Treatment of colorectal cancer by anticancer and antibacterial effects of hemiprotonic phenanthroline-phenanthroline^(+) with nanomicelle delivery

Colorectal cancer(CRC)is a common digestive tract tumor worldwide.Specific microorganisms,including Fusobacterium nucleatum(F.nucleatum)and Escherichia coli(E.coli),are abundant in colonic mucosa and can promote the cancer progression and malignancy.Therefore,a therapeutic strategy is proposed to deliver effective drugs to colorectum for both anticancer and antibacteria.Here we used thin-film dispersionmethod to encapsulate hemiprotonic phenanthroline-phenanthroline^(+)(ph-ph^(+))into nanomicelle.The results showed that the drug-loading nanomicelle had good dispersion,and the particle size was about 28 nm.In vitro assay indicated that the nanomicelle was active against CRC-related obligate and facultative anaerobes.In human CRC cells,the nanomicelle could effectively inhibit cell proliferation and induce apoptosis.In vivo distribution showed that the nanomicelle could release ph-ph^(+) mainly in the colorectum.In CRC model mice,the nanomicelle significantly reduced tumor number and volume,and decreased the bacteria load and colorectal inflammation.Together,the study identifies that the ph-ph^(+) nanomicelle has the potential to apply in treating CRC,and also suggests that anticancer combined with antimicrobial therapy would be a feasible way for CRC therapy.

Preparation Process of Plumbagin Nanomicelle In-situ Gel

[Objectives]To prepare plumbagin nanomicelle(PLB-N)in-situ gel,and optimize the formulation and process.[Methods]PLB-N was prepared by self-assembly method,and the optimal formulation of PLB-N in-situ gel was determined by orthogonal experiment design and single factor method.[Results]The optimal preparation process for PLB-N was a drug to lipid ratio of 1:3,a Tween 80 content of 5%,an ethanol content of 7.5%of the hydration medium,a magnetic stirring speed of 2200 rpm,a stirring time of 30 min,and an ultrasound time of 10 min.The optimal formulation of PLB-N in-situ gel was 22%of poloxamer 407,6%of poloxamer 188,and 1:1 of PLB-N to water.The encapsulation efficiency of PLB-N prepared with the optimal formula was(95.8%±0.4%),and the average particle size was(75.19±1.14)nm,and the Zeta potential was(-20.73±1.19)mv.[Conclusions]PLB-N in-situ gel had stable and reliable preparation process,uniform content,and broad application prospects.

Ocular drug delivery systems:An overview

The major challenge faced by today's pharmacologist and formulation scientist is ocular drug delivery. Topical eye drop is the most convenient and patient compliant route of drug administration,especially for the treatment of anterior segment diseases. Delivery of drugs to the targeted ocular tissues is restricted by various precorneal,dynamic and static ocular barriers. Also,therapeutic drug levels are not maintained for longer duration in target tissues. In the past two decades,ocular drug delivery research acceleratedly advanced towards developing a novel,safe and patient compliant formulation and drug delivery devices/techniques,which may surpass these barriers and maintain drug levels in tissues. Anterior segment drug delivery advances are witnessed by modulation of conventional topical solutions with permeation and viscosity enhancers. Also,it includes development of conventional topical formulations such as suspensions,emulsions and ointments. Various nanoformulations have also been introduced for anterior segment ocular drug delivery. On the other hand,for posterior ocular delivery,research has been immensely focused towards development of drug releasing devices and nanoformulations for treating chronic vitreo-retinal diseases. These novel devices and/or formulations may help to surpass ocular barriers and associated side effects with conventional topicaldrops. Also,these novel devices and/or formulations are easy to formulate,no/negligibly irritating,possess high precorneal residence time,sustain the drug release,and enhance ocular bioavailability of therapeutics. An update of current research advancement in ocular drug delivery necessitates and helps drug delivery scientists to modulate their think process and develop novel and safe drug delivery strategies. Current review intends to summarize the existing conventional formulations for ocular delivery and their advancements followed by current nanotechnology based formulation developments. Also,recent developments with other ocular drug delivery strategies employing in situ gels,implants,contact lens and microneedles have been discussed.

Preparation and Characterization of Florfenicol/Chitosan-stearic Acid Polymer Nanomicelle and Its Antibiotic Properties

Amphiphilic CS-SA polymers were prepared using the SA modified by small molecule water-soluble chitosan, and CS-SA nanomicelles and FF/CS-SA nanomicelles were prepared by using CSSA polymers as the material with dialysis-ultrasound method. CS-SA polymers, CS-SA nanomicelles, and FF/CS-SA nanomicelles were characterized by FT-IR, TGA, and SEM. Results showed that the SA was grafted to the amino of CS by amide bond. CS-SA nanomicelles and FF/CS-SA nanomicelles were spherical, smooth without fold. The influence of different molar ratio of FF to CS-SA polymers on the encapsulation efficiency and drug-loading rate determined the best molar ratio that was 1：1.09. In vitro release experiments, drug release performance study found that hydrophobic drug releasing from FF/CS-SA nanomicelles presented sustainedrelease. In vitro bacteriostasis experiments, when the concentration was higher than 4.98 mg/mL, FF/CSSA nanomicelles had antibacterial action and a positive correlation with concentration. The results revealed that amphiphilic chitosan derivative nanomicelles were carriers with broad prospects, increasing solubility of hydrophobic drugs and presenting sustained release for hydrophobic drugs, which provided a new research idea for drug delivery and targeted drug delivery of hydrohobic drugs.

Thin-film hydration preparation method and stability test of DOX-loaded disulfide-linked polyethylene glycol 5000-lysine-di-tocopherol succinate nanomicelles

A novel redox-responsive PEG-sheddable copolymer of disulfide-linked polyethylene glycol 5000-lysine-di-tocopherol succinate(P_(5k)SSLV)was designed and synthesized.Thin-film hydration method was used to prepare DOX-loaded P_(5k)SSLV nanomicelle.To optimize the preparation technology,we investigate the effects of dosage,type of organic solvent,hydration temperature and time,and cryoprotectant on drug-loading content,encapsulation efficiency,particle size,and zeta potential.The mean particle size and zeta potential were determined by Zetasizer.The morphology of the P_(5k)SSLV-DOX nanomicelles was visualized by transmission electron microscopy.The drug-loading content and encapsulation efficiency of P_(5k)SSLV-DOX nanomicelle were investigated by UV.The drug-loading content,encapsulation efficiency,particle size,and zeta potential of the final optimized nanomicelles were 4.58%,97.20%,30.21 nm and -0.84 mV,respectively.In addition,the stability of nanomicelles was investigated,which included dilution stability and storage stability.The results showed that P_(5k)SSLV-DOX nanomicelle had good dilution stability and storage stability at 4℃.The preparation method of P_(5k)SSLV-DOX nanomicelle with thinfilm hydration method was practical and simple,which was valuable to be further studied.

Construction of AIEgen functionalized nanomicelles and their stability study through‘seesaw-like'fluorescence changes

As nanocarriers,nanomicelles play vital roles in the toolbox of drug delivery.The stability of nanomicelles affects the nanomedicines'bioactivity.Therefore,it is important to understand the stability of nanomicelles for further improvements.Here,we report a strategy to construct new nanomicelles(NM)by introducing aggregation-induced emission(AIE)functional group tetraphenylethylene(TPE)in the component polymer vitamin E(D-α-tocopheryl polyethylene glycol 1000 succinate)(TPGS).The stability of doxorubicin(DOX)loaded nanomicelles DOX@NM in different conditions was studied by fluorescence analysis.The fluorescence changes of DOX@NM are‘seesaw-like'when they transform between assembled and disassembled forms.In the assembled form,TPE gives emission from AIE effect,while in the disassembled form,the fluorescence of DOX is observed due to the disappearance of ACQ effect.

Melphalan-loaded methoxy poly(ethylene glycol)-poly(D,L-lactide)copolymer nanomicelles in the treatment of multiple myeloma

Multiple myeloma(MM)is the second most common hematological tumor characterized by the proliferation of monoclonal plasma cells.Melphalan(MEL)is commonly used in the treatment of MM and is especially essential for patients undergoing autologous stem cell transplantation(ASCT).Although many drugs for MM have been developed in recent years,chemotherapy followed by ASCT remains the optimal option.Melphalan,the backbone of the conditioning regimen,brings severe toxicities at a high dose.Nanodrug delivery systems enable drugs to be highly effective and have low toxicity.In this study,methoxy poly(ethylene glycol)-poly(D,L-lactide)copolymer(MPEG-PDLLA)was chosen to encapsulate melphalan,and the characteristics,effectiveness,and safety of MEL/MPEG-PDLLA in vitro and in vivo were investigated.MEL/MPEG-PDLLA showed slow release and was easily engulfed by MM cells despite a result of the antitumor assay comparable to that of free melphalan in vitro.The in vivo results showed that MEL/MPEG-PDLLA could significantly alleviate tumor burden and prolong survival time without increasing the toxicity to vital organs.In addition,MEL/MPEG-PDLLA could significantly reduce the damage to the intestinal mucosa caused by melphalan.In conclusion,MEL/MPEG-PDLLA shows improved antitumor activity and has the potential to alleviate pains of MM patients undergoing ASCT.

Peptide-assembled siRNA nanomicelles confine MnOx-loaded silicages for synergistic chemical and gene-regulated cancer therapy

Chemodynamic therapy(CDT) is a promising therapeutic approach for in situ cancer treatment, but it is still hindered by inefficient single-modality treatment and the weak targeted delivery of reagents into mitochondria(the main site of intracellular ROS production). Herein, to obtain a multimodal strategy,peptide-assembled si RNA nanomicelles were prepared to confine ultrasmall MnOxin small silica cages(silicages), which is convenient for synergistic chemical and gene-regulated cancer therapy. Given the free energy and versatility of small silicages, as well as the excellent Fenton-like activity of ultrasmall MnOx,MnOx-inside-loaded silicages(10 nm) were prepared for CDT delivery to mitochondria. Subsequently, to obtain a synergistic CDT and gene silencing treatment, the peptide-mediated assembly of si RNA and MnOx-loaded silicages were employed to obtain silicage@MnOx-si RNA nanomicelles(SMS NMs). After multiple modifications, sequential cancer cell-targeted delivery, GSH-controlled reagent release of si RNA and mitochondria-targeted delivery of MnOx-loaded silicages were successfully achieved. Finally, by both in vitro and in vivo experiments, SMS NMs were confirmed to be effective for synergistic chemical and gene-regulated cancer therapy. Our findings expand the applications of silicages and initiate the development of multimodal CDT.

Targeted delivery of hyaluronic acid nanomicelles to hepatic stellate cells in hepatic fibrosis rats

Hepatic fibrosis is one kind of liver diseases with a high mortality rate and incidence.The activation and proliferation of hepatic stellate cells(HSCs)is the most fundamental reason of hepatic fibrosis.There are no specific and effective drug delivery carriers for the treatment of hepatic fibrosis at present.We found that when hepatic fibrosis occurs,the expression of CD44 receptors on the surface of HSCs is significantly increased.Based on this finding,we designed silibinin-loaded hyaluronic acid(SLB-HA)micelles to achieve the treatment of hepatic fibrosis.Meanwhile,we constructed liver fibrosis rat model using Sprague-Dawley rats.We demonstrated that HA micelles had specific uptake to HSCs in vitro while avoiding the distribution in normal liver cells and the phagocytosis of macrophages.Importantly,HA micelles showed a significant liver targeting effect in vivo,especially in fibrotic liver which highly expressed CD44 receptors.In addition,SLB-HA micelles could selectively kill activated HSCs,having an excellent anti-hepatic fibrosis effect in vivo and a significant sustained release effect,and also had a good biological safety and biocompatibility.Overall,HA micelles represented a novel nanomicelle system which showed great potentiality in anti-hepatic fibrosis drugs delivery.

Self-assembly nanomicelle-microneedle patches with enhanced tumor penetration for superior chemo-photothermal therapy

Nanomedicine with high specificity has been a promising tool for cancer diagnosis and therapy.However,the successful application of nanoparticle-based superficial cancer therapy is severely hindered by restricted deep tumor tissue accumulation and penetration.Herein,a self-assembly nanomicelle dissolving microneedle(DMN)patch according to the“nano in micro”strategy was conducted to co-deliver a first-line chemotherapeutic agent paclitaxel(PTX),and a photosensitizer IR780(PTX/IR780-NMs@DMNs)for chemo-photothermal synergetic melanoma therapy.Upon direct insertion into the tumor site,DMNs created a regular and multipoint three-dimensional drug depot to maximize the tumor accumulation.Accompanied by the DMN dissolution,the composition of the needle matrixes self-assembled into nanomicelles,which could efficiently penetrate deep tumor tissue.Upon laser irradiation,the nanomicelles could not only ablate tumor cells directly by photothermal conversion but also trigger PTX release to induce tumor cell apoptosis.In vivo results showed that compared with intravenous injection,IR780 delivered by PTX/IR780-NMs@DMNs was almost completely accumulated at the tumor site.The antitumor results revealed that the PTX/IR780-NMs@DMNs could effectively eliminate tumors with an 88%curable rate without any damage to normal tissues.This work provides a versatile and generalizable framework for designing self-assembly DMN-mediated combination therapy to fight against superficial cancer.

Reduction-sensitive nanomicelles:Delivery celastrol for retinoblastoma cells effective apoptosis

Celastrol,a Chinese herbal medicine,has exhibited anticancer activity in many types of cancer cells.However,the further clinical application of celastrol is restricted by its poor water solubility and serious side effects.Furthermore,the apoptosis mechanism of tumor cells induced by celastrol has not been exhausted yet.In this study,we developed a reduction sensitive polymeric vector for tumor-targeted celastrol delivery.And our researches indicated that the celastrol could be delivered by reductionsensitive nanomedicine(RSNMs)with a controlled release strategy.Meanwhile,the cell uptake results indicated that excellent reduction-sensitive behavior of RSNMs could effectively accelerate celastrol into the human retinoblastoma(RB)cell.The cell cytotoxicity assay demonstrated that celastrol inhibited proliferation of human RB Y79 cells growth in a dose-dependent manner.Furthermore,the results of flow cytometry and terminal dUTP nick-end labeling(TUNEL)staining showed that celastrol induced apoptosis of the RB Y79 cells,and revealed a time-dependent increase in apoptosis induction of RB Y79 cells.The results of western blotting showed that celastrol induced the apoptosis of human RB Y79 cells involving the activation of caspase-3 and caspase-9.In conclusion,our results revealed that RSNMs may be utilized as a novel therapy for retinoblastoma.

Co-delivery of doxorubicin and curcumin via cRGD-peptide modified PEG-PLA self-assembly nanomicelles for lung cancer therapy

Lung cancer is the most common malignancy in the world, with a high mortality rate. Nevertheless,therapies to act effectively against lung cancer remain elusive. So far, chemotherapy is still the frontline treatment of lung cancer. Doxorubicin(DOX) is a broad-spectrum anti-tumor drug. However, DOX often has serious side effects and causes multi-drug resistance, which greatly limits its clinical application.In this work, biodegradable methoxy poly(ethylene glycol)-poly(lactic acid)(MPEG-PLA) and cyclo(ArgGly-Asp-D-Phe-Lys)(c RGD) polypeptide modified PEG-PLA(c RGD-PEG-PLA) copolymers were used for the co-delivery of curcumin(CUR) and DOX(CUR-DOX/c RGD-M). The particle size of the self-assembled drugloaded nanomicelle approximately was 27.4 nm and the zeta potential was -2.7mV. Interestingly, CUR can enhance the uptake of DOX by Lewis lung carcinoma(LL/2) cells. The experimental results in vivo and in vitro showed that CUR-DOX/c RGD-M combination therapy could promote apoptosis of lung cancer cells, and conspicuously inhibit the tumor growth. Our data indicate that CUR-DOX/c RGD-M will be biodegradable and sustainable, which may have potential clinical application value in the treatment of lung cancer.

Hyaluronic acid-based dual-responsive nanomicelles mediated mutually synergistic photothermal and molecular targeting therapies

Precise clinical treatment of triple-negative breast cancer(TNBC)is an obstacle in clinic.Nanotechnology-assisted photothermal therapy(PTT)is a superior treatment modality for TNBC in terms of precision.However,thermoresistance arising from PTT and insufficient drug release from nanocarriers decrease the efficacy of PTT.AT13387 is a novel HSP90 inhibitor that can weaken thermoresistance and undergoing clinic II phase study,showing satisfactory antitumour activity through molecularly targeted therapy(MTT).Whereas,it has poor solubility.Hence hyaluronic acid and stearic acid were connected by hydrazone bonds and disulfide bonds,forming an amphipathic copolymer that could self-assembled into nanomicelles,followed by encapsulating Cypate and AT13387.These nanomicelles exhibited great features,including achieving mutually synergistic PTT/MTT for overcoming thermoresistance and promoting translocation of drugs,increasing the solubility of Cypate and AT13387,showing a pH/redox dual response that contributes to drug release,and having the ability of active targeting.Thus,the nanomicelles developed in this study may be a promising strategy for the precise treatment of TNBC.

Sprayable nanomicelle hydrogels and inflammatory bowel disease patient cell chips for development of intestinal lesion-specific therapy

All-in-one treatments represent a paradigm shift in future medicine.For example,inflammatory bowel disease(IBD)is mainly diagnosed by endoscopy,which could be applied for not only on-site monitoring but also the intestinal lesion-targeted spray of injectable hydrogels.Furthermore,molecular conjugation to the hydrogels would program both lesion-specific adhesion and drug-free therapy.This study validated this concept of all-in-one treatment by first utilizing a well-known injectable hydrogel that underwent efficient solution-to-gel transition and nanomicelle formation as a translatable component.These properties enabled spraying of the hydrogel onto the intestinal walls during endoscopy.Next,peptide conjugation to the hydrogel guided endoscopic monitoring of IBD progress upon adhesive gelation with subsequent moisturization of inflammatory lesions,specifically by nanomicelles.The peptide was designed to mimic the major component that mediates intestinal interaction with Bacillus subtilis flagellin during IBD initiation.Hence,the peptide-guided efficient adhesion of the hydrogel nanomicelles onto Toll-like receptor 5(TLR5)as the main target of flagellin binding and Notch-1.The peptide binding potently suppressed inflammatory signaling without drug loading,where TLR5 and Notch-1 operated collaboratively through downstream actions of tumor necrosis factor-alpha.The results were produced using a human colorectal cell line,clinical IBD patient cells,gut-on-a-chip,a mouse IBD model,and pig experiments to validate the translational utility.

Efficient production of mannosylerythritol lipids by a marine yeast Moesziomyces aphidis XM01 and their application as self‑assembly nanomicelles

Mannosylerythritol lipids(MELs)are one of the most promising biosurfactants because of their excellent physicochemical properties,high environmental compatibility,and various biological functions.In this study,a mangrove yeast strain Moesziomyces aphidis XM01 was identified and used for efficient extracellular MEL production.The MEL titer reached 64.5±0.7 g/L at flask level within 7 days with the optimized nitrogen and carbon source of 2.0 g/L NaNO_(3) and 70 g/L soybean oil.Furthermore,during a 10-L two-stage fed-batch fermentation,the final MEL titer reached 113.6±3.1 g/L within 8 days,with prominent productivity and yield of 14.2 g·L^(−1)·day^(−1)and 94.6 g/g_((glucose and soybean oil)).Structural analysis indicated that the produced MELs were mainly MEL-A and its fatty acid profile was composed of only medium-chain fatty acids(C8–C12),especially C10 acids(77.81%).Further applications of this compound were evaluated as one-step selfassembly nanomicelles.The obtained MEL nanomicelles showed good physicochemical stability and antibacterial activity.In addition,using clarithromycin as a model hydrophobic drug,the MEL nanomicelles exhibited high loading capacity and could be used for the controlled and sustained drug release in low-pH environments.Therefore,M.aphidis XM01 is an excellent candidate for efficient MEL production,and the prepared MEL nanomicelles have broad application prospects in the pharmaceutical and cosmetic fields.

A self-assembling prodrug nanosystem to enhance metabolic stability and anticancer activity of gemcitabine

Self-assembly is a powerful approach in molecular engineering for biomedical applications,in particu-lar for creating self-assembling prodrugs.Here,we report a self-assembling prodrug of the anticancer drug gemcitabine(Gem)based on amphiphilic dendrimer approach.The prodrug reported in this study demonstrates high drug loading(40%)and robust ability to self-assemble into small nanomicelles,which increase the metabolic stability of Gem and enable entry into cells via endocytosis,hence bypassing transport-mediated uptake.In addition,this prodrug nanosystem exhibited an effective pH-and enzyme-responsive release of Gem,resulting in enhanced anticancer activity and reduced toxicity.Harboring ad-vantageous features of both prodrug-and nanotechnology-based drug delivery,this self-assembling Gem prodrug nanosystem constitutes a promising anticancer candidate.This study also offers new perspectives of the amphiphilic dendrimer nanoplatforms for the development of self-assembling prodrugs.

Vapreotide-modified nanomicelle as a targeted nanocarrier for delivering paclitaxel to the tumors with overexpression of somatostatin receptors

Somatostatin receptors （SSTRs） were widely expressed in many tumor cells. As a somatostatin analogue, vapreotide （VAP） can be exploited as a modifier for targeting tumor therapy based on its high affinity to SSTR. In this study, we conjugated α-NH2 of exocyclic n-phenylalanine （D-Phe） of vapreotide to N-hydroxysuccinimidyl-PEG2000-DSPE （NHS-PEG-DSPE）, and the resulted DSPE-PEG-VAP was used as a targeting component to construct the targeted micelles for delivering paclitaxel （VAP-M-PTX） through a thin-film hydration method. Similar particle size, zeta potential, drug encapsulation efficiencies, drug release behaviors and hemolysis effects were observed between the targeted micelles （VAP-M-PTX） and the non-targeted micelles （M-PTX）. In MCF-7 cells, significantly higher intracellular fluorescence intensity （1.5-fold） was determined by flow cytometry after incubation of coumarin-6 loaded targeted micelles （VAP-M-Cou） for 3 h compared with non-targeted mieelles （M-Cou）, and similar finding was observed confocal microscopy. Furthermore, in comparison with non-targeted formulations, higher antitumor efficacy and higher drug accumulation were found in MCF-7 tumors in nude mice after intravenous injection of the targeted micelles. In conclusion, we believed that the vapreotide-modified nanomicelles could be a promising targeted nanocarrier for delivering anticancer drugs to the tumors with overexpression of somatostatin receptors.

Reverse thinking:Tumor nutritional therapy

Tumor cells often exhibit metabolic abnormalities to meet the needs of rapid proliferation,and targeting tumor metabolism has become one of the effective strategies for cancer treatment.However,most of the current methods targeting metabolism focus on inhibiting hyperactivated metabolic pathways,hindering their further application.A recent innovative work,proposed a nutrient-based strategy to reactivate metabolism for tumor therapy by targeting suppressed metabolic pathways.This approach through delivering nutrients to tumor cells directly using nanotechnology indicates that specific nutrients can serve as potent activators of metabolic pathways.As a new direction along the reverse thinking,this study suggests that this nutrient-based metabolism reactivation strategy will inspire broad applications in the treatment of other diseases associated with metabolic disorders,besides tumor.