Triblock copolymers and hydrogels thereof

Abstract

Disclosed herein are block copolymers that can self-assemble into particles, which can be used to crosslink a hydrophilic polymer. An example block copolymer includes a first block that includes pendant groups that can modulate reactive oxygen species responsiveness and drug interactions, a second block that is hydrophilic, and a third block that is hydrophilic and includes a grafted guest-host moiety. Also disclosed are compositions including the block copolymers, and methods of treating diseases.

Description

Attorney Docket No.093386-0041-WO01 TRIBLOCK COPOLYMERS AND HYDROGELS THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No.63 / 610,008 filed on December 14, 2023, which is incorporated fully herein by reference. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with Government support under Federal Grant No. EB028690 awarded by the National Institutes of Health. The Federal Government has certain rights in the invention. TECHNICAL FIELD

[0003] The present disclosure relates to block copolymers and their use in hydrogels for biomedical applications, such as drug delivery. INTRODUCTION

[0004] Chronic wounds are a significant source of mortality and morbidity and are expected to become an even larger public health concern, with an expanding diabetic population which is responsible for a significant portion of healthcare expenditures associated with treating chronic wounds. Over one-third of the global diabetic population will develop a chronic skin wound within their lifetime, resulting in $100 billion per year associated with repeated palliative treatments and wound management. However, up to 15% of these patients will still require transtibial amputation and up to 70% of amputations will result in mortality within five years. Thus, improved therapeutic strategies to heal chronic skin wounds would be beneficial for these patients. SUMMARY

[0005] In one aspect, disclosed are block copolymers comprising: a first block (A), the first block comprising recurring units of formula (I)Attorney Docket No.093386-0041-WO01 wherein: X1, at each occurrence, is methyl, of formula (II)(II), or of formula (III)L1and L2are each independently -C(O)O-alkylene, -C(O)NRb-alkylene, alkylene, alkylene-O-alkylene, alkylene-O-, or bond; R1is a -interacting moiety; R2is a hydrogen donating moiety; Rais hydrogen or C1-3alkyl; and Rbis hydrogen or C1-6alkyl; and a second block (B), the second block comprising a first hydrophilic polymer; and a third block (C), the third block comprising a multi-unit segment of formula (IV)wherein: Z is a recurring hydrophilic unit; G is NH or O; L3is alkylene, heteroalkylene, alkenylene, alkynylene, or bond; R3is a guest-host moiety; Rcis hydrogen or C1-3alkyl; and y is no more than 30 mol% of formula (IV).Attorney Docket No.093386-0041-WO01

[0006] In another aspect, disclosed are networks of a second hydrophilic polymer crosslinked with a plurality of particles, each particle comprising a plurality of self-assembled block copolymers as disclosed herein, wherein the second hydrophilic polymer comprises a guest-host moiety that is capable of forming a guest-host complex with the guest-host moiety of the block copolymer.

[0007] In another aspect, disclosed are methods of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition as disclosed herein.

[0008] In another aspect, disclosed are methods of tissue repair in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition as disclosed herein, wherein the tissue repair comprises treating a volumetric tissue defect in the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] This patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0010] FIG.1. Schematic of integration of synthetic nanoparticles with a natural biopolymer through physical crosslinking mediated by guest-host inclusion complexation results in a hybrid shear-thinning hydrogel system which can serve as a bioresorbable and injectable therapeutic delivery platform.

[0011] FIG.2A. Formation of hybrid NP / HA hydrogels composed of candidate PPS-based triblock copolymers of varying composition and HA-CD20% is facilitated by guest-host pairing mediated aggregation.

[0012] FIG.2B. Representative cryo-TEM imaging of example NP / HA hydrogels.

[0013] FIG.2C. Representative cryo-SEM imaging of example NP / HA hydrogels.

[0014] FIG.2D. Stepwise strain-based time sweep of example NP / HA hydrogels.

[0015] FIG.2E. Composition of HA (from example NP / HA hydrogels) as observed by representative Masson's trichrome stained explanted hydrogel sections at day 7 post-injection.

[0016] FIG.3. Schematic synthesis of hyaluronic acid grafted with -cyclodextrin (HA-CDX) is accomplished through a BOP-mediated amidation reaction between HA-TBA salt and CD-NH2.

[0017] FIG.4. Schematic synthesis of hyaluronic acid grafted with adamantane (HA-ADx) is performed through an esterification reaction between adamantane acetic acid and HA-TBA salt.Attorney Docket No.093386-0041-WO01

[0018] FIG.5A. Schematic of P(PSncoRm) D300-AD20%Structure.

[0019] FIG.5B. Relative ROS-sensitivities of example NP / HA hydrogels. Formation of hybrid NP / HA hydrogels with varying ROS-sensitivities composed of triblock copolymers of varying polysulfide chemistry in core block and hyaluronic acid grafted with 20% -cyclodextrin (HA- CD20%).

[0020] FIG.6 Schematic synthesis of benzyl and (protected) alcohol thiirane polysulfide monomers from epoxides is accomplished using thiourea as sulfur donor.

[0021] FIG.7A. Schematic synthesis of example triblock copolymers comprised ofPSncoPSOHmblock copolymers in core block architecture is accomplished by a combination ofanionic ring opening and RAFT polymerizations.

[0022] FIG.7B. Schematic synthesis of example triblock copolymers comprised of PSncoPSBnmblock copolymers in core block architecture is accomplished by a combination of anionic ring opening and RAFT polymerizations.

[0023] FIG.7C. Schematic synthesis of example triblock copolymers comprised of PSBnncoPSOHmblock copolymers in core block architecture is accomplished by a combination of anionic ring opening and RAFT polymerizations.

[0024] FIG.8A. DLS measurements of nanoparticle size confirms self-assembly of triblock copolymers with varying polysulfide chemistry to form nanoparticles by nanoprecipitation method.

[0025] FIG.8B. DLS measurements of nanoparticle PDI confirms self-assembly of triblock copolymers with varying polysulfide chemistry to form nanoparticles by nanoprecipitation method.

[0026] FIG.8C. DLS measurements of nanoparticle size confirms self-assembly of triblock copolymers with varying polysulfide chemistry to form nanoparticles by thin film rehydration method.

[0027] FIG.8D. DLS measurements of nanoparticle PDI confirms self-assembly of triblock copolymers with varying polysulfide chemistry to form nanoparticles by thin film rehydration method.

[0028] FIG.9A. Mechanical strength of hybrid NP / HA hydrogels can be dependent upon grafting density of CD where grafting density of 20% for HA-CD results in highest mechanical strength of hybrid hydrogel as observed through storage modulus (G’) measurements (n=3). Data are presented as mean ± s.d.

[0029] FIG.9B. Representative cryo-SEM imaging of hybrid hydrogel illustrates porosity of crosslinked network. Data are presented as mean ± s.d.Attorney Docket No.093386-0041-WO01

[0030] FIG.10. Example hybrid NP / HA hydrogels can be formed by mixing nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers with HA-CD20%(7.5 wt.% PBS) results in stable hydrogels as observed by rheological characterization of hybrid NP / HA hydrogels which demonstrates that each hybrid NP / HA hydrogel retains a larger storage modulus (G’) (n=3) compared to its loss modulus (G’’) (n=3) and confirms stable hydrogel formation among all hydrogels of varied polysulfide chemistry. Data are presented as mean ± s.d

[0031] FIG.11. The oxidative degradation profiles of hybrid NP / HA hydrogels (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%) can be modulated by polysulfide chemistry but retain degradation profiles that are dependent on increasing concentrations of H2O2 as observed byincreases in hydrogel tan delta ( ) (n=3) when hybrid NP / HA hydrogels of modified polysulfidechemistry are subjected to incubation with 1 mM H2O2, 10 mM H2O2, and 100 mM H2O2.

[0032] FIG.12A. Triblock copolymers with modified polysulfide P(PSncoPSRm) D300-AD20%triblock copolymers demonstrate increased encapsulation efficiency (n=3) of small molecule PHD2 inhibitors with decreased drug to polymer loading ratios. Data presented as mean ± s.d. (Nanoparticle: D300-AD20%2CD / 1AD, 5.5 w / v%)

[0033] FIG.12B. Triblock copolymers with modified polysulfide P(PSncoPSRm) D300-AD20%triblock copolymers demonstrate loading capacity (n=3) of small molecule PHD2 inhibitors which is dependent on the polysulfide copolymer architecture in core block at increasing drug to polymer loading. Data presented as mean ± s.d. (Nanoparticle: D300-AD20%2CD / 1AD, 5.5 w / v%)

[0034] FIG 13A. PPSBn100%.Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0035] FIG 13B. PPSBn70%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0036] FIG 13C. PPSBn50%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoringAttorney Docket No.093386-0041-WO01 percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0037] FIG 13D. PPSBn20%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0038] FIG 13E. Bn80%OH20%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0039] FIG 13F. PPS. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0040] FIG 13G. Bn40%OH60%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0041] FIG 13H. PPSOH20%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)Attorney Docket No.093386-0041-WO01

[0042] FIG 13I. PPSOH50%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0043] FIG 13J. PPSOH70%. Modified polysulfide chemistries can alter rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels as observed by monitoring percent release (n=3) of the small molecule PHD2 inhibitor MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0044] FIG 14A and FIG.14B. Modification of polysulfide chemistry can alter the rate of oxidative release of small molecule drugs from hybrid NP / HA hydrogels (NP / HA Hydrogel: D300- AD20%2CD / 1AD, 7.5 w / v%) incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers as determined through simple linear regression of kinetic release data in order to estimate the time to release (A) 50% of total small molecule drug or (B) 100% of total small molecule drug MK-8617 from hybrid NP / HA hydrogels incorporating modified polysulfide chemistries following incubation with increasing doses of H2O2. Data presented as mean ± s.d.

[0045] FIG 15. The incorporation of modified polysulfide monomers into nanoparticles self- assembled from P(PSncoPSRm) D300-AD20%triblock copolymers of hybrid NP / HA hydrogels does not significantly impact cytocompatibility of hybrid NP / HA hydrogels (NP / HA Hydrogel: D300- AD20%2CD / 1AD, 7.5 w / v%). The incubation of mouse mesenchymal stem cells (mMSCs) with hybrid NP / HA hydrogels of varied polysulfide chemistry results in signal cell viability comparable to a control shear-thinning hydrogel composed of only hyaluronic acid (HA) (HA / HA Only) (n=8- 10) following incubation of cells in 3D culture with hybrid NP / HA hydrogels for 24 hours. Data presented as mean ± s.d.

[0046] FIG 16A. The modification of polysulfide chemistry alters the antioxidant potential and scavenging capacity of nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers. The incubation of mouse NIH 3T3 fibroblasts in 2D culture with the same concentration of self-assembled nanoparticles with varied polysulfide chemistry reduces levels of hydrogen peroxide (H2O2) (n=5). Data presented as mean ± s.d. (Nanoparticle: D300-AD20%, 0.1 w / v%)Attorney Docket No.093386-0041-WO01

[0047] FIG 16B. The modification of polysulfide chemistry alters the antioxidant potential and scavenging capacity of nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers. The incubation of mouse NIH 3T3 fibroblasts in 2D culture with the same concentration of self-assembled nanoparticles with varied polysulfide chemistry results in increased cell viability (n=5) following exposure to increasing concentrations of H2O2for 24 hours. Data presented as mean ± s.d. (Nanoparticle: D300-AD20%, 0.1 w / v%)

[0048] FIG 17. The incorporation of modified polysulfide monomers P(PSncoPSRm) in self- assembled nanoparticles of NP / HA hydrogels can control the antioxidant potential and scavenging capacity of hybrid NP / HA hydrogels. (NP / HA Hydrogel: D300-AD20%2CD / 1AD 7.5 w / v%.) The incubation of mouse mesenchymal stem cells (mMSCs) with hybrid NP / HA hydrogels of varied polysulfide chemistry results in increased cell viability (n=8-10) following exposure to increasing concentrations of H2O2for 24 hours. Data presented as mean ± s.d.

[0049] FIG 18A. The closure of excisional wounds in diabetic mice can be accelerated following a single treatment of hybrid NP / HA hydrogels incorporating nanoparticles self- assembled from P(PSncoPSRm) D300-AD20%triblock copolymers as illustrated through the percent of wound remaining open over time. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0050] FIG 18B. The closure of excisional wounds in diabetic mice is accelerated following a single treatment of hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers as illustrated as quantified by the area under the curve (AUC) for the time to closure (n=6-12). Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0051] FIG.19. The levels of ROS in excisional wounds of diabetic mice can be reduced to the levels observed in unwounded skin of diabetic mice following a single treatment of hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers as observed through a significant reduction of Amplex red signal for H2O2 in wound tissue over time following treatments (n=3-6). Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0052] FIG.20. The vascular perfusion of excisional wounds in diabetic mice is unaffected over time following a single treatment of hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers as observed by the mean perfusion of the wounds measured through laser doppler perfusion imaging (LDPI) at each timepoint (n=6-12). Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)Attorney Docket No.093386-0041-WO01

[0053] FIG.21A. The treatment of excisional wounds of diabetic mice by hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds but produced varying degrees of collagen deposition in wounds compared to untreated wounds, as observed by histological evaluation of all excisional wounds at 7 days following treatment by Gomori’s trichrome stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0054] FIG.21B. The treatment of excisional wounds of diabetic mice by hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds but produced varying degrees of collagen deposition in wounds compared to untreated wounds, as observed by histological evaluation of all excisional wounds at 14 days following treatment by Gomori’s trichrome stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0055] FIG.22A. The treatment of excisional wounds of diabetic mice by hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds but produced varying degrees of collagen deposition in wounds compared to untreated wounds, as observed by histological evaluation of all excisional wounds at 7 following treatment by H&E stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0056] FIG.22B. The treatment of excisional wounds of diabetic mice by all hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds but produced varying degrees of collagen deposition in wounds compared to untreated wounds, as observed by histological evaluation of all excisional wounds at 14 days following treatment by H&E stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%.)

[0057] FIG.23. The delivery of small molecule PHD2 inhibitor by hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymerscan provide sustained stabilization of HIF1- (n=3) for up to 7 days as observed in proteinlysates collected from NIH 3T3 fibroblasts following incubation with hybrid NP / HA hydrogels with the small molecule PHD2 inhibitor, MK-8617, loaded into the core of the self-assembled nanoparticles. Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK-8617, 1 w / w%)Attorney Docket No.093386-0041-WO01

[0058] FIG.24. The delivery of small molecule PHD2 inhibitor by hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers results in more sustained activation of HRE-regulated genes in human umbilical vein endothelial cells (HUVECs) (n=1) for up to 5 days as observed through the tube formation and proliferation of HUVECs cultured in Matrigel and incubated with hybrid NP / HA hydrogels with the small molecule PHD2 inhibitor, MK-8617, loaded into the core of the self-assembled nanoparticles. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK-8617, 1 w / w%)

[0059] FIG.25A. The closure of excisional wounds in diabetic mice is accelerated following a single treatment of hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from PPS D300-AD20%or PPSBn20%triblock copolymers with the small molecule PHD2 inhibitor, MK- 8617, loaded into the core of the self-assembled nanoparticles as illustrated through the percent of wound remaining open over time (n=6-8). Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK-8617, 1 w / w%.

[0060] FIG.25B. The closure of excisional wounds in diabetic mice is accelerated following a single treatment of hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from PPS D300-AD20%or PPSBn20%triblock copolymers with the small molecule PHD2 inhibitor, MK- 8617, loaded into the core of the self-assembled nanoparticles as quantified by the area under the curve (AUC) for the time to closure (n=6-8). Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK-8617, 1 w / w%.

[0061] FIG.26. The vascularization of excisional wounds in diabetic mice are increased following single treatment of hybrid NP / HA hydrogels incorporating nanoparticles self- assembled from PPS D300-AD20%or PPSBn20%triblock copolymers with the small molecule PHD2 inhibitor, MK-8617, loaded into the core of the self-assembled nanoparticles which is observed by the mean perfusion of the wounds measured through laser doppler perfusion imaging (LDPI) at each timepoint (n=6-8). Data presented as mean ± s.d. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK-8617, 1 w / w%)

[0062] FIG.27A. The treatment of excisional wounds of diabetic mice by all hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds however, delivery of the small molecule PHD2 inhibitor by the same hybrid NP / HA hydrogels produced more mature wound remodelling and greater collagen deposition compared to all other treatments, as observed by histological evaluation of all excisional wounds at 7 days following treatment by Gomori’s trichrome stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK-8617, 1 w / w%)Attorney Docket No.093386-0041-WO01

[0063] FIG.27B. The treatment of excisional wounds of diabetic mice by all hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds however, delivery of the small molecule PHD2 inhibitor by the same hybrid NP / HA hydrogels produced more mature wound remodelling and greater collagen deposition compared to all other treatments, as observed by histological evaluation of all excisional wounds at 14 days following treatment by Gomori’s trichrome stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK-8617, 1 w / w%)

[0064] FIG.28A. The treatment of excisional wounds of diabetic mice by all hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds however, delivery of the small molecule PHD2 inhibitor by the same hybrid NP / HA hydrogels produced more mature wound remodelling and greater collagen deposition compared to all other treatments, as observed by histological evaluation of all excisional wounds at 7 days following treatment by H&E stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK- 8617, 1 w / w%)

[0065] FIG.28B. The treatment of excisional wounds of diabetic mice by all hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%resulted in thicker granulation tissue formation and a thicker epithelium compared to untreated wounds however, delivery of the small molecule PHD2 inhibitor by the same hybrid NP / HA hydrogels produced more mature wound remodelling and greater collagen deposition compared to all other treatments, as observed by histological evaluation of all excisional wounds at 14 days following treatment by H&E stain. (NP / HA Hydrogel: D300-AD20%2CD / 1AD, 7.5 w / v%; + MK- 8617, 1 w / w%) DETAILED DESCRIPTION

[0066] Disclosed herein is a hybrid shear-thinning hydrogel system that incorporates polysulfide chemistries into a synthetic triblock copolymer. Alteration of polysulfide structures can be used to alter the reaction rate at which reactive oxygen species (ROS) oxidizes polysulfide-based polymer units to create its more hydrophilic derivatives. The present disclosure sought to explore whether the addition of a -interacting moiety (e.g., benzyl ring) to the propylene sulfide (PS) monomer can slow the oxidation of the sulfide by ROS and whether the addition of a hydrogen donating moiety (e.g. a hydroxyl) can, conversely, accelerate the oxidation of the sulfide in the presence of ROS.Attorney Docket No.093386-0041-WO01

[0067] To provide polysulfide monomers with different sensitivities to ROS oxidation, benzyl (Bn) or hydroxyl (OH) functional groups were incorporated with PS monomers to create a library of triblock copolymers containing a range of different polysulfide block copolymers in the core block while maintaining consistent block architecture for the second and third blocks. This library of modified PS polymeric nanoparticles (NPs) was created with varied polysulfide core chemistry and surfaces tuned for physical crosslinking with hyaluronic acid (HA) through guest- host complexation. This produced shear-thinning hydrogels with varying sensitivities to ROS. This also provided a library of shear-thinning hydrogels, each with a unique polysulfide composition, that can be used to assess the effect of polysulfide chemistry on drug loading, antioxidant potency, and / or ROS-responsive behaviors of shear-thinning hydrogels. The impact of these functions of shear-thinning hydrogels was also evaluated in stented excisional wounds of diabetic mice. 1. Definitions

[0068] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting. Methods and materials similar or equivalent to those described herein can be used in practice or testing of the disclosed technology. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety.

[0069] The terms “comprise(s),” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms “a,” “and” and “the” include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments “comprising,” “consisting of” and “consisting essentially of,” the embodiments or elements presented herein, whether explicitly set forth or not.

[0070] For the recitation of numeric ranges herein, each intervening number therebetween with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are contemplated, and for the range 1.5-2, the numbers 1.5, 1.6, 1.7, 1.8, 1.9, and 2 are contemplated.Attorney Docket No.093386-0041-WO01

[0071] Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75thEd., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March's Advanced Organic Chemistry, 5thEdition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3rdEdition, Cambridge University Press, Cambridge, 1987; the entire contents of each of which are incorporated herein by reference.

[0072] The modifier “about” used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context (for example, it includes at least the degree of error associated with the measurement of the particular quantity). The modifier “about” should also be considered as disclosing the range defined by the absolute values of the two endpoints. For example, the expression “from about 2 to about 4” also discloses the range “from 2 to 4.” The term “about” may refer to plus or minus 10% of the indicated number. For example, “about 10%” may indicate a range of 9% to 11%, and “about 1” may mean from 0.9-1.1. Other meanings of “about” may be apparent from the context, such as rounding off, so, for example “about 1” may also mean from 0.5 to 1.4.

[0073] The term “alkyl,” as used herein, refers to a straight or branched, saturated hydrocarbon chain containing from 1 to 20 carbon atoms. The term “lower alkyl” or “C1-C6alkyl” means a straight or branched chain hydrocarbon containing from 1 to 6 carbon atoms. The term “C1-C4alkyl” means a straight or branched chain hydrocarbon containing from 1 to 4 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n- propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl and n- dodecyl.

[0074] The term “alkylene,” as used herein, refers to a divalent group derived from a straight or branched chain hydrocarbon of 1 to 50 carbon atoms, for example, of 2 to 10 carbon atoms. Representative examples of alkylene include, but are not limited to, -CH2CH2-, -CH2CH2CH2-, - CH2CH2CH2CH2-, and -CH2CH2CH2CH2CH2-.Attorney Docket No.093386-0041-WO01

[0075] The term “alkenylene,” as used herein, refers to a divalent alkenyl group, examples of which include, but are not limited to -CH=CH-, -CH=CH-CH2-, -CH=CH-CH2-CH2- and -CH2- CH=CH-CH2-. An alkenylene group may be optionally substituted with one or more substituents.

[0076] The term “alkynylene,” refers to a divalent alkynyl group, examples of which include,but are not limited to -C C-, -C C-CH2-,and -CH2-C C-CH2-. An alkynylenegroup may be optionally substituted with one or more substituents.

[0077] The term “aryl,” as used herein, refers to a phenyl group, or a bicyclic or tricyclic fused ring system in which at least one ring is aromatic. Examples of bicyclic fused ring systems include a phenyl group appended to the parent molecular moiety and fused to another phenyl group, a cycloalkyl group, a heteroaryl group, or a heterocyclic group. Representative examples of aryl include, but are not limited to, phenyl, naphthyl, anthracenyl, indolyl, and tetrahydroquinolinyl.

[0078] The term “cycloalkoxy,” as used herein, refers to a cycloalkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom.

[0079] The term “cycloalkyl” or “cycloalkane,” as used herein, refers to a saturated ring system containing all carbon atoms as ring members and zero double bonds. The term “cycloalkyl” is used herein to refer to a cycloalkane when present as a substituent. A cycloalkyl may be a monocyclic cycloalkyl (e.g., cyclopropyl), a fused bicyclic cycloalkyl (e.g., decahydronaphthalenyl), or a bridged cycloalkyl in which two non-adjacent atoms of a ring are linked by an alkylene bridge of 1, 2, 3, or 4 carbon atoms (e.g., bicyclo[2.2.1]heptanyl). Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantyl, and bicyclo[1.1.1]pentanyl.

[0080] The term “carbocyclyl” means a “cycloalkyl” or a “cycloalkenyl.” The term “carbocycle” means a “cycloalkane” or a “cycloalkene.” The term “carbocyclyl” refers to a “carbocycle” when present as a substituent.

[0081] The term “drug” refers to a substance that can act on a cell, virus, tissue, organ, organism, or the like, to create a change in the functioning of the cell, virus, tissue, organ, or organism. Examples of drugs include, but are not limited to, peptide-based drugs, chemotherapeutics, anti-inflammatory drugs, and immunomodulating drugs. A drug is capable of treating and / or ameliorating a condition or disease, or one or more symptoms thereof, in a subject. Drugs of the present disclosure also include prodrug forms of the agent.

[0082] The term “effective dosage” or “therapeutic dosage” or “therapeutically effective amount” or “effective amount,” as used herein, refers to an amount sufficient to effect beneficialAttorney Docket No.093386-0041-WO01 or desirable biological and / or clinical results, to modulate a biological process, and / or treat a disease or one or more of its symptoms and / or to prevent or reduce the risk of the occurrence or reoccurrence of the disease or disorder or symptom(s) thereof. A therapeutically effective amount is also one in which any toxic or detrimental effects of substance are outweighed by the therapeutically beneficial effects. A “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount. In reference to wound healing, an effective or therapeutically effective amount can include an amount sufficient to, among other things, accelerate closure of a wound.

[0083] The term “guest-host moiety,” as used herein, refers to a reversible affinity (e.g., Kd~ 1 x 105M-1) reaction between two molecules typically driven by noncovalent interactions such as hydrophobic interactions, hydrogen bonding, ionic interactions, and / or van der Waals forces. The reversible affinity reaction between two guest-host moieties can form a supramolecular complex (“guest-host complex”) in which the host molecule has a cavity that allows the guest molecule to fit into. Example guest-host pairings include, but are not limited to, -cyclodextrin (host)-cholesterol (guest); cucurbiturils (host)-pyrene (guest); crown ethers (host)-potassium ion(guest); calixarenes (host)-long-chain alkyl ammonium ion (guest); and pillararene (host)- -carotene (guest), Lutein (guest).

[0084] The term “heteroalkylene” as used herein, means an alkylene group, as defined herein, in which one or more of the carbon atoms has been replaced by a heteroatom selected from S, Si, O, P and N.

[0085] The term “heteroaryl” as used herein, refers to an aromatic monocyclic ring or an aromatic bicyclic ring system or an aromatic tricyclic ring system. The aromatic monocyclic rings are five or six membered rings containing at least one heteroatom independently selected from the group consisting of N, O and S (e.g.1, 2, 3, or 4 heteroatoms independently selected from O, S, and N). The five membered aromatic monocyclic rings have two double bonds and the six membered aromatic monocyclic rings have three double bonds. The bicyclic heteroaryl groups are exemplified by a monocyclic heteroaryl ring appended to the parent molecular moiety and fused to a monocyclic cycloalkyl group, as defined herein, a monocyclic aryl group, as defined herein, a monocyclic heteroaryl group, as defined herein, or a monocyclic heterocycle, as defined herein. The tricyclic heteroaryl groups are exemplified by a monocyclic heteroaryl ring appended to the parent molecular moiety and fused to two of a monocyclic cycloalkyl group, as defined herein, a monocyclic aryl group, as defined herein, a monocyclic heteroaryl group, asAttorney Docket No.093386-0041-WO01 defined herein, or a monocyclic heterocycle, as defined herein. Representative examples of monocyclic heteroaryl include, but are not limited to, pyridinyl (including pyridin-2-yl, pyridin-3-yl, pyridin-4-yl), pyrimidinyl, pyrazinyl, thienyl, furyl, thiazolyl, thiadiazolyl, isoxazolyl, pyrazolyl, and 2-oxo-1,2-dihydropyridinyl. Representative examples of bicyclic heteroaryl include, but are not limited to, chromenyl, benzothienyl, benzodioxolyl, benzotriazolyl, quinolinyl, thienopyrrolyl, thienothienyl, imidazothiazolyl, benzothiazolyl, benzofuranyl, indolyl, quinolinyl, imidazopyridine, benzooxadiazolyl, and benzopyrazolyl. Representative examples of tricyclic heteroaryl include, but are not limited to, dibenzofuranyl and dibenzothienyl. The monocyclic, bicyclic, and tricyclic heteroaryls are connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the rings, and can be unsubstituted or substituted.

[0086] The term “halogen” or “halo,” as used herein, means Cl, Br, I, or F.

[0087] The term “haloalkyl,” as used herein, means an alkyl group, as defined herein, in which one, two, three, four, five, six, seven or eight hydrogen atoms are replaced by a halogen.

[0088] The term “hydroxyl” or “hydroxy,” as used herein, means an -OH group.

[0089] The term “hydroxyalkyl,” as used herein, means at least one -OH group, is appended to the parent molecular moiety through an alkylene group, as defined herein.

[0090] The term “substituted” refers to a group that may be further substituted with one or more non-hydrogen substituent groups. Substituent groups include, but are not limited to, halogen, =O (oxo), =S (thioxo), cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, – COOH, ketone, amide, carbamate, and acyl.

[0091] Terms such as "alkyl," "cycloalkyl," "alkylene," etc. may be preceded by a designation indicating the number of atoms present in the group in a particular instance (e.g., "C1-4alkyl," "C3-6cycloalkyl," "C1-4alkylene"). These designations are used as generally understood by those skilled in the art. For example, the representation "C" followed by a subscripted number indicates the number of carbon atoms present in the group that follows. Thus, "C3alkyl" is an alkyl group with three carbon atoms (i.e., n-propyl, isopropyl). Where a range is given, as in "C1-4," the members of the group that follows may have any number of carbon atoms falling within the recited range. A "C1-4alkyl," for example, is an alkyl group having from 1 to 4 carbon atoms, however arranged (i.e., straight chain or branched).Attorney Docket No.093386-0041-WO01

[0092] For compounds described herein, groups and substituents thereof may be selected in accordance with permitted valence of the atoms and the substituents, such that the selections and substitutions result in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

[0093] The terms “subject” or “subject in need thereof,” as used herein, refer to a target of administration, which optionally displays symptoms related to a particular disease, pathological condition, disorder, or the like. The subject of the herein disclosed methods can be a vertebrate, such as a mammal, a fish, a bird, a reptile, or an amphibian. Thus, the subject of the herein disclosed methods can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The term does not denote a particular age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be covered. A patient refers to a subject afflicted with a disease or disorder. The term “patient” includes human and veterinary subjects.

[0094] The terms “treatment” or “treating” refer to the medical management of a patient with the intent to heal, cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. 2. Block Copolymers

[0095] Provided herein are block copolymers having structures that can advantageously form particles, which in turn can be used as part of a hydrogel, e.g., as a crosslinker. The block copolymer can include three different polymer blocks. The first block can be referred to as the A block, the second block can be referred to as the B block, and the third block can be referred to as the C block. In some embodiments, the block copolymer is an A-B-C triblock copolymer. The three different blocks can differ in hydrophilicity and / or hydrophobicity, which can allow the copolymer to self-assemble into different particulate structures.Attorney Docket No.093386-0041-WO01

[0096] The block copolymer can have a varying molecular weight. For example, the block copolymer can have a number average molecular weight of about 10 kilodalton (kDa) to about 300 kDa, such as about 10 kDa to about 250 kDa, about 10 kDa to about 200 kDa, about 10 kDa to about 200 kDa, about 15 kDa to about 150 kDa, about 15 kDa to about 75 kDa, about 10 kDa to about 60 kDa, about 15 kDa to about 50 kDa, about 10 kDa to about 45 kDa, about 10 kDa to about 50 kDa, about 10 kDa to about 30 kDa, about 10 kDa to about 25 kDa, or about 10 kDa to about 15 kDa. In some embodiments, the block copolymer has a number average molecular weight of greater than 10 kDa, greater than 15 kDa, greater than 20 kDa, greater than 25 kDa, greater than 30 kDa, greater than 35 kDa, or greater than 40 kDa. In some embodiments, the block copolymer has a number average molecular weight of less than 300 kDa, less than 250 kDa, less than 200 kDa, less than 150 kDa, less than 100 kDa, less than 75 kDa, or less than 50 kDa.

[0097] The term “molecular weight” in relation to the polymer refers to number average molecular weight (Mn) unless noted otherwise. Molecular weight of the block copolymer and the individual blocks can be measured by techniques used within the art, such as size exclusion chromatography (SEC), SEC combined with multi-angle light scattering, gel permeation chromatography, intrinsic viscosity, nuclear magnetic resonance (NMR), and the like. In some embodiments, the molecular weight is measured by gel permeation chromatography. In some embodiments, the molecular weight is measured by NMR.

[0098] The block copolymer can have a varying structure associated with a varying molecular weight. For example, the block copolymer can have a structure of A5kDa-20kDa-B10kDa-40kDa-C20kDa-40kDa, A1kDa-50kDa-B5kDa-70kDa-C10kDa-60kDa, or A0.5kDa-100kDa-B1kDa-100kDa-C1kDa-100kDa. In some embodiments, the block copolymer has a structure of A5kDa-20kDa-B10kDa-40kDa-C20kDa-40kDa.The foregoing are example structures and any combination of molecular weight and block alignment as disclosed herein can be used in the block copolymers. A. First Block (A)

[0099] The first block is a hydrophobic block or a block that is more hydrophobic than the second and third blocks. The first block can include polymer(s) having a core monomer group (e.g., making up the polymer backbone) and a pendant group(s) attached to the core monomer group. The core monomer can be a sulfide (e.g., thioether) monomer having pendant group as described herein and that can be polymerized through chain-growth polymerization, such as anionic ring opening polymerization and free radical polymerization (e.g., atom transfer radical polymerization (ATRP)). The polymer(s) can include three different pendant groups. The three pendent groups can include a methyl group, a hydrogen bond donating moiety, and / or a -Attorney Docket No.093386-0041-WO01 interacting moiety. The pendent groups can be included in the first block in a random manner and can be included in varying amounts. In other words, the pendant groups can be included in the first block as a random copolymer.

[0100] Hydrogen bond donating moieties can allow the first block and copolymer thereof to participate in hydrogen bonding interactions with, e.g., a drug. As used herein, “hydrogen bonding” refers to a primarily electrostatic force of attraction between a hydrogen atom which is covalently bound to a more electronegative "donor" atom or moiety, and another electronegative atom bearing a lone pair of electrons, which can be called the hydrogen bond acceptor moiety. For example, drugs having a hydrogen bond acceptor moiety, such as amides, pyridines, aldehydes, ketones, esters, carboxylic acids, sulfoxides, and sulfones, can participate in hydrogen bonding interactions with the hydrogen bond donating moiety of the first block. Example hydrogen bond donating moieties include, but are not limited to, hydroxyl, carboxy, amine, ammonium, amide, imide, hydroxylamine, hydrazine, hydrazide, phenol, aniline, urea, thiourea, sulfonylamine, acetamidine, oxime, carbamate, O-thiocarbamate, S-thiocarbmate, and boronic acid. The first block can include the same hydrogen bond donating moiety throughout the first block. In other embodiments, the first block includes at least 2, at least 3, at least 4, or at least 5 different hydrogen bond donating moieties throughout the first block. In some embodiments, the hydrogen bond donating moiety includes hydroxyl, carboxy, amine, amide, or a combination thereof. In some embodiments, the hydrogen bond donating moiety includes hydroxyl, amine, or a combination thereof. In some embodiments, the hydrogen bond donating moiety includes hydroxyl.

[0101] -interacting moieties can allow the first block and copolymer thereof to participate in --interactions with, e.g., a drug. As used herein, “ - -interactions” refer to a particular type ofdispersion force from, e.g., van der Waals forces, which can be established between unsaturated (poly)cyclic moieties. For example, drugs having a -interacting moiety, such asaromatic moieties, can participate in - interactions with the -interacting moiety of the firstblock. Example -interacting moieties include, but are not limited to, phenyl, naphthyl, pyridinyl, pyrrole, imidazolyl, pyrazolyl, indolyl, and furyl – all of which can be optionally substituted. The first block can include the same -interacting moiety throughout the first block. In other embodiments, the first block includes at least 2, at least 3, at least 4, or at least 5 different - interacting moieties throughout the first block. In some embodiments, the -interacting moiety includes phenyl, furyl, pyridinyl, pyrrolyl, imidazolyl, or a combination thereof. In some embodiments, the -interacting moiety includes phenyl, furyl, pyridinyl, or a combination thereof. In some embodiments, the -interacting moiety includes phenyl.Attorney Docket No.093386-0041-WO01

[0102] The sulfide-based polymer of the first block can be responsive to reactive oxygen species (ROS). The ROS reactivity can provide a tunable mechanism for bulk hydrogel degradation, a triggerable mechanism for release of encapsulated drug, and can endow the gel with antioxidant activity; the latter can help to detoxify excessive levels of oxidative stress which can cause cell and tissue damage and inflammation, exacerbating many pathological scenarios. In addition, a reactive oxygen species-triggered release of a drug can be used, e.g., in particulate formulations. See general scheme below:.

[0103] In some embodiments, the first block includes recurring units of formula (I)(I), wherein: X1, at each occurrence, is methyl, of formula (II)(II), or of formula (III)L1and L2are each independently -C(O)O-alkylene, -C(O)NRb-alkylene, alkylene, alkylene-O- alkylene, alkylene-O-, or bond; R1is a -interacting moiety; R2is a hydrogen donating moiety; Rais hydrogen or C1-3alkyl; and Rbis hydrogen or C1-6alkyl. The different options for X1(e.g., pendant group) can arise from different monomers. When the first block includes monomers having different X1pendant groups (e.g., at least one unit where X1is methyl and at least one unit where X1is of formula (II)), such monomers can be polymerized as a random copolymer.

[0104] The first block can include recurring units of formula (I) with varying amounts of X1. For example, the first block can include recurring units of formula (I) where X1is methyl at aboutAttorney Docket No.093386-0041-WO01 0 mol% to about 100 mol% of the first block, such as about 1 mol% to about 90 mol%, about 5 mol% to about 80 mol%, about 10 mol% to about 90 mol%, about 15 mol% to about 75 mol%, about 1 mol% to about 60 mol%, or about 40 mol% to about 90 mol%. In addition, the first block can include recurring units of formula (I) where X1is formula (II) at about 0 mol% to about 100 mol% of the first block, such as about 1 mol% to about 90 mol%, about 5 mol% to about 80 mol%, about 10 mol% to about 90 mol%, about 15 mol% to about 75 mol%, about 1 mol% to about 60 mol%, or about 40 mol% to about 90 mol%.. The first block can also include recurring units of formula (I) where X1is formula (III) at about 0 mol% to about 100 mol% of the first block, such as about 1 mol% to about 90 mol%, about 5 mol% to about 80 mol%, about 10 mol% to about 90 mol%, about 15 mol% to about 75 mol%, about 1 mol% to about 60 mol%, or about 40 mol% to about 90 mol%.

[0105] In some embodiments, the first block includes a multi-unit segment of formula (V)(V), wherein: X2, at each occurrence, is of formula (II)(II), or of formula (III)(III); L1and L2are each independently -C(O)O-alkylene, -C(O)NRb-alkylene, alkylene, alkylene-O- alkylene, alkylene-O-, or bond; R1is a -interacting moiety; R2is a hydrogen donating moiety; Ra’and Ra’’are each independently hydrogen or C1-3alkyl; Rbis hydrogen or C1-6alkyl; n is 0 to 200; m is 0 to 200; if n is 0, m is greater than or equal to 10; and if m is 0, n is greater than or equal to 10.

[0106] As can be seen, the multi-unit segment of formula (V) shows the recurring units of formula (I) as a random copolymer. As used herein, the term “ran” refers to a random copolymerAttorney Docket No.093386-0041-WO01 of the recurring units it is associated with. For example, for formula (V), “ran” refers to a random copolymer of recurring units of.

[0107] In some embodiments, n is 0 to 200, 0 to 100, 0 to 50, 0 to 25, 1 to 200, 1 to 100, 1 to 50, 1 to 25, 50 to 200, or 100 to 200. In some embodiments, n is greater than 0, greater than 1, greater than 2, greater than 3, greater than 4, greater than 5, greater than 10, greater than 20, greater than 50, or greater than 100. In some embodiments, n is less than 200, less than 150, less than 125, less than 100, less than 75, less than 50, less than 20, less than 15, less than 10, or less than 5.

[0108] In some embodiments, m is 0 to 200, 0 to 100, 0 to 50, 0 to 25, 1 to 200, 1 to 100, 1 to 50, 1 to 25, 50 to 200, or 100 to 200. In some embodiments, m is greater than 0, greater than 1, greater than 2, greater than 3, greater than 4, greater than 5, greater than 10, greater than 20, greater than 50, or greater than 100. In some embodiments, m is less than 200, less than 150, less than 125, less than 100, less than 75, less than 50, less than 20, less than 15, less than 10, or less than 5.

[0109] In some embodiments, R1is an aryl or heteroaryl, wherein the aryl and heteroaryl are optionally substituted with 1 or 2 substituents, each independently halogen, cyano, C1-4alkyl, C1-2fluoroalkyl, –OC1-2alkyl, or –OC1-2fluoroalkyl; and R2is hydroxy, amine, or amide.

[0110] In some embodiments, R1is an aryl or heteroaryl, wherein the aryl and heteroaryl are optionally substituted with 1 or 2 substituents, each independently halogen, C1-4alkyl, or –OC1-2alkyl; and R2is hydroxy or amine.

[0111] In some embodiments, L1and L2are each independently -C(O)O-C1-4alkylene, - C(O)NH-C1-4alkylene, C1-10alkylene, C1-6alkylene-O-C1-6alkylene, C1-4alkylene-O-, or bond.

[0112] In some embodiments, L1and L2are each independently -C(O)O-C1-2alkylene, - C(O)NH-C1-2alkylene, C1-6alkylene, C1-4alkylene-O-C1-4alkylene, C1-2alkylene-O-, or bond.

[0113] In some embodiments, L1and L2are each independently C1-10alkylene, C1-6alkylene- O-C1-6alkylene, or bond.

[0114] In some embodiments, L1and L2are each independently C1-10alkylene or C1-6alkylene-O-C1-6alkylene; R1is aryl or heteroaryl; and R2is hydroxy or amine.

[0115] In some embodiments, L1is C1-4alkylene or C1-2alkylene-O-C1-2alkylene; L2is C1-4alkylene; and Ra’and Ra’’are each independently hydrogen.

[0116] In some embodiments, X2is of formula (II-a)Attorney Docket No.093386-0041-WO01(II-a), or formula (III-a)(III-a), wherein L1and L2are defined as described above for formula (I) and formula (V). Formula (II-a) and formula (III-a) can also be applied to X1as described in formula (I).

[0117] In some embodiments, X2, at each occurrence, is of formula (II-a)(II-a), or formula (III-a)(III-a); wherein: L1is C1-4alkylene or C1-2alkylene-O-C1-2alkylene; and L2is C1-4alkylene. This embodiment can also be applied to X1as described in formula (I).

[0118] Varying amounts of recurring units including formula (II) can be included in the multi- unit segment of formula (V). For example, the multi-unit segment of formula (V) can include recurring units including formula (II) at about 0% to about 100% by weight of the multi-unit segment of formula (V), such as about 0% to about 80%, about 5% to about 90%, about 10% to about 80%, about 15% to about 70%, about 1% to about 100%, about 50% to about 100%, about 1% to about 50%, or about 0% to about 50%.

[0119] In some embodiments, the multi-unit segment of formula (V) includes recurring units including formula (II) at greater than 1% by weight of the multi-unit segment of formula (V), greater than 10% by weight of the multi-unit segment of formula (V), greater than 20% by weight of the multi-unit segment of formula (V), greater than 30% by weight of the multi-unit segment ofAttorney Docket No.093386-0041-WO01 formula (V), greater than 40% by weight of the multi-unit segment of formula (V), or greater than 50% by weight of the multi-unit segment of formula (V). In some embodiments, the multi-unit segment of formula (V) includes recurring units including formula (II) at less than 100% by weight of the multi-unit segment of formula (V), less than 90% by weight of the multi-unit segment of formula (V), less than 80% by weight of the multi-unit segment of formula (V), less than 70% by weight of the multi-unit segment of formula (V), less than 60% by weight of the multi-unit segment of formula (V), or less than 50% by weight of the multi-unit segment of formula (V).

[0120] Varying amounts of recurring units including formula (III) can be included in the multi- unit segment of formula (V). For example, the multi-unit segment of formula (V) can include recurring units including formula (III) at about 0% to about 100% by weight of the multi-unit segment of formula (V), such as about 0% to about 60%, about 5% to about 90%, about 10% to about 80%, about 15% to about 70%, about 1% to about 100%, about 50% to about 100%, about 1% to about 50%, or about 0% to about 50%.

[0121] In some embodiments, the multi-unit segment of formula (V) includes recurring units including formula (III) at greater than 0% by weight of the multi-unit segment of formula (V), greater than 1% by weight of the multi-unit segment of formula (V), greater than 10% by weight of the multi-unit segment of formula (V), greater than 20% by weight of the multi-unit segment of formula (V), greater than 30% by weight of the multi-unit segment of formula (V), greater than 40% by weight of the multi-unit segment of formula (V), or greater than 50% by weight of the multi-unit segment of formula (V). In some embodiments, the multi-unit segment of formula (V) includes recurring units including formula (III) at less than 100% by weight of the multi-unit segment of formula (V), less than 90% by weight of the multi-unit segment of formula (V), less than 80% by weight of the multi-unit segment of formula (V), less than 70% by weight of the multi-unit segment of formula (V), less than 60% by weight of the multi-unit segment of formula (V), or less than 50% by weight of the multi-unit segment of formula (V).

[0122] The foregoing description of the weight percentage for recurring units including formulas (II) or (II) as it relates to the multi-segment of formula (V) can also be applied to weight percentages for the first block. For example, the first block can include recurring units including formula (II) at greater than 0% by weight of the first block, greater than 1% by weight of the first block, greater than 10% by weight of the first block, etc. In addition, the foregoing description for formulas (II) and (III) with respect to amounts included in the multi-unit segment of formula (V) or the first block can also be applied to formulas of (II-a) and (III-a).Attorney Docket No.093386-0041-WO01

[0123] Example substituents for X1and X2and its associated recurring unit include, but are not limited to,Attorney Docket No.093386-0041-WO01

[0124] The multi-unit segment of formula (V) can include a varying total degree of polymerization (e.g., total number of monomer units in formula (V)). For example, the multi-unit segment of formula (V) can have a total unit degree of polymerization of 50 to 300, such as 60 to 280, 70 to 250, 80 to 225, 100 to150, 100 to 200, 50 to 200, or 100 to 300. In some embodiments, the multi-unit segment of formula (V) has a total unit degree of polymerization of greater than 50, greater than 60, greater than 70, greater than 80, greater than 90, greater than 100, greater than 150, or greater than 200. In some embodiments, the multi-unit segment of formula (V) has a total unit degree of polymerization of less than 300, less than 275, less than 250, less than 225, less than 200, less than 175, less than 150, or less than 100.

[0125] The foregoing description of the total unit degree of polymerization as it relates to the multi-segment of formula (V) can also be applied to the degree of polymerization of the first block and to the degree of polymerization for the recurring units of formula (I). For example, the first block can have a degree of polymerization of greater than 50, greater than 60, greater than 70, greater than 80, greater than 90, etc. Similarly, the recurring units of formula (I) can have a degree of polymerization of greater than 50, greater than 60, greater than 70, greater than 80, greater than 90, etc.

[0126] The first block can have a varying molecular weight. For example, the first block can have a number average molecular weight of about 0.5 kDa to about 50 kDa, such as about 1 kDa to about 50 kDa, about 1 kDa to about 40 kDa, about 5 kDa to about 30 kDa, about 0.5 kDa to about 25 kDa, or about 20 kDa to about 50 kDa. In some embodiments, the first block has a number average molecular weight of greater than 0.5 kDa, greater than 1 kDa, greater than 5 kDa, greater than 10 kDa, greater than 15 kDa, greater than 20 kDa, or greater than 30 kDa. In some embodiments, the first block has a number average molecular weight of less than 50 kDa, less than 45 kDa, less than 40 kDa, less than 35 kDa, less than 30 kDa, less than 20 kDa, or less than 15 kDa. B. Second Block (B)

[0127] The second block of the copolymer is a hydrophilic block. Accordingly, the second block can include a hydrophilic polymer. The second block can include one type of hydrophilic polymer or can include a plurality of different hydrophilic polymers. For example, in some embodiments, the second block includes at least 2, at least 3, at least 4, or at least 5 different hydrophilic polymers. Different hydrophilic polymers can be included within the same B block or each different hydrophilic polymer can correspond to a different B block within the block copolymer.Attorney Docket No.093386-0041-WO01

[0128] Any suitable hydrophilic polymer known within the art can be used such that the block copolymer can, e.g., self-assemble into particulate structures. Example hydrophilic polymers include, but are not limited to, poly(dimethylacrylamide) (PDMA), poly(ethylene glycol) (PEG), poly(PEG), poly(methyl oxazoline)(PMOX), poly(ethyl oxazoline), polysarcosine, poly(4- acryloylmorpholine), poly(glycerol monomethacrylate), poly(propylene sulfoxide), poly(2- (methylsulfinyl)ethyl acrylate)(PMSEA), poly(vinyl alcohol)(PVA), poly(glycidol), poly(thioglycidyl glycerol)(PTGG), poly(2-methacryloyloxyethyl phosphorylcholine)(PMPC), poly(vinyl pyrrolidone)(PVP), poly(N-(2-hydroxypropyl)methacrylamide)(PHPMA), poly(trimethylamine N- oxide), poly(lysine-methacrylamide), poly(lysine-acrylamide), poly(carboxybetaine), poly(sulfobetaine), heparosan, poly(acrylic acid), poly(acrylamide), and any combination thereof.

[0129] In some embodiments, the second block includes PDMA, PEG, poly(methyl oxazoline), polysarcosine, poly(4-acryloylmorpholine), poly(glycerol monomethacrylate), poly(propylene sulfoxide), or a combination thereof. In some embodiments, the second block includes PDMA, PEG, poly(glycerol monomethacrylate), or a combination thereof. In some embodiments, the second block includes PDMA.

[0130] The second block can have a varying molecular weight. For example, the second block can have a number average molecular weight of about 1 kDa to about 100 kDa, such as about 10 kDa to about 100 kDa, about 15 kDa to about 90 kDa, about 20 kDa to about 80 kDa, about 30 kDa to about 70 kDa, about 10 kDa to about 40 kDa, about 10 kDa to about 60 kDa, or about 50 kDa to about 100 kDa. In some embodiments, the second block has a number average molecular weight of greater than 1 kDa, greater than 10 kDa, greater than 20 kDa, greater than 30 kDa, greater than 40 kDa, greater than 50 kDa, greater than 60 kDa, or greater than 70 kDa. In some embodiments, the second block has a number average molecular weight of less than 100 kDa, less than 90 kDa, less than 80 kDa, less than 70 kDa, less than 60 kDa, less than 50 kDa, or less than 40 kDa. C. Third Block (C)

[0131] The third block of the copolymer includes a recurring hydrophilic unit and a recurring unit that include a guest-host moiety. The third block can include the recurring hydrophilic unit and the recurring unit including a guest-host moiety as a copolymer. The copolymer may be a random copolymer. When the block copolymer is in particulate form, the balance between the recurring hydrophilic unit and the recurring guest-host moiety unit can facilitate that the third block can act as a hydrophilic corona block (along with the second block) that can present the guest-host moiety on a surface of the particle. This can facilitate interaction of the guest-host moiety of the third block with, e.g., a suitable partner guest-host moiety of the composition. TheAttorney Docket No.093386-0041-WO01 third block (along with the second block) may also instill stealth properties to particles formed by the block copolymer.

[0132] In some embodiments, the third block includes a multi-unit segment of formula (IV)wherein: Z is a recurring hydrophilic unit; G is NH or O; L3is alkylene, heteroalkylene, alkenylene, alkynylene, or bond; R3is a guest-host moiety; Rcis hydrogen or C1-3alkyl; and y is no more than 30 mol% of formula (IV). As discussed elsewhere herein, “ran” refers to a random copolymer, where here ran refers to a copolymer between the recurring hydrophilic unit and the guest-host moiety unit.

[0133] The recurring hydrophilic unit can be derived from any suitable hydrophilic monomer that can be polymerized, e.g., via RAFT polymerization, as part of the third block. Example hydrophilic monomers that can be used to provide the recuring hydrophilic include, but are not limited to, dimethylacrylamide (DMA), oligo(ethylene glycol), 2-methacryloyloxyethyl phosphorylcholine, N-(2-hydroxypropyl)methacrylamide, and combinations thereof. In some embodiments, the recurring hydrophilic unit is derived from DMA.

[0134] The guest-host moiety can be any moiety known in the art that can act in guest-host interactions. Examples of guest-host moieties include, but are not limited to, -cyclodextrin, adamantane, cholesterol, cucurbiturils, pyrene, crown ethers, potassium ion, calixarenes, long- chain alkyl ammonium ion, pillararene, -carotene, and lutein. In some embodiments, R3is adamantane.

[0135] The guest-host moieties can be integrated into a polymer through various techniques known within the art. For example, polymerization of guest-host moiety containing acrylamide / acrylate monomers can be done. In addition, post-polymerization grafting of guest- host moieties with functional groups such as amines, carboxylic acid, thiols, and alkenes / alkynes onto the polymer backbone can be done. An example bioconjugation method is carbodiimide coupling (e.g., amidation), maleimide-thiol conjugation, click chemistry, Staudinger ligation, etc. For example, a polymer backbone with pendent functional groups like vinyl azlactone ring, activated NHS esters, aldehyde, and maleimides can be used for grafting of amine functional host and guest moieties.Attorney Docket No.093386-0041-WO01

[0136] In some embodiments, G is NH.

[0137] In some embodiments, L3is alkylene or bond.

[0138] In some embodiments, Rcis hydrogen.

[0139] In some embodiments, G is NH, L is alkylene or bond, Rcis hydrogen, and R3is adamantane.

[0140] The amount of guest-host moiety recurring units can be varied to provide advantageous particle formation, which can aid composition formation as disclosed herein. For example, y can be about 1 mol% to about 35 mol% of formula (IV), such as about 1 mol% to about 30 mol%, about 1 mol% to about 25 mol%, about 1 mol% to about 20 mol%, about 1 mol% to about 15 mol%, about 1 mol% to about 10 mol%, about 10 mol% to about 25 mol%, about 5 mol% to about 20 mol%, or about 15 mol% to about 30 mol%. In some embodiments, y is no more than 35 mol% of formula (IV), no more than 30 mol% of formula (IV), no more than 25 mol% of formula (IV), no more than 20 mol% of formula (IV), no more than 15 mol% of formula (IV), or no more than 10 mol% of formula (IV). In some embodiments, y is no less than 1 mol% of formula (IV), no less than 5 mol% of formula (IV), no less than 10 mol% of formula (IV), no less than 15 mol% of formula (IV), no less than 20 mol% of formula (IV), or no less than 25 mol% of formula (IV). Mol% can be measured via NMR.

[0141] The multi-unit segment of formula (IV) can include a varying total unitdegree of polymerization (e.g., total number of monomer units in formula (IV)). For example, the multi-unit segment of formula (IV) can have a total unit degree of polymerization of 50 to 500, such as 75 to 500, 100 to 500, 80 to 300, 100 to 400, 200 to 400, 50 to 200, or 300 to 500. In some embodiments, the multi-unit segment of formula (IV) has a total unit degree of polymerization of greater than 50, greater than 75, greater than 100, greater than 150, greater than 200, greater than 250, greater than 300, or greater than 400. In some embodiments, the multi-unit segment of formula (IV) has a total unit degree of polymerization of less than 500, less than 450, less than 400, less than 350, less than 300, less than 250, less than 200, or less than 100.

[0142] The foregoing description of the total unit degree of polymerization as it relates to the multi-segment of formula (IV) can also be applied to the degree of polymerization of the third block. For example, the third block can have a degree of polymerization of greater than 50, greater than 75, greater than 100, etc.

[0143] The third block can have a varying molecular weight. For example, the third block can have a number average molecular weight of about 0.5 kDa to about 100 kDa, such as about 1 kDa to about 100 kDa, about 10 kDa to about 90 kDa, about 20 kDa to about 80 kDa, about 30 kDa to about 70 kDa, about 0.5 kDa to about 30 kDa, about 10 kDa to about 40 kDa, about 0.5Attorney Docket No.093386-0041-WO01 kDa to about 50 kDa, or about 50 kDa to about 100 kDa. In some embodiments, the third block has a number average molecular weight of greater than 0.5 kDa, greater than 10 kDa, greater than 20 kDa, greater than 30 kDa, greater than 40 kDa, greater than 50 kDa, or greater than 60 kDa. In some embodiments, the third block has a number average molecular weight of less than 100 kDa, less than 90 kDa, less than 80 kDa, less than 70 kDa, less than 60 kDa, less than 50 kDa, or less than 40 kDa. D. Synthesis of Block Copolymers

[0144] Example block copolymer synthetic details can be found in the Examples herein. Generally, anionic and RAFT polymerization techniques can be combined to synthesize ABC triblock polymers with varying sensitivities to reactive oxygen species (ROS) degradation. The first block of these polymers can be a ROS-degradable polysulfide prepared through ring- opening polymerization. This can be done using either polymerizing three-membered cyclic propylene sulfide or by copolymerizing with functional propylene sulfide containing, e.g., benzyl or hydroxyl groups. These polymers can be functionalized with a RAFT chain transfer agent, such as ECT, to build the second and, subsequently, the third block onto the first block. An individual macro-CTA functional can be used for RAFT polymerization of DMA to synthesize respective diblock copolymers of P(PSn-co-Rm)-b-PDMA300-ECT (P(PSncoRm)-D300). RAFT random copolymerization of hydrophilic DMA and ADA (guest monomer) from respective diblock macro-CTA of P(PSn-co-Rm)-b-PDMA300-ECT can then lead to the formation of triblock copolymers represented as P(PSn-co-Rm)-b-PDMA300-b-PDMA160-co-ADA40(P(PSn-co-Rm) D300-AD20%). Additional description on synthetic methods for block copolymers can be found in Bezold et al., Hybrid Shear-thinning Hydrogel Integrating Hyaluronic Acid with ROS-Responsive Nanoparticles, Adv Funct Mater, 1;33(31):2213368 and U.S. Patent Application No.18 / 698,763, both of which are incorporated by reference herein in their entirety. E. Example Block Copolymers

[0145] In some embodiments, the block copolymer comprises a first block comprising a multi-unit segment of formula (V-a)wherein: X3, at each occurrence, is of formula (II-a)Attorney Docket No.093386-0041-WO01 ,or formula (III-and the second block comprises a hydrophilic polymer selected from the group consisting of PDMA, PEG, poly(glycerol monomethacrylate), and a combination thereof. F. Particles

[0146] The composition and structure of the block copolymer can allow the copolymers to self-assemble into particulate structures. For example, the block copolymer can have a critical micelle concentration (CMC) and at a concentration above the CMC, the block copolymers can self-assemble, e.g., with other block copolymers into a particulate structure. Accordingly, also disclosed herein are particles that can include a plurality of self-assembled block copolymers. Example structures include micelles, vesicles, inverted micelles, spherical polymersomes, tubular polymersomes, and nanofibers. Micelles can be a variety of shapes with varied aspect ratios. For example, the particle can be a filomicelle, a rodlike micelle, a cylindrical micelle, and / or a wormlike micelle.

[0147] The block copolymer can have a varying CMC. For example, the block copolymer can have a CMC of about 0.01 mg / mL to about 0.07 mg / mL, such as about 0.015 mg / mL to about 0.06 mg / mL, about 0.02 mg / mL to about 0.055 mg / mL, about 0.025 mg / mL to about 0.055 mg / mL, or about 0.025 mg / mL to about 0.052 mg / mL. CMC can be determined by atomic force microscopy imaging analysis and / or fluorescence intensity analysis of micelle formation. Further details of determining CMC can be found in J. N. Phillips, Trans. Faraday 1955, 51, 561, which is incorporated by reference herein in its entirety.

[0148] The particle can have a varying diameter. For example, the particle can have a diameter of about 50 nm to about 250 nm, such as about 60 nm to about 225 nm, about 70 nm to about 200 nm, about 50 nm to about 150 nm, or about 100 nm to about 250 nm. In some embodiments, the particle has a diameter of greater than 50 nm, greater than 75 nm, greater than 100 nm, or greater than 150 nm. In some embodiments, the particle has a diameter of less than 250 nm, less than 225 nm, less than 200 nm, or less than 150 nm. Particle size can be measured by techniques known within the art, such as dynamic light scattering and electron microscopy (e.g., TEM). i. Drugs

[0149] In contrast to current systems, which typically exploit H-bond acceptors, the disclosed block copolymers can uniquely leverage OH / H-bond donor combined with aromatic, as well asAttorney Docket No.093386-0041-WO01thioether -electron interacting groups (e.g., in sulfide embodiments). S- interactions are notextensivity explored in the drug delivery field but can be strong. As such, the block copolymers can advantageously load drugs into a particulate form, which can then be included in compositions as disclosed herein.

[0150] Any suitable drug can be used in the disclosed particles and compositions. Drugs having a hydrogen bond acceptor moiety, aromatic moiety, or both can benefit in their loading into the particle as these moieties can beneficially interact with the first block of the disclosed copolymers. In some embodiments, the drug includes a hydrogen bond acceptor moiety, an aromatic moiety, or a combination thereof.

[0151] Drugs with a range of hydrophobicity can be used in the compositions. The drug’s hydrophobicity may be characterized by its octanol-water distribution coefficient (logD), where a larger value indicates greater hydrophobicity. For example, the drug can have a logD of about -1 to about 10, such as about 0 to about 8, about 0.5 to about 7, about -1 to about 8, about -1 to about 7, about 0 to about 6, about 0.5 to about 5.5, or about 1 to about 5.5. In some embodiments, the drug has a logD of greater than -1, greater than -0.5, greater than 0, greater than 0.5, or greater than 1. In some embodiments, the drug has a logD of less than 10, less than 8, less than 7, less than 6, or less than 5.5. The above-listed logD values can be measured at a pH of 7.4 (e.g., logD of greater than 0.5 at a pH of 7.4).

[0152] Examples of drugs include, but are not limited to, peptide-based drugs (e.g., cyclosporin, bortezomib, etc.), chemotherapeutics (e.g., a taxane, a tyrosine kinase inhibitor, a topoisomerase inhibitor, a bcl-family inhibitor, a DNA crosslinking agent, a DNA antimetabolite, a PHD-family inhibitor, etc.); anti-inflammatory drugs (e.g., a steroid, a corticosteroid, a non- steroidal anti-inflammatory drug (NSAID), etc.); immune modulating drugs (e.g., an immunotherapy adjuvant); and a pro-angiogenic drug. In some embodiments, the drug includes a chemotherapeutic, an anti-inflammatory drug, an immune modulating drug, a pro-angiogenic drug, or a combination thereof. In some embodiments, the drug includes a taxane, a steroid, a corticosteroid, a NSAID, a tyrosine kinase inhibitor, a topoisomerase inhibitor, a bcl-family inhibitor, an immunotherapy adjuvant, a DNA crosslinking agent, a DNA antimetabolite, a PHD- family inhibitor, or a combination thereof.

[0153] In some embodiments, the drug includes cyclosporin A, paclitaxel, bortezomib, etoposide, neratinib, osimertinib, chloroquine, GANT58, docetaxel, dexamethasone, carmofur, dexamethasone, dexamethasone palmitate, carfilzomib, afatinib, irinotecan, doxorubicin, doxycycline, camptothecin, imiquimod, MK-8617, ciclopirox, roxadustat, or a combination thereof. In some embodiments, the drug includes cyclosporin A, paclitaxel, bortezomib,Attorney Docket No.093386-0041-WO01 etoposide, neratinib, osimertinib, chloroquine, GANT58, or a combination thereof. In some embodiments, the drug includes paclitaxel, GANT 58, or a combination thereof. 3. Compositions

[0154] Also disclosed herein are compositions that include a second hydrophilic polymer crosslinked with a plurality of self-assembled block copolymer particles as disclosed herein. Crosslinking between the particles and the second hydrophilic polymer is mediated by guest- host interactions. For example, the second hydrophilic polymer can include a guest-host moiety as disclosed herein that is capable of forming a guest-host complex with the guest-host moiety of the block copolymer. In some embodiments, the guest-host moiety of the block copolymer is adamantane and the guest-host moiety of the second hydrophilic polymer is -cyclodextrin. Accordingly, the second hydrophilic polymer can be physically crosslinked, not covalently, which can allow for dynamic shear-thinning properties for the composition. As such, the composition can be shear thinning.

[0155] The second hydrophilic polymer can be any suitable hydrophilic polymer that can be grafted with a guest-host moiety and used in the disclosed compositions. Example hydrophilic polymers include, but are not limited to, hyaluronic acid, gelatin, alginate, carboxymethyl cellulose (CMC), glycol chitosan, carrageenan, dextran, pullulan, and combinations thereof. In some embodiments, the second hydrophilic polymer comprises hyaluronic acid, gelatin, alginate, or carboxymethyl cellulose (CMC). In some embodiments, the second hydrophilic polymer comprises hyaluronic acid.

[0156] The second hydrophilic polymer can have a varying molecular weight. For example, the second hydrophilic polymer can have a number average molecular weight of about 0.5 kDa to about 500 kDa, such as about 1 kDa to about 400 kDa, about 10 kDa to about 300 kDa, about 20 kDa to about 250 kDa, about 50 kDa to about 150 kDa, about 5 kDa to about 300 kDa, about 0.5 kDa to about 250 kDa, or about 200 kDa to about 500 kDa. In some embodiments, the second hydrophilic polymer has a number average molecular weight of greater than 0.5 kDa, greater than 10 kDa, greater than 50 kDa, greater than 70 kDa, greater than 100 kDa, greater than 200 kDa, or greater than 300 kDa. In some embodiments, the second hydrophilic polymer has a number average molecular weight of less than 500 kDa, less than 400 kDa, less than 300 kDa, less than 200 kDa, less than 150 kDa, less than 100 kDa, or less than 50 kDa. In some embodiments, the second hydrophilic polymer is hyaluronic acid having a number average molecular weight of about 50 kDa to about 150 kDa.

[0157] The composition can include a guest-host grafting density that provides advantageous mechanical properties, such as shear thinning. For example, the secondAttorney Docket No.093386-0041-WO01 hydrophilic polymer can have a guest-host grafting density of about 10% to about 40% such as about 15% to about 35%, about 10% to about 30%, about 10% to about 20%, or about 20% to about 40%. In some embodiments, the second hydrophilic polymer has a guest-host grafting density of greater than 10%, greater than 15%, greater than 20%, or greater than 25%. In some embodiments, the second hydrophilic polymer has a guest-host grafting density of less than 40%, less than 35%, less than 30%, or less than 25%. The guest-host grafting density is measured as the mol% of the monomer repeats in the third block or in the multi-unit segment of formula (IV) that include the guest-host moiety (e.g., the guest-host recurring unit). The guest- host grafting density can be measured via NMR.

[0158] The guest-host moieties of the block copolymer and the second hydrophilic polymer can be added in an amount that provides useful mechanical properties for the composition. For example, the composition can have a ratio of the guest-host moiety of the block copolymer to the guest host-moiety of the second hydrophilic polymer of about 1:4 to about 4:1 (block copolymer:second hydrophilic polymer). In some embodiments, the composition has a ratio of the guest-host moiety of the block copolymer to the guest host-moiety of the second hydrophilic polymer of about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, or about 4:1 (block copolymer:second hydrophilic polymer).

[0159] As discussed elsewhere herein, the particle can include a drug. In some embodiments, at least one particle of the composition includes a drug. In some embodiments, each of the particles of the composition includes a drug. Including drug laden particles can be beneficial for methods of treatment as disclosed herein.

[0160] The composition can have mechanical properties that allow it to be shear thinning. For example, the composition can have a storage modulus (G’) of about 500 Pa to about 5,000 Pa, such as about 700 Pa to about 4,500 Pa, about 800 Pa to about 4,000 Pa, about 900 Pa to about 3,500 Pa, about 1,000 Pa to about 5,000 Pa, about 1,500 Pa to about 5,000 Pa, or about 500 Pa to about 3,000 Pa. In some embodiments, the composition has a storage modulus (G’) of greater than 500 Pa, greater than 700 Pa, greater than 1,000 Pa, greater than 1,500 Pa, greater than 2,000 Pa, or greater than 3,000 Pa. In some embodiments, the composition has a storage modulus (G’) of less than 5,000 Pa, less than 4,500 Pa, less than 4,000 Pa, less than 3,500 Pa, less than 3,000 Pa, or less than 2,000 Pa. Storage modulus can be measured as described in the Examples.

[0161] The composition can also be referred to as a hydrogel.

[0162] Compositions (e.g., hydrogels) can be made through mixing of individual triblock polymer NPs grafted with guest-host moiety (e.g., adamantane (AD)) with a hydrophilic polymerAttorney Docket No.093386-0041-WO01 (e.g., hyaluronic acid) grafted with a guest-host moiety (e.g., -cyclodextrin) (HA-CD) (Component B) at varying stoichiometric ratios of AD to CD. Example ratios between CD and AD include, but are not limited to, 1:2, 1:1, and 2:1.

[0163] An example procedure for preparation of a shear-thinning composition including nanoparticles self-assembled from PPS D300-AD20%triblock copolymers and HA-CD20%mixed at a 2CD / 1AD ratio is described as follows. Briefly, in separate Eppendorf tubes (2 mL), lyophilized PPS D300-AD20%(5 w / v%) and HA-CD20%(8.75 w / v%) can be dissolved into a buffer (e.g., PBS) and placed on a vortex mixer for a period of time (e.g., 24 hours). After the period of time, PPS D300-AD20%triblock copolymer nanoparticles (50 L) and HA-CD20%polymer (50 L) can be transferred to the same 2 mL centrifuge tube, where they can spontaneously form a shear- thinning hydrogel upon mixing both components with, e.g., a positive displacement pipette. The final shear-thinning composition can be centrifuged for 10 minutes at 12,000 rpm. The vial inversion method can be used to confirm composition formation, which can be done at 37° Celsius.

[0164] The description of the block copolymers, e.g., first block, second block, third block, particles, and drugs above can be applied to the disclosed compositions. 4. Methods

[0165] Further disclosed are methods of treating a disease or disorder in the subject (e.g., in need thereof). The method can include administering to the subject a therapeutically effective amount of a composition as disclosed herein. Example diseases include, but are not limited to, osteoarthritis, rheumatoid arthritis, ulcerative colitis, and wound healing. In some embodiments, the disease or disorder includes wound healing. In some embodiments, the subject is human.

[0166] Also disclosed are methods of tissue repair in a subject (e.g., in need thereof). The method can include administering to the subject a therapeutically effective amount of a composition as disclosed herein. In addition, the method of tissue repair can include treating a volumetric tissue defect in the subject, which can be human.

[0167] An example of wound healing and / or tissue repair relates to diabetic ulcers. In the acute phases of wound healing in healthy tissue, reactive oxygen species (ROS) can be produced to fight microorganisms, prevent infection, and promote immune cell activation. However, persistent pathologic hyperglycemia in diabetic skin wounds can impair the antioxidant function of incoming immune cells and can produce dysfunctional immune cells, such as pro-inflammatory macrophages and neutrophils, which can sustain overproduction of ROS, resulting in an imbalance of redox signaling and elevating oxidative stress in the wound microenvironment. Biomaterial antioxidants, such as those disclosed herein, are a promisingAttorney Docket No.093386-0041-WO01 therapeutic treatment for addressing sustained levels of oxidative stress in, e.g., diabetic skin wounds. Although few antioxidant-based therapies have been pursued for diabetic wound healing, several preclinical studies in diabetic wound healing models have demonstrated that restoring antioxidant function reduces elevated levels of oxidative stress and can accelerate wound closure of non-healing diabetic wounds.

[0168] The description of the block copolymers, first block, second block, third block, particles, drugs, and compositions above can be applied to the disclosed methods. A. Administration

[0169] The composition can be administered prophylactically or therapeutically. In prophylactic administration, the composition can be administered in an amount sufficient to induce a response. In therapeutic applications, the composition can be administered to a subject in need thereof in an amount sufficient to elicit a therapeutic effect. An amount adequate to accomplish this is defined as “therapeutically effective dose.” Amounts effective for this use will depend on, e.g., the particular composition of the regimen administered, the manner of administration, the stage and severity of the disease, the general state of health of the patient, and the judgment of the prescribing physician.

[0170] The composition may be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub- dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.

[0171] As will be readily apparent to one skilled in the art, the useful in vivo dosage to be administered and the particular mode of administration will vary depending upon the age, weight, the severity of the affliction, and subjects treated, the particular compounds employed, and the specific use for which these compounds are employed. The determination of effective dosage levels, that is the dosage levels necessary to achieve the desired result, can be accomplished by one skilled in the art using routine methods, for example, human clinical trials, in vivo studies and in vitro studies.

[0172] The compositions can be administered via a variety of routes. Typical delivery routes include parenteral administration, e.g., intradermal, intramuscular or subcutaneous delivery. Other routes include intravaginal, intratumoral, intraperitoneal, and epidermal routes. In some embodiments, the composition is administered subcutaneously, intradermally, intramuscularly, or intraperitoneally. In some embodiments, the composition is administered topically.

[0173] Dosage amount and interval may be adjusted individually to provide plasma levels of the drug which are sufficient to maintain the modulating effects, or minimal effectiveAttorney Docket No.093386-0041-WO01 concentration (MEC). The MEC will vary for each agent but can be estimated from in vivo and / or in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, assays well known to those in the art can be used to determine plasma concentrations. Dosage intervals can also be determined using MEC value. Compositions can be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, such as between 30-90% or between 50-90%. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

[0174] It should be noted that the attending physician would know how to and when to terminate, interrupt, or adjust administration due to toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response were not adequate (precluding toxicity). The magnitude of an administrated dose in the management of the disorder of interest will vary with the severity of the symptoms to be treated and the route of administration. Further, the dose, and perhaps dose frequency, will also vary according to the age, body weight, and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.

[0175] The disclosed technology has multiple aspects, illustrated by the following non-limiting examples. 5. Examples Example 1 Synthetic Methods Materials

[0176] Unless stated otherwise, all reagents were purchased from Sigma-Aldrich (Milwaukee, WI, USA) or Fisher Scientific (Pittsburgh, PA, USA). Sodium hyaluronate (HA, 85 KDa) was obtained from Lifecore Biomedical (Chaska, MN). The compound 2,2'-Azobis(4- methoxy-2,4-dimethylvaleronitrile) (V-70) was acquired from Wako Pure Chemical Industries,Ltd. 2 -Azoisobutyronitrile (AIBN) was recrystallized from methanol at 45 degrees Celsius. 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) was dried and distilled over anhydrous calcium hydride(CaH ). Propylene sulfide (PS) and N, N-dimethyl acrylamide (DMA) was purified by vacuumdistillation and stored at -20 °C. The synthesis of 4-Cyano-4-(ethyl sulfanyl thiocarbonyl) sulfanyl pentanoic acid (ECT) and the hyaluronic acid tetra butyl ammonium (HA-TBA) salt and -Attorney Docket No.093386-0041-WO01 cyclodextrin (CD) mono-functionalized with hexyl methylene diamine (CD-HDA) and N-(1- adamantyl)acrylamide (Ad-AAm) monomer were carried out as outlined in the literature. Synthesis of benzyl glycidyl thioether (PSBn) monomer

[0177] The synthesis of the benzyl glycidyl thioether (PSBn) monomer was accomplished as follows (FIG.6). In a dry, 250 mL round-bottom flask equipped with a water condenser, 25 g (152.26 mmol) of benzyl glycidyl ether, 23.2 g (304.52 mmol, two eq) of thiourea, and 8.1 g (152.25 mmol) of ammonium chloride were combined with 250 mL of methanol. The solution was degassed for 5 minutes. The reaction assembly was placed in a preheated oil bath and maintained at 60 degrees Celsius for 90 minutes. Once the reaction was complete, the mixture was allowed to cool to room temperature, and the solvent was evaporated using a rotary evaporator. The crude product was dissolved in a 3:1 mixture of diethyl ether (150 mL) and deionized water (50 mL). The organic layer was washed three times with water, followed by a wash with brine. The separated organic layer was dried over sodium sulfate and concentrated under a vacuum. Finally, the crude mixture was distilled under vacuum to yield benzyl glycidyl thioether. The representative1H NMR spectra is as follows:1H NMR (400 MHz, CDCl3): (ppm) 2.63 (1H, -CH2-S-), 2.85 (1H, S-CH), 3.20 (1H, S-CH2), 3.45 (1H, CH2-O-CH2-C6H5), 3.77 (1H, CH2-O-CH2-C6H5), 4.47 (m, 2H, methylene, O-CH2-C6H5), 7.35 (s, 5H, Phenyl). Synthesis of tert-butyldimethylsilyl glycidyl thioether (PS-OTBDMS) monomer

[0178] The synthesis of the hydroxyl-protected PS-OTBDMS monomer was accomplished using the method described above for Synthesis of benzyl glycidyl thioether (PSBn) monomer, with ter-butyldimethylsilyl glycidyl ether used as starting materials instead place of benzyl glycidyl ether (FIG.6). The representativeNMR spectra is as follows:1H NMR (400 MHz, CDCl3): (ppm) 0.09 (m, 6H, two methyl), 0.91 (m, 9H, t-butyl), 2.20 (1H, -CH2-S-), 2.49 (1H, S-CH), 3.05 (1H, S-CH2), 3.57 (1H, CH2-OSi), 3.87 (1H, CH2-OSi-). Synthesis of N-(1-adamantyl)acrylamide (Ad-AAm)

[0179] 1-Adamantylamine (100 mM, 15.12 g) and triethylamine (110 mM, 11.13 g, 15.34 mL) were dissolved in dry THF (300 mL) in a 500 mL flask and then cooled over an ice bath. Acryloyl chloride (110 mM, 9.95 g, 8.73 mL) was added dropwise to this solution at 0 degrees Celsius while stirring, and then this solution was stirred at room temperature for 24 hours. The next day, the precipitate was filtered, and the supernatant was concentrated through rotatory evaporation. The crude product was purified using 2% methanol in DCM through a silica gel column. TheAttorney Docket No.093386-0041-WO01 representative1H NMR spectra is as follows:1H NMR (400 MHz, CDCl3): 6.22 (dd, 1H, double bond), 6.03 (m, 1H, double bond), 5.55 (dd, 1H, double bond), 5.1-5.3 (bs, 1H, NH), 2.06 (m, 9H, adamantane), 1.59 (m, 6H, adamantane). Synthesis of PPS135-b-PDMA300-b-P(DMA165-co-ADA46)

[0180] The synthesis of the triblock copolymer PPS135-b-PDMA300-b-P(DMA165-co-ADA46) (FIG.7C) was accomplished by combination of anionic ring opening and RAFT polymerizations. The synthesis protocol for each step for triblock copolymers and precursor polymers is summarized below. Synthesis of hydroxyl end-capped poly (propylene sulfide) (PPS135-OH)

[0181] The ring-opening polymerization of three-membered cyclic propylene sulfide (PS) was performed using DBU / 1-butane thiol as an initiator, followed by end-capping of the thiol with 2- iodo ethanol. Briefly, DBU (2 mmol, 0.30 g, 0.299 mL) and anhydrous THF (10 mL) were added to a 25 mL round bottom (RB) flask equipped with rubber septum and then purged with nitrogen for 30 minutes. A separate 100 mL RB flask was oven dried, degassed, and fitted with a rubber septum.1-Butane thiol (0.67 mmol, 0.082 g, 0.092 mL) was transferred in a solution of degassed, anhydrous THF (30 mL) to this 100 mL RB flask and then cooled in an ice bath at 0 degrees Celsius. A DBU solution in THF was added dropwise to this flask and allowed to react for 30 minutes at room temperature. Next, degassed propylene sulfide (135 mmol, 9.98 g, 10.56 mL) monomer was added slowly over the course of 5 minutes to the polymerization mixture in this RB flask, with the temperature of this polymerization mixture maintained at room temperature for the following 2 hours. The active polymer chains were end-capped through the addition of 2-iodoethanol (2.66 mmol, 0.46 g, 0.21 mL) to the polymerization mixture, which was then left overnight, stirring at RT. The precipitated salt in the crude polymerization mixture was removed through filtration, and the final product was concentrated through solvent removal by rotovap. The resulting viscous polymer was dissolved in a minimal amount of CH2Cl2, purified by three precipitations into a ten-fold excess of cold methanol, and dried under high vacuum at 40 degrees Celsius to yield a colorless, viscous polymer. The representative1H NMR spectra of PPS135-OH is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.35- 1.45 (s, CH3, side chain), 2.5-2.8 (s, -CH, main backbone), 2.8-3.1 (s, CH2, main backbone), 3.72 (t, CH2-OH, at the terminal end). To calculate the number of propylene sulfide units in PPS, the integration was set to 2 at 3.72 ppm (CH2-OH, methylene protons at terminal end) to determine the integration areaAttorney Docket No.093386-0041-WO01 for side chain methyl groups from 1.35-1.45 ppm. The resulting number was divided by three, which provided the final degree of polymerization. Synthesis of PPS-based RAFT macro-CTA (PPS135-ECT)

[0182] Chain extension of PPS to create block copolymers by RAFT polymerization was accomplished with PPS135-ECT created by conjugation of PPS135-OH to the RAFT chain transfer agent ECT (4-cyano-4-(ethylsulfanylthiocarbonyl) sulfanylvpentanoic acid) using a DCC / DMAP conjugation method. A solution of N,N'-dicyclohexyl carbodiimide (DCC) (3.27 mmol, 0.67 g) in DCM (3 mL) was added dropwise to a 100 mL RB flask containing a solution of PPS135-OH (0.81 mmol, 8.17 g), ECT (3.27 mmol, 0.85 g), and 4-dimethylaminopyridine (DMAP) (0.33 mmol, 0.039 g) in anhydrous DCM (20 mL) cooled to 0 degrees Celsius. White dicyclohexyl urea (DCU) precipitates out from the solution as the esterification-mediated PPS-ECT reaction proceeds. Following the addition of DCC, the solution was brought to room temperature and stirred in the dark for the next 24 hours. After 24 hours, the yellow polymer solution was filtered to remove any insoluble products, and the resulting solution was concentrated by rotovap. The crude yellow macro-CTA was then dissolved in DCM (10 mL) and precipitated into a ten-fold excess of cold methanol. The precipitation process was repeated three times to ensure the complete removal of small molecule impurities. The recovered polymer was then dried under a high vacuum at 40 degrees Celsius for 2 hours. The representative1H NMR spectra of PPS135-ECT is as follows:(ppm) 1.35 (t, 3H, S CH2 CH3, ECT), 1.35-1.45 (s, 3H, CH3, PPS sidechain), 1.85 (s C(CN) CH3, ECT), 2.4 2.67 (m, CH2 CH2 S,ECT), 2.5-2.8 (broad s, S-CH, PPS backbone), 2.8-3.1 (broad s, 2H, CH2, PPS backbone), 3.42(q, S CH2 CH3, ECT), 4.2 (t, -OCH2-CH2, ester). The conjugation of ECT to PPS135-OH wascalculated as described below. The integration was set to 2 at 3.34 ppm (S-CH2-CH3, methylene near trithiocarbonate group) in order to calculate the integration area for side chain PPS methyl groups from 1.35-1.45 ppm. The resulting number was divided by three, indicating quantitative conjugation of ECT to the polymer. Synthesis of PPS135-b-PDMAx-ECT

[0183] The PPS-ECT macro-RAFT CTA was used to synthesize three different linear diblock copolymers of PPS135-b-PDMAxwhere x = 150, 200, and 300 by RAFT polymerization of DMA using V-70 as an initiator. PPS135-ECT (0.02 mmol, 0.2 g) and DMA (3 mmol, 0.297 g, 0.31 mL) were dissolved in 1,4-dioxane (5 mL) and added to a 100 mL RB flask which was then degassed for 5 minutes. A solution of V-70 (0.002 mmol, 0.62 mg, 67 L from freshly preparedAttorney Docket No.093386-0041-WO01 10 mg / mL solution) in 1,4-dioxane was added to this polymerization mixture which was then purged with high-purity nitrogen for 30 minutes. The polymerization mixture was then submerged in a preheated oil bath at 35 degrees Celsius and allowed to react for 24 hours. Following 24 hours, the crude yellow viscous copolymer was purified through three repeated precipitations into cold diethyl ether. The precipitated polymer was recovered through centrifugation and then dried overnight under a vacuum. The representative1H NMR spectra of PPS135-b-PDMA147-ECT is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.35-1.45 (s, CH3in pendent in PPS block), 1.2-1.75 (-CH2main backbone PDMA block), 2.5-2.7 (-CH backbone PDMA block), 2.5-2.8 (broad s, CH in PPS block), 2.8-3.1 (broad s, CH2, PPS block), 2.9-3.3 (dimethyl group PDMA block). To determine the number of DMA units, the integration of the PPS10K peak at 1.35-1.45 ppm was set to 405 (135 units X 3H from methyl groups). This was then used to calculate the number of units of DMA in the diblock copolymer. After the relative integration of the area from 3.2-2.5 ppm was determined, the integration area of 405 (CH2and CH from the PPS backbone) was subtracted from this integration area from 3.2-2.5 ppm and the resulting number was divided by six, representing the protons of the NMe2group from DMA. This calculation provides the total number of DMA units in the diblock copolymer. Synthesis of PPS135-b-PDMAx-b-P(DMA-co-ADA)z

[0184] The PPS135-b-PDMAxwhere x = 147, 204, and 293 diblock copolymer macro-CTAs were further RAFT chain extended through random copolymerization of DMA and ADA to achieve triblock copolymers of PPS135-b-PDMAx-b-P(DMA-co-ADA)zwherein the poly(DMA-co- ADA) block has a total degree of polymerization of 200. The molar percentage of AD incorporated was varied to be either 10% (equivalent to 20 units of AD) or 20% (equivalent to 40 units of AD) relative to DMA. The representative procedure for synthesizing one candidate triblock copolymer of PPS135-b-PDMA147-b-P(DMA188-co-ADA24) is as follows. A solution of PPS135-b-PDMA147-ECT (0.012 mmol, 0.3 g), DMA (2.13 mmol, 2.16 g, 0.222 mL), adamantyl acrylamide (0.024 mmol, 0.049 g), and V-70 (0.0024 mmol, 0.74 mg) in 1,4-dioxane (5 mL) was placed in a 50 mL RB flask which was then sealed with a rubber septum. The solution was purged with ultra-high purity nitrogen for 30 minutes. The polymerization flask was then submerged in a preheated oil bath at 35 degrees Celsius for 24 hours. Following 24 hours, the viscous polymer solution was dissolved into DCM (5 mL) and purified through precipitation into cold diethyl ether. The polymer was precipitated into cold diethyl ether three times, and the precipitated pale-yellow polymer was then dried under a vacuum. To further purify the precipitated polymer, the product was dissolved into THF and dialyzed against methanol for 24Attorney Docket No.093386-0041-WO01 hours, followed by dialysis against DI water for 48 hours. The DI water was changed at least three times during dialysis. The dialyzed polymer solution was then lyophilized to afford a white, fleecy product. The representative1H NMR spectra of PPS135-b-PDMA291-b-P(DMA165-co-ADA46) is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.19-1.29 (CH2main backbone ADA), 1.35- 1.45 (s, CH3in pendent in PPS block), 1.2-1.75 (-CH2main backbone PDMA block), 1.51-1.78 (b, pendent AD protons), 1.92 (pendent AD protons) 2.5-2.7 (-CH backbone PDMA block), (-CH main backbone ADA), 2.5-2.8 (broad s, CH in PPS block), 2.8-3.1 (broad s, CH2, PPS block), 2.9-3.3 (dimethyl group PDMA block). To determine the number of DMA and AD units in the third DMA-co-ADA copolymer block, the number of DMA units in the DMA-co-ADA block was determined in an identical manner as described above for determination of the total number of DMA units for the diblock copolymer of PPS-b-PDMA. This calculation provided the total number of DMA units present in the triblock from the B and C blocks. Next, the peaks from 2.3- 1.0 ppm area were integrated relative to 3.3-2.5 ppm. This value subtracted the integration from the PPS methyl protons and DMA backbone (methine and methylene backbone protons) based on calculating the DMA units in the 3.2-2.5 ppm region. The resulting number was divided with 18 protons (15 ADA moiety and 3 from the backbone) which provided the number of AD units in the DMA-co-ADA (C) block. Synthesis of PCL87-b-PDMA291-b-P(DMA167-co-ADA47)

[0185] The synthesis of the triblock copolymer PCL87-b-PDMA291-b-P(DMA167-co-ADA47) was accomplished by combination of anionic ring opening and RAFT polymerizations. The synthesis protocol for each step for triblock copolymers and precursor polymers is summarized below. Synthesis of hydroxyl functional poly ( -caprolactone) (PCL87-OH)

[0186] Hydroxyl functional PCL87-OH was synthesized by ring-opening polymerization (ROP) of seven-membered -caprolactone using benzyl alcohol as the initiator and Tin (II) octoate as the catalyst. The representative1H NMR spectra of PCL87-OH is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.37 (m, 2H, -CH2CH2CH2CH2CH2COO-), 1.62 (m, 4H, - CH2CH2CH2CH2CH2COO-), 2.28 (t, 2H, -CH2CH2CH2CH2CH2COO-), 4.04 (t, 2H, - CH2CH2CH2CH2CH2OC(O))-), 5.2 (s, 2H, CH2-C6H5), 7.35 (s, 2H, CH2-C6H5). Synthesis of PCL-based RAFT macro-CTA) (PCL87-ECT)

[0187] To build a triblock copolymer with PCL as the core-forming unit, ECT RAFT CTA was conjugated through an esterification reaction described above for synthesis of PPS135-ECT. TheAttorney Docket No.093386-0041-WO01 representative1H NMR spectra of PCL87-ECT is as follows:1H NMR (400 MHz, CDCl3): (ppm)1.35 (t, 3H, S CH2 CH3, ECT), 1.37 (m, 2H, -CH2CH2CH2CH2CH2COO-), 1.62 (m, 4H, -CH2CH2CH2CH2CH2COO-), 1.85 (s C(CN) CH3, ECT), 2.28 (t, 2H, -CH2CH2CH2CH2CH2COO-), 2.4 2.67 (m, CH2 CH2 S, ECT), 3.42 (q, S CH2 CH3, ECT),4.04 (t, 2H, - CH2CH2CH2CH2CH2OC(O))-), 5.2 (s, 2H, CH2-C6H5), 7.35 (s, 2H, CH2-C6H5). Synthesis of diblock copolymer PCL87-b-PDMA291-ECT

[0188] The precursor diblock copolymer based on PCL was RAFT synthesized similarly to the method reported for the Synthesis of PPS135-b-PDMAx-ECT. The representative1H NMR spectra of PCL87-b-PDMA291-ECT is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.2-1.75 (-CH2 main backbone PDMA block), 1.35 (t, 3H, S CH2 CH3, ECT), 1.37 (m, 2H, -CH2CH2CH2CH2CH2COO-, PCL block), 1.62 (m, 4H, -CH2CH2CH2CH2CH2COO-, PCL block),1.85 (s C(CN) CH3, ECT), 2.28 (t, 2H, -CH2CH2CH2CH2CH2COO- PCL block), 2.4 2.67 (m,CH2CH2S, ECT), 2.5-2.7 (-CH backbone PDMA block), 2.9-3.3 (dimethyl group PDMAblock), 3.42 (q, S CH2 CH3, ECT), 4.04 (t, 2H, - CH2CH2CH2CH2CH2OC(O))-, PCL block).Synthesis of triblock copolymer of PCL87-b-PDMA291-b-P(DMA167-co-ADA47)

[0189] The control PCL-based triblock copolymer was RAFT synthesized by chain extension of PCL87-b-PDMA291-ECT as the macro-CTA according to the method described above for Synthesis of PPS135-b-PDMA293-b-P(DMA164-co-ADA41). The representative1H NMR spectra of PCL87-b-PDMA291-b-P(DMA167-co-ADA47) is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.19-1.29 (CH2main backbone ADA), 1.37 (m, 2H, -CH2CH2CH2CH2CH2COO-, PCL block), 1.2- 1.75 (-CH2main backbone PDMA block), 1.51-1.78 (b, pendent AD protons), 1.62 (m, 4H, - CH2CH2CH2CH2CH2COO-, PCL block), 1.92 (pendent AD protons), 2.28 (t, 2H, - CH2CH2CH2CH2CH2COO-, PCL block), 2.5-2.7 (-CH backbone PDMA block), (-CH main backbone ADA), 2.9-3.3 (dimethyl group PDMA block), 4.04 (t, 2H, - CH2CH2CH2CH2CH2OC(O))-, PCL block). Synthesis of poly(PSn%-co-PSBnm%)-b-PDMA300-b-P(DMA165-co-ADA46)

[0190] The synthesis of the P(PSn%-co-PSBnm%) D300-AD20%triblock copolymers of the form poly(PSn%-co-PSBnm%)-b-PDMA300-b-P(DMA165-co-ADA46) where n=80%, 50%, 30%, and 0% and m=20%, 50%, 70%, and 100% was accomplished by combination of anionic ring opening and RAFT polymerizations. The synthesis protocol for each step for triblock copolymers and precursor polymers is summarized below.Attorney Docket No.093386-0041-WO01 Synthesis of hydroxyl end functional poly(PSn%-co-PSBnm%)

[0191] The random copolymerization of PS and PSBn was performed analogously to PPS135- OH synthesis. Four different molar feed ratios of PSBn relative to PS (20% PSBn, 50% PSBn, 70% PSBn, and 100% PSBn) were used for polymerization using a 1-butanethiol / DBU system to prepare poly(PSn%-co-BnPSm%) where m = 20, 50, 70, and 100% relative to PS monomer. The representative1H NMR spectra of the various poly(PSn%-co-PSBnm%) random copolymer monomers is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.34- 1.46 (s, CH3, pendent), 2.45- 2.82 (s, -CH, polymer backbone), 2.82-3.15 (s, CH2, polymer backbone), 3.75 (t, CH2-OH, at the terminal end), 3.5-3.75 (doublet, pendent CH2between main backbone and OCH2C6H5), 4.47 (m, 2H, methylene, O-CH2-C6H5), 7.35 (s, 5H, C6H5ring). Synthesis of poly(PSn%-co-PSBnm%)-ECT

[0192] The attachment of ECT to respective hydroxyl end functional poly(PSn%-co-PSBnm%) was performed using the protocol outlined for PPS135-ECT where m=20%, 50%, 70%, and 100%, and n=80%, 50%, 30%, and 0%. The representative1H NMR spectra of the poly(PS80%- co-PSBn20%)-ECT monomer is as follows:1H NMR (400 MHz, CDCl3): 1.35 (t, 3H, SCH2 CH3, ECT), 1.3-1.4 (s, 3H, CH3, PPS side chain), 1.85 (s, C(CN) CH3, ECT), 2.4 2.67(m, CH2CH2S, ECT), 2.5-2.8 (broad s, S-CH, PPS backbone), 2.8-3.1 (broad s, 2H, CH2,PPS backbone), 3.42 (q, S CH2 CH3, ECT), and 3.5-3.75 (doublet, pendent CH2 betweenmain backbone and OCH2C6H5), 4.2 (t, -COOOCH2-CH2, ester), 4.47 (m, 2H, methylene, O- CH2-C6H5), 7.35 (s, 5H, C6H5ring). Synthesis of poly(PSn%-co-PSBnm%)-b-PDMA300-ECT

[0193] The respective diblock copolymers with different molar feed ratios of PS and PSBn monomers were prepared through RAFT polymerization of DMA using respective RAFT macro- CTA poly(PSn%-co- BnPSm%)-ECT analogously as reported for synthesizing PPS135-b-PDMA300- ECT where m=20%, 50%, 70%, and 100% and n=80%, 50%, 30%, and 0%. The representative1H NMR of the poly(PS80%-co-PSBn20%)-b-PDMA300diblock copolymer is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.3-1.4 (s, CH3in pendent in PPS ), 1.2-1.75 (-CH2main backbone PDMA), 2.5-2.7 (-CH backbone PDMA), 2.5-2.8 (broad s, CH in PPS backbone), 2.8-3.1 (broad s, CH2, PPS backbone, 2.89-3.31 (pendent N(CH3)2group PDMA), 3.5-3.75 (doublet, pendent CH2between main backbone and OCH2C6H5), 4.47 (m, 2H, methylene, O-CH2-C6H5), 7.35 (s, 5H, C6H5ring).Attorney Docket No.093386-0041-WO01 Synthesis of poly(PSn%-co- PSBnm%)-b-PDMA300-b-P(DMA160-co-ADA40)

[0194] The respective triblock copolymers with different molar feed ratios of PS and PSBn monomers were prepared through RAFT polymerization of DMA and ADA using diblock copolymer poly(PSn%-co-PSBnm%)-b-PDMA300-ECT analogously as reported for synthesizing PPS135-b-PDMA300-b-P(DMA160-co-ADA40). The representative1H NMR for the poly(PS80%-co- PSBn20%-)-b-PDMA300-b-P(DMA160-co-ADA40) triblock copolymer is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.19-1.29 (CH2main backbone ADA), 1.3-1.4 (s, CH3in pendent in PPS block), 1.2-1.75 (-CH2main backbone PDMA block), 1.51-1.78 (b, pendent AD protons), 1.92 (pendent AD protons) 2.5-2.7 (-CH backbone PDMA block), (-CH main backbone ADA), 2.5-2.8 (broad s, CH in PPS block), 2.8-3.1 (broad s, CH2, PPS block), 2.9-3.3 (dimethyl group PDMA block), 3.5-3.75 (doublet, pendent CH2between main backbone and OCH2C6H5), 4.47 (m, 2H, methylene, O-CH2-C6H5), 7.35 (s, 5H, C6H5ring). Synthesis of poly(PSn%-co-PSOHm%)10K-b-PDMA300-b-P(DMA160-co-ADA40)

[0195] The synthesis protocol for each step for P(PSn%-co-PSOHm%) D300-AD20%triblock copolymers of the form poly(PSn%-co- PSOHm%)10K-b-PDMA300-b-P(DMA160-co-ADA40) and precursor polymers is summarized below where n=80%, 50%, and 30% and m=20%, 50%, and 70%. Synthesis of Boc-amine end functional poly(PSn%-co- PS-OTBDMSm%)-Boc

[0196] The random copolymerization of PS and PSOH was performed analogously to PPS135-OH synthesis. A library of three different (hydroxyl-protected poly(PSn%-co- PS- OTBDMSm%) copolymers were prepared with three different molar feed ratios of PSBn relative to PS-OTBDMS (20% PSOH, 50% PSOH, 70% PSOH) using a ring-opening random copolymerization of PS and PS-OTBDMS similar to the methods described above for Synthesis of PPS135-OH. The polymerization was quenched using tert-butyl N-(2-bromoethyl) carbamate to have terminal Boc-protected amine. The representative1H NMR for the various poly(PSn%-co- PS-OTBDMSm%)-Boc random copolymer monomers is as follows:1H NMR (400 MHz, CDCl3): (ppm) 0.09 (m, 6H, two methyl from TBDMS), 0.91 (m, 9H, t-butyl from TBDMS), 1.34- 1.46 (s, CH3, pendent), 2.45-2.82 (s, -CH, PPS and PS-OTBDMS polymer backbone), 2.82-3.15 (s, CH2, PPS and PS-OTBDMS polymer backbone), 3.65-3.85 (2H, doublet, pendent -CH2- OTBDMS). Synthesis the amine functional poly(PSn%-co-PSOHm%)-NH2Attorney Docket No.093386-0041-WO01

[0197] To obtain the various amine-functional poly(PSn%-co-PSOHm%) copolymer monomers from the library of poly(PSn%-co-PS-OTBDMSm%) monomers, the polymers were dissolved in THF and placed over activated Dowex (50WX8-200) ion exchange resin and heated at 50 degrees Celsius for 3 days. After, the deprotected polymers were filtered to remove the resin, and the solution was dialyzed against methanol to eliminate impurities. The dialyzed solution was then concentrated using a rotary evaporator, and the obtained polymer was dried under vacuum overnight in order to deprotect the TBDMS groups and removes the terminal Boc group, resulting in amine-terminated poly(PSn%-co-PSOHm%). The successful deprotection of these polymers was confirmed by the disappearance of the TBDMS peaks at 0.1 ppm and 1.1 ppm. The representative1H NMR for the poly(PS50%-co-PSOH50%)-NH2monomer is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.34- 1.46 (s, CH3, pendent), 2.45-2.82 (s, -CH, PPS and PS-OTBDMS polymer backbone), 2.82-3.15 (s, CH2, PPS and PPS-OTBDMS polymer backbone), 3.65-3.85 (bs, 2H, pendent -CH2-OH). Synthesis of poly(PSn%-co-PSOHm%)10K-ECT

[0198] The RAFT macro-CTA of these polymers was prepared using the reaction of an activated NHS ester with respective an amine end functional polymer. The representative procedure for poly(PS50%-co-PSOH50%)-ECT is as follows. Briefly, poly(PS50%-co-PSOH50%)-NH2(0.50 g, 0.05 mmol) and TEA (0.3 mmol) were dissolved in chloroform. To this solution, NHS- ECT (0.09 g, 0.25 mmol) in CHCl3was added slowly at 0 degrees Celsius. The solution was stirred overnight at room temperature. The next day, the crude mixture was concentrated on rotavapor. The viscous solution was dialyzed against methanol for 48 hours. The dialyzed solution was concentrated, and the yellow viscous polymer was dried under vacuum. The representativeNMR spectra of the poly(PS50%-co-PSOH50%)-ECT monomer is as follows:1HNMR (400 MHz, CDCl3) (ppm): 1.35 (t, 3H, S CH2 CH3, ECT), 1.3-1.4 (s, 3H, CH3, pendentmethyl in PPS), 1.85 (s, C(CN) CH3, ECT), 2.4 2.67 (m, CH2 CH2 S, ECT), 2.5-2.8 (broad s,S-CH, PPS backbone and PPSOH backbone), 2.8-3.1 (broad s, 2H, CH2, PPS backbone andPPSOH backbone), 3.42 (q, S CH2 CH3, ECT), 3.65-3.85 (2H, doublet, PPSOH pendent CH2-OH, ) and 4.2 (t, -COOCH2-CH2, ester). Synthesis of poly(PSn%-co-PSOHm%)10K-b-PDMA300-ECT

[0199] The respective diblock copolymers with different molar feed ratios of PS and PSOH were prepared using respective macro RAFT CTA of poly(PSn%-co-PSOHm%)-ECT as reported for synthesizing PPS135-b-PDMA300-ECT. The representative1H NMR spectra of the poly(PS50%-Attorney Docket No.093386-0041-WO01 co- PSOH50%)-b-PDMA300diblock copolymer is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.3-1.4 (s, CH3in pendent in PPS polymer), 1.2-1.75 (-CH2main backbone PDMA polymer), 2.5-2.7 (-CH backbone PDMA), 2.5-2.8 (broad s, CH in PPS backbone), 2.8-3.1 (broad s, CH2, PPS backbone, 2.89-3.31 (pendent N(CH3)2group, PDMA).2.45-2.82 (s, -CH, PPS, and PPSOH polymer backbone), 2.82-3.15 (s, CH2, PPS, and PPSOH polymer backbone), 3.65- 3.85 (bs, 2H, PPSOH pendent methylene -CH2-OH). Synthesis of poly(PSn%-co-PSOHm%)10K-b-PDMA300-b-P(DMA160-co-ADA40)

[0200] The respective triblock copolymers with different molar ratios of PS and PSOH monomers were prepared through RAFT polymerization of DMA and ADA using diblock copolymer poly(PSn%-co-PSOHm%)-b-PDMA300-ECT analogously as reported for synthesizing PPS135-b-PDMA300-b-P(DMA165-co-ADA46). The representative1H NMR spectra of the poly(PS50%-co-PSOH50%)10K-b-PDMA300-b-P(DMA160-co-ADA40) triblock copolymer is as follows:1H NMR (400 MHz, CDCl3): (ppm) 1.19-1.29 (CH2main backbone ADA), 1.3-1.4 (s, CH3in pendent in PPS block), 1.2-1.75 (-CH2main backbone PDMA block), 1.51-1.78 (b, pendent AD protons), 1.92 (pendent AD protons) 2.5-2.7 (-CH backbone PDMA block), (-CH main backbone ADA), 2.45-2.82 (s, -CH, PPS, and PS-OTBDMS polymer backbone), 2.82-3.15 (s, CH2, PPS, and PS-OTBDMS polymer backbone), 2.9-3.3 (pendent dimethyl group PDMA block), 3.65-3.85 (broad s, 2H, pendent -CH2-OH). Synthesis of HA-AD by grafting of AD on HA backbone

[0201] The functionalization of HA with adamantane was performed through an esterification reaction between 1-adamantane acetic acid and HA-TBA salt. Briefly, HA-TBA salt (4.18 mM, 2.98 g), 1-adamantane acetic acid (12.54 mM 2.43 g, 3 eq), and 4-dimethylamino pyridine (DMAP; 3.13 mM, 0.383 g) were placed in 250 mL flask, degassed three times, and then fully dissolved in 100 mL of anhydrous DMSO. Di-tert-butyl dicarbonate (BOC2O) was added to this reaction mixture via a syringe (1.71 mM, 0.37 g, 0.39 mL) and heated at 45 degrees Celsius for 24 hours. After 24 hours, the crude reaction mixture was cooled to room temperature and dialyzed against DI water for 24 hours. The reaction mixture was further dialyzed against a 0.5 M NaCl solution for two days to remove the TBA salt. After two days, the reaction mixture was dialyzed against DI water for another two days with multiple water changes per day. The dialyzed solution was filtered to remove excess adamantane acetic acid and other insoluble organic impurities before being dried on a lyophilizer. The fluffy powder was suspended in acetone and centrifuged at 2500 rpm for 5 minutes to remove any remaining impurities. TheAttorney Docket No.093386-0041-WO01 acetone wash process was repeated three times after which the powder was dried under a high vacuum. The dried product was dissolved in water and lyophilized to yield a fluffy powder of HA- AD. The percent grafting of AD onto HA was estimated through the relative integration of the HA backbone protons from 3.1-4.2 ppm to the peak of adamantane protons at 1.5 ppm in1H NMR spectra. Synthesis of HA-CD by grafting of -CD on HA backbone

[0202] The functionalization of hyaluronic acid grafted with 20% -cyclodextrin (HA-CD20%) was accomplished by functionalization of HA-TBA salt with different amounts of CD-HDA to prepare hyaluronic acid polymers grafted with varying densities of -cyclodextrin (HA-CDxwhere x = 10%, 20%, and 34%) by using a BOP-mediated amidation reaction between HA and CD. A representative procedure for synthesizing HA-CD20%is as follows (FIG.3). A representative method for the synthesis of HA-CD20%is as follows. Briefly, HA-TBA salt (2.05 mmol, 1.46 g) and CD-HDA (1.025 mmol, 0.43 g) were fully dissolved in anhydrous DMSO at 2 w / v % in a 100 mL RB flask which was then purged with ultra-high purity nitrogen for 15 minutes. A solution of BOP (benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate) (1.02 mmol, 1.27 g) in DMSO (1 mL) was added dropwise to this clear solution which was then stirred for 2 hours. After 2 hours, the solution was dialyzed against DI water for 24 hours , then dialyzed against 1M solution of NaCl salt for 48 hours, and finally dialyzed against DI water for eight days, with DI water changed at least three times a day. The final dialyzed solution was dried on the lyophilizer and then characterized by1H NMR to determine the resulting percent grafting of CD through the calculation of a relative integration ratio between hexyl linker protons (1.22-1.77 ppm, 12H) on CD with the methyl singlet protons of HA (2.1 ppm, 3H). The representative1H NMR spectra of HA-CD20%is as follows:1H NMR (400 MHz, D2O): (ppm) 1.14-1.61 (m, 12 H), 2.1 (s, N-acetyl groups, HA backbone), 3.12-3.45 (m, overlaps with HOD), 3.3-4.1 (m, HA backbone), 3.48-3.78 (m, 28 H), 4.28-4.56 (br s, 6 H), 4.83 (s, 7 H), 5.59-5.88 (br s, 14 H). Characterization of Triblock Copolymers of Varying Polysulfide Chemistry by Proton Nuclear Magnetic Resonance Spectroscopy (¹H NMR)

[0203] The chemical structure and composition of all synthesized organic compounds and polymers (10 mg / mL in deuterated solvents CDCl3, and D2O) were analyzed through proton nuclear magnetic resonance spectroscopy (¹H NMR). Spectra were recorded on a BrukerAttorney Docket No.093386-0041-WO01 AV400 MHz spectrometer at 25 degrees Celsius and acquired spectra were analyzed with Bruker TopSpin 3.6.2 software (Billerica, MA). Characterization of Triblock Copolymers of Varying Polysulfide Chemistry by Gel Permeation Chromatography (GPC)

[0204] The number average molecular weight (Mn) and the polydispersity index (PDI) of the triblock and precursor polymers were confirmed using refractive index (RI) traces from gel permeation chromatography (Tosoh Bioscience, Inc., Tosoh EcoSEC Elite HLC-8420 GPC system, Grove City, OH, USA) equipped with an auto-injector, a dual differential refractive index detector, three sequential columns, and a multi-angle LENS 3 multi-angle laser light scattering detector. Polymers were dissolved at a 2.5 mg / mL concentration and filtered using a 0.22 m syringe filter prior injection into a mobile phase consisting of N, N-dimethylformamide (DMF) with 0.1 M lithium bromide (LiBr) at a flow rate of 1 mL / min and a temperature of 60 degrees Celsius. Example 2 Particles and Hydrogels Preparation and Characterization of Nanoparticles Self-Assembled from Triblock Copolymers of Varying Polysulfide Chemistry

[0205] To prepare self-assembled nanoparticles from P(PSncoPSRm) D300-AD20%triblock copolymers by nanoprecipitation, 1 mg of each triblock polymer was placed in a 4 mL glass vial containing a magnetic stir bar and was dissolved in 100 L of THF. Self-assembly of P(PSncoPSRm) D300-AD20%triblock copolymers was induced through the dropwise addition of phosphate-buffered saline (PBS) (1 mL) via a syringe pump while the triblock polymer solution was stirred at 200 rpm. The resulting suspension was then filtered through a 0.2 M syringe filter.

[0206] To prepare self-assembled nanoparticles from P(PSncoPSRm) D300-AD20%triblock copolymers by thin film rehydration, 1 mg of each triblock copolymer was dissolved in 0.3 mL of chloroform in a glass scintillation vial, and vortexed to ensure the polymer was completely dissolved in the organic phase. Each triblock copolymer in organic solution was heated to 60 degrees Celsius and placed under a constant flow of nitrogen gas to yield a thin film. Each thin film was then placed under high vacuum (<1 mbar) overnight. The thin films were rehydrated by adding an excess of DI water into each glass scintillation vial. The glass scintillations wereAttorney Docket No.093386-0041-WO01 capped, and each solution was heated to 37 degrees Celsius while being stirred vigorously at 1000 RPM for 15 minutes. After 30 minutes, the nanoparticle solutions were transferred to a 2 mL Eppendorf tube and sonicated at 37 degrees Celsius for 30 to 45 minutes. The solutions with rehydrated nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers were freeze-dried and lyophilized, and each sample of lyophilized nanoparticles was reconstituted in PBS (pH 7.4) at 0.1 w / v% (1 mg / mL).

[0207] Nanoparticle size and morphology were evaluated to confirm stable nanoparticles' formation in an aqueous solution (FIG.8). The average hydrodynamic diameter of the self- assembled ABC triblock copolymer nanoparticles (1 mg / mL PBS) was measured with Malvern Zetasizer Nano-ZS based dynamic light scattering (DLS) instrument (Malvern Instruments S6Ltd, Worcestershire, U.K) equipped with a 4 mW He Ne laser operating at = 632.8 nm and at25 degrees Celsius.

[0208] To visualize the morphology of nanoparticles self-assembled from triblock copolymers of varying polysulfide chemistries by cryogenic transmission electron microscopy (cryo-TEM), self-assembled nanoparticles were prepared from P(PSncoPSRm) D300-AD20%triblock copolymers described above to create aqueous nanoparticle solutions (10 mg / mL PBS) (FIG. 2B, FIG.2C and FIG.9B). Samples for cryo-TEM analysis were prepared on copper lacey grids (Electron Microscopy Sciences, Hatfield, PA, USA) and vitrified by Vitrobot Mark IV operating at 100% humidity and 25 degrees Celsius. Grids were prepared for analysis by applying 2.5 L of each nanoparticle solution (10 mg / mL PBS) to the grid surface for 3.5 seconds. The grids were blotted and immediately plunged into liquid ethane, cooled by liquid nitrogen. After vitrification, sample grids remained in liquid nitrogen until analysis by cryo-TEM. Sample grids with vitrified nanoparticle solutions were imaged by a Tecnai TF-20 TEM microscope operating at 200 kV (FIG.2B). For cryo-SEM sample preparation, a copper hole TEM aperture grid (3.0 mm outer diameter) was dipped into 100 L of each hybrid NP / HA hydrogel (7.5 wt% PBS) or 100 L of the control HA / HA hydrogel (7.5 wt% PBS) and then removed quickly to generate a thin layered coating of the hydrogel (FIG.2C and FIG.9B). Next, samples were plunge-frozen into liquid nitrogen slurry and placed onto a cryo-SEM shuttle under liquid nitrogen before placing into a preparation chamber held at -180 degrees Celsius. The samples were sublimated at -90 degrees Celsius for 45 minutes. The frozen sample surface was then sputter coated with platinum to make them more conductive and transferred to the imaging chamber maintained at - 180 degrees Celsius. The images were captured over the surface of the frozen hydrogels. Preparation of Hybrid NP / HA HydrogelsAttorney Docket No.093386-0041-WO01

[0209] To prepare hybrid NP / HA shear-thinning hydrogels, nanoparticles self-assembled from triblock copolymers of varied polysulfide chemistry and grafted with adamantane (AD) were mixed with hyaluronic acid grafted with -cyclodextrin (HA-CD) at a stoichiometric ratio of 1 CD to 1 AD, 1 CD to 2 AD, or 2 CD to 1 AD (FIG.2A). The representative procedure for preparation of one hybrid NP / HA shear-thinning hydrogel composed of nanoparticles self-assembled from PPS D300-AD20%triblock copolymers and HA-CD20%mixed at a 2CD / 1AD ratio are described below (FIG.1). Briefly, in separate Eppendorf tubes (2 mL), lyophilized PPS D300-AD20%(5 w / v%) and HA-CD20%(8.75 w / v%) were dissolved into PBS and placed on a vortex mixer for 24 hours. After 24 hours, PPS D300-AD20%triblock copolymer nanoparticles (50 L) and HA-CD20%polymer (50 L) were transferred to the same 2 mL centrifuge tube, where they spontaneously formed a shear-thinning hydrogel upon mixing both components with a positive displacement pipette. The final shear-thinning hydrogel was centrifuged for 10 minutes at 12,000 rpm. The vial inversion method confirmed the hybrid NP / HA shear-thinning hydrogel formation at 37 degrees Celsius (FIG.1). Rheological Characterization of Hybrid NP / HA Hydrogels

[0210] The mechanical and shear-thinning properties of hybrid NP / HA hydrogels were evaluated by rheological characterization using an AR2000 stress-controlled rheometer (TA Instruments, New Castle, DE) tted with a 20 mm diameter cone and plate geometry, 59 minutes 42 seconds (0.995°) cone angle, and 27 m gap (FIG.2D, FIG.10, ). Rheological properties were examined at 25 degrees Celsius by oscillatory frequency sweeps at 0.5% strain (0.01 to 100 Hz) , percent strain-step-based oscillatory time sweeps at 0.5% for low strain, and300% for high strain, oscillatory strain sweeps at 10 Hz frequency (0.1 500% strain), and owmeasurements in which shear rate linearly ramped from 0.1 to 51 / s. For shear recovery experiments, the shear-thinning behavior of hybrid NP / HA hydrogels was evaluated at 300% strain with recovery at 0.5% strain, each at 10 Hz frequency (FIG.2D). To determine thestability of storage modulus (G ) at a physiological temperature of 37 degrees Celsius,temperature sweep measurements were performed from 20 to 45 degrees Celsius at 0.5% strain and 10 Hz frequency. To observe the effect of variable high percent strains on shear- thinning behavior and recovery of hybrid NP / HA hydrogels, percent strain time sweep measurements were performed at variable high percent strains of 300%, 400%, and 500% while keeping low strain constant at 0.5%. Degradation of Hybrid NP / HA Hydrogels under Oxidative EnvironmentsAttorney Docket No.093386-0041-WO01

[0211] To assess the oxidative degradation profiles of hybrid NP / HA hydrogels of varied polysulfide chemistry, P(PSncoPSRm) D300-AD20%2CD / 1AD hydrogels were incubated with 1 mM, 10 mM, and 100 mM hydrogen peroxide (H2O2) at 37 degrees Celsius for up to 1 day, 2 days, and 7 days at which point the mechanical properties of the remaining hydrogels were assessed by rheology. All rheology measurements were done in triplicate. Hydrogel degradationwas evaluated with respect to an observed decrease in hydrogel storage modulus (G ) withincreasing incubation time with each degradation stimulus. At each time point, the change in Grelative to parent G of the hydrogel was confirmed by time sweep measurements (0.5% strain,10 Hz frequency) at the same time point. Over time, a decrease in G and a progressiveincrease in tan values for each hydrogel were used as benchmark parameters for monitoringhydrogel degradation. The degradation kinetic profiles for each hydrogel were also analyzedwith respect to percentage decrease in G over time.Uronic Acid Assessment of Hybrid NP / HA Hydrogels under Oxidative, Enzymatic, and Combined Oxidative and Enzymatic Environments

[0212] In addition to assessing degradation profiles by rheological measurements of hydrogel modulus, protection of HA in NP / HA hydrogels (PPS D300-AD20%1CD / 1AD, PCL D300- AD20%1CD / 1AD, and HA / HA 1CD / 1AD) was further evaluated by assessing levels of uronic acid present following degradation of hydrogels with 10 mM of H2O2for 48 hours, 7 U / mL of hyaluronidase for 48 hours, and both 1 mM H2O2and 7 U / mL of hyaluronidase for 24 hours at 37 degrees Celsius. Following incubation, the remaining hydrogels were transferred and sealed in benzoylated dialysis tubing (2K MWCO, Sigma-Aldrich, Cat. No. D2272) and dialyzed against 100 mL DI water for 24 hours. The dialysate containing the lower molecular weight HA degradation products was lyophilized and resuspended in 2 mL sodium acetate buffer (20 mM). To perform the uronic acid assay, 0.2 mL of each degradation sample was mixed with 1.2 mL of sodium tetraborate (12.5 mM) in concentrated H2SO4and heated to 100 degrees Celsius for 5 minutes. Each reaction mixture was cooled on an ice bath for 5 minutes, and a 20 L solution of m-hydroxy diphenyl (0.15 wt% in 0.5% NaOH in DI water) was added to each sample. The prepared solutions were placed in a 96-well plate, and their absorbance at 520 nm was measured using a microplate reader (Infinite M1000 Pro, Tecan Group Ltd., Mannedorf, Switzerland). The carbohydrate itself demonstrates absorbance in the presence of H2SO4; therefore, 0.2 mL of HA only (0.2 mg / mL) was mixed with 1.2 ml of sodium tetraborate (12.5 mM) in concentrated H2SO4and heated at 100 degrees Celsius for 5 minutes to measure the absorbance of HA without the addition of m-hydroxydiphenyl (0.15 wt% in 0.5% NaOH). TheAttorney Docket No.093386-0041-WO01 concentration of uronic acid in each degradation sample was quantified by a standard curve of the absorbances of solutions with known concentrations of HA (82 KDa). Small Molecule PHD2 Inhibitor Loading and Encapsulation Efficiency in Hybrid NP / HA Hydrogels

[0213] The loading and encapsulation efficiency of the small molecule PHD2 inhibitor, MK- 8617, was evaluated by encapsulating the small molecule at a range of weight ratios of small molecule drug to polymer (defined as the weight of the small molecule to the weight of polymer) into the core of nanoparticles self-assembled from triblock copolymers with various polysulfide chemistry in the nanoparticle core. Various amounts of small molecule PHD2 inhibitor, MK- 8617, was dissolved in a minimum amount of solvent equal parts chloroform and methanol to afford solutions of MK-8617 to achieve loading of MK-8617 at a range of weight ratios of drug to polymer including 1, 2.5, 5, 10, and 20 w / w%.

[0214] A total of 50 mg of each triblock copolymer P(PSncoPSRm) D300-AD20%was dissolved in 0.3 mL of chloroform in a glass scintillation vial, and the total volume of MK-8617 in organic solution was transferred to the glass scintillation vial to co-dissolve MK-8616 with the polymer at the correct weight ratio of small molecule drug to polymer. This was repeated for each sample to be loaded with MK-8617. Each solution was vortexed to ensure MK-8617 and polymer were completely dissolved in the organic phase. To load MK-8617 into the hydrophobic core of self- assembled nanoparticles, each polymer and drug in organic solution was heated to 60 degrees Celsius and was placed under a constant flow of nitrogen gas to yield a thin film. Each thin film was then placed under high vacuum (<1 mbar) overnight. The thin films were rehydrated by adding an excess of DI water into each glass scintillation vial. The glass scintillations were capped, and each solution was heated to 37 degrees Celsius while being stirred vigorously at 1000 RPM for 15 minutes. After 30 minutes, the nanoparticle solutions loaded with MK-8617 was transferred to a 2 mL Eppendorf tube and sonicated at 37 degrees Celsius for 30 to 45 minutes. To remove unloaded small molecule PHD2 inhibitor, the 2 mL Eppendorf tubes were centrifuged at 8000 x g for 5 minutes, and each supernatant containing the nanoparticles with MK-8617 loaded stably into the hydrophobic core was transferred to a 2 mL Eppendorf tube. The supernatants were freeze-dried and lyophilized, and each sample of lyophilized nanoparticles loaded with MK-8617 was reconstituted in PBS (pH 7.4) at 5 w / v% (50 mg / mL).

[0215] The amount of MK-8617 loaded was determined through reverse phase high performance liquid chromatography (RP-HPLC) with a Luna C18 column and an isocratic gradient of 70% HPLC grade acetonitrile (ACN) and 30% water (H2O). The nanoparticleAttorney Docket No.093386-0041-WO01 samples were diluted 1:100 into 1 mL of the mobile phase and injected into the column at a flowrate of 1 mL / min. The amount of MK-8617 loaded was determined by integrating the MK- 8617 peak within the unknown samples relative to a standard curve generated by injection a series of samples with a known concentration of MK-8617. The encapsulation efficiency (EE) was defined as the ratio of encapsulated MK-8617 to the amount of MK-8617 initially added to the triblock copolymers in solution multiplied by 100. The loading capacity was defined as the ratio of encapsulated MK-8617 to the total concentration of polymer and encapsulated MK-8617 (polymer assumed to be 50 mg / mL) multiplied by 100. Oxidative Release of Small Molecule PHD2 Inhibitors from Nanoparticles Self-Assembled from Triblock Copolymers of Varying Polysulfide Chemistry

[0216] The rate of release of small molecule PHD2 inhibitors from the core of nanoparticles self-assembled from triblock copolymers of varied polysulfide composition in response to reactive oxygen species (ROS) was evaluated by loading the small molecule PHD2 inhibitor, MK-8617, into the core of P(PSncoPSRm) D300-AD20%triblock copolymers. A total of 50 mg of each triblock copolymer P(PSncoPSRm) D300-AD20%was dissolved in 0.3 mL of chloroform in a glass scintillation vial, and the total volume of MK-8617 in organic solution was transferred to the glass scintillation vial to co-dissolve MK-8617 with the polymer at 1 w / w% weight ratio of small molecule drug to polymer. This was repeated for each sample to be loaded with MK-8617. Each solution was vortexed to ensure MK-8617 and polymer were completely dissolved in the organic phase. To load MK-8617 into the hydrophobic core of self-assembled nanoparticles, each polymer and drug in organic solution was heat to 60 degrees Celsius and was placed under a constant flow of nitrogen gas to yield a thin film. Each thin film was then placed under high vacuum (<1 mbar) overnight. The thin films were rehydrated by adding DI water into each glass scintillation vial. The glass scintillations were capped, and each solution was heated to 37 degrees Celsius while being stirred vigorously at 1000 RPM for 15 minutes. After 30 minutes, the nanoparticle solutions loaded with MK-8617 was transferred to a 2 mL Eppendorf tube and sonicated at 37 degrees Celsius for 30 to 45 minutes. To remove unloaded small molecule PHD2 inhibitor, the 2 mL Eppendorf tubes were centrifuged at 8000 x g for 5 minutes, and each supernatant containing the nanoparticles with MK-8617 loaded stably into the hydrophobic core was transferred to a 2 mL Eppendorf tube. The supernatants were freeze-dried and lyophilized, and each sample of lyophilized P(PSncoPSRm) D300-AD20%nanoparticles loaded with MK-8617 was reconstituted in PBS (pH 7.4) at 5 w / v% (50 mg / mL). To assess oxidative release of the small molecule PHD2 inhibitor, MK-8617, from the self-assembled nanoparticles alone, 50 L ofAttorney Docket No.093386-0041-WO01 each nanoparticle composed of a different polysulfide chemistry with loaded MK-8617 was transferred to a 2 mL Eppendorf tube, after which each aliquot was incubated with 1 mL of 0 mM, 100 mM, or 500 mM of hydrogen peroxide (H2O2). All 2 mL Eppendorf tubes containing all aliquots were incubated at 37 degrees Celsius and 5% CO2. At each timepoint, 2 mL Eppendorf tubes containing all aliquots were removed from the incubator and 1 mL of releasate from each 2 mL Eppendorf tube was collected into a new 2 mL Eppendorf tube and replaced with 1 mL of new buffer containing 0 mM, 100 mM, or 500 mM of hydrogen peroxide (H2O2). All releasates were centrifuged at 8000 x g for 5 minutes to remove all unloaded free MK-8617 from the releasate. The supernatant containing nanoparticles with MK-8617 loaded into the hydrophobic core was removed from each 2 mL Eppendorf tube and transferred to a new 2 mL Eppendorf tube. After removing the supernatant, 1000 L of DMSO was transferred to each of the 2 mL Eppendorf tube containing unloaded free MK-8617 in order to fully solubilize the free MK-8617 from the releasate. The release of MK-8617 from nanoparticles composed of different polysulfide chemistry was evaluated by determining the amount of MK-8617 present in each releasate collected over time. The amount of MK-8617 in each releasate was determined through reverse phase high performance liquid chromatography (RP-HPLC) with a Luna C18 column and an isocratic gradient of 70% HPLC grade acetonitrile (ACN) and 30% water (H2O). The releasate samples were diluted 1:100 into 1 mL of the mobile phase and injected into the column at a flowrate of 1 mL / min. The amount of MK-8617 in each sample was determined by integrating the MK-8617 peak within the unknown samples relative to a standard curve generated by injection a series of samples with a known concentration of MK-8617. The percent release of MK-8617 at each timepoint was defined as the cumulative amount of MK-8617 in all releasates up to that timepoint normalized by the initial amount of MK-8617 loaded into the core of each P(PSncoPSRm) D300-AD20%nanoparticle prior to initiation of oxidative release study. Oxidative Release of Small Molecule PHD2 Inhibitors from Hybrid NP / HA Hydrogels

[0217] The rate of release of small molecule PHD2 inhibitors from hybrid NP / HA hydrogels incorporating nanoparticles of varied polysulfide composition in response to reactive oxygen species (ROS) was evaluated by loading the small molecule PHD2 inhibitor, MK-8617, into the core of P(PSncoPSRm) D300-AD20%triblock copolymers which were then combined with HA- CD20%polymer (8.75 wt% PBS) at a 2CD / 1AD ratio to form hybrid NP / HA hydrogels (FIG.14A, FIG.14B). A total of 50 mg of each triblock copolymer P(PSncoPSRm) D300-AD20%was dissolved in 0.3 mL of chloroform in a glass scintillation vial, and the total volume of MK-8617 in organic solution was transferred to the glass scintillation vial to co-dissolve MK-8617 with the polymer atAttorney Docket No.093386-0041-WO01 1 w / w% weight ratio of small molecule drug to polymer. This was repeated for each sample to be loaded with MK-8617. Each solution was vortexed to ensure MK-8617 and polymer were completely dissolved in the organic phase. To load MK-8617 into the hydrophobic core of self- assembled nanoparticles, each polymer and drug in organic solution was heat to 60 degrees Celsius and was placed under a constant flow of nitrogen gas to yield a thin film. Each thin film was then placed under high vacuum (<1 mbar) overnight. The thin films were rehydrated by adding DI water into each glass scintillation vial. The glass scintillations were capped, and each solution was heated to 37 degrees Celsius while being stirred vigorously at 1000 RPM for 15 minutes. After 30 minutes, the nanoparticle solutions loaded with MK-8617 was transferred to a 2 mL Eppendorf tube and sonicated at 37 degrees Celsius for 30 to 45 minutes. To remove unloaded small molecule PHD2 inhibitor, the 2 mL Eppendorf tubes were centrifuged at 8000 x g for 5 minutes, and each supernatant containing the nanoparticles with MK-8617 loaded stably into the hydrophobic core was transferred to a 2 mL Eppendorf tube. The supernatants were freeze-dried and lyophilized, and each sample of lyophilized P(PSncoPSRm) D300-AD20%nanoparticles loaded with MK-8617 was reconstituted in PBS (pH 7.4) at 5 w / v% (50 mg / mL) and were then placed on a vortex mixer for 24 hours. After 24 hours, the HA-CD20%polymer (8.75 wt% PBS) was mixed with each of the P(PSncoPSRm) D300-AD20%nanoparticles loaded with MK-8617 at a 2CD / 1AD ratio to form hybrid NP / HA hydrogels each of a varied polysulfide chemistry but loaded with MK-8617 at the same weight ratio. To assess oxidative release of the small molecule PHD2 inhibitor, MK-8617, from the final hybrid NP / HA hydrogels, 50 L of each hybrid NP / HA hydrogel with MK-8617 was transferred to the transwell of a 12-well transwell plate using a positive displacement pipette, after which each aliquot was incubated with 2 mL of 0 mM, 100 mM, or 500 mM of hydrogen peroxide (H2O2). All transwell plates with hybrid NP / HA hydrogels were incubated at 37 degrees Celsius and 5% CO2. At each timepoint, the transwell insert containing each hybrid NP / HA hydrogel was removed and 1 mL of releasate from the bottom of each well was collected into a 2 mL Eppendorf tube and replaced with 1 mL of new buffer containing 0 mM, 100 mM, or 500 mM of hydrogen peroxide (H2O2). All releasates were centrifuged at 8000 x g for 5 minutes to remove all unloaded free MK-8617 from the releasate. The supernatant containing nanoparticles with MK-8617 loaded into the hydrophobic core was removed from each 2 mL Eppendorf tube and transferred to a new 2 mL Eppendorf tube. After removing the supernatant, 1000 L of DMSO was transferred to each of the 2 mL Eppendorf tube containing unloaded free MK-8617 in order to fully solubilize the free MK-8617 from the releasate. The release of MK-8617 from hybrid NP / HA hydrogels was evaluated by determining the amount of MK-8617 present in each releasate collected over time. The amount of MK-8617Attorney Docket No.093386-0041-WO01 in each releasate was determined through reverse phase high performance liquid chromatography (RP-HPLC) with a Luna C18 column and an isocratic gradient of 70% HPLC grade acetonitrile (ACN) and 30% water (H2O). The releasate samples were diluted 1:100 into 1 mL of the mobile phase and injected into the column at a flowrate of 1 mL / min. The amount of MK-8617 in each sample was determined by integrating the MK-8617 peak within the unknown samples relative to a standard curve generated by injection a series of samples with a known concentration of MK-8617. The percent release of MK-8617 at each timepoint was defined as the cumulative amount of MK-8617 in all releasates up to that timepoint normalized by the initial amount of MK-8617 loaded into the core of each P(PSncoPSRm) D300-AD20%nanoparticle prior to hybrid NP / HA hydrogel formation and initiation of oxidative release study (FIG.14A, FIG.14B). A simple linear regression was performed on the kinetic release data in order to estimate the time to release 50% of total small molecule drug MK-8617 and 100% of total small molecule drug MK-8617 from hybrid NP / HA hydrogels incorporating nanoparticles self-assembled from P(PSncoPSRm) D300-AD20%triblock copolymers following incubation with increasing doses of H2O2. Cytoprotective Capabilities of Nanoparticles Self-Assembled from Triblock Copolymers of Varying Polysulfide Chemistry

[0218] The antioxidant potential and cytoprotective capabilities of nanoparticles self- assembled from triblock copolymers of varying polysulfide chemistry were evaluated by incubating cells with nanoparticles composed of different polysulfide chemistries and then treating cells with increasing doses of hydrogen peroxide (H2O2) (FIG.17). To perform 2D cell culture with treatment by nanoparticles of varied polysulfide chemistries, NIH 3T3 fibroblasts were seeded at a density of 5000 cells per well in a 96-well plate and cultured with standard cell growth media at 37 degrees Celsius and 5% CO2for 3 days, at which point 80% confluency was observed. Once 80% confluency was observed, the remaining media was aspirated from each well, and each well of cells was treated with a different type of nanoparticles of a specific type of polysulfide chemistry by transferring a total volume of 50 L of each nanoparticle at a concentration of 0.1 w / v% to each well for each increasing dose of H2O2(n=10). After all cells received the appropriate nanoparticle treatment, all cells were incubated with standard mMSC growth media supplemented with either 0 M, 62.5 M, 125 M, 250 M, 500 M, or 1000 M of H2O2. The entire 96-well plate with cells was incubated at 37 degrees Celsius and 5% CO2for 24 hours. After 24 hours, an aliquot from the cell media was collected so that the concentrationAttorney Docket No.093386-0041-WO01 of H2O2remaining could be quantified by Amplex Red assay, and the cells were harvested so that cell viability was quantified by Cell-Titer Glo luminescent readout. Cytocompatibility and Cytoprotective Capabilities of Hybrid NP / HA Hydrogels

[0219] The cytocompatibility of hybrid NP / HA hydrogels was evaluated by encapsulation of mouse mesenchymal stem cells (mMSCs) in a 3D suspension in hybrid hydrogels. The hybrid NP / HA hydrogels of varied polysulfide chemistry as well control HA / HA hydrogels, were formulated at 7.5 w / v% polymer in PBS (pH 7.4). To perform 3D cell culture with hydrogel encapsulation, mMSCs were cultured at 37 degrees Celsius and 5% CO2in standard mMSC growth media until 80% confluency was observed, at which point mMSCs were encapsulated in each hydrogel at a density of 4 x 106cells per mL. After confirming cell encapsulation by visualizing hydrogels under a microscope, 50 L of each hydrogel was transferred to the wells of a 96-well plate (n = 10) and incubated with cell culture growth media. The 96-well plate with hydrogels and cells was incubated at 37 degrees Celsius and 5% CO2for 24 hours. After 24 hours, cell viability was quantified by Cell-Titer Glo luminescent readout.

[0220] The antioxidant potential and cytoprotective capabilities of hybrid NP / HA hydrogels was evaluated by encapsulation of mMSCs in a 3D suspension in hybrid hydrogels. The hybrid NP / HA hydrogels of varied polysulfide chemistry as well control HA / HA hydrogels, were formulated at 7.5 w / v% polymer in PBS (pH 7.4). For performing 3D cell culture with hydrogel encapsulation, mMSCs were cultured at 37 degrees Celsius and 5% CO2in standard cell growth media until 80% confluency was observed, at which point mMSCs were encapsulated in each hydrogel at a density of 4 x 106cells per 1 mL of hydrogel. After confirming cell encapsulation by visualizing hydrogels under a microscope, 50 L of each hydrogel with encapsulated cells was transferred to the bottom of a well of a 96-well plate and was repeated for each increasing dose of H2O2(n=10). After each hybrid NP / HA hydrogel with encapsulated cells was transferred to the wells of the 96-well plate, all hydrogels with encapsulated cells were incubated with standard mMSC growth media supplemented with either 0 M, 62.4 M, 125 M, 250 M, 500 M, or 1000 M of H2O.. The 96-well plate with hydrogels and encapsulated cells was incubated at 37 degrees Celsius and 5% CO2for 24 hours. After 24 hours, cell viability was quantified by Cell-Titer Glo luminescent readout.Stabilization of HIF1- following Small Molecule PHD2 Inhibitor Delivery by Hybrid NP / HAHydrogelsAttorney Docket No.093386-0041-WO01

[0221] The stabilization of HIF1- following delivery of small molecule PHD2 inhibitors fromhybrid NP / HA hydrogels incorporating nanoparticles of varied polysulfide composition was evaluated through treating NIH 3T3 mouse fibroblasts and collecting protein lysates for Western blot analysis. The small molecule PHD2 inhibitor, MK-8617, was loaded into the core of P(PSncoPSRm) D300-AD20%triblock copolymers at a weight ratio of 1 w / w% drug to polymer. A total of 50 mg of each triblock copolymer P(PSncoPSRm) D300-AD20%was dissolved in 0.3 mL of chloroform in a glass scintillation vial, and the total volume of MK-8617 in organic solution was transferred to the glass scintillation vial to co-dissolve MK-8617 with the polymer at 1 w / w% weight ratio of small molecule drug to polymer. This was repeated for each sample to be loaded with MK-8617. Each solution was vortexed to ensure MK-8617 and polymer were completely dissolved in the organic phase. To load MK-8617 into the hydrophobic core of self-assembled nanoparticles, each polymer and drug in organic solution was heat to 60 degrees Celsius and was placed under a constant flow of nitrogen gas to yield a thin film. Each thin film was then placed under high vacuum (<1 mbar) overnight. The thin films were rehydrated by adding DI water into each glass scintillation vial. The glass scintillations were capped, and each solution was heated to 37 degrees Celsius while being stirred vigorously at 1000 RPM for 15 minutes. After 30 minutes, the nanoparticle solutions loaded with MK-8617 was transferred to a 2 mL Eppendorf tube and sonicated at 37 degrees Celsius for 30 to 45 minutes. To remove unloaded small molecule PHD2 inhibitor, the 2 mL Eppendorf tubes were centrifuged at 8000 x g for 5 minutes, and each supernatant containing the nanoparticles with MK-8617 loaded stably into the hydrophobic core was transferred to a 2 mL Eppendorf tube. The supernatants were freeze- dried and lyophilized, and each sample of lyophilized P(PSncoPSRm) D300-AD20%nanoparticles loaded with MK-8617 was reconstituted in PBS (pH 7.4) at 5 w / v% (50 mg / mL) and were then placed on a vortex mixer for 24 hours. After 24 hours, the HA-CD20%polymer (8.75 wt% PBS) was mixed with each of the P(PSncoPSRm) D300-AD20%nanoparticles loaded with MK-8617 at a 2CD / 1AD ratio to form hybrid NP / HA hydrogels each of a varied polysulfide chemistry but loaded with MK-8617 at the same weight ratio.

[0222] To perform 2D cell culture with treatment by small molecule PHD2 inhibitor delivery by hybrid NP / HA hydrogels, NIH 3T3 fibroblasts were seeded at a density of 600,000 cells per well in the bottom wells of a 6-well transwell plate and cultured with standard NIH 3T3 fibroblast growth media at 37 degrees Celsius and 5% CO2for 3 days, at which point 80% confluency was observed. Once 80% confluency was observed, the remaining media was aspirated from the bottom well and 50 L of each hybrid NP / HA hydrogel with MK-8617 was transferred to the transwell of a 6-well transwell plate using a positive displacement pipette, after which each wellAttorney Docket No.093386-0041-WO01 was incubated with 2 mL of standard NIH 3T3 fibroblast growth media. All transwell plates with cells in the bottom well and hybrid NP / HA hydrogels with MK-8617 in the transwells were incubated at 37 degrees Celsius and 5% CO2. At each timepoint, the transwell insert containing each hybrid NP / HA hydrogel with MK-8617 was removed from each well and the transwell plate with cells was placed on ice. The remaining media was aspirated from well after which 200 L of 1X RIPA lysis buffer supplemented with protease inhibitor cocktail and phosphatase I and II cocktails was added to each well. The transwell plate with cells was incubated at 4 degrees Celsius for 15 minutes while placed on a rocking platform. After 15 minutes, cells from each well were lifted from their 2D culture on the bottom of each well using a cell scaper to scrape the lysate in the bottom of each well in a downward motion. The lysate was lifted from the well with a pipette and mixed by the pipette to homogenize each lysate. The lysate with cells from each well was transferred to a pre-chilled 2 mL Eppendorf tube and was centrifuged at 14000 x g for 15 minutes at 4 degrees Celsius to clear the lysates. The supernatant containing lysates were transferred to a new pre-chilled 2 mL Eppendorf tube, and protein concentration was quantified through use of the bicinchoninic acid (BCA) assay according to the manufacturer’s instructions.The samples were stored at -80 degrees Celsius prior to analysis of stabilization of HIF1- byWestern blots. Activation of Angiogenesis in Endothelial Cells following Small Molecule PHD2 Inhibitor Delivery by Hybrid NP / HA Hydrogels

[0223] The activation of angiogenesis in human umbilical vein endothelial cells (HUVECs) following delivery of small molecule PHD2 inhibitors from hybrid NP / HA hydrogels incorporating nanoparticles of varied polysulfide composition was evaluated through a tube formation assay (FIG.25A, FIG.25B). The small molecule PHD2 inhibitor, MK-8617, was loaded into the core of P(PSncoPSRm) D300-AD20%triblock copolymers at a weight ratio of 1 w / w% drug to polymer. A total of 50 mg of each triblock copolymer P(PSncoPSRm) D300-AD20%was dissolved in 0.3 mL of chloroform in a glass scintillation vial, and the total volume of MK-8617 in organic solution was transferred to the glass scintillation vial to co-dissolve MK-8617 with the polymer at 1 w / w% weight ratio of small molecule drug to polymer. This was repeated for each sample to be loaded with MK-8617. Each solution was vortexed to ensure MK-8617 and polymer were completely dissolved in the organic phase. To load MK-8617 into the hydrophobic core of self-assembled nanoparticles, each polymer and drug in organic solution was heat to 60 degrees Celsius and was placed under a constant flow of nitrogen gas to yield a thin film. Each thin film was then placed under high vacuum (<1 mbar) overnight. The thin films were rehydrated by adding DIAttorney Docket No.093386-0041-WO01 water into each glass scintillation vial. The glass scintillations were capped, and each solution was heated to 37 degrees Celsius while being stirred vigorously at 1000 RPM for 15 minutes. After 30 minutes, the nanoparticle solutions loaded with MK-8617 was transferred to a 2 mL Eppendorf tube and sonicated at 37 degrees Celsius for 30 to 45 minutes. To remove unloaded small molecule PHD2 inhibitor, the 2 mL Eppendorf tubes were centrifuged at 8000 x g for 5 minutes, and each supernatant containing the nanoparticles with MK-8617 loaded stably into the hydrophobic core was transferred to a 2 mL Eppendorf tube. The supernatants were freeze- dried and lyophilized, and each sample of lyophilized P(PSncoPSRm) D300-AD20%nanoparticles loaded with MK-8617 was reconstituted in PBS (pH 7.4) at 5 w / v% (50 mg / mL) and were then placed on a vortex mixer for 24 hours. After 24 hours, the HA-CD20%polymer (8.75 wt% PBS) was mixed with each of the P(PSncoPSRm) D300-AD20%nanoparticles loaded with MK-8617 at a 2CD / 1AD ratio to form hybrid NP / HA hydrogels each of a varied polysulfide chemistry but loaded with MK-8617 at the same weight ratio.

[0224] To perform 2D cell culture with treatment by small molecule PHD2 inhibitor delivery by hybrid NP / HA hydrogels, HUVECs were harvested at 80% confluency and were resuspended in serum-free HUVEC basal media with NucBlue Live Ready Probe to fluorescently label the cells prior to seeding the cells for the tube formation assay. The HUVECs were then seeded at a density of 15,000 cells per well on top of Matrigel basement membrane which was pre-coated on the bottom of each well of a 96-well plate. Immediately following, 50 L of each hybrid NP / HA hydrogel with MK-8617 was transferred to the wells seeded with HUVECs using a positive displacement pipette after which each well was incubated with 100 L of serum-free HUVEC basal media. Immediately after the cells were treated with each hydrogel, the 96-well plate with hydrogels and cells set up for the tube formation assay was imaged by scanning each well of the plate at 10X at a 405 nm laser ling with a confocal scanning laser microscope to obtain a baseline image of fluorescently labelled-cells prior to the start of the assay. The 96-well plate with hydrogels and cells set up for the tube formation assay was incubated in a sterile incubator at 37 degrees Celsius and 5% CO2. At each time point, the 96- well plate was removed from sterile incubator and was imaged by scanning each well of the plate at 10X at a 405 nm laser ling with a confocal scanning laser microscope to monitor proliferation, migration, and formation of tubules by HUVECs (FIG.24). Infiltration, Degradation, and Resorption of Hybrid NP / HA Hydrogels in Subcutaneous Mouse ModelAttorney Docket No.093386-0041-WO01

[0225] This study randomly divided twenty male C57BL / 6 mice aged 12 weeks into two groups. Mice were anesthetized with 2% isoflurane and maintained at 37 ºC. The ventral region of each mouse was shaved and prepped with betadine and ethanol. Four 1 mm incisions were created, two on either side of the midline. Incisions allowed for the creation of four subcutaneous pockets per mouse and 50 L of each hybrid NP / HA hydrogel was injected from sterile 1 mL Luer lock syringes into the subcutaneous pockets via an 18G needle (FIG.2E). Each incision was closed with nonabsorbent sutures and sterilized with alcohol swabs. Mice from each group were euthanized on day 7 and day 14 post-procedure, and tissue samples containing candidate NP / HA hydrogels were explanted for histological analysis (FIG.2E). Histological sections were evaluated by ImageJ Software to quantify hydrogel degradation and cellular infiltration of surrounding tissue (data not shown). Cellular infiltration was defined as the percent area occupied by nuclei within the remaining hydrogel across three fields of view per sample. Hydrogel degradation was defined as the percent area occupied by hydrogel material across the same three fields of view per sample. Infiltration, Degradation, and Resorption of Hybrid NP / HA Hydrogels in Excisional Wound Healing Model in Diabetic Mice

[0226] This study randomly divided forty-two male db / db mice and forty-two db / db female mice aged 12-16 weeks into three different groups. Mice were anesthetized with 2% isoflurane and maintained at 37 degrees Celsius. The dorsal region of each mouse was shaved and prepped with betadine and ethanol. Two full thickness 6 mm excisional wounds were created, with one on either side of the midline, with use of a sterile biopsy punch. Bilateral 8 mm stents made of silicon rubber were sterilized and placed alongside the edges of each wound and secured with nonabsorbent sutures and sterilized with alcohol swabs in order to prevent wound contraction. Immediately after wound induction and stent placement, 50 L of each treatment hybrid NP / HA hydrogel or 50 L of the control HA / HA shear-thinning hydrogel was applied to the top of the wounds, or the wounds were left untreated (FIG.18, FIG.19, FIG.20, FIG.21A, FIG.21B, FIG.22A, FIG.22B). Immediately following wound treatment, each wound was covered with Tegaderm transparent dressing which was secured with nonabsorbent sutures in order to prevent any artificial manipulation of the wound. All mice were returned to individual sterile cages once removed from isoflurane and monitored while recovering from anesthesia. To assess resulting wound healing outcomes, wound closure and wound vascular perfusion were evaluated longitudinally through digital photographs (FIG.25A-B) and laser doppler imaging (LDPI). Equal numbers of both male and female mice from each group were euthanized on dayAttorney Docket No.093386-0041-WO01 3, day 7, and day 14 post-procedure. Following euthanasia on day 3, all wounds were bisected, and half of the tissue was prepared to assess ROS load of hydrogen peroxide (H2O2) by Amplex Red (FIG.20) while the other half of the tissue was prepared for protein extraction for an enzyme linked immunosorbent assays (ELISAs) to quantify key protein biomarkers in inflammatory pathways as well as tissue repair and remodelling pathways (data not shown). Following euthanasia on day 7, half of the total number of wounds were prepared for histology to assess wound structural qualities and the rate of hydrogel degradation (FIG.21A and FIG. 22A), while the other half of the total number of wounds were bisected so that half of the tissue could be prepared to assess ROS load of H2O2by Amplex Red (FIG.19) while the other half of the tissue could be prepared for protein extraction for ELISAs to quantify key protein biomarkers in inflammatory pathways as well as tissue repair and remodelling pathways (data not shown). Following euthanasia on day 14, half of the total number of wounds were prepared for histology to assess wound structural qualities and the rate of hydrogel degradation (FIG.21B and FIG. 22B), while the other half of the total number of wounds was prepared to assess ROS load of H2O2by Amplex Red (FIG.19). Therapeutic Delivery of Small Molecule PHD2 Inhibitors by Hybrid NP / HA Hydrogels in Excisional Wound Healing Model in Diabetic Mice

[0227] This study randomly divided thirty-six male db / db mice and thirty-six db / db female mice aged 12-16 weeks into two different groups. Mice were anesthetized with 2% isoflurane and maintained at 37 ºC. The dorsal region of each mouse was shaved and prepped with betadine and ethanol. Two full thickness 6 mm excisional wounds were created, with one on either side of the midline, with use of a sterile biopsy punch. Bilateral 8 mm stents made of silicon rubber were sterilized and placed alongside the edges of each wound and secured with nonabsorbent sutures and sterilized with alcohol swabs in order to prevent wound contraction. Immediately after wound induction and stent placement, 50 L of each treatment hybrid NP / HA hydrogel with or without the small molecule PHD2 inhibitor, MK-8617, loaded into the core of the self-assembled nanoparticles, or 50 L of MK-8617 alone was applied to the top of the wounds, or the wounds were left untreated (FIG.25, FIG.26, FIG.27A, FIG.27B, FIG.28A, and FIG. 28B) Immediately following wound treatment, each wound was covered with Tegaderm transparent dressing which was secured with nonabsorbent sutures in order to prevent any artificial manipulation of the wound. All mice were returned to individual sterile cages once removed from isoflurane and monitored while recovering from anesthesia. To assess resulting wound healing outcomes, wound closure and wound vascular perfusion were evaluatedAttorney Docket No.093386-0041-WO01 longitudinally through digital photographs and laser doppler imaging (LDPI) (FIG.26). Equal numbers of both male and female mice from each group were euthanized on day 7 and day 14 post-procedure. Following euthanasia on day 7 and day 14, one third of the total number of wounds were prepared for histology to assess to assess wound structural qualities as well as the quality of wound healing through wound scoring (FIG.27A, FIG.27B, FIG.28A, and FIG. 27B), while the second third of the total number of wounds were bisected so that half of the tissue could be prepared for quantitative reverse transcription polymerase chain reaction (RT- qPCR) to assess gene expression by a comprehensive panel (data not shown) while the other half of the tissue could be prepared for protein extraction for ELISAs to quantify key protein biomarkers in inflammatory pathways as well as tissue repair and remodelling pathways (data not shown). Following euthanasia on day 7 and day 14, the final third of the total number of wounds were explanted so that the tissue could be frozen in liquid nitrogen and prepared forWestern blots to confirm protein stabilization of HIF1- (data not shown).Histological Evaluation

[0228] Explanted wound samples were placed in histology cassettes and fixed in 10% neutral buffered formalin (NBF) for 24 hours. After 24 hours, all wound samples in cassettes were removed from 10% NBF, each wound was bisected, and each half of the wound was placed cut side down in the same cassette prior to processing. Bisected wound samples in cassettes were placed back into 10% NBF for 24 hours. Tissue samples were then dehydrated in a gradient of ethanol, exposed to xylene, and finally embedded in paraffin. Tissue samples in paraffin blocks were sectioned to create 5 m sections for further histological and immunohistochemical analysis. For histological analysis, tissue sections were deparaffinized and rehydrated in Tris-buffered Saline 0.1% Tween 20 (TBST) buffer and stained with hematoxylin and eosin (H&E) (FIG.22A, FIG.22B and FIG.28A, FIG.28B) and Gomori’s trichrome strain according to the manufacturer’s protocol (FIG.21A, FIG.21B and FIG.27A, FIG.27B). In vitro Evaluation of ROS Load of Hydrogen Peroxide (H2O2) by Amplex Red Assay

[0229] The aliquots collected from in vitro analysis of nanoparticles composed of varied polysulfide chemistry were transferred to 2 mL Eppendorf tubes, and 50 L of each sample aliquot was transferred to the wells of a 96-well plate. The working Amplex Red reaction buffer was prepared according to the manufacturer’s instructions, and 50 L of working Amplex Red reaction buffer was added to each sample in the wells of a 96-well plate. All samples wereAttorney Docket No.093386-0041-WO01 incubated for 30 minutes at room temperature while rocking on a platform. After 30 minutes, the fluorescence intensity of the sample supernatants was measured at an excitation wavelength of 570 nm and an emission wavelength of 585 nm. The average fluorescence measure from each sample was normalized to the mass of tissue to determine the normalized fluorescence of each sample. In vivo Evaluation of ROS Load of Hydrogen Peroxide (H2O2) by Amplex Red Assay

[0230] The tissue from explanted wound samples was weighed in 2 mL Eppendorf tubes and then minced into fine pieces using stainless steel dissecting scissors. All tissue samples were placed on ice. The working Amplex Red reaction buffer was prepared according to the manufacturer’s instructions, and 500 L of working Amplex Red reaction buffer was added to each sample in the 2 mL Eppendorf tubes. All samples were incubated for 1 hour at room temperature while on a vortex. After 1 hour, all samples were centrifuged at 13000 x g for 5 minutes to clear samples, and the supernatants were transferred to new 2 mL Eppendorf tubes. A total of 100 L of each supernatant was transferred to the wells of a 96-well plate, and all samples were incubated for 30 minutes at room temperature while rocking on a platform. After 30 minutes, the fluorescence intensity of the sample supernatants was measured at an excitation wavelength of 570 nm and an emission wavelength of 585 nm. The average fluorescence measure from each sample was normalized to the mass of tissue to determine the normalized fluorescence of each sample. In vivo Protein Extraction for Enzyme Linked Immunosorbent Assays (ELISAs)

[0231] The tissue from explanted wound samples was weighed in 2 mL Eppendorf tubes. All tissue samples were placed on ice and 300 L of 1X T-PER lysis buffer supplemented with protease inhibitor cocktail and phosphatase I and II cocktail was added to each sample in the 2 mL Eppendorf tube. A pre-chilled 5 mm stainless steel bead was added to each 2 mL Eppendorf tube, and all samples were homogenized in lysis buffer at 30 Hz for 5 minutes. After homogenization, pre-chilled 10% NP-40 was added to each sample to afford a maximum of 0.5 w / v% non-ionic detergent solution in the lysis buffer with each sample. All samples were incubated at 4 degrees Celsius for 1 hour while placed on a rocking platform. After 1 hour, all samples in 2 mL Eppendorf tubes were centrifuged at 2000 x g for 15 minutes at 4 degrees Celsius to clear the lysates. The supernatant containing lysates were transferred to a new pre- chilled 2 mL Eppendorf tube, and protein concentration was quantified through use of the bicinchoninic acid (BCA) assay according to the manufacturer’s instructions. The samples wereAttorney Docket No.093386-0041-WO01 stored at -80 degrees Celsius prior to analysis by enzyme linked immunosorbent assays (ELISAs) to quantify key protein biomarkers in inflammatory pathways as well as tissue repair and remodelling pathways (data not shown).In vivo Protein Extraction for Western Blot for Stabilization of HIF1-

[0232] The tissue from explanted wound samples was weighed in 2 mL Eppendorf tubes. All tissue samples were placed on ice and 200 L of 1X RIPA lysis buffer supplemented with protease inhibitor cocktail and phosphatase I and II cocktail was added to each sample in the 2 mL Eppendorf tube. A pre-chilled 5 mm stainless steel bead was added to each 2 mL Eppendorf tube, and all samples were homogenized in lysis buffer at 30 Hz for 5 minutes. All samples were incubated at 4 degrees Celsius for 1 hour while placed on a rocking platform. After 1 hour, all samples in 2 mL Eppendorf tubes were centrifuged at 14000 x g for 15 minutes at 4 degrees Celsius to clear the lysates. The supernatant containing lysates were transferred to a new pre- chilled 2 mL Eppendorf tube, and protein concentration was quantified through use of the bicinchoninic acid (BCA) assay according to the manufacturer’s instructions. The samples werestored at -80 degrees Celsius prior to analysis of stabilization of HIF1- by Western blots (datanot shown).Western Blots for Stabilization of HIF1-

[0233] Equal amounts (20 g) of proteins were added to Laemmli sample buffer and heated for 5 minutes at 100 degrees Celsius to denature proteins for analysis by gel. The proteins were separated by adding to a 10% SDS polyacrylamide gel and separating at 100 V for approximately 1 hour. Proteins from the gels were then transferred onto nitrocellulose membranes which were each cut in half at ~100 kDa, and each half was transferred to its own individual black staining container. All membranes with separated proteins were blocked with blocking buffer for 1 hour at room temperature. The blocking buffer was decanted, and the membranes containing the top portion of the gel with separated proteins (>100 kDa) wasincubated at 4 degrees Celsius for 24 hours with antisera against HIF-1 (1:500), and themembranes containing the bottom portion of the gel with separated proteins (<100 kDa) was incubated at 4 degrees Celsius for 24 hours with antisera against -Actin (1:1000). After 24 hours, the membranes were washed three times with TBS containing Tween 20 (0.1%) (TBST) and incubated with 680 nm and 800 nm infrared-labeled secondary antibodies for 1 hour at room temperature. The membranes were subsequently washed with TBST, and protein-Attorney Docket No.093386-0041-WO01 antibody complexes were visualized and quantified using the Odyssey direct infrared fluorescence imaging system (FIG.23).

[0234] It is understood that the foregoing detailed description and accompanying examples are merely illustrative and are not to be taken as limitations upon the scope of the disclosure.

[0235] Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications, including without limitation those relating to the chemical structures, substituents, derivatives, intermediates, syntheses, compositions, formulations, or methods of use of the disclosed technology, may be made without departing from the spirit and scope thereof.

[0236] For reasons of completeness, the following Embodiments are provided.

[0237] Clause 1. A block copolymer comprising: a first block (A), the first block comprising recurring units of formula (I)wherein: X1, at each occurrence, is methyl, of formula (II)(II), or of formula (III)(III); L1and L2are each independently -C(O)O-alkylene, -C(O)NRb-alkylene, alkylene, alkylene-O-alkylene, alkylene-O-, or bond; R1is a -interacting moiety; R2is a hydrogen donating moiety; Rais hydrogen or C1-3alkyl; and Rbis hydrogen or C1-6alkyl; and a second block (B), the second block comprising a first hydrophilic polymer; andAttorney Docket No.093386-0041-WO01 a third block (C), the third block comprising a multi-unit segment of formula (IV)wherein: Z is a recurring hydrophilic unit; G is NH or O; L3is alkylene, heteroalkylene, alkenylene, alkynylene, or bond; R3is a guest-host moiety; Rcis hydrogen or C1-3alkyl; and y is no more than 30 mol% of formula (IV).

[0238] Clause 2. The block copolymer of clause 1, wherein the first block comprises a multi- unit segment of formula (V)wherein: X2, at each occurrence, is of formula (II)(II), or of formula (III)Attorney Docket No.093386-0041-WO01 L1and L2are each independently -C(O)O-alkylene, -C(O)NRb-alkylene, alkylene, alkylene-O-alkylene, alkylene-O-, or bond; R1is a -interacting moiety; R2is a hydrogen donating moiety; Ra’and Ra’’are each independently hydrogen or C1-3alkyl; Rbis hydrogen or C1-6alkyl; n is 0 to 200; m is 0 to 200; if n is 0, m is greater than or equal to 10; and if m is 0, n is greater than or equal to 10. Clause 3. The block copolymer of clause 1 or 2, wherein R1is aryl or heteroaryl, wherein the aryl and heteroaryl are optionally substituted with 1 or 2 substituents, each independently halogen, cyano, C1-4alkyl, C1-2fluoroalkyl, –OC1-2alkyl, or –OC1-2fluoroalkyl; and R2is hydroxy, amine, or amide.

[0239] Clause 4. The block copolymer of any one of clauses 1-3, wherein: L1and L2are each independently -C(O)O-C1-4alkylene, -C(O)NH-C1-4alkylene, C1-10alkylene, C1-6alkylene-O-C1-6alkylene, C1-4alkylene-O-, or bond.

[0240] Clause 5. The block copolymer of any one of clauses 1-4, wherein: L1and L2are each independently C1-10alkylene or C1-6alkylene-O-C1-6alkylene; R1is aryl or heteroaryl; and R2is hydroxy or amine.

[0241] Clause 6. The block copolymer of any one of clauses 2-5, wherein: X2, at each occurrence, is of formula (II-a)(II-a), or formula (III-a)(III- L1is C1-4alkylene or C1-2alkylene-O-C1-Attorney Docket No.093386-0041-WO01

[0242] Clause 7. The block copolymer of any one of clauses 2-6, wherein the multi-unit segment of formula (V) comprises recurring units including formula (II) at about 0% to about 100% by weight of the multi-unit segment of formula (V).

[0243] Clause 8. The block copolymer of any one of clauses 2-7, wherein the multi-unit segment of formula (V) comprises recurring units including formula (III) at about 0% to about 100% by weight of the multi-unit segment of formula (V).

[0244] Clause 9. The block copolymer of any one of clauses 2-8, wherein the multi-unit segment of formula (V) has a total unit degree of polymerization of 50 to 300.

[0245] Clause 10. The block copolymer of any one of clauses 1-8, wherein the hydrophilic polymer comprises poly(dimethylacrylamide)(PDMA), poly(ethylene glycol) (PEG), poly(PEG), poly(methyl oxazoline)(PMOX), poly(ethyl oxazoline), polysarcosine, poly(4-acryloylmorpholine), poly(glycerol monomethacrylate), poly(propylene sulfoxide), poly(2-(methylsulfinyl)ethyl acrylate)(PMSEA), poly(vinyl alcohol)(PVA), poly(glycidol), poly(thioglycidyl glycerol)(PTGG), poly(2-methacryloyloxyethyl phosphorylcholine)(PMPC), poly(vinyl pyrrolidone)(PVP), poly(N-(2- hydroxypropyl)methacrylamide)(PHPMA), poly(trimethylamine N-oxide), poly(lysine- methacrylamide), poly(lysine-acrylamide), poly(carboxybetaine), poly(sulfobetaine), heparosan, poly(acrylic acid), poly(acrylamide), or a combination thereof.

[0246] Clause 11. The block copolymer of any one of clauses 1-10, wherein the hydrophilic polymer has a number average molecular weight of about 10 kDa to about 100 kDa as measured by gel permeation chromatography.

[0247] Clause 12. The block copolymer of any one of clauses 1-11, wherein the hydrophilic unit of formula (IV) is derived from dimethylacrylamide, oligo(ethylene glycol), 2- methacryloyloxyethyl phosphorylcholine, N-(2-hydroxypropyl)methacrylamide, or a combination thereof.

[0248] Clause 13. The block copolymer of any one of clauses 1-12, wherein the guest-host moiety is selected from the group consisting of -cyclodextran, adamantane, cholesterol, cucurbiturils, pyrene, crown ethers, potassium ion, calixarenes, long-chain alkyl ammonium ion, pillararene, -carotene, and lutein.

[0249] Clause 14. The block copolymer of any one of clauses 1-13, wherein y is about 1 mol% to about 25 mol% of formula (IV).

[0250] Clause 15. The block copolymer of any one of clauses 1-14, wherein the multi-unit segment of formula (IV) has a total unit degree of polymerization of 100 to 500.Attorney Docket No.093386-0041-WO01

[0251] Clause 16. The block copolymer of any one of clauses 1-15, wherein the third block has a number average molecular weight of about 0.5 kDa to about 100 kDa as measured by gel permeation chromatography.

[0252] Clause 17. The block copolymer of any one of clauses 1-16, wherein the block copolymer is an A-B-C triblock copolymer.

[0253] Clause 18. The block copolymer of any one of clauses 1-17, wherein the block copolymer has a number average molecular weight of about 10 kDa to about 300 kDa as measured by gel permeation chromatography.

[0254] Clause 19. The block copolymer of any one of clauses 1-18, wherein the block copolymer is A5kDa-20kDa-B10Da-40kDa-C20kDa-40Da.

[0255] Clause 20. The block copolymer of any one of clauses 1-19, wherein: the first block comprises a multi-unit segment of formula (V-a) wherein: the first block comprises a multi-unit segment of formula (V-a)(V-a), wherein: X3, at each occurrence, is of formula (II-a)(II-a), or formula (III-a)(III- L1is C1-4alkylene or C1-2alkylene-O-C1-the second block comprises a hydrophilic polymer selected from the group consisting of PDMA, PEG, poly(glycerol monomethacrylate), and a combination thereof.Attorney Docket No.093386-0041-WO01

[0256] Clause 21. A composition comprising: a network of a second hydrophilic polymer crosslinked with a plurality of particles, each particle comprising a plurality of self-assembled block copolymers according to any one of clauses 1-20, wherein the second hydrophilic polymer comprises a guest-host moiety that is capable of forming a guest-host complex with the guest- host moiety of the block copolymer.

[0257] Clause 22. The composition of clause 21, wherein the second hydrophilic polymer comprises hyaluronic acid, gelatin, alginate, carboxymethyl cellulose (CMC), glycol chitosan, carrageenan, dextran, pullulan, or a combination thereof.

[0258] Clause 23. The composition of clause 21 or 22, wherein the second hydrophilic polymer is hyaluronic acid.

[0259] Clause 24. The composition of any one of clauses 21-23, wherein the second hydrophilic polymer has a number average molecular weight of about 0.5 kDa to about 500 kDa as measured by gel permeation chromatography.

[0260] Clause 25. The composition of any one of clauses 21-24, wherein the particle has a diameter of about 50 nm to about 250 nm as measured by dynamic light scattering.

[0261] Clause 26. The composition of any one of clauses 21-25, wherein the second hydrophilic polymer has a guest-host grafting density of about 10% to about 40%.

[0262] Clause 27. The composition of any one of clauses 21-26, wherein the composition has a ratio of the guest-host moiety of the block copolymer to the guest-host moiety of the second hydrophilic polymer of about 1:4 to about 4:1.

[0263] Clause 28. The composition of any one of clauses 21-27, wherein at least one particle comprises a drug.

[0264] Clause 29. The composition of clause 28, wherein the drug comprises a hydrogen bond acceptor moiety, an aromatic moiety, or a combination thereof.

[0265] Clause 30. The composition of clause 28 or 29, wherein the drug comprises a peptide-based drug, a chemotherapeutic, an anti-inflammatory drug, an immune modulating drug, a pro-angiogenic drug, or a combination thereof.

[0266] Clause 31. The composition of any one of clauses 28-30, wherein the drug comprises cyclosporin A, paclitaxel, bortezomib, etoposide, neratinib, osimertinib, chloroquine, GANT58, docetaxel, dexamethasone, carmofur, dexamethasone, dexamethasone palmitate, carfilzomib, afatinib, irinotecan, doxorubicin, doxycycline, camptothecin, imiquimod, MK-8617, ciclopirox, roxadustat, or a combination thereof.

[0267] Clause 32. The composition of any one of clauses 28-31, wherein the drug has a logD of about -1 to about 10.Attorney Docket No.093386-0041-WO01

[0268] Clause 33. The composition of any one of clauses 21-32, wherein the composition is shear thinning.

[0269] Clause 34. The composition of any one of clauses 21-33, wherein the composition has a storage modulus (G') of about 500 Pa to about 5,000 Pa.

[0270] Clause 35. The composition of any one of clauses 21-34, wherein the crosslinked network is a hydrogel.

[0271] Clause 36. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to any one of clauses 21-35.

[0272] Clause 37. A method of tissue repair in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to any one of clauses 21-35, wherein the tissue repair comprises treating a volumetric tissue defect in the subject.

[0273] Clause 38. The method of clause 36, wherein the disease or disorder is osteoarthritis, rheumatoid arthritis, ulcerative colitis, or wound healing.

[0274] Clause 39. The method of any one of clauses 36-38, wherein the subject is human.

Claims

Attorney Docket No.093386-0041-WO01 CLAIMS What is claimed is:

1. A block copolymer comprising: a first block (A), the first block comprising recurring units of formula (I)wherein: X1, at each occurrence, is methyl, of formula (II)of formula (III)L1and L2are each independently -C(O)O-alkylene, -C(O)NRb-alkylene, alkylene, alkylene-O-alkylene, alkylene-O-, or bond; R1is a -interacting moiety; R2is a hydrogen donating moiety; Rais hydrogen or C1-3alkyl; and Rbis hydrogen or C1-6alkyl; and a second block (B), the second block comprising a first hydrophilic polymer; and a third block (C), the third block comprising a multi-unit segment of formula (IV)Attorney Docket No.093386-0041-WO01 (IV) wherein: Z is a recurring hydrophilic unit; G is NH or O; L3is alkylene, heteroalkylene, alkenylene, alkynylene, or bond; R3is a guest-host moiety; Rcis hydrogen or C1-3alkyl; and y is no more than 30 mol% of formula (IV).

2. The block copolymer of claim 1, wherein the first block comprises a multi-unit segment of formula (V)(V), wherein: X2, at each occurrence, is of formula (II)(II), or of formula (III)L1and L2are each independently -C(O)O-alkylene, -C(O)NRb-alkylene, alkylene, alkylene-O-alkylene, alkylene-O-, or bond; R1is a -interacting moiety; R2is a hydrogen donating moiety; Ra’and Ra’’are each independently hydrogen or C1-3alkyl; Rbis hydrogen or C1-6alkyl; n is 0 to 200;Attorney Docket No.093386-0041-WO01 m is 0 to 200; if n is 0, m is greater than or equal to 10; and if m is 0, n is greater than or equal to 10.

3. The block copolymer of claim 1, wherein: R1is aryl or heteroaryl, wherein the aryl and heteroaryl are optionally substituted with 1 or 2 substituents, each independently halogen, cyano, C1-4alkyl, C1-2fluoroalkyl, –OC1-2alkyl, or – OC1-2fluoroalkyl; and R2is hydroxy, amine, or amide.

4. The block copolymer of claim 1, wherein: L1and L2are each independently -C(O)O-C1-4alkylene, -C(O)NH-C1-4alkylene, C1-10alkylene, C1-6alkylene-O-C1-6alkylene, C1-4alkylene-O-, or bond.

5. The block copolymer of claim 1, wherein: L1and L2are each independently C1-10alkylene or C1-6alkylene-O-C1-6alkylene; R1is aryl or heteroaryl; and R2is hydroxy or amine.

6. The block copolymer of claim 2, wherein: X2, at each occurrence, is of formula (II-a)Attorney Docket No.093386-0041-WO01 7. The block copolymer of claim 2, wherein the multi-unit segment of formula (V) comprises recurring units including formula (II) at about 0% to about 100% by weight of the multi-unit segment of formula (V).

8. The block copolymer of claim 2, wherein the multi-unit segment of formula (V) comprises recurring units including formula (III) at about 0% to about 100% by weight of the multi-unit segment of formula (V).

9. The block copolymer of claim 2, wherein the multi-unit segment of formula (V) has a total unit degree of polymerization of 50 to 300.

10. The block copolymer of claim 1, wherein the hydrophilic polymer comprises poly(dimethylacrylamide)(PDMA), poly(ethylene glycol) (PEG), poly(PEG), poly(methyl oxazoline)(PMOX), poly(ethyl oxazoline), polysarcosine, poly(4-acryloylmorpholine), poly(glycerol monomethacrylate), poly(propylene sulfoxide), poly(2-(methylsulfinyl)ethyl acrylate)(PMSEA), poly(vinyl alcohol)(PVA), poly(glycidol), poly(thioglycidyl glycerol)(PTGG), poly(2-methacryloyloxyethyl phosphorylcholine)(PMPC), poly(vinyl pyrrolidone)(PVP), poly(N-(2- hydroxypropyl)methacrylamide)(PHPMA), poly(trimethylamine N-oxide), poly(lysine- methacrylamide), poly(lysine-acrylamide), poly(carboxybetaine), poly(sulfobetaine), heparosan, poly(acrylic acid), poly(acrylamide), or a combination thereof.

11. The block copolymer of claim 1, wherein the hydrophilic polymer has a number average molecular weight of about 10 kDa to about 100 kDa as measured by gel permeation chromatography.

12. The block copolymer of claim 1, wherein the hydrophilic unit of formula (IV) is derived from dimethylacrylamide, oligo(ethylene glycol), 2-methacryloyloxyethyl phosphorylcholine, N- (2-hydroxypropyl)methacrylamide, or a combination thereof.

13. The block copolymer of claim 1, wherein the guest-host moiety is selected from the group consisting of -cyclodextran, adamantane, cholesterol, cucurbiturils, pyrene, crown ethers, potassium ion, calixarenes, long-chain alkyl ammonium ion, pillararene, -carotene, and lutein.Attorney Docket No.093386-0041-WO01 14. The block copolymer of claim 1, wherein y is about 1 mol% to about 25 mol% of formula (IV).

15. The block copolymer of claim 1, wherein the multi-unit segment of formula (IV) has a total unit degree of polymerization of 100 to 500.

16. The block copolymer of claim 1, wherein the third block has a number average molecular weight of about 0.5 kDa to about 100 kDa as measured by gel permeation chromatography.

17. The block copolymer of claim 1, wherein the block copolymer is an A-B-C triblock copolymer.

18. The block copolymer of claim 1, wherein the block copolymer has a number average molecular weight of about 10 kDa to about 300 kDa as measured by gel permeation chromatography.

19. The block copolymer of claim 1, wherein the block copolymer is A5kDa-20kDa-B10Da-40kDa- C20kDa-40Da.

20. The block copolymer of claim 1, wherein: the first block comprises a multi-unit segment of formula (V-a)(V-a), wherein: at each occurrence, is of formula (II-a)(II-a), or formula (III-a)Attorney Docket No.093386-0041-WO01(III-a); L1is C1-4alkylene or C1-2alkylene-O-C1-2alkylene; and L2is C1-4alkylene; and the second block comprises a hydrophilic polymer selected from the group consisting of PDMA, PEG, poly(glycerol monomethacrylate), and a combination thereof.

21. A composition comprising: a network of a second hydrophilic polymer crosslinked with a plurality of particles, each particle comprising a plurality of self-assembled block copolymers according to claim 1, wherein the second hydrophilic polymer comprises a guest-host moiety that is capable of forming a guest-host complex with the guest-host moiety of the block copolymer.

22. The composition of claim 21, wherein the second hydrophilic polymer comprises hyaluronic acid, gelatin, alginate, carboxymethyl cellulose (CMC), glycol chitosan, carrageenan, dextran, pullulan, or a combination thereof.

23. The composition of claim 21, wherein the second hydrophilic polymer is hyaluronic acid.

24. The composition of claim 21, wherein the second hydrophilic polymer has a number average molecular weight of about 0.5 kDa to about 500 kDa as measured by gel permeation chromatography.

25. The composition of claim 21, wherein the particle has a diameter of about 50 nm to about 250 nm as measured by dynamic light scattering.

26. The composition of claim 21, wherein the second hydrophilic polymer has a guest-host grafting density of about 10% to about 40%.

27. The composition of claim 21, wherein the composition has a ratio of the guest-host moiety of the block copolymer to the guest-host moiety of the second hydrophilic polymer of about 1:4 to about 4:1.Attorney Docket No.093386-0041-WO01 28. The composition of claim 21, wherein at least one particle comprises a drug.

29. The composition of claim 28, wherein the drug comprises a hydrogen bond acceptor moiety, an aromatic moiety, or a combination thereof.

30. The composition of claim 28, wherein the drug comprises a peptide-based drug, a chemotherapeutic, an anti-inflammatory drug, an immune modulating drug, a pro-angiogenic drug, or a combination thereof.

31. The composition of claim 28, wherein the drug comprises cyclosporin A, paclitaxel, bortezomib, etoposide, neratinib, osimertinib, chloroquine, GANT58, docetaxel, dexamethasone, carmofur, dexamethasone, dexamethasone palmitate, carfilzomib, afatinib, irinotecan, doxorubicin, doxycycline, camptothecin, imiquimod, MK-8617, ciclopirox, roxadustat, or a combination thereof.

32. The composition of claim 28, wherein the drug has a logD of about -1 to about 10.

33. The composition of claim 21, wherein the composition is shear thinning.

34. The composition of claim 21, wherein the composition has a storage modulus (G’) of about 500 Pa to about 5,000 Pa.

35. The composition of claim 21, wherein the crosslinked network is a hydrogel.

36. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to claim 21.

37. A method of tissue repair in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to claim 21, wherein the tissue repair comprises treating a volumetric tissue defect in the subject.

38. The method of claim 36, wherein the disease or disorder is osteoarthritis, rheumatoid arthritis, ulcerative colitis, or wound healing.Attorney Docket No.093386-0041-WO01 39. The method of claim 36, wherein the subject is human.

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