Koji Nagahama

Biodegradable Polymeric Assemblies for Biomedical Materials

Recently, self-assembled systems using biodegradable polymers at the nanometer scale, such as microspheres, nanospheres, polymer micelles, nanogels, and polymersomes, have attracted much attention especially in biomedical fields. To construct such self-assembled systems, it is extremely important to have precise control of intermolecular noncovalent interactions, such as hydrophobic interactions based on their amphiphilic molecular structures. Biodegradable polymers, especially aliphatic polyesters such as polylactide, polyglycolide, poly(e-caplolactone) and their copolymers, have been used as biomedical materials for a long time. This chapter is mainly focused on aliphatic polyesters and related polymers, and reviews the synthetic methods for amphiphilic biodegradable polymers containing aliphatic polyesters as components. Moreover, the application of various types of self-assembly systems using amphiphilic biodegradable copolymers such as micro- or nanosized particles (microspheres, nanospheres, polymer micelles, nanogels, polymersomes), supramolecular physically interlocked systems, and stimuli-responsive systems for biomedical use such as drug delivery systems are also reviewed.

Self-assembling polymer micelle/clay nanodisk/doxorubicin hybrid injectable gels for safe and efficient focal treatment of cancer.

The purpose of this study was to fabricate a safe and effective doxorubicin (DOX)-delivery system for focal cancer chemotherapy. A novel biodegradable injectable gel was developed through self-assembly of poly(D,L-lactide-co-glycolide)-b-poly(ethylene glycol)-b-poly(D,L-lactide-co-glycolide) (PLGA-PEG-PLGA) copolymer micelles, clay nanodisks (CNDs), and DOX. We discovered that DOX loaded in the hybrid gels acts as an anticancer drug and as a building block to organize new gel networks. Accordingly, long-term sustained release of DOX from hybrid injectable gels without initial burst release was achieved. Moreover, it was revealed that the DOX incorporated into gel networks controls its own release profile. This hybrid injectable gel is a self-controlled drug release system, which is a novel concept in controlled drug release. Importantly, a single injection of PLGA-PEG-PLGA/CND/DOX hybrid gel provides long-term sustained antitumor activity in vivo against human xenograft tumors in mice, suggesting the potential of hybrid gels as a valuable local DOX-delivery platform for cancer focal therapy.

Anticancer drug-based multifunctional nanogels through self-assembly of dextran-curcumin conjugates toward cancer theranostics.

Curcumin (CCM) has been received much attention in cancer theranostics because CCM exhibits both anticancer activity and strong fluorescence available for bio-imaging. However, CCM has never been utilized in clinical mainly due to its extremely low water solubility and its low cellular uptake into cancer cells. We fabricated novel CCM-based biodegradable nanoparticles through self-assembly of amphiphilic dextran-CCM conjugates. Significantly high CCM loading contents in the nanoparticles and the high water solubility were achieved. Importantly, the dextran-CCMs nanoparticles were effectively delivered into HeLa cells and exhibited strong fluorescence available for live-cell imaging, although the nanoparticles were not delivered into normal cells. Thus, the dextran-CCMs nanoparticles could be a promising for creation of novel CCM-based cancer theranostics with high efficacy.

Polyrotaxane Composed of Poly-l-lactide and α-Cyclodextrin Exhibiting Protease-Triggered Hydrolysis

Biodegradable polyrotaxanes (PRXs) were synthesized from bis-amino-terminated poly(L-lactide) (PLLA) and alpha-cyclodextrin (alpha-CD), combined with capping by bulky end groups at the amino-termini of PLLA through enzymatically cleavable peptide linkages. The crystalline structure of the PRXs in the solid state was investigated by wide-angle X-ray diffraction (WAXRD), and the results suggested that PRX forms a column-type crystalline structure. Hydrolysis of ester bonds of the PLLA in PRX was prevented due to the supramolecular structure. However, in the presence of a protease (papain), the hydrolysis of PLLA in PRX was induced. The removal of the bulky end groups by the protease acted as a trigger for the release of alpha-CD and allowed hydrolysis of the PLLA ester bonds. Such unique hydrolysis behavior, that is, proteinase-triggered degradation of a polyester, was achieved by the combination of the supramolecular architecture, biodegradable PLLA, and enzymatically cleavable end groups.

Biodegradable Nanogels Prepared by Self‐Assembly of Poly(L‐lactide)‐Grafted Dextran: Entrapment and Release of Proteins

We showed previously that poly(L-lactide)-grafted dextran could form biodegradable nanogels in water. In this paper, various properties of Dex-g-PLLA nanogels were compared with Dex-Chol (dextran-cholesterol conjugate) nanogels to investigate the effects of hydrophobic units. Dex-g-PLLA nanogels exhibited significantly lower CAC and higher colloidal stability, indicating a strong tendency to form nanogels. We prepared lysozyme-loaded Dex-g-PLLA nanogels, and they exhibited a sustained release of lysozyme for 1 week without denaturation in PBS at 37 degrees C. The Dex-g-PLLA nanogels therefore have great potential as a delivery vehicle for therapeutic protein.

The Synthesis and Biodegradability of Poly(lactide-random-depsipeptide)-PEGPoly(lactide-random-depsipeptide) ABA-type Triblock Copolymers

Amphiphilic ABA-type triblock copolymers were synthesized to develop a biodegradable anti-adhesive membrane. In this particular synthesis, poly[L-lactide(LA)-co-depsipeptide] (poly[LA-co-(Glc-Leu)]: PLGL) was used as the A segment, and the poly(ethylene glycol)s (PEG)s, Mn 10,000 and Mn 20,500 were used as the B segment. The synthesis of the triblock copolymer (PLGL-PEG-PLGL) was carried out via a ring-opening copolymerization of L-lactide and cyclo(Glc-Leu) in the presence of hydroxytelechelic poly(ethylene glycol) using tin 2-ethylhexanoate as a catalyst. To evaluate the copolymer films as candidates for biodegradable anti-adhesive membranes, physicochemical properties such as degradation on behavior under physiological conditions and water absorption were investigated. The degradation rate of the PLGLPEG-PLGL films varied with changes in the molecular architecture; specifically, the molecular weight of the hydrophilic B segment and the depsipeptide unit content in the A segment were more prominent. The biocompatibility and resorption of the PLGL-PEG-PLGL films were also evaluated. The PLGLPEG-PLGL films were degraded and depleted gradually in vivo without inflammation.

Discovery of a new function of curcumin which enhances its anticancer therapeutic potency

Curcumin has received immense attention over the past decades because of its diverse biological activities and recognized as a promising drug candidate in a large number of diseases. However, its clinical application has been hindered due to extremely low aqueous solubility, chemical stability, and cellular uptake. In this study, we discovered quite a new function of curcumin, i.e. pH-responsive endosomal disrupting activity, derived from curcumin’s self-assembly. We selected anticancer activity as an example of biological activities of curcumin, and investigated the contribution of pH-responsive property to its anticancer activity. As a result, we demonstrated that the pH-responsive property significantly enhances the anticancer activity of curcumin. Furthermore, we demonstrated a utility of the pH-responsive property of curcumin as delivery nanocarriers for doxorubicin toward combination cancer therapy. These results clearly indicate that the smart curcumin assemblies act as promising nanoplatform for development of curcumin-based therapeutics.

A nanocomposite approach to develop biodegradable thermogels exhibiting excellent cell-compatibility for injectable cell delivery

A new class of injectable nanocomposite thermogels having excellent cell-compatibility were developed through cooperative self-assembly of biodegradable poly(lactide-co-glycolide)-b-poly(ethylene glycol)-b-poly(lactide-co-glycolide) copolymer micelles and clay nanosheets for effective cell delivery. This study will be valuable for the establishment of injectable cell delivery technology.

Biodegradable Nano-aggregates of Star-Shaped 8-arm PEG-PLLA Block Co-polymers for Encapsulation of Water-Soluble Macromolecules

A series of amphiphilic 8-arm PEG-b-PLLA co-polymers with star-shaped structure was synthesized through ring-opening polymerization of L-lactide (L-LA) in the presence of 8-arm PEG as a macroinitiator by varying feeding molar rations of L-LA to 8-arm PEG. 8-arm PEG-b-PLLA co-polymers having certain PEG content and PEG length were found to self-assemble into nano-aggregates in aqueous solutions. The size and the morphology of the nano-aggregates were investigated by dynamic light scattering and 1H-NMR in CDCl3 and D2O. The results indicate that the average diameter was ca. 150 nm, the surface of the nano-aggregates was covered by PEG chains and the PLLA cores formed by hydrophobic interaction are located inside of the nano-aggregates. FITC-dextran molecules, as model for water-soluble macromolecular drugs, were successfully encapsulated into 8-arm PEG-b-PLLA nano-aggregates by simple addition of FITC-dextran to the aqueous phase during the self-assembly process. This result suggests that the nanoaggregates have a vesicle-like morphology. The nano-aggregates dissociated gradually in the order of weeks in PBS (pH 7.4, ionic strength 140 mM) at 37°C. Thus, the novel nano-aggregates of 8-arm PEG-b-PLLA can be expected to have advantages, such as long circulation times, as drug carriers which show sustained release of loaded macromolecular drugs such as antibodies and DNA vaccines in the blood stream.

Enhanced stereocomplex formation of enantiomeric polylactides grafted on a polyrotaxane platform

We report (i) synthesis of novel graft copolymers of a α-CD/PEG polyrotaxane (PRX) backbone and enantiomeric PLA side chains, PRX-g-PLLA and PRX-g-PDLA, (ii) selective stereocomplex formation on a movable PRX in the bulk state, and (iii) preparation of soft materials with continuous anisotropic phases by blending of the L- and D-isomers.