Biomimetic nanoparticles deliver mRNAs encoding costimulatory receptors and enhance T cell mediated cancer immunotherapy

Antibodies targeting costimulatory receptors of T cells have been developed for the activation of T cell immunity in cancer immunotherapy. However, costimulatory molecule expression is often lacking in tumor-infiltrating immune cells, which can impede antibody-mediated immunotherapy. Here, we hypothesize that delivery of costimulatory receptor mRNA to tumor-infiltrating T cells will enhance the antitumor effects of antibodies. We first design a library of biomimetic nanoparticles and find that phospholipid nanoparticles (PL1) effectively deliver costimulatory receptor mRNA (CD137 or OX40) to T cells. Then, we demonstrate that the combination of PL1-OX40 mRNA and anti-OX40 antibody exhibits significantly improved antitumor activity compared to anti-OX40 antibody alone in multiple tumor models. This treatment regimen results in a 60% complete response rate in the A20 tumor model, with these mice being resistant to rechallenge by A20 tumor cells. Additionally, the combination of PL1-OX40 mRNA and anti-OX40 antibody significantly boosts the antitumor immune response to anti-PD-1 + anti-CTLA-4 antibodies in the B16F10 tumor model. This study supports the concept of delivering mRNA encoding costimulatory receptors in combination with the corresponding agonistic antibody as a strategy to enhance cancer immunotherapy.

Assessing the effect of ultra-small phospholipid nanoparticles on biochemical parameters of carbohydrate and fat metabolism in patients with combined hyperlipidemia

Background and aims Based on the developed technology of preparation of ultra-small size phospholipid (PL) nanoparticles without the use of detergents/surfactants and stabilizers drug preparation, exhibiting hypolipidemic properties has been obtained. Preclinical and clinical studies have shown that phospholipid nanoparticles of 20–30 nm (PLN) have activated reverse cholesterol transport and reduced non-high-density lipoprotein cholesterol levels. The aim of the study was to compare the effect of PLN relative to placebo on the selected parameters of carbohydrate and fat metabolism in patients with combined hyperlipidemia. Methods This Phase III, randomized, double-blind, placebo-controlled, parallel-group study was conducted at 4 centers. The patients (pts) received PLN or placebo (1:1), powder for oral solution preparation (500 mg PL/dose) orally 2 times per day for 12 weeks. Biochemical parameters of carbohydrate and fat metabolism were measured. The data is presented in M (s). Statistical analysis was performed using the Mann-Whitney U-test and the Wilcoxon sign rank test. P-value <0.05 was considered statistically significant. Results A total of 100 pts with combined hyperlipidemia were randomized (age 35–70 years, males 58%). The mean age of patients was 56.4 (10.2) years, body-mass index (BMI) – 30.4 kg/m (2). Mean glucose, triglyceride (TG), fasting insulin (INS) and adiponectin (ADN) serum levels at baseline were 5.58 (0.9) mmol/l, 2.38 (0.6) mmol/l, 13.71 (6.03) mU/ml and 7.07 (2.25) μg/ml, respectively. At Week 12, PLN significantly reduced TG and insulin (p=0.03 and p=0.02, respectively). At the same time, the mean adiponectin level was increased (ns). The drug was well tolerated, and no clinically significant laboratory abnormalities were detected. Conclusions PLN therapy is well tolerated with a favorable effect on the chosen parameters of carbohydrate and fat metabolism in patients with moderate combined hyperlipidemia. FUNDunding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Ministry of industry and trade, Ministry of Healthcare of the Russian Federation

The effect of lipid derivative of anti-tumor drug sarcolysin embedded in phospholipid nanoparticles in the experiments in vivo

To improve the therapeutic properties of the antitumor agent Sarcolysin, we have previously developed and characterized a dosage form representing its ester conjugate with decanol embedded in ultra-small phospholipid nanoparticles less than 30 nm in size (“Sarcolysin-NP”). The effect of the resulting composition was investigated in vivo in comparison with the free substance of sarcolysin. The composition intravenous administration to mice showed an improvement in the pharmacokinetic parameters of sarcolysin associated with its initial higher (by 22%) level in the blood and prolonged circulation, which was also observed in mice with P388 tumor. In mice with three types of tumors — lymphocytic leukemia P388, lymphocytic leukemia L1210, and adenocarcinoma of the mammary gland Ca755 — administration of two doses of sarcolysin over a period of 7 days showed its predominant antitumor effect. The maximum tumor growth inhibition was noted for lymphocytic leukemia L1210 and adenocarcinoma of the mouse mammary gland Ca755 (at a dose of Sarcolysin-NP — 8,4 mg/kg), which was higher in comparison with free substance by more than 24% and 17%, respectively. Differences in the life span of the treated animals were revealed significantly at a dose of 10 mg/kg and amounted to 25% and 17,4% for lymphocytic leukemia P388 and L1210, respectively, and 11% for adenocarcinoma Ca755. In an experiment on rats, acute toxicity of Sarcolysin-NP administered intravenously showed that an average LD50 value 2-3 times exceeded a similar parameter for commercial preparations of free sarcolysin (Melphalan and Alkeran), which indicates its lower toxicity.

Usage of a new pharmacological agent of ultra-small phospholipid nanoparticles (micelles) in the treatment of combined hyperlipidemia

Background and aims Based on the developed technology of prerparation of ultra small size phospholipid nanoparticles without the use of detergents/surfactants and stabilizers drug preparation, exhibiting hypolipidemic properties has been obtained. Preclinical studies have shown that phospholipid nanoparticles of 20–30 nm (PLN) activate reverse cholesterol transport. PLN is now at the stage of clinical trials (Phase II study completed). The aim of the study was to compare the efficacy, safety and tolerability of PLN relative to placebo in the treatment of combined hyperlipidemia. Methods This Phase III, multicenter, randomized, double-blind, placebo-controlled, parallel-group study was conducted at 4 centers. The patients (pts) received PLN or placebo (1:1), powder for oral solution preparation (in an aqueous environment is a nanoemulsion with a particle size of 20–30 nm), 500 mg (sachet) orally 2 times a day for 12 weeks. Lipid parameters, safety and tolerability were measured. The data is presented in M (s), Me (lq; hq). Mann-Whitney U Test was to compare the significant difference of groups. P value <0.05 was considered statistically significant. Results A total of 100 pts with combined hyperlipidemia were randomized (age 35–70 years, males 58%).The mean age of patients was 56.4 (10.2) years. Mean total cholesterol, non-high-density lipoprotein cholesterol (non-HDL-C), TG, and apolipoprotein B 100 (apoB) levels at baseline were 5.59 (0.8) mmol/l, 4.51 (0.75) mmol/l, 2.38 (0.6) mmol/l and 114.26 (24.96) mg/dl, respectively (n=100). Mean high-density lipoprotein cholesterol (HDL-C) and Lp(a) levels at baseline were 1.0 (0.25) mmol/l and 21.1 (3.9; 20.3)mg/dl, respectively (n=100). At Week 12, PLT significantly reduced total cholesterol, non-HDL-C levels, TG and apo B compared to placebo (p=0.02, p=0.03, p=0.001 and p=0.02, respectively). At the same time, the mean HDL-C level was significantly increased (p=0.03). The drug was well tolerated and no clinically significant laboratory abnormalities were detected. Conclusions This study demonstrates that therapy with PLT is well tolerated in additional beneficial effects on key lipid parameters in patients with moderate combined hyperlipidemia. Funding Acknowledgement Type of funding source: Public Institution(s). Main funding source(s): Ministry of industry and trade