, refractive surgery, corneal surgery, vitreoretinal surgery, oculomuscular surgery, oculoplastic surgery, and surgery involving the lacrimal punctum, lacrimal canaliculus, and lacrimal sac. The ophthalmic formulations comprising the derivatized polyvalent polymers of the present disclosure may further include excipients. The derivatized polyvalent polymers of the present disclosure may be formulated as a solution or suspension, which is then administered to the eye. The ophthalmic formulations comprising the derivatized polyvalent polymers of the present disclosure may further include bioactive agents. The bioactive agents may be present as part of a solution, or in the form of a suspension or emulsion. The derivatized polyvalent polymers of this disclosure can be formulated as a solution or suspension, which is then administered to the eyes.

[0190] In another embodiment, the derivatized polyvalent polymer compositions of the present disclosure can be prepared for use in lubricating and wetting contact lenses. Contact lenses can be immersed in or stored in a solution containing the derivatized polyvalent polymer of the present disclosure. The solution may contain one or more excipients. The solution may further contain boric acid or sodium borate. The solution may be formulated to be preservative-free.

[0191] In one embodiment, the derivatized polyvalent polymer of the present disclosure can be formed as a formulation to be inserted into the lacrimal punctum, lacrimal canaliculi, or lacrimal sac. The derivatized polyvalent polymer of the present disclosure may be in the form of a solution, a swollen hydrogel, or a dehydrated hydrogel. In one embodiment, the derivatized polyvalent polymer composition may further comprise excipients. In another embodiment, the derivatized polyvalent polymer is crosslinked. In yet another embodiment, the derivatized polyvalent polymer composition may further comprise bioactive agents. In one embodiment, the bioactive agent may be, but is not limited to, a corticosteroid (e.g., dexamethasone, mometasone fuorate, triamcinolone acetonide, triamcinolone hexaacetonide, triamcinolone acetate, betamethasone, fluoromethalone, hydrocortisone, medlison, or prednisolone), a prostaglandin (e.g., latanoprost, travoprost, or bimatoprost), a beta-blocker (e.g., timolol or betaxolol), an alpha-adrenergic agonist (e.g., apraclonidine or brimonidine), a carbonic anhydrase inhibitor (e.g., dorzolamide or brinzolamide), a mitotic agent, or a cholinergic agent (e.g., pilocarpine).

[0192] In another embodiment, a derivatized polyvalent polymer and / or composition is crosslinked in the presence of a bioactive agent and then dried. In another embodiment, a derivatized polyvalent polymer is crosslinked, dried, re-swelled in the presence of a bioactive agent, and then dried. In another embodiment, a biological agent is incorporated into a non-crosslinked derivatized polyvalent polymer in solution. In another embodiment, a derivatized polyvalent polymer is dried, re-swelled in the presence of a bioactive agent, and then dried. The dried formulations may be of suitable dimensions so as to be placed in the lacrimal punctum. Upon contact with tears and tears, the final dried formulation hydrates and swells in a manner that allows it to be physically held in the lacrimal punctum. In another embodiment, the dried formulation can be inserted into the lacrimal canaliculi. Upon contact with tears and tears, the dried formulation hydrates and swells in a manner that allows it to be physically held in the lacrimal canaliculi. The formulation may then release the bioactive agent it contained over a period of 24 hours to 3 weeks. In one embodiment, the bioactive agent is released continuously for 7 days. In one embodiment, the bioactive agent is released continuously for 4 weeks. In one embodiment, the dry formulation can be introduced into the vitreous humor so that the bioactive agent is delivered to the vitreous humor of the eye. In one embodiment, the dry formulation is inserted into the anterior chamber.

[0193] In one embodiment, the derivatized polyvalent polymers and compositions of the present disclosure are selected to provide punctal plugs. The punctal plugs may contain a bioactive agent, such as a steroid or an analgesic.

[0194] In one embodiment, the derivatized polyvalent polymer compositions of the present disclosure can be used to treat mucositis. Examples of mucositis include oral and vaginal mucositis. During cancer treatment, rapidly dividing epithelial cells lining the gastrointestinal tract (extending from the mouth to the anus) break down, leaving the mucosal tissue susceptible to ulceration and infection. This results in mucositis. Oral mucositis can be commonly seen after chemotherapy and radiation therapy. It can cause pain and increase the risk of infection. This can lead to nutritional problems due to these symptoms, which reduce the ability to eat and the desire to eat. Pain and the likelihood of infection can be reduced by providing a coating to cover these lesions. The derivatized polyvalent polymers of the present disclosure can be formulated so that the derivatized polyvalent polymer composition acts as a tissue adhesive and adheres to oral or vaginal mucosal tissue for a period longer than two hours. The derivatized polyvalent polymer composition may further comprise one or more excipients. The derivatized polyvalent polymer composition may further comprise a bioactive agent. In one embodiment, the bioactive agent may be, but is not limited to, a local anesthetic, an anti-infective agent, an anti-inflammatory agent, or a combination thereof. Local anesthetics may include, but are not limited to, benzocaine, bupivacaine, ametokine, lignocaine, lidocaine, cocaine, cincocaine, dibucaine, mepivacaine, prinocaine, etidocaine, veratridine (a specific C-fiber blocker), and procaine. For oral mucositis, the derivatized polyvalent polymer composition of this disclosure can be formulated for use as a mouthwash or as a gel. For vaginal mucositis, the derivatized polyvalent polymer composition of this disclosure can be formulated for intravaginal application to the surface of vaginal tissue.

[0195] In one embodiment, the derivatized polyvalent polymer composition of the present disclosure may be used to treat the surgical site during and after any of the following procedures related to the ear: canalplasty, tympanoplasty, tympanoplasty, stapedectomy, mastoidectomy, or any other ear-related procedure. The derivatized polyvalent polymer composition may be used to regulate wound healing and control bleeding. The derivatized polyvalent polymer composition of the present disclosure may be in the form of a freeze-dried sponge, an electrospun matrix, a film, a gel, or a combination thereof. The derivatized polyvalent polymer composition of the present disclosure may contain excipients. In another embodiment, the derivatized polyvalent polymer composition of the present disclosure may contain bioactive agents.

[0196] In another embodiment, the derivatized polyvalent polymer compositions of the present disclosure may be used to treat otitis media, acute otitis externa, balance disorders (e.g., Meniere's disease, tinnitus, and sensory nervous system hearing loss). The derivatized polyvalent polymer compositions of the present disclosure may be in the form of solutions, suspensions, lyophilized sponges, electrospun matrices, films, gels, solid cylindrical forms, or combinations thereof. The derivatized polyvalent polymer compositions of the present disclosure may contain excipients. In another embodiment, the derivatized polyvalent polymer compositions of the present disclosure may contain bioactive agents. For treating ear infections, the derivatized polyvalent polymer may contain antibiotics, antimicrobial agents, antiviral agents, antifungal agents, or combinations thereof. In one embodiment, a derivatized polyvalent polymer composition containing at least one bioactive agent may include, but is not limited to, amoxicillin, clavulanate, cefuroxime acetyl, ceftriaxone, levofloxacin, cephalosporin The composition includes phosphorus, trimethoprim-sulfamethoxazole, macrolides, ofloxacin, gentamicin sulfate, tobramycin sulfate, and cyprophylaxin, and in another embodiment, the derivatized polyvalent polymer composition may include a corticosteroid. The corticosteroids include, but are not limited to, betamethasone, betamethasone valerate, cortisone, dexamethasone, dexamethasone 21-phosphate, fludrocortisone, flumetasone, and flu The composition may include ocinonide, fluocinonide, desonide, fluocinolone, fluocinolone acetonide, fluocortolone, halcinonide, halopredone, hydrocortisone, hydrocortisone 17-valeric acid, hydrocortisone 17-butyric acid, hydrocortisone 21-acetic acid, methylprednisolone, prednisolone, prednisolone 21-phosphate, prednisone, triamcinolone, triamcinolone acetonide, and mometasone fuorate. In another embodiment, a combination of antibiotics and corticosteroids may be added to the derivatized polyvalent polymer composition of the Disclosure. In one embodiment, the derivatized polyvalent polymer composition of the Disclosure may be applied to a treatment area by administering it through a needle or catheter using an infusion device or syringe, or by introducing the derivatized polyvalent polymer composition into the body.

[0197] In one embodiment, the derivatized polyvalent polymer composition of the Disclosure may contain a bioactive agent. The derivatized polyvalent polymer of the Disclosure can be used as a matrix from which the bioactive agent can be delivered. In one embodiment, the release characteristics of the bioactive agent into phosphate-buffered saline are slower than the normal dissolution characteristics of the bioactive agent. In one embodiment, the derivatized polyvalent polymer composition of the Disclosure may be in the form of a crosslinked gel.

[0198] In one embodiment, the treatment using the drug delivery formulation may be a single injection or two or more injections spaced apart. The composition may be injected subcutaneously, intradermally, or intramuscularly. The derivatized polyvalent polymer composition may be injected via a needle, trowel, catheter, tube, or cannula.

[0199] In another embodiment, the contents of the filled syringe or vial are sterile. In another embodiment, the contents of the filled syringe or vial are stable for at least 6 months, preferably 12 months, and most preferably 24 months, at 2-8°C or 20-25°C. In another embodiment, the drug delivery formulation can be applied topically or by intravenous infusion.

[0200] In one embodiment, a crosslinked derivatized polyvalent polymer of the present disclosure can be used as a device for filling defects after tissue fragment removal or needle marks after a biopsy procedure. In another embodiment, a crosslinked derivatized polyvalent polymer composition can be prepared and then dried. The dried derivatized polyvalent polymer composition can be delivered to a needle mark or site where tissue fragment has been removed. The dried derivatized polyvalent polymer composition can absorb moisture from the surrounding tissue, rehydrate and swell, so that its swollen size is greater than its initial size. The swollen derivatized polyvalent polymer composition is then retained at the site where it is placed. In another embodiment, a crosslinked dried derivatized polyvalent polymer composition can be used to seal holes in tissue, where the crosslinked derivatized polyvalent polymer composition is placed in the hole and swells to seal the hole. This example could be for sealing lung tissue after lung puncture following a biopsy or surgery. In another embodiment, the crosslinked dried derivatized polyvalent polymer composition may contain elements such as metal pieces visible under X-ray or fluoroscopy. The metal pieces can take various forms, but are not limited to, flat pieces, cylinders, coils, loops, hoops, hooks, numbers, and letters of the alphabet. In one embodiment, the crosslinked dried derivatized polyvalent polymer composition may contain a bioactive agent. In another embodiment, the bioactive agent may have hemostatic properties. In one embodiment, the crosslinked dried derivatized polyvalent polymer composition may contain collagen, chitosan, or thrombin.

[0201] In one embodiment, the derivatized polyvalent polymers and compositions of the present disclosure are formulated for and useful in female contraceptive plugs. Female contraception can be achieved by inserting a plug into the fallopian tube. This plug can provide a physical barrier to the passage of the egg into the uterus and to the arrival of sperm to the egg. Laparoscopic methods are the primary method of achieving female contraception, in which the fallopian tubes are cut and then ligated. In other variations of this method, the fallopian tubes can be closed by closing and tightening them with clips or rings. Cauterization has also been used to seal the fallopian tubes. These methods are generally classified as major surgery, usually requiring general anesthesia, and the patient requires a recovery period. Vaginal contraception became an alternative to laparoscopy due to its less invasive nature. The first transvaginal methods used chemical agents such as sodium molunate, or quinacrine, methyl cyanacrylate, and silver nitrate, but their success rates and side effects limited their use. Hysteroscopic tubal sterilization emerged as a minimally invasive alternative to conventional tubal ligation. Hysteroscopic tubal sterilization can be performed in a clinic in approximately 10 minutes without the use of general or even local anesthesia.

[0202] Two types of tubal contraception methods were commercialized, but both were removed from the US market at the end of 2018. The Essure system consisted of a device insert loaded into a single-use delivery system. This device consisted of an inner coil of stainless steel and polyethylene terephthalate (PET) fiber and an outer coil of nickel-titanium (nitinol). The metal components held the device in place while the PET fiber allowed tissue to implant into the device, thereby leading to occlusion of the fallopian tube. Because this implantation process takes time, the patient had to use other forms of contraception for three months. At this stage, a hysterosalpingogram was performed to confirm implantation and occlusion of the fallopian tube. This device is permanent and remains in the patient's body for life. This product received a black box warning due to potential safety concerns and was subsequently removed from the US market. This device has since been removed from overseas markets as well.

[0203] Another method of contraception was developed by Hologic. The Adiana® method used high-frequency energy to cause controlled thermal damage to the inner lining of the fallopian tube lumen. After thermal damage to the fallopian tube, a porous, non-degradable silicone plug was inserted into the damaged tube. Within a few weeks, fallopian tube occlusion occurred due to tissue infiltration into the porous plug. At three months, a hysterosalpingogram was performed to confirm fallopian tube occlusion. The silicone plug is a permanent implant. The Adiana® system has been removed from the market.

[0204] Both the Essure system and the Adiana® system leave permanent devices in the patient. This could potentially pose long-term safety problems for the patient. A system containing a biodegradable plug component would be beneficial in that it leaves little to no permanent residue of the derivatized polyvalent polymer and / or composition in the patient. The method and apparatus described herein provide a means of occlusion of the fallopian tube that results in a reduction of female fertility. The method involves mechanically damaging the inner lining of the fallopian tube and then inserting a biodegradable plug.

[0205] A method of mechanically damaging the fallopian tube involves inserting a device containing a roughened surface into the fallopian tube and then physically moving this device in a rotational motion, a linear motion along the fallopian tube, or a combination thereof. This motion can be repeated two or more times. This physical motion is continued until the endothelial layer of the fallopian tube in which the physical motion occurs is partially or completely removed.

[0206] The apparatus used to exfoliate the endothelial layer of the fallopian tube may include a series of fibers radiating from a central core. In one embodiment, the apparatus is structurally similar to a bottle brush, for example, a rod with bristles (fibers) extending perpendicularly from the rod.

[0207] In one embodiment, these fibers can be arranged continuously at regular intervals. In one embodiment, these fibers can be arranged in multiple rows with spaces between them. In one embodiment, these fibers can be oriented helically along the axis of the device. In one embodiment, these fibers can be oriented as one or more linear rows aligned substantially parallel to the axis from which they diverge. In another embodiment, these fibers are arranged in one or more rows such that the rows are substantially perpendicular to the axis from which they diverge.

[0208] In one embodiment, these fibers can be made from non-degradable polymers. Polymers that can be used to make the fibers include, but are not limited to, polyethylene, polypropylene, polyethylene terephthalate (PET), nylon, polyurethane, polyether ether ketone (PEEK), polyaryl ether ketone (PAEK), fluorocarbon polymers, such as polytetrafluoroethylene, silk, and combinations thereof.

[0209] In one embodiment, these fibers can be made from metals. Metals that can be used to make the fibers include, but are not limited to, stainless steel, titanium, nitinol, magnesium, alloys Co-Cr-Mo, Cr-Ni-Cr-Mo, CP-Ti, Ti-Al-V, Ti-Al-Nb, Ti-13Nb-13Zr, Ti-Mo-Zr-Fe, or combinations thereof.

[0210] In one embodiment, the central core (rod) of the peeling apparatus may include a core made from two or more metal strands twisted together so that fibers are trapped between the metal strands. Metals that can be used to make the central core include, but are not limited to, stainless steel, titanium, nitinol, magnesium, alloys Co-Cr-Mo, Cr-Ni-Cr-Mo, CP-Ti, Ti-Al-V, Ti-Al-Nb, Ti-13Nb-13Zr, Ti-Mo-Zr-Fe, or combinations thereof.

[0211] In one embodiment, the end of the central core (rod) initially introduced into the fallopian tube may include a non-traumatic tip that does not damage tissue as the device is guided to a desired location in the fallopian tube. This non-traumatic tip may be a rounded end cap, a domed end, or a conical end with a rounded tip. The surface of the non-traumatic tip may have a smooth surface. The non-traumatic tip can be manufactured from a non-degradable polymer or metal. Non-degradable polymers that can be used to manufacture the non-traumatic tip include, but are not limited to, polyethylene, polypropylene, polyethylene terephthalate (PET), nylon, polyurethane, polyether ether ketone (PEEK), polyaryl ether ketone (PAEK), fluorocarbon polymers, such as polytetrafluoroethylene, silk, and combinations thereof. Metals that can be used to create non-traumatic tips include, but are not limited to, stainless steel, titanium, nitinol, magnesium, alloys Co-Cr-Mo, Cr-Ni-Cr-Mo, CP-Ti, Ti-Al-V, Ti-Al-Nb, Ti-13Nb-13Zr, Ti-Mo-Zr-Fe, or combinations thereof.

[0212] The non-traumatic tip can be bonded to the core by a crimping process, a molding process, a process using an adhesive to bond the tip to the core, or a heat process.

[0213] The plug may contain a hydrogel. In one embodiment, the hydrogel is prepared using one or more crosslinked derivatized polyvalent polymers and / or compositions of the present disclosure. A hydrogel containing the polyvalent polymer composition is prepared in the form of a rod larger than the size of the fallopian tube. The hydrogel rod is then dried. The hydrogel can be dried under normal or reduced pressure. In one embodiment, the hydrogel can be freeze-dried. Upon delivery to the desired site, the hydrogel plug absorbs moisture from the fallopian tube and swells. This swelling of the hydrogel plug allows it to be retained in the site where it is placed.

[0214] In one embodiment, the hydrogel further comprises a pologen to assist in the formation of pores within the hydrogel. This pologen may comprise particles. These particles may comprise a biodegradable polymer. Biodegradable polymers usable as pologens include, but are not limited to, biodegradable polyesters, polyanhydrides, polyurethanes, polyether esters, polycarbonates, polyether carbonates, polyether ester carbonates, polyhydroxyalkanoates, polyamides, and polymers synthesized from one or more monomers from the group consisting of l-lactide, dl-lactide, glycolide, ε-caprolactone, trimethylene carbonate, morpholine dione, p-dioxanone, and 1,5-dioxapan-2-one.

[0215] In one embodiment, pologens may leak from the hydrogel during the device manufacturing process. This can be achieved by incubating hydrogen-containing pologens in a solvent in which pologens dissolve. The solvent is preferably a water-miscible solvent. In another embodiment, pologens may remain in the device during the manufacturing process and decompose and leak out when the hydrogel plug is inserted into the patient.

[0216] In one embodiment, the plug comprises a biodegradable polymer. Biodegradable polymers usable in the plug include, but are not limited to, biodegradable polyesters, polyanhydrides, polyurethanes, polyether esters, polycarbonates, polyether carbonates, polyether ester carbonates, polyhydroxyalkanoates, polyamides, and polymers synthesized from one or more monomers from the group consisting of l-lactide, dl-lactide, glycolide, ε-caprolactone, trimethylene carbonate, morpholine dione, p-dioxanone, and 1,5-dioxapan-2-one.

[0217] The plug may include a monofilament structure, a multifilament structure, or a braided structure. In one embodiment, the plug can be made by obtaining particles or chopped fibers of a biodegradable polymer and compression molding them into a certain shape. Heat can be used to thermally fuse the particles together to obtain a porous structure. In one embodiment, the shape may be in the form of a rod. This porous rod can then be cut to a predetermined length.

[0218] In one embodiment, the plug can be made from an electrospun biodegradable polymer. In one embodiment, the plug is made from a thin film of an electrospun derivatized polyvalent polymer and / or composition. The plug can be cut directly from a sheet of electrospun composition. In one embodiment, the plug can be made by rotating an electrospun film to form a roll. The electrospun plug or roll rod molded structure can be coated with a second biodegradable polymer so as to maintain the roll structure. In one embodiment, the polymer used to make the roll structure has a longer decomposition time than the polymer used to coat the roll structure. This makes the plug stiffer for easier handling during manufacturing, but allows the faster-degrading material to begin decomposing upon delivery to the desired site, aiding tissue endografting, while the first, longer-lasting polymer provides a scaffold for the endografting tissue.

[0219] In another embodiment, the electrospun plug may be coated with a solution of a water-soluble polymer or immersed in a solution of a water-soluble polymer. The plug is then dried under ambient pressure or reduced pressure. The plug may also be dried by freeze-drying. The presence of the water-soluble polymer makes the electrospun composition more rigid and therefore easier to handle during manufacturing and delivery to the intended site. Where it is located at the intended site, the polymer begins to dissolve and leak out of the electrospun composition. Tissue from the mechanically damaged oviduct can then be implanted into the electrospun composition. The electrospun composition decomposes over time, leaving behind the occluded oviduct. In one embodiment, the water-soluble polymer can be selected from the group consisting of polyethylene oxide, polyethylene glycol, block copolymers of polyethylene glycol and polypropylene glycol (e.g., Pluronic F126 and Pluronic F68, Sigma-Aldrich Corp., St. Louis, MO, USA), dextran, hyaluronic acid, or hyaluronic acid derivatives of the present disclosure.

[0220] The biodegradable polymer used to form the plug may further contain pologens. Pologens may include inorganic salts, small organic molecules, or polymers. The pologens are selected so that the biodegradable polymer used to make the plug is soluble in a solvent having limited solubility.

[0221] Inorganic salts that can be used as pologens include, but are not limited to, sodium salts, potassium salts, calcium salts, magnesium salts, aluminum salts, copper salts, barium salts, and iron salts. Examples of these salts include, but are not limited to, sodium chloride, sodium bromide, sodium iodide, sodium sulfate, sodium phosphate, sodium hydrogen phosphate, or combinations thereof.

[0222] The porous plug can be fabricated by 3D printing the plug. A degradable polymer can be used for 3D printing the plug. In one aspect, degradable polymers that can be used in the plug include, but are not limited to, degradable polyesters, polyanhydrides, polyurethanes, polyether esters, polycarbonates, polyether carbonates, polyether ester carbonates, polyhydroxyalkanoates, polyamides, and polymers synthesized from one or more monomers from the group of l-lactide, dl-lactide, glycolide, ε-caprolactone, trimethylene carbonate, morpholine-dione, p-dioxanone, and 1,5-dioxapan-2-one.

[0223] The plug can include position-retaining features. These features can include an asymmetric shape, a valve, a ridge, a pore, a slit, a slot, or a combination thereof. These valves can be unidirectional in that they all point in the same direction, or these valves can also point in two or more different directions. These valves can be arranged at regular intervals on the plug, or the valves can be present only in a specific portion of the plug.

[0224] In one aspect, the plug can be immersed in a solution of the derivatized polyvalent polymer of the present disclosure. Next, the solution can be activated such that the solution crosslinks and the pores of the plug contain the crosslinked derivatized polyvalent polymer. The crosslinking process can be activated by adjusting the pH of the solution, adding a crosslinking agent, increasing the temperature, adding an initiator, or a combination of one or more of these.

[0225] In one embodiment, the derivatized polyvalent polymer composition of the present disclosure can be used as a scaffold enabling tissue or bone endoproliferation. In one embodiment, the derivatized polyvalent polymer of the present disclosure can be prepared as a crosslinked matrix, which is then freeze-dried. The freeze-dried derivatized polyvalent polymer composition can then be rehydrated in the presence of cells so that the hydrated matrix acts as a scaffold enabling cell endoproliferation on and within the scaffold. In another embodiment, the derivatized polyvalent polymer of the present disclosure having vinyl sulfone residues can be electrospun to form a porous matrix. The electrospun fibers can then be crosslinked using heat, ultraviolet light, an electron beam, or gamma rays. In another embodiment, the derivatized polyvalent polymer of the present disclosure containing vinyl sulfone residues may further contain a photocrosslinker. A solution of this composition can be electrospun, and the electrospun matrix can then be exposed to ultraviolet light so that the photocrosslinker results in crosslinking of the derivatized polyvalent polymer. The resulting matrix can then be rehydrated in the presence of cells so that it acts as a scaffold for tissue growth. In another embodiment, a derivatized polyvalent polymer of the Disclosure containing a carboxylic acid can be electrospun to form a matrix by mixing a solution of the derivatized polyvalent polymer of the Disclosure with a solution of polyvalent cations immediately before electrospinning. In one embodiment, a solution of the composition of the Disclosure containing a carboxylic acid can be placed in one syringe, and a solution of polyvalent cations or a cationic polymer can be placed in another syringe. These syringes can be connected via a Y-connector, and a needle can be connected to the final arm of the Y-connector. These two solutions can then be drawn up with the needle, and this mixture can be electrospun to form a surface such that the derivatized polyvalent polymer of the Disclosure is ionically crosslinked. Examples of polyvalent cations include calcium, magnesium, ferric ions, ferrous ions, aluminum, and chromium.

[0226] Examples of cationic polymers that can be used include, but are not limited to, chitosan and its derivatives, polyvinylpyrrolidone, peptides containing two or more lysine groups, and polyethyleneimine.

[0227] In another aspect, the solution of the derivatized multivalent polymer composition of the present disclosure can be used to coat a degradable or non-degradable scaffold matrix. In one aspect, the derivatized multivalent polymer of the present disclosure modified with an alkyl group or an aryl group can be used to coat a scaffold for tissue growth. The alkyl group or aryl group interacts with the scaffold via hydrophobic bonds, and between them, the hydrophilic portion of the derivatized multivalent polymer enables cell growth on the coated scaffold surface. In another aspect, the derivatized multivalent polymer of the present disclosure having a vinyl sulfone residue can be coated on a scaffold. The coated scaffold can be exposed to heat, thereby producing a derivatized multivalent polymer that is converted into a cross-linked derivatized multivalent polymer.

[0228] In another aspect, the derivatized multivalent polymer and / or composition of the present disclosure may contain a sulfonic acid group. In another aspect, the derivatized multivalent polymer and / or composition of the present disclosure may contain both a hydrophobic group and a sulfonic acid group. The hydrophobic group can be alkyl or aromatic.

[0229] In another aspect, the tissue scaffold support structure can be 3D printed or electrospun using a degradable polymer. Examples of degradable polymers that can be used include, but are not limited to, degradable polyesters, polyanhydrides, polyurethanes, polyether esters, polycarbonates, polyether carbonates, polyether ester carbonates, polyhydroxyalkanoates, polyamides, and polymers synthesized from one or more monomers from the group of l-lactide, dl-lactide, glycolide, ε-caprolactone, trimethylene carbonate, morpholine-dione, p-dioxanone, and 1,5-dioxapan-2-one.

[0230] In another embodiment, one or more polymer solutions may be prepared for the electrospinning scaffold support structure. The polymers used may be biodegradable polymers, but are not limited to, polyester, polyanhydride, polyorthoester, polycarbonate, polyester-co-carbonate, polyhydroxybutyrate, or combinations thereof. Biodegradable polymers may include polylactice-co-glycolide copolymers, polydioxanones, polylactice-trimethylene carbonate copolymers, and copolymers containing repeating units derived from at least one of the following monomers: l-lactide, dl-lactide, glycolide, trimethylene carbonate, ε-caprolactone, p-dioxanone, and morpholindione.

[0231] The solvent used may be an organic solvent, water, or a combination thereof. For example, HFIP, DMSO, NMP, chloroform, acetic acid, ethanol, dimethylformamide (DMF) solvent, or a mixture of solvents can be used. Solutions with a concentration of 0.5–25% (w / v) can be prepared. The solution to be electrospun can be placed in a syringe with a needle. This syringe is then placed in a syringe pump. The needle may have a blunt end and an inner diameter in the range of 0.25–2.5 mm. The needle and recovery plate are attached to a high voltage source. In some applications, two or more needles can be used to produce a single sheet. The needles can be configured so that the same polymer solution flows through all needles, different solutions flow through different needles, or a combination thereof. These needles can be configured so that adjacent needles allow different polymer solutions to flow through them. This alternating pattern can be repeated. Then, a voltage is applied to this system. The applied voltage may be 10kV–45kV. A syringe pump can push out the solution. The flow rate of the syringe pump may range from 0.0001 uL / min to 423 mL / min. The collector plate may be stationary, rotating, or moving in a specific linear direction to give the fibers some directional orientation. The shape of the collector plate is not limited to but may vary, and may include the following shapes: flat surface, woven surface, curved surface, regular prism, prismatic, round core, elliptical core, semicircular core, or a combination of these shapes. The distance between the needle tip and the collector plate is variable. The distance between the needle tip and the collector plate may range from 2 to 50 cm. The collection plate may also be immersed in a solvent that aids in the precipitation of the newly spun fibers, or the solvent may be sprayed onto it. For example, an ethanol bath may be used when electrospinning the hyaluronic acid-based derivatized polyvalent polymer of this disclosure. The derivatized polyvalent polymer of this disclosure can be incorporated by solution coating or immersion of the electrospinning matrix.

[0232] In one embodiment, the polymer composition used for 3D printing or electrospinning the scaffold may further comprise an inorganic filler or a combination of inorganic fillers. In one embodiment, the inorganic filler can be selected from the group consisting of calcium carbonate, calcium phosphate, tricalcium phosphate, hydroxyapatite, bioglass, or combinations thereof.

[0233] In one embodiment, a 3D printed or electrospun scaffold can be coated with a solution of the derivatized polyvalent polymer of the Disclosure. This derivatized polyvalent polymer can be coated onto the scaffold by a dip coating method or a spray coating method. In another embodiment, the derivatized polyvalent polymer can be dispersed onto the scaffold by applying compression. In another embodiment, the derivatized polyvalent polymer can be dispersed onto the scaffold by immersion in a solution, which may or may not include sonic treatment to aid dispersion. In another embodiment, the coated scaffold can be dried. This drying step may include drying at high temperature, drying under reduced pressure, or freeze-drying. In another embodiment, the solution of the derivatized polyvalent polymer composition of the Disclosure may further contain a bioactive agent.

[0234] In another embodiment, the scaffold can be immersed in a solution of the derivatized polyvalent polymer of the Disclosure. The solution can then be activated so that it crosslinks and the pores of the scaffold contain the crosslinked derivatized polyvalent polymer. The crosslinking step can be activated by adjusting the pH of the solution, adding a crosslinking agent, increasing the temperature, adding an initiator, or one or more of these.

[0235] In another embodiment, the scaffold can be immersed in a solution of the derivatized polyvalent polymer of the Disclosure and dried or freeze-dried. The derivatized polyvalent polymer in the substrate can then be immersed in a crosslinking solution so that the solution crosslinks and the pores of the scaffold contain the crosslinked derivatized polyvalent polymer. The crosslinking step can be activated by adjusting the pH of the solution, adding a crosslinking agent, increasing the temperature, adding an initiator, or one or more of these.

[0236] In another embodiment, a scaffold can be immersed in a solution of the derivatized polyvalent polymer and crosslinking agent of the Disclosure. The rate of the crosslinking reaction can be controlled so that the scaffold is coated with the derivatized polyvalent polymer and / or composition before the derivatized polyvalent polymer is fully crosslinked. In one embodiment, a bioactive agent can be incorporated into the derivatized polyvalent polymer and / or composition before or immediately after the start of the crosslinking reaction. The scaffold can then be coated with this composition, and once applied to the scaffold, the crosslinking reaction is completed so that the apparatus contains the crosslinked derivatized polyvalent polymer together with the bioactive agent essentially encapsulated by the crosslinked derivatized polyvalent polymer composition.

[0237] In another embodiment, the derivatized polyvalent polymers and / or compositions of the Disclosure used for constructing or coating scaffolds may include a bioactive agent. In one embodiment, the bioactive agent may promote cell growth. In one embodiment, the bioactive agent may be one or more growth factors or peptides that promote cell growth and cell adhesion. In another embodiment, the derivatized polyvalent polymers and / or compositions of the Disclosure used for constructing or coating scaffolds may further include excipients. In one embodiment, the derivatized polyvalent polymer composition of the Disclosure may include one or more extracellular matrix components. Extracellular matrix components may include, but are not limited to, heparan sulfate, chondroitin sulfate, keratin sulfate, hyaluronic acid, collagen, elastin, fibronectin, and laminin.

[0238] In one embodiment, cells that can be added to a scaffold containing the derivatized polyvalent polymer composition of the present disclosure include embryonic stem cells, mesenchymal stem cells, adipose-derived stem cells, endothelial stem cells, dental pulp stem cells, tumor cells, chondrocytes, osteoblasts, dermal fibroblasts, myofibroblasts, myofibroblasts, hepatocytes, smooth muscle cells, endothelial cells, epithelial cells, adipose tissue, adipocytes, and cardiac cells.

[0239] In one embodiment, the derivatized polyvalent polymers and compositions of the present invention contain free vinyl sulfone functional groups and are usable for 3D printing of structures. The derivatized polyvalent polymers can be prepared as solutions having a viscosity that allows them to be 3D printed. In one embodiment, a solution of a derivatized polyvalent polymer having vinyl sulfone residues can be prepared. A second solution containing a derivatized polyvalent polymer having at least two free thiol groups can be prepared. In one embodiment, the first solution and the second solution can be mixed together. Immediately before printing, the pH of the mixture can be adjusted to a pH greater than 8, preferably greater than 9, so that the mixture can be printed and then cured after printing. In one embodiment, the pH can be adjusted by mixing the mixture with a buffer solution having a pH greater than 8. Mixing occurs immediately before the print head so that the mixture does not gel in the print head and therefore the printer does not solidify, and in another embodiment, the solution of the derivatized polyvalent polymer containing vinyl sulfone residues may have a pH that is adjusted to a pH greater than 8 by mixing it with a buffer solution. Next, this solution can be mixed with solution 2 just before the print head so that the mixture is printed and then completely gelled where it is printed.

[0240] The viscosity of the mixture can be used to control the retention of the printed structure until gelation is complete. In another embodiment, the thermal gelling material can be added to either the first or second buffer solution. Thus, the mixture can be printed, and then, while the crosslinking process is moving towards completion, the temperature of the printing environment can be different from that of the solution before printing, so that the printed solution undergoes thermal gelation after the printing process to preserve the initially printed structure.

[0241] The thermal gelling material may include, but is not limited to, polyethylene-block-polypropylene copolymers, such as Pluronic F127 or F68 (Sigma-Aldrich Corp., St. Louis, MO, USA) or polyester-polyethylene glycol block copolymers. The polyester-polyethylene glycol copolymers may include 2-block and 3-block copolymers. The polyester component is a polymer synthesized from at least one monomer from the group consisting of l-lactide, dl-lactide, glycolide, ε-caprolactone, morpholine-dione, p-dioxanone, and 1,5-dioxapan-2-one. In another embodiment, a thermal gelling polymer containing trimethylene carbonate may be used.

[0242] After the gelation process is complete, the printed structure can be rinsed to neutralize the pH of the printed gel. In another embodiment, the printed structure can be dried to a residual moisture content of less than 10%. In yet another embodiment, the printed structure can be freeze-dried.

[0243] The printed structures can be used as tissue scaffolds for wound healing applications, occluding lumens, biopsy sites, or needle marks.

[0244] Holes are often made in the skull for procedures such as neuroendoscopy, intracranial decompression, and treatment of chronic subdural hematomas. These are often called burr holes. In many cases, these burr holes are left untreated after surgery, and the scalp takes over them. This can result in scalp depressions at the burr holes. These scalp depressions can lack mechanical strength. To prevent this, burr hole plugs can be inserted into the burr holes, which can promote and assist bone regrowth. Autologous bone can be used to fill the burr holes, but this requires bone harvesting. Synthetic materials can be used as burr hole plugs. Degradable burr hole plugs that degrade while promoting bone endografting allow for burr hole healing without leaving any residual material. Polycaprolactone (PCL) burr hole plugs are commercially available. The drawbacks of PCL are that it degrades slowly and the interface between the polymer and the endografting tissue is usually not optimal due to the hydrophobicity of the polymer.

[0245] The derivatized polyvalent polymers and their compositions disclosed herein can be prepared as burr hole plugs. A solution of the derivatized polyvalent polymer can be placed in a mold, and the derivatized polyvalent polymer composition can then be freeze-dried to produce a porous structure that can be inserted into a burr hole. In another embodiment, the derivatized polyvalent polymer composition disclosed herein can be electrospun, and then cut to form a plug that can be inserted into a burr hole. In another embodiment, a solution of the derivatized polyvalent polymer disclosed herein can be placed in a mold, and the solution can be crosslinked. The crosslinked plug can then be used as is. In another embodiment, the crosslinked derivatized polyvalent polymer composition can be freeze-dried to obtain a porous crosslinked structure that can be used as a burr hole plug.

[0246] In another aspect, the trephine plug can be 3D printed or electrospun using a degradable polymer. Examples of degradable polymers that can be used include, but are not limited to, degradable polyesters, polyanhydrides, polyurethanes, polyether esters, polycarbonates, polyether carbonates, polyether ester carbonates, polyhydroxyalkanoates, polyamides, and polymers synthesized from one or more monomers from the group consisting of l-lactide, dl-lactide, glycolide, ε-caprolactone, trimethylene carbonate, morpholine-dione, p-dioxanone, and 1,5-dioxapan-2-one.

[0247] In one aspect, the polymer used to 3D print or electrospin the trephine plug can further include an inorganic filler or a combination of inorganic fillers. In one aspect, the inorganic filler can be selected from the group consisting of calcium carbonate, calcium phosphate, tricalcium phosphate, and hydroxyapatite.

[0248] In one aspect, the 3D printed or electrospun trephine plug can further include an extracellular matrix material. In one aspect, the extracellular matrix material can be selected from the group consisting of collagen, hyaluronic acid, chondroitin sulfate, heparan sulfate, keratin sulfate, elastin, fibronectin, and laminin.

[0249] In one aspect, the 3D printed or electrospun plug can be coated with a solution of the derivatized polyvalent polymer of the present disclosure. This derivatized polyvalent polymer composition can be coated onto the plug by a dip coating method or a spray coating method. In another aspect, the coated plug can be dried. The drying process can include drying at high temperature, drying under reduced pressure, or freeze drying.

[0250] In another embodiment, a polymer-degradable plug can be immersed in a solution of the derivatized polyvalent polymer of the Disclosure. The solution can then be activated so that it crosslinks and the pores of the plug contain the crosslinked derivatized polyvalent polymer composition. The crosslinking step can be activated by adjusting the pH of the solution, adding a crosslinking agent, increasing the temperature, adding an initiator, or one or more of these.

[0251] In another embodiment, the polymeric biodegradable plug can be immersed in a solution of the derivatized polyvalent polymer of the disclosure containing vinyl sulfone residues. The coated device can be dried at a high temperature to remove the solvent and to crosslink the coating so that the pores of the plug contain the crosslinked derivatized polyvalent polymer composition.

[0252] In one embodiment, the crosslinked derivatized polyvalent polymers and / or compositions of the present disclosure can be used to form nerve guides. Optionally, the nerve guides can be prepared by lyophilization. In one embodiment, collagen, gelatin, chitosan, heparan sulfate, or a combination thereof can be further added to the derivatized polyvalent polymers and / or compositions of the present disclosure to form nerve guides. In another embodiment, Schwann cells can be incorporated into the derivatized polyvalent polymer composition during nerve guide formation.

[0253] In one embodiment, the derivatized polyvalent polymer of this disclosure can be prepared as a solution having a viscosity greater than 50 cP. In one embodiment, this solution can be applied to tissue to reduce the coefficient of friction with the tissue surface. In one embodiment, the derivatized polyvalent polymer composition can be used as a vaginal lubricant. In another embodiment, the solution can be applied to a device inserted into an opening, hole, or lumen so as to facilitate the passage of the device into the opening, hole, or lumen. In one embodiment, the device may be an endoscope.

[0254] In one embodiment, the derivatized polyvalent polymers and compositions of the present disclosure can be used to coat medical devices. Medical devices that can be coated include, but are not limited to, catheters, needles, biopsy needles, tissue markers, guidewires, and intraluminal sheaths, sutures, braids, trocanic needles, hernia meshes, surgical meshes, contact lenses, intraocular lenses, stents (e.g., vascular stents, esophageal stents, biliary stents, coronary stents, renal stents, peripheral vascular stents), nasal splints, vascular grafts, stent grafts, aneurysm coils, introducer sheaths, balloon catheters, vascular occlusion devices, inferior vena cava filters, and hydrocephalus shunts.

[0255] In one embodiment, the derivatized polyvalent polymer of the Disclosure may be prepared as a solution and then applied by spray coating or dip coating. The solvent can then be removed to leave a coating of the derivatized polyvalent polymer composition of the Disclosure on the surface of the device. In one embodiment, the solution may be an aqueous solution. In another embodiment, the solution may contain an organic solvent. In another embodiment, the solution may contain water and a water-miscible organic solvent. In one embodiment, the derivatized polyvalent polymer of the Disclosure may be functionalized with aliphatic or aromatic groups such that hydrophobic interactions exist between these groups and the surface of the device. In one embodiment, the derivatized polyvalent polymer of the Disclosure having vinyl sulfone residues may be coated onto a medical device by dip coating or spray coating. The coating is then dried. The coating may be exposed to heat, gamma rays, electron beams or ultraviolet light to crosslink the derivatized polyvalent polymer. In another embodiment, the coating may further contain a bioactive agent. In another embodiment, the coating, upon hydration, increases the lubricity of the coated device. The increase in lubricity of the coated device can be measured by a decrease of at least 20° in the water contact angle. In another embodiment, the increase in lubricity can be measured as a reduction of at least 20% in the coefficient of friction. In another embodiment, the apparatus can be partially coated, leaving some parts of the apparatus uncoated. In another embodiment, the apparatus can be pre-coated with a binding polymer coating that enhances the binding of the coating derivatized polyvalent polymer composition of the present disclosure. In another embodiment, the coating may further contain heparin to impart antithrombotic properties to the coating.

[0256] The process for producing derivative polymers of polyvalent polymers includes the following: a) Reacting the hydroxyl group of a polyvalent polymer with divinyl sulfone (DVS) to obtain a first polyvalent derivative; and b) The first polyvalent polymer derivative and formula X'-R 1 -Y, or X'-R 2-Y, or both, are reacted as nucleophiles to obtain a second polyvalent polymer derivative; where R 1 and R 2 These are different, and each is either substituted or non-substituted C1-C. 20 The polyvalent polymer is an aliphatic or aromatic moiety, where X' contains a nucleophilic group of SH or NH2, Y is the same or different, and Y is one or more of H, a carboxylic acid group or its salt or ester, a hydroxyl group, a sulfonic acid group or its salt, or an amine group. In this process, the polyvalent polymer is hyaluronic acid (HA). This process may further include step c) derivatizing a second polyvalent polymer derivative by repeating step a) or steps a) and b) one or more times.

[0257] The following examples are provided for illustrative purposes only and are not limiting. In these examples, DI represents distilled water, PEG represents polyethylene glycol, and IV represents intrinsic viscosity. [Examples]

[0258] Example 1 DVS modified HA (DVS2) 2.5 g of sodium hyaluronate (900 kDa) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. The solution was then stirred at approximately 200 rpm. 250 g of deionized water was added to the kettle. The solution was stirred for approximately 18 hours. 166.5 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.69. Next, a freshly prepared solution of 66 g of DI water and 10.6 g of divinyl sulfone was rapidly added to the stirred solution. After 75 seconds, 50 g of 1 M HCl solution was added to the reaction mixture. Next, 1 M NaOH was added dropwise until the solution pH was 5-7. Next, 6 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.25 L of acetone was slowly added over 20 minutes. The suspension was stirred for approximately 3 hours. 200 mL of denatured ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel through a 0.22 μm PTFE filter membrane. Once the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 100 mL of ethanol. Next, the ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried under vacuum at room temperature in a vacuum furnace.

[0259] Approximately 10-20 mg of the dried sample was added to the vial. D2O was added to the sample to bring the final concentration of the solution to approximately 6 mg / mL. The sample was shaken on an orbital shaker until dissolved. Once dissolved, the sample was transferred to an NMR tube, and the sample was analyzed. 1 The 1H-NMR spectrum was recorded using an NMR spectrometer. The spectrum was output, and specific peaks were incorporated in the regions of 6.3–6.5 ppm (two peaks originating from two CH2= protons from the vinyl sulfone residue), 6.8–7.0 ppm (the CH peak of the vinyl group), and 1.8–2.5 ppm (a single line originating from three CH3 protons from the N-acetyl group of HA). The modification percentage was calculated using the molar ratio of vinyl CH protons (6.8–7 ppm) to acetamide protons (1.8–2.5 ppm). The substitution percentage was found to be approximately 8.9%. 1The 1H-NMR spectrum is shown in Figure 1.

[0260] Example 2 DVS modified HA (DVS13) 3.5 g of sodium hyaluronate (approximately 800 kDa; 1.4 m³ / Kg IV) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 350 g of deionized water was added to the kettle. The solution was then stirred at approximately 300 rpm. The solution was stirred for approximately 18 hours. The stirring speed was then increased to 750 rpm, and approximately 233 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.95. Next, a freshly prepared solution of 92.4 g of DI water and 15.5 g of divinyl sulfone was rapidly added to the stirred solution. After 4.5 minutes, 63 g of 1 M HCl solution was added to the reaction mixture. Next, 1 M NaOH was added dropwise until the pH of the solution was 5-7. Finally, 8.4 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.5 L of acetone was slowly added over 30 minutes. This suspension was stirred for approximately 3 hours. 300 mL of denatured ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and a 0.22 μm PTFE filter membrane. After the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 150 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried under vacuum at room temperature in a vacuum furnace. The substitution percentage was determined to be approximately 25% by the procedure described in Example 1.

[0261] Example 3 DVS modified HA (DVS14) The reaction described in Example 2 was carried out with a reaction time of 6 minutes. The substitution percentage was determined to be approximately 31% according to the procedure described in Example 1.

[0262] Example 4 Reaction of HA-DVS with 3-mercaptopropionic acid (HA-DVS2-MPA) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone-derived HA (approximately 9%, according to Example 1) was added to 50 g of DI water. This solution was stirred overnight until the materials were dissolved. Next, the flask was purged with nitrogen. 0.022 g of 3-mercaptopropionic acid (MPA) was added to the solution. After the MPA was dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 1.25 g of NaCl was added to the reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 25 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MPA substitution was evident from peaks at 2.3–2.4 ppm (triplet) and 2.6–2.8 ppm (triplet). The MPA substitution was calculated to be 7.6% from the integrals at 2.3–2.4 ppm (MPA-CH2) and 1.7–2 ppm (HA-acetamide).

[0263] Example 5 Reaction of HA-DVS with 1-octanthiol (HA-DVS2-oct) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone-derived HA (approximately 9%, according to Example 1) was added to 50 g of DI water. This solution was stirred at room temperature for about 4 hours. Approximately 15.8 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen. Next, 0.023 g of 1-octanthiol was added to this derivatized HA solution in 7.9 g of ethanol. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. 0.5 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol to prevent the filter funnel from drying out. The precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of octanthiol substitution was evident from peaks at 0.8-0-9 ppm (-CH3), 1.2-1.6 ppm (-CH2-), 2.6-2.7 ppm (-CH2-S-), and 2.9-3.0 ppm (-S-CH2-). The molar substitution of octanthiol was calculated to be 5.4% from the integrals at 2.6-2.7 ppm (Oct-CH2-S-) and 1.7-2 ppm (HA-acetamide).

[0264] Example 6 Reaction of HA-DVS with 1-octanthiol (HA-DVS2-oct-DMF) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone derivatized HA (approximately 9%, according to Example 1) was added to 50 g of DI water. This solution was stirred at room temperature for about 4 hours. Approximately 18.88 g of dimethylformamide (DMF) was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen. Next, 0.029 g of 1-octanthiol was added to this derivatized HA solution in 9.4 g of DMF. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. Approximately 0.25 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 25 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated using vacuum filtration. This precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The molar substitution of octanthiol was calculated to be 5.5% from the integrals at 2.4-2.5 ppm (Oct-CH2-S-) and 1.7-2 ppm (HA-acetamide).

[0265] Example 7 Reaction of HA-DVS with 1-dodecanethiol (HA-DVS2-dod) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone derivatized HA (approximately 9%, according to Example 1) was added to 50 g of DI water. This solution was stirred at room temperature for about 4 hours. Approximately 15.8 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen. Next, 0.04 g of 1-dodecanethiol was added to this derivatized HA solution in 7.9 g of ethanol. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. 0.25 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol to prevent the filter funnel from drying out. The precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of octanthiol substitution was evident from peaks at 0.8-0-9 ppm (CH3-), 1.2-1.6 ppm (-CH2-), 2.6-2.7 ppm (-CH2-S-), and 2.9-3.0 ppm (-S-CH2-). The molar substitution of octanthiol was calculated to be 5.2% from the integrals at 2.6-2.7 ppm (Oct-CH2-S-) and 1.7-2 ppm (HA-acetamide).

[0266] Example 8 DVS-modified HA-reaction 2 (DVS3) 3.5 g of sodium hyaluronate (approximately 900 kDa, 17 dL / g) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 350 g of deionized water was added to the kettle. The solution was stirred at approximately 200 rpm for approximately 18 hours. 233 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.6. Next, a freshly prepared solution of 92.4 g of DI water and 14.8 g of divinyl sulfone was rapidly added to the stirred solution. After 1.25 minutes, 70 g of 1 M HCl was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed until the solution pH was 5-7. Finally, approximately 6 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.25 L of acetone was slowly added over 20 minutes. This suspension was stirred for approximately 3 hours. 200 mL of ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and through a 0.22 μm PTFE filter membrane. The precipitate was rinsed with 100 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried under vacuum at room temperature in a vacuum furnace. The substitution percentage, determined as described in the examples, was found to be 8.6%.

[0267] Example 9 DVS-modified HA-(DVS5-800kDa) 3.5 g of sodium hyaluronate (1.4 m³ / kg, approximately 800 kDa) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 350 g of deionized water was added to the kettle. The solution was stirred at approximately 200 rpm for approximately 18 hours. 233 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.85. Next, a freshly prepared solution of 92.4 g of DI water and 14.8 g of divinyl sulfone was rapidly added to the stirred solution. After 75 seconds, 63 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed until the solution pH was 5-7. Finally, 8.4 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.5 L of acetone was slowly added over 30 minutes. This suspension was stirred for approximately 3 hours. 200 mL of ethanol (ethanol, alcohol reagent, denatured anhydride 94-96%) was added, and this solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and through a 0.22 μm PTFE filter membrane. The precipitate was rinsed with 150 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried under vacuum at room temperature in a vacuum furnace. The substitution percentage, determined as described in Example 1, was found to be approximately 8.9%.

[0268] Example 10 Reaction of HA-DVS with 1-pentanethiol (HA-DVS5-pent2) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone derivatized HA (approximately 9%, according to Example 9) was added to 27.5 g of DI water. This solution was stirred at room temperature for about 4 hours. Approximately 16 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen. Next, 0.042 g of 1-pentanethiol was added to this derivatized HA solution in approximately 1.8 g of ethanol. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. Approximately 0.5 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol to prevent the filter funnel from drying out. The precipitate was dried under vacuum at room temperature. The sample was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of pentanethiol substitution was evident from peaks at 0.6-0-8 ppm (CH3-), 1.2-1.6 ppm (-CH2-), 2.4-2.6 ppm (-CH2-S-), and 2.7-2.9 ppm (-S-CH2-). The molar substitution of pentanethiol was calculated to be 7.3% from the integrals at 2.3-2.7 ppm (pent-CH2-S-) and 1.7-2 ppm (HA-acetamide).

[0269] Example 11 Reaction of HA-DVS with 1-decanethiol (HA-DVS5-dec) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone-derived HA (approximately 9%, according to Example 9) was added to 20 DI water. This solution was stirred at room temperature for about 4 hours. 19.7 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen. Next, 0.035 g of 1-decanethiol was added to this derivatized HA solution in 4 g of ethanol. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. 0.25 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol to prevent the filter funnel from drying out. The precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of decanethiol substitution was evident from peaks at 0.6-0-8 ppm (CH3-), 1.1-1.6 ppm (-CH2-), 2.4-2.6 ppm (-CH2-S-), and 2.7-2.9 ppm (-S-CH2-). The molar substitution of decanethiol was calculated to be 6.5% from the integrals at 2.3-2.7 ppm (pent-CH2-S-) and 1.7-2 ppm (HA-acetamide).

[0270] Example 12 Reaction of HA-DVS with 3-mercapto-1-propanesulfonate (HA-DVS5-SMPS) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone-derived HA (approximately 9%, according to Example 9) was added to 50 g of DI water. This solution was stirred overnight until the materials were dissolved. Next, the flask was purged with nitrogen. 0.036 g of sodium-3-mercapto-1-propanesulfonate (SMPS) was added to the solution. After the SMPS had dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 1.25 g of NaCl was added to the reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 25 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of SMPS substitution was evident from peaks at 2.0–2.1 ppm (-CH2-), 2.5–2.7 ppm (-CH2-S-), and 2.8–3.0 ppm (-S-CH2-). The SPMS substitution was calculated to be 6.4% from the integrals at 2.5–2.7 ppm (SMPS-CH2-S-) and 1.7–2 ppm (HA-acetamide).

[0271] Example 13 Reaction of HA-DVS with cysteine ​​(HA-DVS5-cys) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone-derived HA (approximately 9%, according to Example 9) was added to 50 g of DI water. The solution was stirred overnight until the materials were dissolved. Next, the flask was purged with nitrogen. 0.024 g of L-cysteine ​​was added to the solution. After the cysteine ​​had dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. The solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 1.25 g of NaCl was added to the reaction solution. The solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to the solution. The reaction mixture was stirred for 1.5 hours. 25 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. The precipitate was dried under vacuum at room temperature. Dissolve a sample of this material in D2O, 1 1H-NMR spectra were measured. The presence of cysteine ​​substitution was evident from a peak at 2.8–3.0 ppm (-S-CH2-). The cysteine ​​substitution was calculated to be 4.9% from the integrals at 2.8–3.0 ppm (-CH2-S-) and 1.7–2 ppm (HA-acetamide).

[0272] Example 14 DVS-modified HA-reaction 5 (DVS10-800kDa) 5 g of sodium hyaluronate (1.4 m³ / kg, approximately 800 kDa) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 500 g of deionized water was added to the kettle. The solution was stirred at approximately 200 rpm for approximately 18 hours. 333 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.93. Next, a freshly prepared solution of 66 g of DI water and 11 g of divinyl sulfone was rapidly added to the stirred solution. After 2.5 minutes, 90 g of 1 M HCl solution was added to the reaction mixture. Either 1 M NaOH or 1 M HCl was added dropwise as needed until the pH of the solution was 5-7. Next, approximately 12 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.75 L of acetone was slowly added over 30 minutes. This suspension was stirred for approximately 3 hours. 300 mL of ethanol (ethanol, alcohol reagent, denatured anhydride 94-96%) was added, and this solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and through a 0.22 μm PTFE filter membrane. After the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 200 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated two more times. The product was dried under vacuum at room temperature in a vacuum furnace. The substitution percentage was determined to be 8.1% according to the procedure described in Example 1.

[0273] Example 15 Reaction of HA-DVS with 2-mercaptobenzoic acid (HA-DVS10-MBA) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone-derived HA (approximately 8%, according to Example 14) was added to 27.5 g of DI water. This solution was stirred at room temperature for about 4 hours. 16 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen and then placed in a water bath (temperature = 30 ± 2 °C). Next, 0.092 g of 2-mercaptobenzoic acid (MBA) was added to this derivatized HA solution in 1.78 g of ethanol. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. Approximately 0.25 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated using vacuum filtration. This precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MBA substitution was evident from a peak at 7.1–7.5 ppm (Ar-H). The molar substitution of MBA was calculated to be 10% from the integrals at 7.1–7.5 ppm (Ar-H) and 1.7–2 ppm (HA-acetamide).

[0274] Example 16 Reaction of HA-DVS with 4-methylbenzenethiol (HA-DVS10-MBT) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone derivatized HA (approximately 8%, according to Example 14) was added to 27.5 g of DI water. This solution was stirred at room temperature for about 4 hours. Approximately 15.98 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen and then placed in a water bath (temperature = 30 ± 2 °C). Next, 0.074 g of 4-methylbenzenethiol (MBT) was added to this derivatized HA solution in approximately 1.78 g of ethanol. The pH of the reaction mixture was adjusted to approximately 9.5 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 using 0.25 M HCl. Approximately 0.25 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. The reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. The precipitate was dried at room temperature under vacuum. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MBT substitution was evident from peaks at 2.3–2.5 ppm (Ar-CH3) and 7.2–7.6 ppm (Ar-H). The molar substitution of MBT was calculated to be 5.0% from the integrals at 7.1–7.5 ppm (Ar-H) and 1.7–2 ppm (HA-acetamide).

[0275] Example 17 Reaction of HA-DVS with 4-methoxy-α-toluenethiol (HA-DVS10-MTT) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone-derived HA (approximately 8%, according to Example 14) was added to 27.5 g of DI water. This solution was stirred at room temperature for about 4 hours. 16 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen and then placed in a water bath (temperature = 30 ± 2 °C). To this derivatized HA solution, approximately 0.092 g of 4-methoxy-α-toluenethiol (MTT) was added in 1.78 g of ethanol. The pH of the reaction mixture was adjusted to about 9.5 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. 0.25 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated using vacuum filtration. This precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MTT substitution was evident from peaks at 6.7–7.0 ppm (Ar-H) and 7.1–7.3 ppm (Ar-H). The molar substitution of MTT was calculated to be 10.2% from the integrals at 7.1–7.3 ppm (Ar-H) and 1.7–2 ppm (HA-acetamide).

[0276] Example 18 Reaction of HA-DVS with thiophenol (HA-DVS10-thiophenol) In a 250 mL round-bottom flask, 0.5 g of vinyl sulfone derivatized HA (approximately 8%, according to Example 14) was added to 27.5 g of DI water. This solution was stirred at room temperature for about 4 hours. 16 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, this flask was purged with nitrogen and then placed in a water bath (temperature = 30 ± 2 °C). Next, 0.066 g of thiophenol was added to this derivatized HA solution in 1.78 g of ethanol. The pH of the reaction mixture was adjusted to about 9.5 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. Approximately 0.25 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated using vacuum filtration. This precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of thiophenol substitution was evident from a peak at 7.1–7.5 ppm (Ar-H). The molar substitution of thiophenol was calculated to be 7.6% from the integrals at 7.1–7.5 ppm (Ar-H) and 1.7–2 ppm (HA-acetamide).

[0277] Examples 19A and 19B Effect of pH on the reaction of HA-DVS with 1-pentanethiol (HA-DVS10-pent) In both Examples 19A and 19B, 0.5 g of vinyl sulfone derivatized HA (approximately 7.5%, according to Example 14) was added to 27.5 g of DI water in a 250 mL round-bottom flask. This solution was stirred at room temperature for about 4 hours. 16 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, the flask was purged with nitrogen and then placed in a water bath (temperature = 30 ± 2 °C). To this derivatized HA solution, approximately 0.062 g of pentanethiol was added in approximately 1.78 g of ethanol. In Example 19A, the pH of the reaction mixture was adjusted to approximately 8.5 using 0.25 M NaOH, and in Example 19B, the pH was adjusted to approximately 9.4. In both reactions, the solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 using 0.25 M HCl. 0.25 g of NaCl was added to this reaction solution. This solution was stirred until NaCl dissolved. 150 mL of cold acetone was slowly added to this solution. The reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. This precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried at room temperature under vacuum. A sample of this material was dissolved in D2O, 1 1H-NMR spectra were measured. The presence of pentanethiol substitution was evident from peaks at 0.6-0-8 ppm (CH3-), 1.2-1.6 ppm (-CH2-), 2.4-2.6 ppm (-CH2-S-), and 2.7-2.9 ppm (-S-CH2-). The molar substitution of pentanethiol was calculated from integrals at 2.3-2.7 ppm (pent-CH2-S-) and 1.7-2 ppm (HA-acetamide) to be 3.9% in the reaction at pH 8.5 and 6.7% in the reaction at pH 9.5.

[0278] Example 20 DVS-modified HA-(DVS12-800kDa) 3.5 g of sodium hyaluronate (1.4 m³ / kg, approximately 800 kDa) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 350 g of deionized water was added to the kettle. The solution was stirred at approximately 750 rpm for approximately 18 hours. Approximately 233 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.91. Next, a freshly prepared solution of 92 g of DI water and 15.5 g of divinyl sulfone was rapidly added to the stirred solution. After 3.25 minutes, 63 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed until the pH of the solution was 5-7. Next, approximately 8.4 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.5 L of acetone was slowly added over 30 minutes. This suspension was stirred for approximately 3 hours. 300 mL of ethanol (ethanol, alcohol reagent, denatured anhydride 94-96%) was added, and this solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and a 0.22 μm PTFE filter membrane. After the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 150 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried under vacuum at room temperature in a vacuum furnace. The substitution percentage was determined to be approximately 22.2% according to the procedure described in Example 1.

[0279] Example 21 Reaction of HA-DVS with 3-mercaptopropionic acid (HA-DVS12-MPA) In a 250 mL round-bottom flask, 1.0 g of vinyl sulfone-derived HA (approximately 22%, according to Example 20) was added to 100 g of DI water. This solution was stirred overnight until the materials were dissolved. Next, the flask was purged with nitrogen. 0.106 g of 3-mercaptopropionic acid (MPA) was added to the solution. After the MPA was dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 2.4 g of NaCl was added to the reaction solution. This solution was stirred until the NaCl was dissolved. 300 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 50 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 50 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MPA substitution was evident from peaks at 2.4–2.6 ppm (-CH2-COOH), 2.7–2.8 ppm (-CH2-S-), and 2.9–3.1 ppm (-S-CH2-). The MPA substitution was calculated to be 20.6% from the integrals at 2.4–2.6 ppm (MPA-CH2) and 1.7–2 ppm (HA-acetamide).

[0280] Example 22 DVS-modified HA-(DVS14-800kDa) 3.5 g of sodium hyaluronate (1.4 m³ / kg, approximately 800 kDa) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 350 g of deionized water was added to the kettle. The solution was stirred at approximately 750 rpm for approximately 18 hours. 233 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.87. Next, a freshly prepared solution of 92 g of DI water and 15.5 g of divinyl sulfone was rapidly added to the stirred solution. After 6 minutes, 63 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed until the solution pH was 5-7. Finally, approximately 8.4 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.5 L of acetone was slowly added over 30 minutes. This suspension was stirred for approximately 3 hours. 300 mL of ethanol (ethanol, alcohol reagent, denatured anhydride 94-96%) was added, and this solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and a 0.22 μm PTFE filter membrane. The precipitate was rinsed with 150 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried under vacuum at room temperature in a vacuum furnace. The substitution percentage was determined to be 31.4% by the procedure described in Example 1.

[0281] Example 23 Reaction of HA-DVS with 2-mercaptobenzoic acid (HA-DVS14-MBA) In a 500 mL round-bottom flask, 1.0 g of vinyl sulfone derivatized HA (approximately 31%, according to Example 22) was added to 55 g of DI water. This solution was stirred at room temperature for about 1 hour. 32 g of denatured ethanol was added, and the mixture was stirred for about 18 hours, at which point the material was dissolved. Next, the flask was purged with nitrogen and then placed in a water bath (temperature = 30 ± 2 °C). Next, 0.554 g of 2-mercaptobenzoic acid (MBA) was added to this derivatized HA solution in 3.55 g of ethanol. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. 1.32 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 300 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated using vacuum filtration. This precipitate was washed three times with 50 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MBA substitution was evident from a peak at 7.1–7.5 ppm (Ar-H) [Figure 4]. The molar substitution of MBA was calculated to be 35% from the integrals at 7.1–7.5 ppm (Ar-H) and 1.7–2 ppm (HA-acetamide).

[0282] Example 24 Reaction of HA-DVS with mercaptosuccinate (HA-DVS14-MSA) In a 500 mL round-bottom flask, 1.0 g of vinyl sulfone derivatized HA (approximately 31%, according to Example 22) was added to 100 g of DI water. This solution was stirred for about 18 hours, at which point the material was dissolved. Next, the flask was purged with nitrogen. Next, 0.18 g of mercaptosuccinic acid (MSA) was added to this derivatized HA solution. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to about 7 using 0.25 M HCl. Approximately 2.4 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 300 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 50 mL of ethanol was added, and the mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. This precipitate was washed four times with 50 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MSA substitution was evident from peaks at 2.3–3.1 ppm. The molar substitution of MSA was calculated to be 33% from the integrals at 3.0 ppm and 1.7–2 ppm (HA-acetamide).

[0283] Example 25 Reaction of HA-DVS with 9-mercapto-1-nonanol (HA-DVS14-nonanol) In a 500 mL round-bottom flask, 1.0 g of vinylsulfone-derivatized HA (approximately 31%, according to Example 22) was added to 55 g of DI water. This solution was stirred at room temperature for about 1 hour. 32 g of denatured ethanol was added and the mixture was stirred for about 18 hours, at which point the material had dissolved. Next, the flask was purged with nitrogen and then placed in a water bath (temperature = 30 ± 2 °C). Next, 0.63 g of 9-mercapto-1-nonanol in 3.55 g of ethanol was added to the derivatized HA solution. The pH of the reaction mixture was adjusted to about 9 using 0.25 M NaOH. This solution was stirred for 4 hours and then the pH was adjusted to about 7 using 0.25 M HCl. 1.32 g of NaCl was added to the reaction solution. This solution was stirred until the NaCl had dissolved. 300 mL of cold acetone was slowly added to this solution. The reaction mixture was stirred for 1.5 hours. The precipitate was isolated using vacuum filtration. The precipitate was washed three times with 50 mL of ethanol so that the filter funnel did not dry. The precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O, 1 1H-NMR spectrum was measured. The presence of nonanol substitution was revealed by peaks at 1.1 - 1.8 ppm (-CH2-), 2.5 - 2.8 ppm (-CH2-S-) and 2.9 - 3.1 ppm (-S-CH2-). The molar substitution of nonanol was 37.5% as calculated from the integrals at 2.5 - 2.8 ppm (-CH2-S-) and 1.7 - 2 ppm (HA-acetamide).

[0284] Example 26 Reaction of DVS with 3-mercaptopropionic acid-derivatized HA [HA-DVS12-MPA] (HA-MPA-DVS) 0.75 g of MPA-derivativeized sodium hyaluronate (see Example 21) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 75 g of deionized water was added to the kettle. The solution was stirred at approximately 750 rpm for approximately 18 hours. 50 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured to be 12.85. Next, a freshly prepared solution of 20 g of DI water and 3.3 g of divinyl sulfone was rapidly added to the stirred solution. After 3.25 minutes, 13.5 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed until the pH of the solution was 5-7. Next, approximately 1.8 g of NaCl was added to the solution. Once the NaCl was dissolved, 300 mL of acetone was slowly added over 30 minutes. The suspension was stirred for approximately 3 hours. 50 mL of ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel. Once the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 150 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried in a vacuum furnace at room temperature under vacuum. The vinyl sulfone substitution percentage was determined to be 10.1% according to the procedure described in Example 1.

[0285] Example 27 Reaction of HA-MPA-DVS with 3-mercaptopropionic acid (HA-MPA^2-DVS^2) In a 250 mL round-bottom flask, 0.5 g of MPA / vinyl sulfone derivatized HA (see Example 26) was added to 50 g of DI water. The solution was stirred overnight until the materials were dissolved. Next, the flask was purged with nitrogen. 0.048 g of 3-mercaptopropionic acid (MPA) was added to the solution. After the MPA had dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. The solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 1.2 g of NaCl was added to the reaction solution. The solution was stirred until the NaCl was dissolved. 150 mL of cold acetone was slowly added to the solution. The reaction mixture was stirred for 1.5 hours. 25 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MPA substitution was evident from peaks at 2.4–2.6 ppm (-CH2-COOH), 2.7–2.8 ppm (-CH2-S-), and 2.9–3.1 ppm (-S-CH2-). The MPA substitution was calculated to be 33% from the integrals at 2.4–2.6 ppm (MPA-CH2) and 1.7–2 ppm (HA-acetamide).

[0286] Example 28 Reaction of DVS with thiophenol-derivatized HA [HA-10-thiophenol-DVS^2] 0.323 g of MPA-derivative sodium hyaluronate (see Example 18) was added to a 250 mL round-bottom flask. An overhead stirrer and anchor impeller were installed. 4.62 g of deionized water was added to the kettle. The solution was stirred at approximately 750 rpm for approximately 18 hours. 23.48 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate derivative. The pH of the solution was measured to 12.82. Next, freshly prepared solutions of 4.62 g of DI water and 0.775 g of divinyl sulfone were rapidly added to the stirred solution. After 2.5 minutes, 6.15 g of 1 M HCl solution was added to the reaction mixture. Next, either 0.25 M NaOH or 1 M HCl was added dropwise as needed until the pH of the solution was 5-7. Next, approximately 0.858 g of NaCl was added to the solution. Once the NaCl was dissolved, 125 mL of acetone was slowly added over 5 minutes. This suspension was stirred for approximately 3 hours. 25 mL of ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel. After the solution was filtered, the precipitate was rinsed with 100 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried in a vacuum furnace at room temperature under vacuum. The vinyl sulfone substitution percentage was determined to be 13.6% according to the procedure described in Example 1. A thiophenol peak was observed at 7.2–7.5 ppm.

[0287] Example 29 Crosslinking with PEG-dithiol compounds Approximately 108 mg of HA-10-thiophenol-DVS^2 [see Example 28] was weighed into a 20 mL glass scintillation vial. 7.2 mL of deionized water was added, and the sample was dissolved overnight. Approximately 65.64 mg of PEG

[3400] -(SH)2 [Sigma Aldrich] was added to the glass scintillation vial. 2.6 mL of deionized water was added to this vial, and the mixture was mixed until the sample was dissolved. The PEG

[3400] -(SH)2 solution was added to the HA-10-thiophenol-DVS^2 solution, and the pH of the resulting solution was adjusted to above pH 10 (pH=10.71) using 0.25 M NaOH. The solution became a gel.

[0288] Example 30 Crosslinking with trimethylolpropanetris(3-mercaptopropionate)[TMP-SH] Approximately 109 mg of HA-10-thiophenol-DVS^2 [see Example 28] was weighed into a 20 mL glass scintillation vial. 7.29 mL of deionized water was added, and the sample was dissolved overnight. Approximately 6.1 mg of trimethylolpropanetris (3-mercaptopropionate) [TMP-SH] [Sigma Aldrich] was added to the glass scintillation vial. 213 mL of deionized water was added to this vial and the sample. The TMP-SH mixture was added to the HA-10-thiophenol-DVS^2 solution, and the pH of the resulting solution was adjusted to above pH 11 (pH=11.31) using 0.25 M NaOH. The solution became a gel.

[0289] Example 31 DVS reaction with HA [HA-DVS-16-2] 10 g of sodium hyaluronate (1.4 m³ / kg, approximately 800 kD) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 1000 g of deionized water was added to the kettle. The solution was stirred at approximately 300 rpm for approximately 18 hours. 166.5 g of 1 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be greater than 12.5. Next, a freshly prepared solution of 250 g of DI water and 44.2 g of divinyl sulfone was rapidly added to the stirred solution. After 8 minutes, 170 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed until the solution pH was 5-7. Next, approximately 24 g of NaCl was added to the solution. Once the NaCl was dissolved, 3 L of acetone was slowly added over 40 minutes. The suspension was stirred for approximately 3 hours. 500 mL of ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and a 0.22 μm PTFE filter membrane. Once the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 200 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried under vacuum at room temperature in a vacuum furnace. The substitution percentage was determined to be 72% by the procedure described in Example 1.

[0290] Example 32 Reaction of HA-DVS with 3-mercapto-1-propanesulfonate [HA-DVS-16-2-SMPS] In a 1 L reaction vessel, 200 g of DI water was added to 2.0 g of vinyl sulfone derivatized HA (according to Example 31). The solution was stirred overnight until the materials were dissolved (approximately 300 rpm). Next, the flask was purged with nitrogen. 1.28 g of sodium-3-mercapto-1-propanesulfonate (SMPS) was added to the solution. After the SMPS had dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. The solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 4.8 g of NaCl was added to the reaction solution. The solution was stirred until the NaCl was dissolved. 300 mL of cold acetone was slowly added to the solution. The reaction mixture was stirred for 1.5 hours. 75 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 75 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of SMPS substitution was evident from peaks at 2.0–2.1 ppm (-CH2-), 2.5–2.7 ppm (-CH2-S-), and 2.8–3.0 ppm (-S-CH2-). The SPMS substitution was calculated to be 82% from the integrals at 2.5–2.7 ppm (SMPS-CH2-S-) and 1.7–2 ppm (HA-acetamide).

[0291] Example 33 Divinyl sulfone - single-component crosslinking Approximately 105 mg of HA-DVS-16-2-SMPS [see Example 32] was weighed into a 20 mL glass scintillation vial. Approximately 2.3 mL of deionized water was added to the vial, and the sample was dissolved overnight. 0.26 g of DI water was then added, and the sample was mixed. The pH of the solution was adjusted to approximately pH 13 using 1 M NaOH. Approximately 26 μL of divinyl sulfone was added to the reaction mixture. The sample was mixed by vortexing, and the reaction mixture was left at room temperature until a gel formed. The gel was removed from the vial and placed in approximately 500 mL of deionized water for 1 hour. After an incubation time of 40–70 minutes, the water was changed twice. The excess water was removed, the gel was transferred to a plastic container, and the gel was frozen at -80°C and then freeze-dried to produce a porous foam structure.

[0292] Example 34 Divinyl sulfone - Crosslinking with two components 100 mg of HA-DVS-16-2-SMPS [see Example 32] and 100 mg of HA-DVS14-nonanol (see Example 25) were weighed into a 20 mL glass scintillation vial. 5 mL of deionized water was added to the vial, and the sample was dissolved overnight. The pH of the solution was adjusted to approximately pH 13 using 1 M NaOH. 52 μL of divinyl sulfone was added to the reaction mixture. The sample was mixed by vortexing, and the reaction mixture was left at room temperature until a gel formed. The gel was removed from the vial and placed in 500 mL of deionized water for 1 hour. After an incubation time of approximately 40–70 minutes, the water was changed twice. The excess water was removed, and the gel was transferred to a plastic container. A portion of this gel was frozen at -80°C and then freeze-dried to produce a porous foam structure.

[0293] Example 35 crosslinked particles Cross-linked hydrogel particles (including, as representative examples, Examples 29, 30, 33, and 34) are prepared by passing a cross-linked derivatized polyvalent polymer gel composition through a mesh. The cross-linked gel is transferred to a 20 mL syringe. 5 mL of physiological saline or 10 mg / mL of hyaluronic acid (approximately 800 kDa) is added to the syringe. A plunger is inserted into the syringe, and the gel is extruded through a mesh (a mesh with a pore size of approximately 500 μm held in a 25 mm polycarbonate filter holder). The extrusion process is repeated to produce gel particles.

[0294] Example 36 DVS-modified HA-(DVS18-800kDa) 3.5g sodium hyaluronate (IV=1.4mg) 3A 4 L glass reaction kettle (approximately 800 kD / kg, MW) was added. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 350 g of deionized water was added to the kettle. The solution was stirred at approximately 750 rpm for approximately 18 hours. 233 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.92. Next, a freshly prepared solution of 92 g of DI water and 15.5 g of divinyl sulfone was rapidly added to the stirred solution. After 15 minutes, 63 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed until the solution pH was 5-7. Next, approximately 8.4 g of NaCl was added to the solution. Once the NaCl was dissolved, 1.5 L of acetone was slowly added over 30 minutes. The suspension was stirred for approximately 3 hours. 300 mL of ethanol (ethanol, alcohol reagent, 94-96% denatured anhydride) was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel and a 0.22 μm PTFE filter membrane. Once the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 150 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried in a vacuum furnace at room temperature under vacuum. The substitution percentage was determined to be 71.3% by the procedure described in Example 1.

[0295] Example 37 Reaction of HA-DVS with 3-mercaptopropionic acid (HA-DVS18-MPA) In a 500 mL round-bottom flask, 100 g of DI water was mixed with 1.0 g of vinyl sulfone-derived HA (approximately 71%, according to Example 36). The solution was stirred overnight at 300 rpm until all the materials were dissolved. Next, the flask was purged with nitrogen. 0.25 g of 3-mercaptopropionic acid (MPA) was added to the solution. After the MPA was dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. The solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 2.4 g of NaCl was added to the reaction solution. The solution was stirred until the NaCl was dissolved. 300 mL of cold acetone was slowly added to the solution. The reaction mixture was stirred for 1.5 hours. 50 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 50 mL of ethanol, taking care not to let the filter funnel dry out. The precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O, and the 1H-NMR spectrum was measured. The presence of MPA substitution was evident from peaks at 2.4–2.6 ppm (-CH2-COOH), 2.7–2.8 ppm (-CH2-S-), and 2.9–3.1 ppm (-S-CH2-). The MPA substitution was calculated to be 79.4% from the integrals at 2.4–2.6 ppm (MPA-CH2) and 1.7–2 ppm (HA-acetamide).

[0296] Example 38 Reaction of DVS with 3-mercaptopropionic acid derivatized HA [HA-18-MPA-DVS] 0.75 g of MPA-derivativeized sodium hyaluronate (see Example 37) was added to a 4 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 75 g of deionized water was added to the kettle. The solution was stirred at approximately 200 rpm for approximately 18 hours. 50 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured to 12.70. Next, a freshly prepared solution of 20 g of DI water and 3.3 g of divinyl sulfone was rapidly added to the stirred solution. After 15 minutes, 13.5 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed to bring the solution pH to 5-7. Next, approximately 1.8 g of NaCl was added to the solution. Once the NaCl was dissolved, 300 mL of acetone was slowly added over 30 minutes. The suspension was stirred for approximately 3 hours. 50 mL of ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel. Once the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 150 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried in a vacuum furnace at room temperature under vacuum. The vinyl sulfone substitution percentage was determined to be 66% according to the procedure described in Example 1.

[0297] Example 39 Reaction of HA-MPA-DVS with 3-mercaptopropionic acid (HA-MPA^2-DVS^2) In a 250 mL round-bottom flask, 0.5 g of MPA / vinyl sulfone derivatized HA (see Example 38) was added to 50 g of DI water. The solution was stirred overnight, during which time the materials dissolved. Next, the flask was purged with nitrogen. 0.175 g of 3-mercaptopropionic acid (MPA) was added to the solution. After the MPA dissolved, the pH was adjusted to approximately 9 with 0.25 M NaOH. The solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 with 0.25 M HCl. 1.2 g of NaCl was added to the reaction solution. The solution was stirred until the NaCl dissolved. 150 mL of cold acetone was slowly added to the solution. The reaction mixture was stirred for 1.5 hours. 25 mL of ethanol was added, and the resulting mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. The precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. A sample of this material was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MPA substitution was evident from peaks at 2.4–2.6 ppm (-CH2-COOH), 2.7–2.8 ppm (-CH2-S-), and 2.9–3.1 ppm (-S-CH2-). The MPA substitution was calculated to be 133% from the integrals at 2.4–2.6 ppm (MPA-CH2) and 1.7–2 ppm (HA-acetamide).

[0298] Example 40 HA-based formulations Various formulations are prepared as shown in Table 1. The numbers listed in Table 1 represent weight percentages of the total weight of the formulation. The following HA derivatives can be used as the "HA derivatives" listed in Table 1: (a) thiophenol derivatives (representative example 28), (b) 2-mercaptobenzoic acid derivatives (representative example 23), (c) mercaptosuccinic acid derivatives (representative example 24), (d) sodium 3-mercapto-1-propanesulfonate derivatives (representative example 32), and (e) decanethiol derivatives (representative example 11). A portion of the gel sample is cast onto a glass sheet or release liner piece, dried at room temperature, and then vacuum-dried for at least 12 hours. This creates a film of the formulation. A second portion of the gel sample is placed in a scintillation vial, frozen at -80°C, and then freeze-dried. The formulations listed in Table 1 may also contain 1% (w / w) collagen or gelatin as well as other excipients and buffers. Water or injected water can be used as a substitute for saline solution.

[0299] [Table 1] HPMC = Hydroxypropyl Methylcellulose CMC = Carboxymethylcellulose sodium Poly-AA = Polyacrylic acid

[0300] Example 41 HA derivative formulation Various formulations of different HA derivatives are prepared as shown in Table 2. The values ​​listed below each component are weight percentages of the total weight of the formulation. A portion of the sample is cast onto a glass sheet or release liner using a Gardner knife, dried at room temperature, and then vacuum-dried for at least 12 hours. This creates a film of the formulation. A second portion of the gel sample is placed in a scintillation vial, frozen at -80°C, and then freeze-dried to produce a porous solid matrix. A third portion is maintained in solution / gel form. The formulations listed in Table 2 may also include 1% (w / w) collagen or gelatin, as well as other excipients and buffers. Water or injection water can be used instead of saline.

[0301] [Table 2]

[0302] Example 42 Incorporation of bioactive agents The bioactive agents listed in Table 3 are directly incorporated into each of the formulations prepared in Examples 40 and 41. These formulations are prepared in gel, film, or lyophilized form. Each formulation contains the active ingredients listed in Table 3 in the amounts indicated by weight / weight (w / w), units / gram (U / g), or μg / mL.

[0303] [Table 3]

[0304] Example 43 Incorporation of antimicrobial agents Clindamycin phosphate (1% w / w) or metronidazole (1.3% w / w) is directly incorporated into the formulations prepared in Examples 40 and 41. In another set of formulations, a combination of clindamycin phosphate (0.5% w / w) or metronidazole (0.5% w / w) is directly incorporated into the formulations prepared in Examples 40 and 41. In another set of formulations, such as those prepared in Examples 40 and 41, containing either clindamycin phosphate (1% w / w) or metronidazole (1.3% w / w), saline is replaced with citrate / saline buffer to ensure that the pH is maintained in the pH range of 4.5–6. These formulations are prepared in either gel, film, or lyophilized form.

[0305] Example 44 Rehydration with biological activators The lyophilized product can be prepared from the formulations described in Examples 29, 30, 33, 34, 40, and 41. The lyophilized derivatized polyvalent polymer composition is rehydrated with either BMP-7 (5 μg / mL) or Botox (5 U / mL). The resulting gel is used as a gel and applied to the target tissue by topical application or injection.

[0306] Example 45 Cell integration into gel Weigh approximately 100 mg of derivatized hyaluronic acid containing vinyl sulfone residues (a representative example as described in Example 28) into a 20 mL glass scintillation vial. Add 7 mL of physiological saline and dissolve the sample overnight. Add approximately 64 mg of PEG

[3400] -(SH)2 (Sigma Aldrich, St. Louis, MO, US) to the glass scintillation vial. Add 1.3 mL of physiological saline to this vial and mix until the sample is dissolved. Add this PEG

[3400] -(SH)2 solution to the derivatized HA sample solution and adjust the pH of the resulting solution to pH 8.5 using 0.25 M NaOH. Aliquots of freshly trypsin-treated hMSC cell suspensions are prepared at final cell concentrations of 1, 5, 10, and 20 × 10⁻⁶. 6Mix with HA / PEG solution to a concentration of cells / mL. Dispense the samples into 12-well plates. Gelate these samples in a 37°C incubator for 20 minutes. Then, add 2 mL of fresh medium to each well.

[0307] Example 46 Incorporation of gel into scaffolding Approximately 100 mg of derivatized hyaluronic acid containing vinyl sulfone residues (a representative example as described in Example 28) is weighed into a 20 mL glass scintillation vial. 7 mL of physiological saline is added, and the sample is dissolved overnight. Approximately 64 mg of PEG

[3400] -(SH)2 [Sigma Aldrich] is added to the glass scintillation vial. 1.3 mL of physiological saline is added to the vial, and the mixture is mixed until the sample is dissolved. This PEG

[3400] -(SH)2 solution is added to the derivatized HA sample solution, and the pH of the resulting solution is adjusted to pH 8.5 using 0.25 M NaOH. The resulting solution is dispensed into a porous scaffold and allowed to permeate the scaffold. Once the scaffold is saturated, the scaffold is placed in an incubator (37°C) until the crosslinking reaction is complete. The porous scaffolds used for integration into the gel are electrospun polydioxanone fabric and 3D printed polylactide scaffolds with pores of approximately 200-500 μm.

[0308] Example 47 Incorporation of gel / cell matrix into scaffolds The gel / cell matrix, as prepared in Example 45, is dispensed onto a porous scaffold before gelation and allowed to permeate the scaffold. Once the scaffold is saturated, it is placed in an incubator (37°C) until the crosslinking reaction is complete. The porous scaffolds used for incorporation into the gel are electrospun polydioxanone cloth and 3D printed polylactide-co-glycolide scaffolds with pore sizes of approximately 200-500 μm. The porous scaffold / hydrogel / cell complex is placed in the wells of a 12-well culture plate containing fresh medium.

[0309] Example 48 HA-derivative electrospinning A 15 mg / mL hyaluronic acid derivative solution is prepared by adding approximately 90 mg of hyaluronic acid derivative to 3 mL of deionized water. The sample is dissolved overnight. 3 mL of dimethylformamide (DMF) is added to the sample, and the sample is vortexed several times for 30 minutes. This solution is transferred to a 5 mL syringe equipped with a 0.3 mm inner diameter needle tip. The syringe pump is set to 60 mL / min. The applied voltage is set to 22 kV, and the distance from the tip to the collector is 15 cm. Heavy pieces of aluminum foil are connected to the ground wire, and these aluminum foils are immersed in a shallow bath containing ethanol. The resulting electrospun derivatized polyvalent polymer composition is carefully removed, and the sample is placed in a vacuum furnace to remove any remaining solvent.

[0310] Example 49 Formation of molded hydrogels Approximately 100 mg of derivatized hyaluronic acid containing vinyl sulfone residues (a typical example as described in Example 28) is weighed into a 20 mL glass scintillation vial. 7 mL of physiological saline is added, and the sample is dissolved overnight. Approximately 64 mg of PEG

[3400] -(SH)2 [Sigma Aldrich] is added to the glass scintillation vial. 1.3 mL of physiological saline is added to the vial, and the mixture is mixed until the sample is dissolved. The PEG

[3400] -(SH)2 solution is added to the derivatized HA sample solution, and the pH of the resulting solution is adjusted to pH 8 using 0.25 M NaOH. 30 mg of the drug (e.g., dexamethasone, triamcinolone acetonide, budesonide, flunisolide, cyprofluxin) is mixed into the solution. The solution is drawn into a syringe equipped with a known diameter cylastic tube attached to a needle. When the tube is almost full, fold the open end and secure it with a clamp. Place the tube in an incubator (37°C) overnight to complete the crosslinking reaction. Next, remove the clamp and dry the gel inside the tube in a vacuum oven. Remove the dried crosslinked derivatized polyvalent polymer composition by carefully slicing the tube. Next, cut the dried crosslinked derivatized polyvalent polymer composition to the desired length (e.g., 2-4 mm and 1-2 cm). 60% (v / v) water and 40% (ethanol) can be used instead of 100% water to prepare the solution. A drug-free dried crosslinked derivatized polyvalent polymer composition is also prepared.

[0311] Example 50 Synthesis of divinyl sulfone derivatized HA with different molecular weights The synthesis of divinyl sulfone-derived HA using different starting HA molecular weights was carried out using the same method as described in Example 1. The specific molecular weights, reaction conditions, and vinyl sulfone substitutions obtained are shown in Table 4.

[0312] [Table 4]

[0313] Example 51 DVS interaction with HA-MSA (HA-DVS12-MSA-DVS) 0.75 g of MPA-derivativeized sodium hyaluronate (see Example 24) was added to a 250 ml round-bottom flask. An overhead stirrer and anchor impeller were attached to the reaction flask. 75 g of deionized water was added to the kettle. The solution was stirred at approximately 750 rpm for approximately 18 hours. 50 g of 0.25 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured to approximately 12.8. Next, freshly prepared solutions of 20 g of DI water and 3.3 g of divinyl sulfone were rapidly added to the stirred solution. After 3.25 minutes, 13.5 g of 1 M HCl solution was added to the reaction mixture. Next, either 1 M NaOH or 1 M HCl was added dropwise as needed to bring the solution pH to 5-7. Next, approximately 1.8 g of NaCl was added to the solution. Once the NaCl was dissolved, 300 mL of acetone was slowly added over 30 minutes. The suspension was stirred for approximately 3 hours. 50 mL of ethanol was added, and the solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel. Once the solution was filtered, the vacuum device was removed, and the precipitate was rinsed with 50 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times. The product was dried in a vacuum furnace at room temperature under vacuum. The vinyl sulfone substitution percentage was determined to be 47.8% according to the procedure described in Example 1.

[0314] Example 52 MPA's response to HA-MSA-DVS (HA-DVS12-MSA-MPA) In a 250 mL round-bottom flask, 0.5 g of HA-MSA-DVS (Example 51) was added to 50 g of DI water. This solution was stirred for approximately 18 hours, at which point the derivatized polyvalent polymer was dissolved. Next, the flask was purged with nitrogen. Then, 0.191 g of 3-mercaptopropionic acid (MPA) was added to this derivatized HA solution. The pH of the reaction mixture was adjusted to approximately 9 using 0.25 M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 using 0.25 M HCl. Approximately 1.8 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 200 mL of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 50 mL of ethanol was added, and the mixture was stirred for 15 minutes. The precipitate was isolated by vacuum filtration. This precipitate was washed four times with 25 mL of ethanol, taking care not to let the filter funnel dry out. This precipitate was dried under vacuum at room temperature. The derivatized polyvalent polymer sample was dissolved in D2O. 1 1H-NMR spectra were measured. The presence of MPA and MSA substitutions was evident from peaks in the 2.1–3.2 ppm range.

[0315] Example 53 DVS modified HA-HA-DVS-37 11.33 g of sodium hyaluronate (1.4 m3 / kg, approximately 800 kDa) was added to a 5 L glass reaction kettle. A lid, an overhead stirrer, and an anchor impeller were attached to the reaction kettle. 1133 g of deionized water was added to this kettle. The temperature controller of the bioreactor heater (Chemglass CLS-1380-19V) was set to 25 °C. This solution was stirred at approximately 300 rpm for approximately 18 hours. The stirring speed was increased to 750 rpm. Next, 35 g of 1 M NaOH solution was added to the dissolved sodium hyaluronate. When the pH of the solution was measured after 2 minutes, it was found to be 12.5. The pH was adjusted to 12.32 using 1 M HCl solution. Next, a solution of 50 g of divinyl sulfone in 282.5 g of freshly prepared DI water was quickly added to this stirred solution. The pH was monitored and adjusted with 1 M NaOH to maintain the pH range between 12.2 and 12.3 during the 10-minute reaction time. After 10 minutes, 35 g of 1 M HCl solution was added to this reaction mixture to adjust the pH of the reaction mixture to a value between 5 and 7. Next, approximately 19.5 g of NaCl was added to this solution. When the NaCl had dissolved, 2 L of acetone was slowly added in less than 30 minutes. This suspension was stirred for approximately 3 hours. 400 mL of ethanol was added and this solution was stirred for approximately 30 minutes. The precipitate was removed under vacuum using a sintered glass funnel. When the solution had been filtered, the vacuum apparatus was removed and the precipitate was rinsed with 200 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times with each aliquot being allowed to stand in static ethanol for 5 minutes before the application of vacuum. This product was used in the following reaction. The percent substitution was found to be 31% by the following NMR method. Approximately 10 - 20 mg of the dry sample was added to a vial. D2O was added to this sample such that the final concentration of the solution was approximately 6 mg / mL. The sample was shaken on an orbital shaker until dissolved. When dissolved, the sample was transferred to an NMR tube and the 1The 1H-NMR spectrum was recorded using an NMR spectrometer. The resulting spectrum included specific peaks in the 6.0–6.4 ppm (two peaks originating from two CH2= protons from the vinyl sulfone residue), 6.6–7.0 ppm (the CH peak of the vinyl group), and 1.7–2.0 ppm (a single line originating from three CH3 protons from the N-acetyl group of HA). The modification percentage was calculated using the molar ratio of vinyl CH protons (6.8–7 ppm) to acetamide protons (1.7–2.0 ppm).

[0316] Example 54 Reaction of HA-DVS with mercaptosuccinic acid (MSA) (HA-DVS-37-MSA) In a 5L reaction kettle, vinyl sulfone-derived HA obtained from Example 53 was added to 1200g of DI water. The temperature controller of the Bioreactor heater (Chemglass CLS-1380-19V) was set to 25°C. This solution was stirred at 300 rpm for approximately 18 hours, at which point the material was dissolved. The stirring speed was then increased to 500 rpm. Next, 2.696g of mercaptosuccinic acid (MSA) was added to this derivatized HA solution and stirred for 10 minutes. The pH of the reaction mixture was adjusted to approximately 9 using 1M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 using 1M HCl. Approximately 20.64g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 2L of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 400mL of ethanol was added and the mixture was stirred for 15 minutes. The precipitate was isolated using vacuum filtration. After the solution was filtered, the vacuum apparatus was removed and the precipitate was rinsed with 200 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times, with each aliquot allowed to stand in still ethanol for 5 minutes before applying vacuum. The product was dried at room temperature under vacuum. A sample of this material was dissolved in D2O, 11H-NMR spectra were measured. The presence of MSA substitution was evident from peaks at 2.3–3.1 ppm. The molar substitution of MSA was calculated to be 20.6% from integrals at 3.0 ppm and 1.7–2 ppm (HA-acetamide). The rheological results for the 2% (w / v) solution were as follows for fluid viscosity (0.1–1000 1 / s) and frequency test (1–10 Hz). See Tables 5 and 6, and Figure 5.

[0317] [Table 5] [Table 6]

[0318] Example 55 Reaction of HA-DVS with thiophenol (HA-DVS-37-THIO) In a 5 L reaction kettle, 660 g of DI water was added to the vinyl sulfone derivatized HA reaction product (prepared in the same manner as in Example 53). This solution was stirred at 300 rpm at 30°C for approximately 1 hour. Next, 426.06 g of ethanol was added, and this solution was stirred at 300 rpm at 30°C for approximately 18 hours. After that, the stirring speed was increased to 500 rpm. Next, 5.935 g of thiophenol was added to this derivatized HA solution, and it was stirred for 10 minutes. The pH of the reaction mixture was monitored and adjusted to approximately 9 using 1 M NaOH. This solution was stirred for 2 hours, and then the pH was adjusted to approximately 7 using 1 M HCl. Approximately 9 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl dissolved. 1 L of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. After the solution was filtered, the vacuum apparatus was removed and the precipitate was rinsed with 200 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times, with each aliquot allowed to stand in still ethanol for 5 minutes before applying vacuum. The product was dried at room temperature under vacuum. A sample of this material was dissolved in D2O, 1¹H-NMR spectra were measured. A thiophenol peak was observed at 7.2–7.5 ppm. The molar substitution of thiophenol was calculated to be 32.8%. The rheological results for the 2% (w / v) solution were as follows for fluid viscosity (0.1–1000 ¹ / s) and frequency test (1–10 Hz). See Tables 7 and 8 and Figure 6.

[0319] [Table 7] [Table 8]

[0320] Example 56 Reaction of HA-DVS with mercaptobenzoic acid (MBA) (HA-DVS-37-MBA) In a 5 L reaction kettle, 660 g of DI water was added to the vinyl sulfone derivatization HA reaction (prepared in the same manner as in Example 53). This solution was stirred at 300 rpm at 30°C for approximately 1 hour. Next, 426.06 g of ethanol was added, and this solution was stirred at 300 rpm at 30°C for approximately 18 hours. After that, the stirring speed was increased to 500 rpm. Next, 8.305 g of MBA was added to this derivatized HA solution and stirred for 10 minutes. The pH of the reaction mixture was monitored and adjusted to approximately 9 using 1 M NaOH. This solution was stirred for 2 hours, and then the pH was adjusted to approximately 7 using 1 M HCl. Approximately 9 g of NaCl was added to this reaction solution. This solution was stirred until the NaCl was dissolved. 1 L of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. The precipitate was isolated by vacuum filtration. Once the solution was filtered, the vacuum apparatus was removed, and the precipitate was rinsed with 200 mL of ethanol. Ethanol was removed by vacuum filtration. This process was repeated three more times, with each aliquot allowed to stand in still ethanol for 5 minutes before applying vacuum. The product was dried at room temperature under vacuum. A sample of this material was dissolved in D2O, 1¹H-NMR spectra were measured. The presence of MBA substitution was evident from the peak at 7.1–7.5 ppm (Ar-H). The molar substitution of MBA was calculated to be 23.3% from the integrals at 7.1–7.5 ppm (Ar-H) and 1.7–2 ppm (HA-acetamide). The rheological results for the 2% (w / v) solution were as follows for fluid viscosity (0.1–1000 ¹ / s) and frequency test (1–10 Hz). See Tables 9 and 10, and Figure 7.

[0321] [Table 9] [Table 10]

[0322] Example 57 Reaction of HA-DVS with 3-mercapto-1-propanesulfonate (HA-DVS-37-SMPS) In a 5L reaction kettle, 500g of DI water was mixed with 5g of vinyl sulfone derivatized HA (prepared in the same manner as in Example 53). The temperature controller of the Bioreactor heater (Chemglass CLS-1380-19V) was set to 25°C. This solution was stirred at 300 rpm for approximately 18 hours, at which point the material was dissolved. The stirring speed was then increased to 500 rpm. Next, 1.244g of 3-mercapto-1-propanesulfonate was added to this derivatized HA solution and stirred for 10 minutes. The pH of this reaction mixture was adjusted to approximately 9 using 1M NaOH. This solution was stirred for 4 hours, and then the pH was adjusted to approximately 7 using 1M HCl. Approximately 8.6g of NaCl was added to this reaction solution. This solution was stirred until the NaCl dissolved. 750ml of cold acetone was slowly added to this solution. This reaction mixture was stirred for 1.5 hours. 150mL of ethanol was added, and the mixture was stirred for 15 minutes. The precipitate was isolated using vacuum filtration. After the solution was filtered, the vacuum apparatus was removed and the precipitate was rinsed with 100 mL of ethanol. The ethanol was removed by vacuum filtration. This process was repeated three more times, with each aliquot allowed to stand in still ethanol for 5 minutes before applying vacuum. The product was dried at room temperature under vacuum. A sample of this material was dissolved in D2O, 1 1H-NMR spectra were measured. The presence of SMPS substitution was evident from peaks at 2.0–2.1 ppm (-CH2-), 2.5–2.7 ppm (-CH2-S-), and 2.8–3.0 ppm (-S-CH2-). The SMPS substitution was calculated to be 26.18% from the integrals at 2.5–2.7 ppm (SMPS-CH2-S-) and 1.7–2 ppm (HA-acetamide).

[0323] Example 58 DVS modified HA-HA-DVS-36 11.33 g of sodium hyaluronate (1.4 m³ / kg, approximately 800 kDa) was added to a 5 L glass reaction kettle. The lid, overhead stirrer, and anchor impeller were attached to the reaction kettle. 1133 g of deionized water was added to the kettle. The temperature controller of the Bioreactor heater (Chemglass CLS-1380-19V) was set to 25°C. The solution was stirred at approximately 300 rpm for approximately 18 hours. The stirring speed was increased to 750 rpm. Next, 35 g of 1 M NaOH solution was added to the dissolved sodium hyaluronate. The pH of the solution was measured after 2 minutes and found to be 12.5. The pH was adjusted to 12.32 using 1 M HCl solution. Ne...

Claims

1. A hyaluronic acid polymer derivative comprising one or more modified hydroxyl groups, wherein the hyaluronic acid polymer derivative has the formula: H-(OCH 2 CH 2 SO 2 CH 2 CH 2 -X-R 1 -Y)n During the ceremony, HA stands for hyaluronic acid. X is S or NH, R 1 This refers to a straight or branched C1-C20 hydrocarbon chain consisting only of carbon atoms and hydrogen atoms, without any unsaturation. Y is H, and n is the number of modified hydroxyl groups, where n ≥ 1. It has, A hyaluronic acid polymer derivative wherein 0.25 to 50% of the hydroxyl groups of hyaluronic acid constituting the hyaluronic acid polymer derivative are modified hydroxyl groups.

2. A crosslinked polymer synthesized from one or more of the hyaluronic acid polymer derivatives described in Claim 1, A crosslinked polymer in which the hydroxyl group, vinyl group, or ionic group of the hyaluronic acid polymer derivative was crosslinked at the site of the hyaluronic acid polymer derivative.

3. A crosslinked polymer synthesized from one or more of the hyaluronic acid polymer derivatives described in Claim 1, A crosslinked polymer, including photocrosslinking.

4. A process for producing a hyaluronic acid polymer derivative according to Claim 1, a) To provide a first HA derivative by reacting the hydroxyl group of a hyaluronic acid (HA) polymer with divinyl sulfone (DVS); and b) To provide a second HA derivative by reacting a first HA derivative with a nucleophile of formula X'-R1-Y; where R1 is a straight or branched C1-C20 hydrocarbon chain consisting only of carbon and hydrogen atoms and not containing unsaturated atoms, X' is a nucleophile of SH or NH2, and Y is H. A process that includes this.

5. The process according to claim 4, further comprising step c) derivatizing a second HA derivative polymer by repeating step a) or step a) and step b) one or more times.

6. The process according to claim 4, wherein the second HA derivative is HA-(OCH2CH2SO2CH2CH2-X-R1-Y)n(HA-DVS-N), where HA is hyaluronic acid, X is S or NH, R1 is a straight or branched C1-C20 hydrocarbon chain consisting only of carbon and hydrogen atoms and not containing unsaturated atoms; Y is H, and n≧1.

7. The process according to claim 4, wherein 0.25 to 50% of the hydroxyl groups present on the HA polymer are derivatized to -OCH2CH2SO2CH2CH2-X-R1-Y groups, where X is S or NH; R1 is a straight or branched C1-C20 hydrocarbon chain consisting only of carbon and hydrogen atoms and not containing unsaturated atoms; and Y is H.

8. The process according to claim 4, wherein 0.25 to 50% of the hydroxyl groups present on the HA polymer are converted to oxyethylethenylsulfone groups of the formula -OCH₂CH₂-SO₂CH=CH₂.

9. The process according to claim 4, wherein 0.25 to 50% of the hydroxyl groups present on the HA polymer are converted to oxyethylethenylsulfone groups of the formulas -OCH₂CH₂-SO₂CH=CH₂ and -OCH₂CH₂SO₂CH₂CH₂-X-R₁-Y, where R₁ is a straight or branched C₁-C₂O hydrocarbon chain consisting only of carbon and hydrogen atoms and not unsaturated, X is S or NH, and Y is H.

10. The process according to claim 4, wherein the first HA derivative is an oxyethyl ethenyl sulfone derivative of hyaluronic acid having the formula: HA-(OCH₂CH₂SO₂CH=CH₂)n(HA-DVS), n≧1.

11. A process for producing a crosslinked polymer, comprising reacting a hyaluronic acid polymer derivative produced by the process described in any one of claims 4 to 10 with a crosslinking agent to provide a crosslinked polymer.

12. A composition comprising the hyaluronic acid polymer derivative described in Claim 1, or the crosslinked polymer described in Claim 2 or 3.

13. The composition according to claim 12, further comprising a pharmaceutically acceptable excipient, a synthetic polymer, a thermoreversible polymer, a biodegradable polymer, a buffer, a complexing agent, a tonicity modifier, an ionic strength modifier, a solvent, an antioxidant, a preservative, a viscosity modifier, a pH modifier, a surfactant, an emulsifier, a phospholipid, a stabilizer, or a pologen.

14. The composition according to claim 12 or 13, further comprising a bioactive agent.

15. The composition according to any one of claims 12 to 14 for use as a wound healing matrix, nasal stent, adhesion prevention barrier, hemostatic agent, filler, skin filler, intra-articular grafting agent, tissue sealant, eye drop, punctal plug, antimicrobial composition, biopsy plug, female contraceptive plug, tissue scaffold, burr hole plug, nerve guide, vaginal lubricant, or coating agent for a device.

16. The composition according to any one of claims 12 to 14 for use in treating wounds in an object, filling cavities in an object, relieving joint pain in an object, preventing postoperative adhesions in an object, sealing tissue in an object, treating bacterial vaginosis in an object, treating ocular conditions in an object, treating mucositis in an object, treating ocular conditions in an object, or assisting tissue growth in an object.

17. A medical device comprising a hyaluronic acid polymer derivative according to claim 1, a crosslinked polymer according to claim 2 or 3, and / or a composition according to any one of claims 12 to 14.

18. An additive manufacturing method comprising manufacturing an article using an additive manufacturing machine, comprising a hyaluronic acid polymer derivative according to claim 1, a crosslinked polymer according to claim 2 or 3, a hyaluronic acid polymer derivative manufactured by a process according to any one of claims 4 to 10, or a crosslinked polymer manufactured by a process according to claim 11.

19. A method for producing an electrospun material or electrospun product, comprising producing an electrospun material or electrospun product comprising a hyaluronic acid polymer derivative according to claim 1, or a crosslinked polymer according to claim 2 or 3, or a hyaluronic acid polymer derivative produced by a process according to any one of claims 4 to 10, or a crosslinked polymer produced by a process according to claim 11, using an electrospun apparatus.

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