Percutaneous Intratumoral Immunoadjuvant Gel Increases the Abscopal Effect of Cryoablation for Checkpoint Inhibitor Resistant Cancer.

Percutaneous cryoablation is a common clinical therapy for metastatic and primary cancer. There are rare clinical reports of cryoablation inducing regression of distant metastases, known as the "abscopal" effect. Intratumoral immunoadjuvants may be able to augment the abscopal rate of cryoablation, but existing intratumoral therapies suffer from the need for frequent injections and inability to confirm target delivery, leading to poor clinical trial outcomes. To address these shortcomings, an injectable thermoresponsive gel-based controlled release formulation is developed for the FDA-approved Toll-like-receptor 7 (TLR7) agonist imiquimod ("Imigel") that forms a tumor-resident depot upon injection and contains a contrast agent for visualization under computed tomography (CT). The poly-lactic-co-glycolic acid-polyethylene glycol-poly-lactic-co-glycolic acid (PLGA-PEG-PLGA)-based amphiphilic copolymer gel's underlying micellar nature enables high drug concentration and a logarithmic release profile that is additive with the neo-antigen release from cryoablation, requiring only a single injection. Rheological testing demonstrated the thermoresponsive increase in viscosity at body temperature and radio-opacity via microCT. Its ability to significantly augment the abscopal rate of cryoablation is demonstrated in otherwise immunotherapy resistant metastatic tumors in two aggressive colorectal and breast cancer dual tumor models with an all or nothing response, responders generally demonstrating complete regression of bilateral tumors in 90-day survival studies.

Engineered insulin-polycation complexes for glucose-responsive delivery with high insulin loading.

Glucose-responsive insulin delivery systems have the potential to improve quality of life for individuals with diabetes by improving blood sugar control and limiting the risk of hypoglycemia. However, systems with desirable insulin release kinetics and high loading capacities have proven difficult to achieve. Here, we report the development of electrostatic complexes (ECs) comprised of insulin, a polycation, and glucose oxidase (GOx). Under normoglycemic physiological conditions, insulin carries a slight negative charge and forms a stable EC with the polycation. In hyperglycemia, the encapsulated glucose-sensing enzyme GOx converts glucose to gluconic acid and lowers the pH of the microenvironment, causing insulin to adopt a positive charge. Thus, the electrostatic interactions are disrupted, and insulin is released. Using a model polycation, we conducted molecular dynamics simulations to model these interactions, synthesized ECs with > 50% insulin loading capacity, and determined in vitro release kinetics. We further showed that a single dose of ECs can provide a glycemic profile in streptozotocin-induced diabetic mice that mimics healthy mice over a 9 h period with 2 glucose tolerance tests.

Ion Exchange Nanoparticles for Ophthalmic Drug Delivery.

We report here on ion-exchange polymeric nanoparticles from a linear copolymer of maleic anhydride methyl vinyl ether esterified with 30% octadecanol. The side chains for the polymer structure were optimized through metadynamics simulations, which revealed the use of octadecanol esters generates ideal free energy surfaces for drug encapsulation and release. Nanoparticles were synthesized using a solvent evaporation-precipitation method by mixing the polymer solution in acetone into water; upon acetone evaporation, a nanodispersion with an average particle size of ∼150 nm was obtained. Gentamicin sulfate, possessing five amino groups, was spontaneously entrapped in the nanocarrier by ionic interactions. Encapsulation efficiency increases significantly with the increase in pH and ionic strength. In vivo results demonstrate high gentamicin (GM) content in the enteric chamber (AUC 8207 ± 1334 (µg min)/mL) compared to 3% GM solution (AUC 2024 ± 438 (µg min)/mL). The formulation was also able to significantly extend the release of gentamicin when applied to rabbit cornea. These anionic nanoparticles can be used for extended-release of other cationic drugs.

Biodegradable Poly(Acetonide Gluconic Acid) for Controlled Drug Delivery.

We report here on the synthesis, characterization, degradation, and drug release of acetal-protected gluconic acid-based poly(α-hydroxy ester). This polyester was synthesized by ring-opening polymerization of O-carboxyanhydride of acetal-protected gluconic acid. The polymer undergoes hydrolytic degradation under mild acidic media, whereas minimal degradation takes place under physiological pH. Under acidic conditions, the acetal-protecting groups are hydrolyzed, resulting in a water-soluble polyester with saccharide side chains that erodes from the surface, leaving the bulk of the polymer matrix intact. At pH 3.5, zero-order kinetics was maintained for 50 days accounting for ∼75% drug release. These biodegradable, pH-responsive, sustained zero-order release kinetics of the polymer have application as drug carriers for oral drug delivery or medical implants or also for nonmedical applications.

Recent Advances in Polyanhydride Based Biomaterials.

This review focusses on recent developments of polyanhydrides, a class of degradable synthetic biopolymers. Polyanhydrides have been used as carriers for controlled delivery of drugs. A polyanhydride copolymer of carboxyphenoxy propane and sebacic acid has been used in Gliadel brain tumor implants for the controlled delivery of carmustine or bis-chloroethylnitrosourea. They are easy and inexpensive to synthesize (especially scale up). However, polyanhydrides possess a short shelf-life. Hydrolytic cleavage and anhydride interchanges lower their molecular weights during storage. One of the highlights in recent developments of polyanhydride chemistry is the discovery of alternating copolymers having extended shelf-life. Other highlights include their applications in biomedical electronics, vaccine delivery, and nano/micro particulate delivery systems. This review examines approaches for polyanhydride synthesis followed by their recent developments in biomedical applications.

Stable polyanhydride synthesized from sebacic acid and ricinoleic acid.

Poly(anhydride) are unstable and prone to hydrolytic degradation and depolymerisation via anhydride interchange. They are stored at -20°C, packed under inert atmosphere until use. We synthesized a new poly(anhydride) from ricinoleic (RA) and sebacic (SA) acid with alternating ester-anhydride structure that is stable at 25°C for over 18months. The copolymer is also stable in chloroform solution and under γ-irradiation. The polymer hydrolyses through anhydride cleavage lasting ~7days to form oligoesters, which are stable for >30days. The release of gentamycin from the synthesized alternate polymer matrix is sustained compared to the random copolymer.

Comparative evaluation of polycyanoacrylates.

Cyanoacrylate esters (CA) and their corresponding polymers (PolyCA) are used as general and medical adhesives, biodegradable carriers for controlled drug delivery, and as agents for fingerprint development in forensic science. Most reports of cyanoacrylate are on ethyl or 2-octyl cyanoacrylate ester with little attention to other esters. It is the objective of this study to determine the differences amongst cyanoacrylate esters regarding their synthesis, chemical characterization, hydrolytic degradation, and thermal and mechanical properties. Cyanoacrylate polymers of short and long alkyls or oxy-alkyls, cyclic, and aromatic esters have been synthesized and evaluated. All monomers form polymers when exposed to triethylamine vapours possessing molecular weights in the range of 15,000-150,000Da, where the alkyl esters form high MW polymers. A wide range of hydrolytic degradation rates has been found, as monitored by the release of formaldehyde over time. Alkoxy CAs show faster hydrolytic degradation compared to alkyl CAs. Regarding mechanical properties, CAs are classified into primarily viscous (G'>G″) and primarily elastic (G″>G'). Alkoxy CA polymers have a higher loss modulus (G') than storage modulus (G″). Octyl CA polymer possess a G'≈G″ (phase angle ∼45°) providing appropriate balance between mechanical strength and plasticity. Most alkyl CAs are compact and brittle. Alkoxy CAs show enhanced plasticity, but they lack mechanical strength. In general, the Tg for alkoxy CAs is less than alkyl CAs. Alkoxy CAs depolymerise rapidly at temperatures >200°C. Overall, ester sidechains of CA esters strongly affect the polymer property. STATEMENT OF SIGNIFICANCE: Polycyanoacrylates are an important class of biodegradable polymers mainly used as bioadhesives. The study describes comparative evaluation of different cyanoacrylate polymers with respect to their chemistry, degradation, safety, mechanical, and thermal properties. The study forms the basis for choosing appropriate combination of cyanoacrylate esters for various biomedical uses. Moreover, this study reveals properties of a few new polycyanoacrylates for the first time.

Discovering Novel and Diverse Iron-Chelators in Silico.

Specific iron chelation is a validated strategy in anticancer drug discovery. However, only a few chemical classes (4-5 categories) have been reported to date. We discovered in silico five new structurally diverse iron-chelators by screening through models based on previously known chelators. To encompass a larger chemical space and propose newer scaffolds, we used our iterative stochastic elimination (ISE) algorithm for model building and subsequent virtual screening (VS). The ISE models were developed by training a data set of 130 reported iron-chelators. The developed models are statistically significant with area under the receiver operating curve greater than 0.9. The models were used to screen the Enamine chemical database of ∼1.8 million molecules. The top ranked 650 molecules were reduced to 50 diverse structures, and a few others were eliminated due to the presence of reactive groups. Finally, 34 molecules were purchased and tested in vitro. Five compounds were identified with significant iron-chelation activity in Cal-G assay. Intracellular iron-chelation study revealed one compound as equivalent in potency to the iron chelating "gold standards" deferoxamine and deferiprone. The amount of discovered positives (5 out of 34) is expected by the realistic enrichment factor of the model.

Poly(α-hydroxy acid)s and poly(α-hydroxy acid-co-α-amino acid)s derived from amino acid.

Polyesters derived from the α-hydroxy acids, lactic acid, and glycolic acid, are the most common biodegradable polymers in clinical use. These polymers have been tailored for a range of applications that require a physical material possessing. The physical and mechanical properties of these polymers fit the specific application and also safely biodegrade. These polymers are hydrophobic and do not possess functional side groups. This does not allow hydrophilic or hydrophobic manipulation, conjugation of active agents along the polymer chain, etc. These manipulations have partly been achieved by block copolymerization with, for example, poly(ethylene glycol), to obtain an amphiphilic copolymer. The objective of this review is to survey PLA functional copolymers in which functional α-hydroxy acids derived from amino acids are introduced along the polymer chain, allowing endless manipulation of PLA. Biodegradable functional polyesters are one of the most versatile biomaterials available to biomedical scientists. Amino acids with their variable side chains are ideal candidates for synthesizing such structural as well as stereochemically diverse polymers. They render control over functionalization, conjugation, crosslinking, stimulus responsiveness, and tunable mechanical/thermal properties. Functionalized amino acid derived polyesters are widely used, mainly due to advancement in ring opening polymerization (primarily O-carboxyanhydride mediated). The reaction proceeds under milder conditions and yields high molecular weight polymers. We reviewed on advances in the synthetic methodologies for poly-α-hydroxy esters derived from amino acids with appropriate recent examples.

Biodegradable inflatable balloons for tissue separation.

Confining radiation to a specific region (during radiation therapy) minimizes damage to surrounding tissues. Biodegradable inflatable balloons (bio-balloons) were developed. The device protects the normal tissues by increasing the gap between radiation source and critical structures. The radiation fades away while passing through the inflated balloon preventing the surrounding tissues from harmful radiation. These bio-balloons have also found clinical use to treat massive rotator cuff tear. This review summarizes the chemistry, engineering, and clinical development of these biomedical devices. These balloons are made of biodegradable polymers folded into the edge of a trocar and inserted between the tissues to be separated, and inflated by normal saline in the site of the application. The inserted balloon protects the tissues from radiation or mechanical stress. They remain inflated on site for two months and are finally eliminated within 12 months.

Computational Discovery and Experimental Confirmation of TLR9 Receptor Antagonist Leads.

Toll-like receptors (TLR) are receptors of innate immunity that recognize pathogen associated molecular patterns. They play a critical role in many pathological states, in acute and chronic inflammatory processes. TLR9 is a promising target for drug discovery, since it has been implicated in several pathologies, including defense against viral infections and psoriasis. Immune-modulators are promising molecules for therapeutic intervention in these indications. TLR9 is located in the endosome and activated by dsDNA with CpG motives encountered in microbial DNA. Here we report on a combined approach to discover new TLR9 antagonists by computational chemistry and cell based assays. We used our in-house iterative stochastic elimination (ISE) algorithm to create models that distinguish between TLR9 antagonists ("actives") and other molecules ("inactives"), based on molecular physicochemical properties. Subsequent screening and scoring of a data set of 1.8 million commercially available molecules led to the purchasing of top scored molecules, which were tested in a new cell based system based on human pattern recognition receptors (PRRs) stably expressed in NIH3T3 fibroblasts. As described previously, this cell line shows a very low endogenous PRR-activity and contains a reporter gene which is selectively activated by the integrated human PRR enabling rapid screening of potential ligands. IC50 values of each of these top scored molecules were determined. Out of 60 molecules tested, 56 showed antagonistic activity. We discovered 21 new highly potential antagonists with IC50 values lower than 10 µM, with 5 of them having IC50 values under 1 µM.

Poly(lactic acid) based hydrogels.

Polylactide (PLA) and its copolymers are hydrophobic polyesters used for biomedical applications. Hydrogel medicinal implants have been used as drug delivery vehicles and scaffolds for tissue engineering, tissue augmentation and more. Since lactides are non-functional, they are copolymerized with hydrophilic monomers or conjugated to a hydrophilic moiety to form hydrogels. Copolymers of lactic and glycolic acids with poly(ethylene glycol) (PEG) provide thermo-responsive hydrogels. Physical crosslinking mechanisms of PEG-PLA or PLA-polysaccharides include: lactic acid segment hydrophobic interactions, stereocomplexation of D and L-lactic acid segments, ionic interactions, and chemical bond formation by radical or photo crosslinking. These hydrogels may also be tailored as stimulus responsive (pH, photo, or redox). PLA and its copolymers have also been polymerized to include urethane bonds to fabricate shape memory hydrogels. This review focuses on the synthesis, characterization, and applications of PLA containing hydrogels.

Injectable formulations of poly(lactic acid) and its copolymers in clinical use.

Poly(lactic acid) and its copolymers have revolutionized the field of drug delivery due to their excellent biocompatibility and tunable physico-chemical properties. These copolymers have served the healthcare sector by contributing many products to combat various diseases and for biomedical applications. This article provides a comprehensive overview of clinically used products of poly(lactic acid) and its copolymers. Multi-dimension information covering product approval, formulation aspects and clinical status is described to provide a panoramic overview of each product. Moreover, leading patented technologies and various clinical trials on these products for different applications are included. This review focuses on marketed injectable formulations of PLA and its copolymers.

Alternating Poly(ester-anhydride) by Insertion Polycondensation.

We report on a synthetic method where polyanhydride is used as starting material and the ester monomers are inserted through complete esterification, leading to an alternating ester-anhydride copolymer. The molar ratio of ricinoleic acid (RA) and sebacic acid (SA) was optimized until polysebacic acid is completely converted to carboxylic acid-terminated RA-SA and RA-SA-RA ester-dicarboxylic acids. These dimers and trimers were activated with acetic anhydride, polymerized under heat and vacuum to yield alternating RA-SA copolymer. The resulting alternating poly(ester-anhydride) have the RA at regular intervals. The regular occurrences of RA side chains prevent anhydride interchange, enhancing hydrolytic stability, which allows storage of the polymer at room temperature.

Tyrosine Residue in the TRPV1 Vanilloid Binding Pocket Regulates Deactivation Kinetics.

Vanilloids are pain evoking molecules that serve as ligands of the "heat and capsaicin receptor" TRPV1. Binding of either endogenous or exogenous vanilloids evokes channel and subsequent neuronal activation, leading to pain sensation. Despite its pivotal physiological role, the molecular basis of TRPV1 activation and deactivation is not fully understood. The highly conserved tyrosine in position 511 (Tyr(511)) of the rat TRPV1 (rTRPV1) was the first residue to be identified as a necessary participant in the vanilloid-mediated response. rTRPV1 cryo-EM structures implicated rotation of this residue in the vanilloids bound state. Therefore, we hypothesize that the rTRPV1 Tyr(511) residue entraps vanilloids in their binding site, prolonging channel activity. To test our hypothesis, we generated an array of rTRPV1 mutants, containing the whole spectrum of Tyr(511) substitutions, and tested their response to both exo- and endovanilloids. Our data show that only substitutions of Tyr(511) to aromatic amino acids were able to mimic, albeit partially, the vanilloid-evoked activation pattern of the wt receptor. Although these substitutions reduced the channel sensitivity to vanilloids, a maximal open-channel lifetime could be achieved. Moreover, whereas their current activation rate remains intact, receptors with Tyr(511) substitutions exhibited a faster current deactivation. Our findings therefore suggest that the duration of channel activity evoked by vanilloids is regulated by the interaction between Tyr(511) and the agonist. To conclude, we suggest that Tyr(511)-mediated anchoring of vanilloids in their binding pocket is pivotal for TRPV1 activation and subsequent pain sensation.

PEG-Biscyanoacrylate Crosslinker for Octyl Cyanoacrylate Bioadhesive.

PEG400 (polyethylene glycol, MW 400) biscyanoacrylate is synthesized and copolymerized with 2-octyl cyanoacrylate for potential use as bioadhesive. PEG400 biscyanoacrylate is synthesized from the esterification of anthracenyl cyanoacrylic acid where the anthracene unit serves as vinyl-protecting group. Copolymerization increases the plasticity, mechanical strength, and resilience of the resulted polymer as determined by dynamic mechanical analysis. Peeling test confirms its superior bioadhesive properties. Surface morphology is characterized by SEM imaging. The formulations are cytocompatible and safe. This cyanoacrylate composition may provide improved bioadhesive cyanoacrylates.

Castor Oil-Based Biodegradable Polyesters.

This Review compiles the synthesis, physical properties, and biomedical applications for the polyesters based on castor oil and ricinoleic acid. Castor oil has been known for its medicinal value since ancient times. It contains ∼90% ricinoleic acid, which enables direct chemical transformation into polyesters without interference of other fatty acids. The presence of ricinoleic acid (hydroxyl containing fatty acid) enables synthesis of various polyester/anhydrides. In addition, castor oil contains a cis-double bond that can be hydrogenated, oxidized, halogenated, and polymerized. Castor oil is obtained pure in large quantities from natural sources; it is safe and biocompatible.

Polysaccharide-Based Conjugates for Biomedical Applications.

Polysaccharides contain different functional groups (such as hydroxyl, amino, carboxylic acid, aldehydes) that make them ideal for conjugation. They are biodegradable, biocompatible, and hydrophilic. Polysaccharide conjugates have been used in drug, gene, and macromolecule delivery, tissue engineering, and other biomedical applications. Polysaccharide conjugates have also been used primarily for solubilization and controlled release of hydrophobic moieties. The advent of nanotechnology, gene therapy, and tissue engineering influenced the way these conjugates are now used. Modern day conjugates are modulated to be thermoresponsive, pH-responsive, photoresponsive, or target-specific (receptor mediated targeting). This Review briefly introduces different polysaccharides followed by different synthetic strategies used for conjugation; finally, recent applications were compiled.