Conversion of an Injectable MMP-Degradable Hydrogel into Core-Cross-Linked Micelles

Abstract

In this study, a new type of injectable hydrogel called “HyMic” that can convert into core cross-linked (CCL) micelles upon exposure to matrix metalloproteinases (MMP’s), was designed and developed for drug delivery applications. HyMic is composed of CCL micelles connected via an enzyme cleavable linker. To this end, two complementary ABA block copolymers with polyethylene glycol (PEG) as B block were synthesized using atom transfer radical polymerization (ATRP). The A blocks were composed of a random copolymer of N-isopropylacrylamide (NIPAM) and either N-(2-hydroxypropyl)­methacrylamide-cysteine (HPMA-Cys) or N-(2-hydroxypropyl) methacrylamide-ethylthioglycolate succinic acid (HPMA-ETSA). Mixing the aqueous solutions of the obtained polymers and rising the temperature above the cloud point of the PNIPAM block resulted in the self-assembly of these polymers into flower-like micelles composed of a hydrophilic PEG shell and hydrophobic core. The micellar core was cross-linked by native chemical ligation between the cysteine (in HPMA-Cys) and thioester (in HPMA-ETSA) functionalities. A slight excess of thioester to cysteine groups (molar ratio 3:2) was used to allow further chemical reactions exploiting the unreacted thioester groups. The obtained micelles displayed a Z-average diameter of 80 ± 1 nm (PDI 0.1), and ζ-potential of −4.2 ± 0.4 mV and were linked using two types of pentablock copolymers of P­(NIPAM-co-HPMA-Cys)-PEG-peptide-PEG-P­(NIPAM-co-HPMA-Cys) (Pep-NC) to yield hydrogels. The pentablock copolymers were synthesized using a PEG-peptide-PEG ATRP macroinitiator and the peptide midblock (lysine-glycine-proline-glutamine-isoleucine-phenylalanine-glycine-glutamine-lysine (Lys-Gly-Pro-Gln-Gly-Ile-Phe-Gly-Gln-Lys)) consisted of either l- or d-amino acids (l-Pep-NC or d-Pep-NC), of which the l-amino acid sequence is a substrate for matrix metalloproteases 2 and 9 (MMPs 2 and 9). Upon mixing of the CCL micelles and the linker (l/d-Pep-NC), the cysteine functionalities of the l/d-Pep-NC reacted with remaining thioester moieties in the micellar core via native chemical ligation yielding a hydrogel within 160 min as demonstrated by rheological measurements. As anticipated, the gel cross-linked with l-Pep-NC was degraded in 7–45 days upon exposure to metalloproteases in a concentration-dependent manner, while the gel cross-linked with the d-Pep-NC remained intact even after 2 months. Dynamic light scattering analysis of the release medium revealed the presence of nanoparticles with a Z-average diameter of ∼120 nm (PDI < 0.3) and ζ-potential of ∼−3 mV, indicating release of core cross-linked micelles upon HyMic exposure to metalloproteases. An in vitro study demonstrated that the released CCL micelles were taken up by HeLa cells. Therefore, HyMic as an injectable and enzyme degradable hydrogel displaying controlled and on-demand release of CCL micelles has potential for intracellular drug delivery in tissues with upregulation of MMPs, for example, in cancer tissues.

Recent Advances in Injectable Hydrogels for Controlled and Local Drug Delivery

“…Since MMPs are upregulated by cancer tissues, this injectable hydrogel system possesses potential for intracellular drug delivery in cancer cells. [199] In another study, Chwalek et al designed an MMP-degradable hydrogel as a tumor angiogenesis model (Figure 8). Angiogenesis, the growth of new blood vessels, is a necessary process in wound healing.…”

A Novel Temperature‐Dependent Hydrogel Emulsion with Sol/Gel Reversible Phase Transition Behavior Based on Polystyrene‐co‐poly(N‐isopropylacrylamide)/Poly(N‐isopropylacrylamide) Core–Shell Nanoparticle

“…Environmental stimulus‐responsive hydrogels are a class of polymers whose solubility, volume, shape, and configuration can change reversibly in response to external physical or chemical stimuli. [ 1–46 ] External stimuli normally alter ionic interaction, hydrogen bonding, or hydrophobic forces of the polymer systems, leading to invertible microphase separation or self‐assembly. Such smart, sensitive, and multifunctional materials have been applied in drug release, biomedical materials, protein purification, therapeutic agents, industrial coatings, sensors, optical devices, and emulsion stabilization.…”

“…Such smart, sensitive, and multifunctional materials have been applied in drug release, biomedical materials, protein purification, therapeutic agents, industrial coatings, sensors, optical devices, and emulsion stabilization. [ 12–21,23,47–49 ] For example, Li et al. have discussed the structures, performances, mechanisms of action, loading and release behaviors, and applications of various antibacterial hydrogel formulations, as well as their prospects in biomedical research and clinical applications.…”

“…Among all environmentally sensitive hydrogels, temperature‐responsive hydrogels, such as poly( N ‐isopropylacrylamide) (PNIPAM) and its related hybrid copolymers, have received the most attention due to their smart tunable properties. [ 1–44 ] PNIPAM hydrogel polymer chains of low critical solution temperature (LCST, usually around 32 °C) stretch with hydrophilicity at low temperature (below LCST) and shrink with hydrophobicity at high temperature (above LCST). The reason is that in PNIPAM hydrogels, the side group NHCO is hydrophilic, but the main chain is hydrophobic.…”

“…Such smart, sensitive, and multifunctional materials have been applied in drug release, biomedical materials, protein purification, therapeutic agents, industrial coatings, sensors, optical devices, and emulsion stabilization. [12][13][14][15][16][17][18][19][20][21]23,[47][48][49] For example, Li et al have discussed the structures, performances, mechanisms of action, loading and release behaviors, and applications of various antibacterial hydrogel formulations, as well as their prospects in biomedical research and clinical applications. [47] Wang et al reported the application of three kinds of thermogels from the polypeptide methoxy poly(ethylene glycol)−polyalanine (mPEG−PAla) with various chiralities to regulate the levels of inflammatory responses in vivo.…”