Liposomal Delivery System Research Articles

Delivery of Rapamycin by Liposomes Synergistically Enhances the Chemotherapy Effect of 5-Fluorouracil on Colorectal Cancer.

BackgroundRapamycin is a promising agent for treating tumors, but clinical applications of rapamycin are limited due to its poor water solubility and low bioavailability. This paper constructs a liposome delivery system for rapamycin to improve the effect in treating colorectal cancer.MethodsWe prepared the rapamycin liposomes using the ethanol injection method. The cellular uptake and biodistribution were detected by LC-MS and in vivo imaging system. MTT assay, transwell migration experiment, flow cytometry, and Western blot analysis evaluated the antitumor effect of rapamycin liposomes in vitro. Furthermore, HCT-116 tumor-bearing mice were used to assess the therapeutic efficacy of rapamycin liposomes in vivo.ResultsThe prepared rapamycin liposomes had a particle size of 100±5.5 nm and with a narrow size distribution. In vitro cellular uptake experiments showed that the uptake of rapamycin liposomes by colorectal cells was higher than that of free rapamycin. Subsequently, in vivo imaging experiments also demonstrated that rapamycin liposomes exhibited higher tumor accumulation. Therefore, the ability of rapamycin liposomes to inhibit tumor proliferation, migration and to induce tumor apoptosis is superior to that of free rapamycin. We also demonstrated in vivo good antitumor efficacy of the rapamycin liposomes in HCT-116 xenograft mice. In addition, rapamycin liposomes and 5-FU can synergistically improve the efficacy of colorectal cancer via the Akt/mTOR and P53 pathways.ConclusionCollectively, rapamycin liposomes are a potential treatment for colorectal cancer, as it not only improves rapamycin’s antitumor effect but also synergistically enhances 5-FU’s chemotherapy effect.

LIPOSOMES – A REVIEW

The function of belayed vesicles as productive transporters for drugs, immunizations, indicative specialists, and other bioactive operators has prompted a fast headway in the liposomal drug conveyance system. The pharmacy- elements and pharmacokinetics properties are altered for the liposomal delivery system,which on thewholeleads to an increased the rapeutic index with decreased toxicity. The liposome can be named multilamellar vesicles or unilamellar vesicles, which can be additionally named large unilamellar vesicles (LUV) or small unilamellar vesicles (SUV). The part of liposome as a medication conveyance framework is to convey drug in a controlled way, diminishing unfortunate reactions improving it is in vitro and in vivo action, just as lessening the harmfulness of the medication and upgrading the viability of the exemplified drug. This article gives a review of techniques to the planning of liposome, just as diagnostic strategies for control physical, concoction, and organic boundaries for various kinds of medications.

Fabrication of A Folic Acid-Modified Arsenic Trioxide Prodrug Liposome and Assessment of its Anti-Hepatocellular Carcinoma Activity

ObjectiveTo reduce the toxicity and side effects of arsenic trioxide (ATO) and provide a new approach for the treatment of primary liver cancer, a folic acid-modified calcium arsenite liposomal “target-controlled” drug delivery system (FA-LP-CaAs) was fabricated using the reverse microemulsion method. MethodsA Malvern particle size analyzer and a transmission electron microscope were employed to determine the particle size, distribution, zeta potential and morphology of FA-LP-CaAs. Further, inductively coupled plasma emission spectrometry was employed to determine the drug loading capacity, entrapment efficiency, and in vitro release behavior of FA-LP-CaAs. To determine its toxicity in human hepatoma cells (HepG2) and human normal hepatocytes (LO2) and its effect on HepG2 cell cycle and apoptosis, the MTT method was used. Laser confocal and flow cytometry were also employed to determine the uptake of FA-LP-CaAs by cells. After establishing a mouse liver cancer model, the in vivo distribution of the drug included in the formulation was investigated using in vivo fluorescence. To evaluate the liver cancer targeting and anti-tumor effects of FA-LP-CaAs in vivo, the distribution of ATO in tissues and changes in tumor volume and body weight after liposomal administration were investigated using hematoxylin-eosin (HE)-stained tumor sections. ResultsThe particle size, zeta potential and PDI of FA-LP-CaAs were (122.67 ± 2.18) nm, (12.81 ± 0.75) mV and 0.22 ± 0.01, respectively, while its drug loading capacity was 18.49% ± 1.14%. In vitro experimental results revealed that FA-LP-CaAs had a strong killing effect on HepG2 cells. Further, the cell uptake capacity of this formulation was found to improve. Based on in vivo assessments, FA-LP-CaAs could significantly increase the distribution of ATO in tumor sites and inhibit tumor growth. ConclusionsHerein, an FA-LP-CaAs formulation was successfully fabricated. This liposomal drug delivery system had a round appearance, uniform particle size, good polydispersity coeffi-cient, evident “core-shell” structure, high drug loading capacity and pH response, tumor targeted drug delivery and sustained drug release. These findings support further research and the application of ATO as an anti-liver cancer prodrug and provide a new method for the treatment of liver cancer.

Liposomal doxorubicin as targeted delivery platform: Current trends in surface functionalization.

Liposomal delivery systems have significantly enhanced the efficacy and safety of chemotherapeutic agents compared to free (non-liposomal) formulations. Liposomes are vesicles made up of lipophilic bilayer and a hydrophilic core which provides perfect opportunity for their application as transport vehicle for various therapeutic and diagnostic agents. Doxorubicin is the most exploited chemotherapeutic agent for evaluation of different liposomal applications, as its physicochemical properties permit high drug entrapment and easy remote loading in pre-formulated liposomes. Pegylated liposomal doxorubicin clinically approved and, on the market, Doxil®, exemplifies the benefits offered upon the surface modification of liposome with polyethylene glycol. This unique formulation prolonged the drug residence time in the circulation and increased accumulation of doxorubicin in tumor tissue via passive targeting (enhanced permeability and retention effect). However, there is ample scope for further improvement in the efficiency of targeting tumors by coupling biological active ligands onto the liposome surface to generate intelligent drug delivery systems. Small biomolecules such as peptides, fraction of antibodies and carbohydrates have the potential to target receptors present on the surface of the malignant cells. Hence, active targeting of malignant cells using functionalised nanocarrier (liposomes encapsulated with doxorubicin) have been attempted which is reviewed in this article.

Design and Construction of pH-Selective Self-Lytic Liposome System

Liposomes are well-investigated drug or gene delivery vehicles for chemotherapy, used by taking advantage of their biocompatibility and biodegradability. A central question on the construction of intracellular liposomal delivery systems is to entrap the liposomes of interest in the highly acidic and proteolytic endosomal environment. In the other words, it is essential that the liposomes release a therapeutic drug into the cytosol before they are degraded in the endosome. As a strategy to enhance the endosome escape, the self-lytic liposomes with acidic pH-selective membrane active polypeptide are considered highly effective. Here, an acidic pH-selective membrane-lytic polypeptide (LPE) and its retro isomer (rLPE) were designed, and then their membrane-lytic activities against EggPC liposomes were determined. It was noticed that the rLPE polypeptide showed an increase in activity compared with the LPE polypeptide. Furthermore, the rLPE polypeptide was conjugated to liposomes via a flexible Gly-Gly-Gly-Gly linker to facilitate the pH-selective content release. The rLPE anchoring liposomes exhibited distinctly different contents release behavior at physiological and endosomal pHs, namely typical contents release from liposomes was positively observed at acidic pH range. The overarching goal of this paper is to develop efficient pH-selective therapeutic delivery systems by using our findings.

Spatial and Temporal Control of CRISPR-Cas9-Mediated Gene Editing Delivered via a Light-Triggered Liposome System.

The CRISPR-Cas9 and related systems offer a unique genome-editing tool allowing facile and efficient introduction of heritable and locus-specific sequence modifications in the genome. Despite its molecular precision, temporal and spatial control of gene editing with the CRISPR-Cas9 system is very limited. We developed a light-sensitive liposome delivery system that offers a high degree of spatial and temporal control of gene editing with the CRISPR-Cas9 system. We demonstrated its efficient protein release by respectively assessing the targeted knockout of the eGFP gene in human HEK293/GFP cells and the TNFAIP3 gene in TNFα-induced HEK293 cells. We further validated our results at a single-cell resolution using an in vivo eGFP reporter system in zebrafish (77% knockout). These findings indicate that light-triggered liposomes may have new options for precise control of CRISPR-Cas9 release and editing.

Tumor-Tropic Liposome-Mediated Therapeutic Delivery of mRNA for T Cell Malignancies

Introduction : Treatment options for relapsed T cell leukemia are limited, and the prognosis remains dismal. Therefore, the development of new therapies is crucial. A liposomal delivery system has been increasingly recognized as a promising strategy for delivering both reagents and nucleic acids. However, little is known about whether the liposomal delivery of mRNA could be employed for cancer treatment. Herein, we propose a novel strategy for the treatment of T cell malignancies using tumor-tropic liposomes that can selectively deliver mRNAs of interest to leukemia cells. Methods: We tested liposomes with various lipid compositions. Leukemia cells or peripheral blood mononuclear cells (PBMCs) were cultured with rhodamine-labeled liposomes and then analyzed for rhodamine expression to investigate the selective uptake of the liposomes. To examine the selective translation of the encapsulated mRNA, the cells were incubated with liposomes loaded with firefly luciferase (ffLuc) mRNA and examined for luciferase activity. The cells were also treated with liposomes loaded with inducible caspase 9 (iC9) mRNA, with or without B/B homodimerizer (chemical-induced dimerization, CID), to examine the in vitro anti-leukemic effects. To investigate the in vivo anti-tumor effects, NSG mice were inoculated with Jurkat-ffLuc cells, and then they were intravenously treated with either liposomes loaded with iC9 or control liposomes on days 4 and 15 after tumor inoculation. Every 24, 48, and 72 h after liposome infusion, the mice were treated intraperitoneally with CID. Bioluminescence imaging was performed twice weekly to track the leukemia cell burden. Results: The screening analysis identified two types of liposomes (No. 43 and No. 79) that could selectively deliver mRNA to the cancer cells. Flow cytometry analysis showed that these liposomes labeled with rhodamine were efficiently taken up into Jurkat cells, whereas they were minimally taken up into PBMCs. Furthermore, when these two liposomes were loaded with ffLuc mRNA (43-ffLuc and 79-ffLuc), they could efficiently deliver mRNA to the cancer cells, with enhanced luciferase activity, but they minimally delivered ffLuc mRNA to PBMCs. Consistently, when these two liposomes were loaded with a suicide gene iC9 (43-iC9 and 79-iC9) in combination with CID, they effectively killed Jurkat cells and CCRF-CEM cells but minimally killed PBMCs in vitro. Furthermore, in a xenograft model of Jurkat-ffLuc, the mice treated with 79-iC9 showed significantly suppressed tumor growth compared with the mice treated with control liposome in combination with CID (Figure 1). These results suggested that the tumor-tropic liposomal delivery of iC9 mRNA could be employed for the treatment of T cell leukemia. Conclusions: Tumor-tropic liposomes can selectively deliver mRNAs of interest to leukemia cells. Moreover, tumor-tropic liposomes loaded with iC9 in combination with CID showed anti-leukemic activity both in vitro and in vivo. Thus, liposomal delivery could be a promising alternative for the treatment of T cell malignancies. Disclosures Saito: Toshiba Corporation: Research Funding. Nakashima:Toshiba Corporation: Research Funding. Hasegawa:Toshiba Corporation: Research Funding. Akahoshi:Toshiba Corporation: Current Employment. Ishihara-Sugano:Toshiba Corporation: Current Employment. Yagyu:Toshiba Corporation: Research Funding. Nakazawa:Toshiba Corporation: Research Funding.

The New Frontiers in Neurodegenerative Diseases Treatment: Liposomal-Based Strategies.

In the last decade, the onset of neurodegenerative (ND) diseases is strongly widespread due to the age increase of the world population. Despite the intensive investigations boosted by the scientific community, an efficacious therapy has not been outlined yet. The drugs commonly used are only able to relieve symptom severity; following their oral or intravenous administration routes, their effectiveness is strictly limited due to their low ability to reach the Central Nervous System (CNS) overcoming the Blood Brain Barrier (BBB). Starting from these assumptions, the engineered-nanocarriers, such as lipid-nanocarriers, are suitable agents to enhance the delivery of drugs into the CNS due to their high solubility, bioavailability, and stability. Liposomal delivery systems are considered to be the ideal carriers, not only for conventional drugs but also for neuroprotective small molecules and green-extracted compounds. In the current work, the LP-based drug delivery improvements in in vivo applications against ND disorders were carefully assessed.

Compositional inhomogeneity of drug delivery liposomes quantified at the single liposome level

Liposomes are the most used drug delivery vehicle and their therapeutic function is closely linked to their lipid composition. Since most liposome characterization is done using bulk techniques, providing only ensemble averages, the lipid composition of all liposomes within the same formulation are typically assumed to be identical. Here we image individual liposomes using confocal microscopy to quantify that liposomal drug delivery formulations, including multiple component mixtures mimicking Doxil, display more than 10-fold variation in their relative lipid composition. Since liposome function is tightly regulated by the physicochemical properties bestowed by the lipid composition, such significant variations could render only a fraction of liposomes therapeutically active. Additionally, we quantified how this degree of compositional inhomogeneity was modulated by liposome preparation method, the saturation state of the membrane lipid, and whether anti-fouling polyethylene glycol (PEG) conjugated lipids were added to the initial lipid mix or inserted after liposome formation. We believe the insights into the factors governing the degree of inhomogeneity offers the possibility for producing more uniform liposomal drug delivery systems, potentially increasing their therapeutic efficacy.

Active Transportation of Liposome Enhances Tumor Accumulation, Penetration, and Therapeutic Efficacy.

Liposomes are the first and mostly explored nanocarriers for cancer drug delivery, which have shown great promise in clinical applications, but their limited accumulation and penetration into the tumor interstitial space, significantly reduce the therapeutic efficacy. Here, a γ-glutamyltranspeptidase (GGT)-triggered charge-switchable approach is reported that can trigger the fast endocytosis and transcytosis of the liposome in tumor microenvironments to overcome the harsh biological barriers in tumor tissues. The active transporting liposomal nanocarrier (GCSDL) is prepared by surface modification with a glutathione (GSH) moiety and encapsulated with doxorubicin (DOX). When the GCSDL contacts with tumor vascular endothelial cells, the overexpressed GGT enzyme on cytomembrane catalyzes the hydrolysis of GSH to generate cationic primary amines. The cationic GCSDL triggers fast caveolae-mediated endocytosis and vesicle-mediated transcytosis, resulting in sequential transcytosis to augment its tumor accumulation and penetration. Along with continual intercellular transportation, GCSDL can release DOX throughout the tumor to induce cancer cell apoptosis, resulting in complete eradication of hepatocellular carcinoma and cessation of pancreatic ductal adenocarcinoma's progression. This study develops an efficient strategy to realize high tumor accumulation and deep penetration for the liposomal drug delivery system via active transcytosis.

In Vitro and In Vivo Evaluation of SP94 Modified Liposomes Loaded with N-14NCTDA, a Norcantharimide Derivative for Hepatocellular Carcinoma-Targeting.

The purpose of this research is to develop a liposomal drug delivery system, which can selectively target hepatocellular carcinoma (HCC) to deliver the antitumor agent N-14NCTDA, a C14 alkyl chain norcantharimide derivative of norcantharidin. N-14NCTDA-loaded liposomes were successfully prepared by lipid membrane hydration and extrusion methods. SP94, a targeting peptide for HCC cells, was attached to the liposomes loaded with N-14NCTDA by the post-insertion method to obtain SP94 modified liposomes (SP94-LPs). SP94-LPs had a significant cytotoxicity against Hep G2 cells with the IC50 of 15.395 ± 0.89μg/mL, which is lower than that of NCTD-S (IC50 = 20.863 ± 0.56μg/mL) and GAL-LPs (IC50 = 24.589 ± 1.02μg/mL). Compared with conventional liposomes (Con-LPs), SP94-LPs showed greater cellular uptake in Hep G2 cells. Likewise, significant tumor suppression was achieved in H22 tumor-bearing mice which were treated with SP94-LPs. The tumor inhibition rate (IRw) of SP94-LPs was 82 ± 0.98%, obviously higher than that of GAL-LPs (69 ± 1.39%), Con-LPs (60 ± 2.78%), and NCTD-S (51 ± 3.67%). SP94-LPs exhibited a significant hepatocellular carcinoma-targeting activity in vitro and in vivo, which will provide a new alternative for hepatocellular carcinoma treatment in future. Graphical Abstract.

Co-delivery of a RanGTP inhibitory peptide and doxorubicin using dual-loaded liposomal carriers to combat chemotherapeutic resistance in breast cancer cells

ABSTRACT Background Multidrug resistance (MDR) limits the beneficial outcomes of conventional breast cancer chemotherapy. Ras-related nuclear protein (Ran-GTP) plays a key role in these resistance mechanisms, assisting cancer cells to repair damage to DNA. Herein, we investigate the co-delivery of Ran-RCC1 inhibitory peptide (RAN-IP) and doxorubicin (DOX) to breast cancer cells using liposomal nanocarriers. Research design A liposomal delivery system, co-encapsulating DOX, and RAN-IP, was prepared using a thin-film rehydration technique. Dual-loaded liposomes were optimized by systematic modification of formulation variables. Real-Time-Polymerase Chain Reaction was used to determine Ran-GTP mRNA expression. In vitro cell lines were used to evaluate the effect of loaded liposomes on the viability of breast and lung cancer cell lines. In vivo testing was performed on a murine Solid Ehrlich Carcinoma model. Results RAN-IP reversed the Ran-expression-mediated MDR by inhibiting the Ran DNA damage repair function. Co-administration of RAN-IP enhanced sensitivity of DOX in breast cancer cell lines. Finally, liposome-mediated co-delivery with RAN-IP improved the anti-tumor effect of DOX in tumor-bearing mice when compared to single therapy. Conclusions This study is the first to show the simultaneous delivery of RAN-IP and DOX using liposomes can be synergistic with DOX and lead to tumor regression in vitro and in vivo.

Engineering Stem Cell Derived Biomimetic Vesicles for Versatility and Effective Targeted Delivery

AbstractTo date, only few a types of nanomedicines have been made available for clinical use even in the most common liposomal delivery systems, due to significant constraints, including poor targeting of specific cells, hindered penetration, and accelerated clearance. Various intricate designs have been generated and show improvements, while also bringing new problems. Biological cells, possessing advantages like low immunogenicity, long circulation time, receptors integration, and innate targeting capability. Herein, liposomes are innovatively functionalized with a stem cell membrane through a facile approach. The biomimetic vesicles achieve controlled release, exhibit better stability, longer circulation time, and targeted delivery. Loading with curcumin leads to enhanced survival rate of ischemic stroke mice with a single injection (from ≈30% to over 90%). The versatility of the method is verified by differently charged liposomes modification and erythrocyte membrane derived vesicles preparation. Overall, the cell membrane derived biomimetic vesicles achieve targeted delivery and versatility prepared by a facile approach, as well as effortless preparation.

Tumor repolarization by an advanced liposomal drug delivery system provides a potent new approach for chemo-immunotherapy.

Immunosuppressive cells in the tumor microenvironment allow cancer cells to escape immune recognition and support cancer progression and dissemination. To improve therapeutic efficacy, we designed a liposomal oxaliplatin formulation (PCL8-U75) that elicits cytotoxic effects toward both cancer and immunosuppressive cells via protease-mediated, intratumoral liposome activation. The PCL8-U75 liposomes displayed superior therapeutic efficacy across all syngeneic cancer models in comparison to free-drug and liposomal controls. The PCL8-U75 depleted myeloid-derived suppressor cells and tumor-associated macrophages in the tumor microenvironment. The combination of improved cancer cell cytotoxicity and depletion of immunosuppressive populations of immune cells is attractive for combination with immune-activating therapy. Combining the PCL8-U75 liposomes with a TLR7 agonist induced immunological rejection of established tumors. This combination therapy increased intratumoral numbers of cancer antigen-specific cytotoxic T cells and Foxp3- T helper cells. These results are encouraging toward advancing liposomal drug delivery systems with anticancer and immune-modulating properties into clinical cancer therapy.

Differential Immunogenicity and Protective Efficacy Elicited by MTO- and DMT-Adjuvanted CMFO Subunit Vaccines against Mycobacterium tuberculosis Infection.

Tuberculosis (TB) remains a major and global problem of public health. An effective TB subunit vaccine is urgently needed. Proper selection of the delivery system for the vaccine is crucial for inducing an appropriate immune response tailored to control the target pathogen. In this study, we compared the immunogenicity and protective efficacy of CMFO subunit vaccines against primary progressive TB in two different adjuvant systems: the MTO oil-in-water (O/W) emulsion composed of monophosphoryl lipid A (MPL), trehalose-6,60-dibehenate (TDB), and oil in water emulsion MF59 and the DMT liposome containing dimethyldioctadecylammonium bromide (DDA), monophosphoryl lipid A (MPL), and trehalose-6,60-dibehenate (TDB). Our results demonstrated that the DMT-adjuvanted CMFO could confer more significant protection against M. tuberculosis infection than the CMFO/MTO did in mice. In particular, the adjuvant DMT showed a stronger ability than the O/W emulsion to adjuvant CMFO subunit vaccine and enhanced protection, attributed to elicit Th1-biased responses, strong Th1/Th17 cytokine responses, and IFN-γ+ or IL-2+ T cell responses. Therefore, our findings demonstrate that the liposome delivery system shows more effectiveness to adjuvant TB subunit vaccine than O/W emulsion and highlight the importance of adjuvant formulation for the better efficacy of a protein vaccine.

HL-284: First Documented Use of Liposomal Doxorubicin in a Pregnant Patient with Hodgkin Lymphoma

Context Hodgkin Lymphoma (HL) patients tend to be younger and may become pregnant concomitantly. We present a pregnant, heavily-pretreated HL patient who received liposomal doxorubicin (doxil) and delivered a healthy boy. Setting Patient is a 29 y/o woman diagnosed with classical HL in 2016. She completed six cycles of ABVD in October, 2016. In May 2017, she received radiation to areas of progression. She was referred to us in November 2017 with disease progression confirmed with inguinal biopsy. She received two cycles of ICE with partial response. She received brentuximab in March, April, and May 2018. In June 2018, nivolumab was added to brentuximab. The combination continued until October 2018. Given better response, she had stem cells collected. She became pregnant in October 2018. She had an abortion and proceeded to transplant November 19, 2018. PET showed remission 3 months later, and she started brentuximab maintenance. Follow-up imaging showed relapse, verified on biopsy 8/9/19. We discussed giving nivolumab, but she became pregnant again. Nivolumab and bendamustine are category D with limited to no safety in pregnancy. We offered gemcitabine, vinorelbine, and doxil (GND). After a baseline MRI and echocardiogram demonstrated ejection fraction of 59%, she started GND in the second trimester. She received three cycles of chemotherapy (January to March 2020). Results Improved lymphadenopathy was noted on exam with shrinking right inguinal mass, fewer fevers, and improved appetite. Patient developed COVID-19 in April 2020 and so discontinued further chemotherapy. She delivered a healthy boy 5/1/20 weighing 2.93 kg. Post-delivery PET-CT showed progression of disease, and palpable nodes had begun to grow again. Bendamustine started 6/3/20. Conclusions ABVD is found to be safe in pregnant HL patients starting in the second trimester. A literature search found no cases of the use of doxil in pregnancy in any malignancy setting. Experimental models, including murine, have demonstrated the potential safety of a liposomal delivery system. GND is a well-recognized chemotherapy combination for HL. While data support the use of ABVD starting in the second trimester, limited data are available for relapsed pregnant HL patients. GND may be a safe regimen in such a setting. Hodgkin Lymphoma (HL) patients tend to be younger and may become pregnant concomitantly. We present a pregnant, heavily-pretreated HL patient who received liposomal doxorubicin (doxil) and delivered a healthy boy. Patient is a 29 y/o woman diagnosed with classical HL in 2016. She completed six cycles of ABVD in October, 2016. In May 2017, she received radiation to areas of progression. She was referred to us in November 2017 with disease progression confirmed with inguinal biopsy. She received two cycles of ICE with partial response. She received brentuximab in March, April, and May 2018. In June 2018, nivolumab was added to brentuximab. The combination continued until October 2018. Given better response, she had stem cells collected. She became pregnant in October 2018. She had an abortion and proceeded to transplant November 19, 2018. PET showed remission 3 months later, and she started brentuximab maintenance. Follow-up imaging showed relapse, verified on biopsy 8/9/19. We discussed giving nivolumab, but she became pregnant again. Nivolumab and bendamustine are category D with limited to no safety in pregnancy. We offered gemcitabine, vinorelbine, and doxil (GND). After a baseline MRI and echocardiogram demonstrated ejection fraction of 59%, she started GND in the second trimester. She received three cycles of chemotherapy (January to March 2020). Improved lymphadenopathy was noted on exam with shrinking right inguinal mass, fewer fevers, and improved appetite. Patient developed COVID-19 in April 2020 and so discontinued further chemotherapy. She delivered a healthy boy 5/1/20 weighing 2.93 kg. Post-delivery PET-CT showed progression of disease, and palpable nodes had begun to grow again. Bendamustine started 6/3/20. ABVD is found to be safe in pregnant HL patients starting in the second trimester. A literature search found no cases of the use of doxil in pregnancy in any malignancy setting. Experimental models, including murine, have demonstrated the potential safety of a liposomal delivery system. GND is a well-recognized chemotherapy combination for HL. While data support the use of ABVD starting in the second trimester, limited data are available for relapsed pregnant HL patients. GND may be a safe regimen in such a setting.

Improved antitumor activity and tolerability of cabazitaxel derived remote-loading liposomes.

The value of the clinical application of chemotherapeutic drugs is dependent on both systemic toxicity and treatment efficacy. Dose intensification and high tolerability suggest the potential for clinical cancer therapy. In this study, we developed a novel strategy for reconstructing a drug molecule into remote-loading liposomes. Two weak-base cabazitaxel derivatives were synthesized, and named CN and CN2. CN exhibited higher cytotoxic effects compared to CN2, and was selected for further study. CN was remotely loaded into nano-size liposomes (CN-LPs) via an ammonium sulfate gradient with high loading and encapsulation efficiency. When compared to the commercial formulation of cabazitaxel, JEVTANA®, CN-LPs showed less systemic toxicity and enhanced tolerability, with at least a 24-fold increase in the tolerated dose. Furthermore, CN-LPs significantly inhibited tumor growth in mice bearing 4T1 and RM-1 xenograft tumors. After intravenous injection, CN-LPs exhibited an extremely high drug concentration in blood, with a 757-fold increase in the area under the curve (AUC). Moreover, 48h after a single intravenous injection, CN-LPs promoted higher drug accumulation in tumors compared to JEVTANA®. In summary, our liposome delivery system exhibits favorable pharmacologic efficacy and an improved safety profile.

Developing a Model for a siRNA Delivery System by Cancer Implantation into Zebrafish Circulation.

In recent decades, zebrafish (Danio rerio) has become a major in vivo model for the evaluation of drug efficacies and toxicities. In the field of drug delivery research, zebrafish larvae are a suitable model for the use of fluorescent-labeled chemicals, nanoparticle, liposome, or micelle-mediated delivery systems because of their transparent body wall. In the current chapter, we describe the method to perform micelle-based siRNA delivery using cancer cells implanted into the circulation of zebrafish.

Spontaneous In Situ Formation of Liposomes from Inert Porous Microparticles for Oral Drug Delivery.

Despite the wide-spread use of liposomal drug delivery systems, application of these systems for oral purposes is limited due to their large-scale formulation and storage issues. Proliposomes are one of the formulation approaches for achieving solid powders that readily form liposomes upon hydration. In this work, we investigated a dry powder formulation of a model low-soluble drug with phospholipids loaded in porous functionalized calcium carbonate microparticles. We characterized the liposome formation under conditions that mimic the different gastrointestinal stages and studied the factors that influence the dissolution rate of the model drug. The liposomes that formed upon direct contact with the simulated gastric environment had a capacity to directly encapsulate 25% of the drug in situ. The emerged liposomes allowed complete dissolution of the drug within 15 min. We identified a negative correlation between the phospholipid content and the rate of water uptake. This correlation corroborated the results obtained for the rate of dissolution and liposome encapsulation efficiency. This approach allows for the development of solid proliposomal dosage formulations, which can be scaled up with regular processes.

Design and Validation of Liposomal ApoE2 Gene Delivery System to Evade Blood-Brain Barrier for Effective Treatment of Alzheimer's Disease.

Targeting gene-based therapeutics to the brain is a strategy actively sought to treat Alzheimer's disease (AD). Recent findings discovered the role of apolipoprotein E (ApoE) isoforms in the clearance of toxic amyloid beta proteins from the brain. ApoE2 isoform is beneficial for preventing AD development, whereas ApoE4 is a major contributing factor to the disease. In this paper, we demonstrated efficient brain-targeted delivery of ApoE2 encoding plasmid DNA (pApoE2) using glucose transporter-1 (glut-1) targeted liposomes. Liposomes were surface-functionalized with a glut-1 targeting ligand mannose (MAN) and a cell-penetrating peptide (CPP) to enhance brain-targeting and cellular internalization, respectively. Among various CPPs, rabies virus glycoprotein peptide (RVG) or penetratin (Pen) was selected as a cell-penetration enhancer. Dual (RVGMAN and PenMAN)-functionalized liposomes were cytocompatible at 100 nM phospholipid concentration and demonstrated significantly higher expression of ApoE2 in bEnd.3 cells, primary neurons, and astrocytes compared to monofunctionalized and unmodified (plain) liposomes. Dual-modified liposomes also showed ∼2 times higher protein expression than other formulation controls in neurons cultured below the in vitro BBB model. These results translated well to in vivo efficacy study with significantly higher transfection of pApoE2 in the C57BL/6 mice brain following single tail vein administration of RVGMAN and PenMAN functionalized liposomes without any noticeable signs of toxicity. These results illustrate the potential of surface-modified liposomes for safe and brain-targeted delivery of the pApoE2 gene for effective AD therapy.