Advanced Delivery Systems Research Articles

Internal cross-linked polymeric nanoparticles with dual sensitivity for combination therapy of muscle-invasive bladder cancer

BackgroundChemotherapy is a standard cancer treatment which uses anti-cancer drugs to destroy or slow the growth of cancer cells. However, chemotherapy has limited therapeutic effects in bladder cancer. One of the reasons of this resistance to chemotherapy is that higher levels of glutathione in invasive bladder cancer cells. We have fabricated nanoparticles that respond to high concentrations of glutathione and near-infrared laser irradiation in order to increase the drug accumulation at the tumor sites and combine chemotherapy with photothermal therapy to overcome the challenges of bladder cancer treatment.MethodsThe DOX&IR780@PEG-PCL-SS NPs were prepared by co-precipitation method. We investigated the tumor targeting capability of NPs in vitro and in vivo. The orthotopic bladder cancer model in C57BL/6 mice was established for in vivo study and the photothermal effects and therapeutic efficacy of NPs were evaluated.ResultsThe DOX&IR780@PEG-PCL-SS NPs were synthesized using internal cross-linking strategy to increase the stability of nanoparticles. Nanoparticles can be ingested by tumor cells in a short time. The DOX&IR780@PEG-PCL-SS NPs have dual sensitivity to high levels of glutathione in bladder cancer cells and near-infrared laser irradiation. Glutathione triggers chemical structural changes of nanoparticles and preliminarily releases drugs, Near-infrared laser irradiation can promote the complete release of the drugs from the nanoparticles and induce a photothermal effect, leading to destroying the tumor cells. Given the excellent tumor-targeting ability and negligible toxicity to normal tissue, DOX&IR780@PEG-PCL-SS NPs can greatly increase the concentration of the anti-cancer drugs in tumor cells. The mice treated with DOX&IR780@PEG-PCL-SS NPs have a significant reduction in tumor volume. The DOX&IR780@PEG-PCL-SS NPs can be tracked by in vivo imaging system and have good tumor targeting ability, to facilitate our assessment during the experiment.ConclusionA nanoparticle delivery system with dual sensitivity to glutathione and near-infrared laser irradiation was developed for delivering IR780 and DOX. Chemo-photothermal synergistic therapy of both primary bladder cancer and their metastases was achieved using this advanced delivery system.

Update on newer approaches to prevent or treat COVID-19 infection: What we all need the most right now!!

COVID-19 convalescent plasma (CCP) therapy involves the use of circulating antibodies administration from recovered COVID 19 patients as a practical strategy to provide immediate passive immunity in susceptible recipients in need. Global concern over the potential for “second” or “third” waves of infection to occur before effective vaccines or drug therapies are available has many looking at other biological sources for large-scale production of neutralizing SARS-CoV-2 antibodies. This report summarizes some of the novel strategies for developing alternative safe sources of therapeutic autologous antibodies from COVID ‐19 infected patients, and provides some original thoughts on how to rapidly implement a safe passive immunity in those COVID-19 patients who are most in need of intervention. COVID-19 antibodies can be isolated or delivered using a number of other techniques including: plasmapheresis, plasma cryoprecipitate reduced (cryosupernatant), antibody hyperconcentrates and advanced cell-based delivery systems. While these proposed technological options may, in some cases, be theoretical, the growing concern over the rapid spread of the SARS-CoV-2 virus has prompted many to pursue innovative and creative solutions to reduce the mortality and morbidity resulting from the current global pandemic. A comparative analysis of various strategies currently in use deserved exploring and this highlighted separately as the essential part of this concise theme.

Abstract 6593: PRGN-3005 UltraCAR-T™: multigenic CAR-T cells generated using non-viral gene delivery and rapid manufacturing process for the treatment of ovarian cancer

Abstract Ovarian cancer is the most lethal of gynecologic cancers. Chimeric antigen receptor (CAR)-T cells have improved clinical outcomes in hematological malignancies; however similar responses have not yet been achieved for solid tumors. Furthermore, traditional methods for CAR-T manufacturing utilize viral vectors and weeks of ex vivo expansion to achieve clinically relevant cell numbers. This leads to CAR-T cells with an exhausted T cell phenotype, high manufacturing costs, and treatment delays. We have developed the UltraCAR-T™ platform to overcome these limitations by utilizing an advanced non-viral gene delivery system and a rapid, decentralized manufacturing process. Our non-viral gene delivery platform is based on Sleeping Beauty transposon system precision engineered for optimal delivery of multiple genes of UltraCAR-T. UltraCAR-T cells are manufactured at medical center's cGMP facility using autologous T cells and administered back to patient only one day after gene transfer. PRGN-3005 UltraCAR-T cells simultaneously co-express (i) a CAR specifically targeting MUC16 which is highly expressed on ovarian tumors with limited normal tissue expression; (ii) membrane bound IL-15 (mbIL15) for enhanced in vivo expansion and persistence; and (iii) a kill switch for improved therapeutic control. Healthy donor T cells were transfected with non-viral system encoding CAR, mbIL15 and kill switch to generate PRGN-3005 cells. Expression of transgenes was confirmed by flow cytometry and western blot analyses. PRGN-3005 demonstrated a robust MUC16-specific cytotoxicity of ovarian cancer cells in vitro. PRGN-3005 exhibited a stem-cell like memory phenotype, significant improvement in survival even in absence of exogenous cytokines and lacked autonomous proliferation in vitro in the absence of MUC16 compared to conventional CAR-T cells, demonstrating beneficial effects of mbIL15 on phenotype and persistence of UltraCAR-T cells. Furthermore, we demonstrated that PRGN-3005 cells were selectively and effectively eliminated by kill switch activator antibody via ADCC. For in vivo anti-tumor efficacy of PRGN-3005, an ovarian cancer model of MUC16+ SK-OV-3 cell line was evaluated in NSG mice. In tumor-bearing mice, a single administration of PRGN-3005 one day after gene transfer resulted rapid in vivo expansion and significant increase in UltraCAR-T cells, in particular T stem cell memory (TSCM) population compared to mbIL15neg CAR-T cells. PRGN-3005 demonstrated significantly higher anti-tumor effect (p<0.01) compared to mbIL15neg CAR-T and led to complete tumor regression in all treated mice. PRGN-3005 UltraCAR-T administered one day after non-viral gene transfer has superior anti-tumor efficacy and persistence compared traditional CAR-T cells and represent a promising opportunity for ovarian cancer treatment. Based on these results, FDA has approved an IND application and the first in human PRGN-3005 Phase I clinical trial for advanced ovarian cancer is currently under evaluation (NCT03907527). Citation Format: Tim Chan, Marion Chakiath, Lindsey Shepard, Simon Metenou, Fernando Carvajal-Borda, Jose Velez, Adeline Govekung, Jacques Plummer, Carol Poortman, Padma Kamineny, Xiaohong Ma, Rutul R. Shah, Mary L. Disis, Helen Sabzevari. PRGN-3005 UltraCAR-T™: multigenic CAR-T cells generated using non-viral gene delivery and rapid manufacturing process for the treatment of ovarian cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6593.

In Situ Biomimetic Mineralization on ZIF-8 for Smart Drug Delivery.

The exploration of metal-organic frameworks (MOFs) with good biocompatibility and physiological stability as carrier platforms for biomedical applications is of great importance but remains challenging. Herein, we developed an in situ biomimetic mineralization strategy on zeolitic imidazolate framework (ZIF) nanocrystals to construct a drug release system with favorable cytocompatibility, improved stability, and pH responsiveness. With lysozyme (Lys) wrapped on the surface of Zn-based ZIF (ZIF-8), Lys/ZIF-8 could strongly bond metal ions to promote nucleation and growth of bone-like hydroxyapatite (HAp), leading to formation of HAp@Lys/ZIF-8 composites. In vitro investigations indicate that the composites with a hollow Lys/ZIF-8 core and a HAp shell exhibited a high drug-loading efficiency (56.5%), smart pH-responsive drug delivery, cytocompatibility, and stability under physiological conditions. The proposed biomimetic mineralization strategy for designing MOFs-based composites may open a new avenue to construct advanced delivery systems in the biomedical field.

Bite-sized Review on Advance Delivery System, Support in the management of Covid-19

COVID-19 are an unprecedented challenge for healthcare providers; there are no approved treatments for this disease, nor are there no approved vaccines. The sources of infection of novel coronavirus detected mainly from animals to humans or infected human to healthy human through respiratory droplets and long contact period are the most prominent way of transmission. Coronaviruses are structurally large-sized single-stranded RNA viruses. The capability of regular therapies is constantly fading away exactly in case of coronavirus due to the modiϑication of new strain, which could be certainly due to speedy adaptation in a protein sequence. The scientist should understand the pandemic situation of virus infection, also as they ought to identify the newest pathway research to regulate the virus infection. Developing new and smart strategies for drugs already in the development pipeline or already exiting drug can be treating diseases in patients could be useful to ϑight against COVID-19. In the last few years, several new smart drug delivery approaches made the changeover from the laboratory development to clinical applications. At present technology, researchers provide effective and low toxicity drug delivery when compared with classical delivery. This approach will be a great opportunity for the scientist to work and update preclinical research of advance drug delivery systems to cloneable and convertible production to the human trial success rate. In conclusion, by understanding the new drug delivery research approaches for antiviral therapy are increases to produce safe and high-quality therapies at reasonable costs.

Treatment with azelastine hydrochloride and fluticasone propionate in a single delivery device of young children and adolescents with allergic rhinitis.

Background: Allergic rhinitis (AR) is the most common chronic noncommunicable disease worldwide that affects any age during the human life span but raises particular concerns among the parents and caregivers of the children who are affected. H1-receptor antagonists and corticosteroids provide the most effective way of bringing the condition under control. Intranasal application of these medications has the advantage of a faster onset of action and avoids systemic unwanted adverse effects. The only combination treatment, which comprises azelastine hydrochloride and fluticasone propionate, so far in a single advanced delivery system has proven its efficacy and safety in clinical trials of adult patients with AR. Objective: To critically review and identify gaps in the existing data for children in all different strata of pediatric ages. Methods: We searched the specialized medical literature for publications on the efficacy and safety of the combined formulation of azelastine and fluticasone in a single delivery device in adolescents (ages < 18 years) and children (ages < 12 years). Results: Altogether, 12 peer-reviewed articles have been published about trials that also involved subjects in different strata of the pediatric ages, seven of the articles pooled adolescents and adults. Three articles presented the results of studies in children ages 4 to 11 years specifically designed to overcome the difficulties that children experience in expressing themselves verbally. Conclusion: All the trials with the novel combination product that involved young children and adolescents documented its efficacy, effectiveness, and safety. However, the numbers of the youngest children (ages 4 and 5 years) were low, which suggested that further data about safety and efficacy in this age group are needed.

Immune effect of a Newcastle disease virus DNA vaccine with IL-12 as a molecular adjuvant delivered by electroporation.

Newcastle disease (ND), caused by virulent Newcastle disease virus (NDV) strains, has been one of the most problematic diseases affecting the poultry industry worldwide. Conventional vaccines provide effective protection for birds to survive ND outbreaks, but they may not completely suppress NDV shedding. NDV strains circulate on farms for a long time after the initial infection and cause potential risks. A new vaccine with fast clearance ability and low viral shedding is needed. In this study, we used interleukin-12 (IL-12) as an adjuvant and electroporation (EP) as an advanced delivery system to improve a DNA vaccine candidate. The fusion (F) protein gene from an NDV strain of the prevalent genotype VII.1.1 was cloned to prepare the vaccine. Chickens immunized with the F gene DNA vaccine co-delivered with an IL-12-expressing plasmid DNA showed higher neutralizing antibody levels and stronger concanavalin-A-induced lymphocyte proliferation than those treated with the F gene DNA vaccine alone. The co-delivered vaccine provided 100% protection, and less viral shedding and a shorter release time were observed in challenged chickens than when the F gene DNA vaccine was administered alone. The use of F gene DNA combined with IL-12 delivered by electroporation is a promising approach for vaccination against ND.

4145 Stepping Stones for Success in T3-T4 Translation: Building Collective Impact

OBJECTIVES/GOALS: We will investigate the influence of multisector partnerships in T3-T4 research associated with advances in delivery systems, patient/population outcomes and health policy and the translational processes linked to these improvements. METHODS/STUDY POPULATION: We are using both quantitative and qualitative data to measure and analyze partnership characteristics linked to successful translation into practice &amp; policy. We aim to complete 100 surveys of investigators who have conducted CTSA-supported T3-T4 research to examine partnerships, conditions of collective impact, and quantifiable changes in delivery systems, health outcomes, and policy. Using rigorous criteria, we will select projects for more in-depth interviews to understand the practices of successful translation and roadblocks and barriers that challenge translation. RESULTS/ANTICIPATED RESULTS: The anticipated research products include: (i) an analytic report on partnership structure and processes and the statistical associations to stages of change outcomes, (ii) a series of vignettes to describe the impact stories and translational processes, (iii) cross-project analysis of the data and vignettes to produce generalizable information to improve T3-T4 translation, and (iv) peer-reviewed manuscript(s) for publication. DISCUSSION/SIGNIFICANCE OF IMPACT: The study will inform and improve researcher competencies and accelerate translation in CTSA hubs that emphasize T3-T4 research. We will develop novel definitions of the T3-T4 research impact. Ultimately, the results will inform research training to better address real-world priorities and needs.

Cyclodextrin-based delivery systems for in vivo-tested anticancer therapies.

Cyclodextrins (CDs) are naturally occurring macromolecules widely used as excipients on pharmaceutical formulations, evidencing a large spectrum of applications in the pharmaceutical industry. Their unique ability to act as molecular containers by entrapping a wide range of guest molecules in their internal cavity makes them a remarkable excipient to improve drug apparent solubility, stability, and bioavailability, and a valuable tool for the assembly of new drug delivery systems. These features are especially useful when it comes to chemotherapy, as most of the anticancer drugs present both low permeability and reduced water solubility. Therefore, guest-host inclusion complexes offer several potential advantages not only regarding the improvement of pharmaceutical formulations characteristics but also considering the reduction of drug toxic side effects. The combination of CDs with additional technologies and materials constitutes a potential strategy towards the development of advanced and multifunctional CD-based delivery systems. Paclitaxel, curcumin, camptothecin, doxorubicin, and cisplatin are among the most studied molecules with anticancer activities and have been successfully incorporated in such nanosystems. Exciting results using CDs and CD-based delivery systems have been obtained so far, paving the way towards the attainment of intelligent delivery systems to possibly address cancer therapeutics' unmet needs. In this review, a comprehensive exposition concerning in vivo-tested CD and CD-based delivery systems for anticancer therapy is undertaken. Additionally, the authors address the multivalent functionalities of CD-based delivery systems, namely the incorporation of active target ligands, stimuli-responsiveness components, surface functionalization, or further associations with other delivery systems, aiming at improved in vivo anticancer therapies. Graphical abstract.

Nanostructure generation during milk digestion in presence of a cell culture model simulating the small intestine

HypothesisThe development of advanced oral delivery systems for bioactive compounds requires the fundamental understanding of the digestion process within the gastrointestinal tract. Towards this goal, dynamic invitro digestion models, capable of characterising the molecular as well as colloidal aspects of food, together with their biological interactions with relevant invitro cell culture models, are essential. ExperimentsIn this study, we demonstrate a novel digestion model that combines flow-through time resolved small angle X-ray scattering (SAXS) with an invitro Caco-2/HT-29 cell co-culture model that also contained a mucus layer. This set-up allows the dynamic insitu characterisation of colloidal structures and their transport across a viable intestinal cell layer during simulated digestion. FindingsAn integrated online SAXS – invitro cell co-culture model was developed and applied to study the digestion of nature’s own emulsion, milk. The impact of the invitro cell culture on the digestion-triggered formation and evolution of highly ordered nanostructures in milk is demonstrated. Reported is also the crucial role of the mucus layer on top of the cell layer, protecting the cells from degradation by digestive juice components such as lipase. The novel model can open unique possibilities for the dynamic investigation of colloidal structure formation during lipid digestion and their effect on the uptake of bioactive molecules by the cells.

Use of Lifetech™ Konar-MF, a device for both perimembranous and muscular ventricular septal defects: A multicentre study

BackgroundThe objective of this study was to evaluate the safety and efficacy of transcatheter closure of ventricular septal defects (VSD) using the LifeTech™ multifunctional occluder device (MF-Konar). MethodsClinical features and demographic characteristics and follow-up findings were evaluated retrospectively from three centers. ResultsMF-Konar was used in 98 patients. The median age and weight of the patients were 3.8 years (range 5.4 months–50 years) and 15.3 kg (range 5.5–80 kg), respectively. The mean fluoroscopy time was 13.7 ± 8.2 min (range 3.4–42.6 min). Procedural success was obtained for 96 out of 98 patients (98%). In 54 out of 98 patients, closure was performed via the antegrade route. Major complications occurred in four patients (embolization in two, complete heart block in one, and device dislocation needing surgical treatment in one). All of the complications were treated successfully, and there was no mortality. Mild residual flow in eight patients (8%), new onset tricuspid valve insufficiency in one (moderate), and new onset aortic valve insufficiency in one (mild) were observed during a mean follow-up duration of 224 ± 149 (10–515) days. Minor rhythm disturbances were observed in eight patients. ConclusionsTranscatheter closure of VSDs in selected patients using the LifeTech MF-Konar device seems effective. Its advantages are softer design, use of both an antegrade and retrograde approach, and an advanced smaller delivery system. Increasing the number of usage and the experience will provide more accurate data and low complication rates.

No‐reference quality assessment for live broadcasting videos in temporal and spatial domains

Nowadays, live broadcasting video has become increasingly popular and high-quality live broadcasting video is highly needed. In practice, live broadcasting videos usually undergo several processing stages, which inevitably introduce multiple distortions, e. g. frame freezing and intensity mutation, causing the degraded quality of experience. However, little work has been done to the quality evaluation of live broadcasting videos, which may hinder the further development of more advanced live broadcasting video delivery systems. Motivated by this, this study presents a no-reference quality evaluation model for live broadcasting videos (LBVQA) in temporal and spatial domains. In the temporal domain, statistic features are extracted to measure the frame freezing and intensity mutation, and the entropy-based feature is extracted to describe the global jitter. In the spatial domain, blurring is measured based on phase coherence, and abnormal exposure ratio is calculated based on an adaptive threshold. Finally, all features are fed into a backpropagation neural network to train the quality prediction model. Experimental results on the Live Broadcasting Video Database demonstrate the advantages of the proposed metric over the state-of-the-art image and video quality metrics.

Thermal analysis studies on the compatibility of furosemide with solid state and liquid crystalline excipients

In the context of the present study, the thermal behavior of furosemide and the solid state excipients, sodium alginate, poly(ethylene oxide), poly(vinylpyrrolidone), lactose mono-hydrate and magnesium stearate, using Differential Scanning Calorimetry (DSC), was probed. It was found that the thermal behavior of these solid-state pharmaceutical excipients and furosemide correlates nicely with the literature relevant data. Regarding the furosemide-excipients mixtures, the DSC scans appear as a compilation of the thermal curves of each excipient. This suggests that the formulations containing these mixtures, may retain their stability over time. This information, which arises from the cooperativity of materials, their thermal stability and behavioris very helpful for the research and development of safe and effective pharmaceutical formulations. DSC experiments were also carried out with chimeric bilayers (called ?liposomes?), composed of hydrogenated soy phosphatidylcholine (HSPC) and poly(n-butylacrylate)-b-poly(acrylic acid) block copolymer with 70 % content of poly(acrylic acid (PnBA-b-PAA 30/70) with the addition of furosemide at the molar ratio of 9:0.1:1.0 in the system HSPC:PnBA-b-PAA 30/70:furosemide. Chimeric liposomal systems were characterized as ?fluid-like? by their DSC curves, which may be potentially translated as an easy way for release of furosemide from the advanced delivery system.

Repairing the heart: State-of the art delivery strategies for biological therapeutics.

Myocardial infarction (MI) is one of the leading causes of mortality worldwide. It is caused by an acute imbalance between oxygen supply and demand in the myocardium, usually caused by an obstruction in the coronary arteries. The conventional therapy is based on the application of (a combination of) anti-thrombotics, reperfusion strategies to open the occluded artery, stents and bypass surgery. However, numerous patients cannot fully recover after these interventions. In this context, new therapeutic methods are explored. Three decades ago, the first biologicals were tested to improve cardiac regeneration. Angiogenic proteins gained popularity as potential therapeutics. This is not straightforward as proteins are delicate molecules that in order to have a reasonably long time of activity need to be stabilized and released in a controlled fashion requiring advanced delivery systems. To ensure long-term expression, DNA vectors-encoding for therapeutic proteins have been developed. Here, the nuclear membrane proved to be a formidable barrier for efficient expression. Moreover, the development of delivery systems that can ensure entry in the target cell, and also correct intracellular trafficking towards the nucleus are essential. The recent introduction of mRNA as a therapeutic entity has provided an attractive intermediate: prolonged but transient expression from a cytoplasmic site of action. However, protection of the sensitive mRNA and correct delivery within the cell remains a challenge. This review focuses on the application of synthetic delivery systems that target the myocardium to stimulate cardiac repair using proteins, DNA or RNA.

Recent Advances in Nanoencapsulation Systems Using PLGA of Bioactive Phenolics for Protection against Chronic Diseases.

Plant-derived polyphenolic compounds have gained widespread recognition as remarkable nutraceuticals for the prevention and treatment of various disorders, such as cardiovascular, neurodegenerative, diabetes, osteoporosis, and neoplastic diseases. Evidence from the epidemiological studies has suggested the association between long-term consumption of diets rich in polyphenols and protection against chronic diseases. Nevertheless, the applications of these phytochemicals are limited due to its low solubility, low bioavailability, instability, and degradability by in vivo and in vitro conditions. Therefore, in recent years, newer approaches have been attempted to solve the restrictions related to their delivery system. Nanoencapsulation of phenolic compounds with biopolymeric nanoparticles could be a promising strategy for protection and effective delivery of phenolics. Poly(lactic-co-glycolic acid) (PLGA) is one of the most successfully developed biodegradable polymers that has attracted considerable attention due to its attractive properties. In this review, our main goal is to cover the relevant recent studies that explore the pharmaceutical significance and therapeutic superiority of the advance delivery systems of phenolic compounds using PLGA-based nanoparticles. A summary of the recent studies implementing encapsulation techniques applied to polyphenolic compounds from plants confirmed that nanoencapsulation with PLGA nanoparticles is a promising approach to potentialize their therapeutic activity.

Nano Sponges

The convenient and recent drug delivery system always likely to have some ideal and unique characters, particularly for safety, desired actions, accurate delivery, enrich with a therapeutic index with minimal adverse occurrence. In this regard, Nano sponges serve as a recent advance new drug delivery. Many researchers are attracted and concerned on Nano sponges presently. The present twofold objective of the review clarifies why Nano sponges are recent and advanced delivery system along with challenges related to solubility, stability and controlled release of Nano sponge and proven techniques to overcome. Since the nano-sized drug free carriers are recently formulated and suggested for drug delivery, which can be loaded with the numerous drugs, hence dealing with an appropriate technique for drug release is the need of the present time. This article states the most convenient method for drug release, and at concluding part gives awareness on recent work done and result obtained on Nano sponge worldwide along with the unique summary on the US granted patents for Nano sponges. Thus, a complete scenario on recent work, methods for evaluations, unique and multiple characters of Nano sponge has been discussed and summarized.

Preclinical Characterization of Prgn-3006 Ultracar-T™ for the Treatment of AML and MDS: Non-Viral, Multigenic Autologous CAR-T Cells Administered One Day after Gene Transfer

Relapsed/refractory (r/r) acute myeloid leukemia (AML) is an aggressive malignancy with poor prognosis and limited treatment options underscoring the need for new therapies. Traditional methods for generating CAR-T cells require extensive ex vivo expansion following viral vector transduction, adding time and high cost to manufacturing with a centralized manufacturing process and prolonging the waiting period between apheresis and administration of CAR-T therapy to a patient. Time is of the essence for advanced cancer patients including patients with r/r AML. The UltraCAR-T™ platform addresses shortcomings of current CAR-T technologies by streamlined rapid manufacturing process that can be performed in a local hospital's GMP facility following apheresis from the patient providing a potential solution for broader adoption of CAR-T therapies. The UltraCAR-T™ platform utilizes Precigen's advanced non-viral multi-gene delivery system and the rapid manufacturing process with high cell viability that allows for administration of autologous CAR-T cells one day after gene transfer. PRGN-3006 UltraCAR-T™ cells, utilizing Precigen's UltraVector® platform, co-express CD33 CAR, membrane bound IL-15 (mbIL15) and a kill switch. Expression of mbIL15 provides CAR-T cells with a source of cytokine without the need for exogenous cytokines to further enhance the expansion potential in the presence of tumor antigen and the overall persistence for PRGN-3006 in. The expression of mbIL15 on PRGN-3006 eliminates any need for ex vivo expansion of T cells prior to cell administration. Co-expression of kill switch provides a mechanism for selective depletion of PRGN-3006 post administration and improves therapeutic control. We manufactured PRGN-3006 UltraCAR-T™ cells using non-viral gene transfer and rapid streamlined manufacturing process from multiple donor T cells. Co-expression of CAR, mbIL15 and kill switch was confirmed by flow cytometry and western blot analyses. PRGN-3006 demonstrated robust expansion in presence of CD33 antigen, lack of autonomous expansion in absence of CD33, and prolonged persistence in absence of exogenous cytokines in in vitro tests. The redirected specificity was demonstrated by the specific killing of CD33+ tumor cells as well as significant release of inflammatory cytokines such as IFNγ upon co-culture with AML tumor cells. We also demonstrated specific elimination of PRGN-3006 cells by kill switch activator antibody treatment. In vivo, a single administration of PRGN-3006 UltraCAR-T™ cells only one day after gene transfer effectively eliminated tumor burden and significantly improved overall survival (p&lt;0.01) of tumor bearing mice compared to CAR-T cells lacking mbIL15 expression (conventional CAR-T) in an aggressive xenograft model of AML in immunocompromised (NSG) mice. PRGN-3006 demonstrated engraftment and significantly higher expansion and persistence in mice compared to CAR-T cells lacking mbIL15 (conventional CAR-T). These pre-clinical data provide a strong rationale to evaluate PRGN-3006 UltraCAR-T™ in a clinical trial for treatment of AML. The FDA has approved an investigational new drug application for PRGN-3006 UltraCAR-T™ and the first in human Phase 1 clinical trial for treatment of patients with relapse/refractory AML and high-risk myeloid dysplastic syndrome is currently ongoing (ClinicalTrials.gov Identifier: NCT03927261). Figure: Antitumor activity of PRGN-3006 UltraCAR-T™ cells in an established model of AML. MOLM-13 tumor cells were engrafted into NSG mice on Day 0 and PRGN-3006 were administered on Day 6 (arrow). Data shown is tumor burden (expressed as total flux in photons/sec) as the mean±SEM for each group of mice shown, with n=6-7 mice mice/group at the start of the study. Figure Disclosures Chan: Precigen: Employment. Ma:Precigen, Inc.: Employment. Carvajal-Borda:Precigen, Inc.: Employment. Velez:Precigen, Inc.: Employment. Plummer:Precigen, Inc.: Employment. Shepard:Precigen, Inc.: Employment. Shah:Precigen, Inc.: Employment; Intrexon Corp.: Equity Ownership. Sallman:Celyad: Membership on an entity's Board of Directors or advisory committees. Sabzevari:Precigen, Inc.: Employment.

Targeted Release of Probiotics from Enteric Microparticulated Formulations.

The development of advanced probiotic delivery systems, which preserve bacteria from degradation of the gastrointestinal tract and achieve a targeted release mediated by pH-independent swelling, is of great interest to improve the efficient delivery of probiotic bacteria to the target tissue. Gram-positive and Gram-negative bacteria models (Lactobacillus acidophilus (Moro) Hansen and Mocquot (ATCC® 4356™) and Escherichia coli S17, respectively) have been successfully encapsulated for the first time in pH-independent microparticulate polymethacrylates (i.e., Eudraguard biotic) used for the targeted delivery of nutraceuticals to the colon. These bacteria have also been encapsulated within the mucoadhesive polymethacrylate Eudragit RS 100 widely used as targeted release formulation for active pharmaceutical ingredients. The enteric microparticles remained unaltered under simulated gastric conditions and released the contained viable microbial cargo under simulated intestinal conditions. Buoyancies of 90.2% and 57.3% for Eudragit and Eudraguard microparticles, respectively, and long-term stability (5 months) for the encapsulated microorganisms were found. Cytotoxicity of the microparticles formulated with both polymers was evaluated (0.5–20 mg/mL) on Caco-2 cells, showing high cytocompatibility. These results underline the suitability of the synthesized materials for the successful delivery of probiotic formulations to the target organ, highlighting for the first time the potential use of Eudraguard biotic as an effective enteric coating for the targeted delivery of probiotics.

Drug Delivery Systems for Phthalocyanines for Photodynamic Therapy.

The focus of this review is to describe the state-of-art in the development of innovative drug delivery systems for phthalocyanines as photosensitizers for photodynamic therapy (PDT). PDT is a medical treatment combining photosensitizers (PSs) activated by visible light of a specific wavelength to selectively destroy targeted cells, tumor tissues and its surrounding vasculature. In the last decades, PDT has been under intense investigation, first as a promising alternative approach for improved cancer treatment, later against microbial infection and nowadays, mainly in aesthetic medicine, against age-related degeneration. The success of PDT is restricted because of difficulties with administration and skin permeation of PSs. As PDT importance raises, there is high interest for advanced formulations and delivery systems (DDS) for PS, especially formulations based on nanotechnology. Accordingly, this review deals with the innovations pertaining to DDS for PDT as disclosed in recent patents and literature.

Synthesis of Thermosensitive Conjugated Triblock Copolymers by Sequential Click Couplings for Drug Delivery and Cell Imaging.

The elegant integration of an excellent light-emitting segment and a biorelevant signal-responsive moiety could generate advanced polymeric delivery systems with simultaneously favorable diagnostic and therapeutic functions with respect to cancer theranostics. Although polymeric delivery systems based on fluorescent polyfluorene (PF) or thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm) have been extensively developed, the preparation of a ternary polymer formulation composed of a PF block, a PNIPAAm sequence, and a hydrophilic moiety remains rarely explored likely because of the difficulty in integrating different synthesis strategies for polymer synthesis. To this end, herein we reported the design and controlled synthesis of a PF- and PNIPAAm-based amphiphilic triblock copolymer, PF11-b-PNIPAAm120-b-poly(oligo(ethylene glycol) monomethyl ether methacrylate)17 (PF11-b-PNIPAAm120-b-POEGMA17), with a well-defined structure by a strategy of sequential click couplings between Suzuki-coupling-generated PF and atom-transfer radical polymerization (ATRP)-produced PNIPAAm and POEGMA. The as-prepared triblock copolymers can self-assemble into micelles with a core-shell-corona (CSC) structure that is composed of an inner hydrophobic core of the PF moiety for fluorescent tracking and drug encapsulation, a thermosensitive middle shell of PNIPAAm block for thermomodulated drug loading and release, and a hydrophilic outer corona of the POEGMA segment for micelle stabilization. Interestingly, the doxorubicin (DOX)-loaded micelles prepared at 25 °C had a greater drug loading capacity than the analogues fabricated at 37 °C due to the better stability of the former formulation, leading to its higher in vitro cytotoxicity in HeLa cells. Together with the integration of a localized hyperthermia-triggered drug release profile and efficiently intracellular trafficking of the nanocarriers by monitoring the fluorescence of the PF moiety, this formulation demonstrates a great potential for cancer theranostics.