Journal: ACS nano

Article Title: Nanocarrier Co-formulation for Delivery of a TLR7 Agonist plus an Immunogenic Cell Death Stimulus Triggers Effective Pancreatic Cancer Chemo-immunotherapy.

doi: 10.1021/acsnano.2c06300

Figure Lengend Snippet: Figure 1. Schematic explaining the key design features for co-formulated drug delivery by lipid bilayer coated silicasomes and liposomes. The basic approach to drug co-formulation is to use the hydrophilic interior of these carriers for remote loading of amphiphilic drugs, such as irinotecan, while employing the lipophilic bilayer to incorporate synthetic lipid moieties and prodrugs. The lipid moieties include synthetic agents with immune stimulatory activity, such as 3M-052 (aka Telratolimod). 3M-052 contains a C18 lipid tail that facilitates bilayer incorporation, which was tested in a liposome to obtain optimal LB composition before applying that to the design of the MSNP bilayer. The schematic also shows that, in addition to 3M-052, the LB can be used to incorporate a list of lipid-conjugated prodrugs that can provide immune checkpoint blockade or interfere in the immunometabolic IDO-1 pathway, as previously described by us.22 Irinotecan remote loading is accomplished by using ammonium sulfate or sucralose octasulfate for import into the aqueous interiors in the liposome or silicasome, respectively. These trapping agents allow amphipathic weak basic molecules (such as irinotecan, doxorubicin, and mitoxantrone) to cross the LB for protonation inside these carriers, where they collect as slowly dissolving drug precipitates. The hypothesis for the experimentation carried out in this article is that the generation of immunogenic cell death by irinotecan will be boosted by co-delivery of 3M-052, leading to improved dendritic cell activation. Adapted with permission from ref 15. Copyright 2022 American Chemical Society.

Article Snippet: 3M-052 (aka Telratolimod) and Cell Counting Kit-8 (CCK-8) were purchased from MedChemExpress, USA.

Techniques: Liposomes, Formulation, Activity Assay, Activation Assay

Journal: ACS nano

Article Title: Nanocarrier Co-formulation for Delivery of a TLR7 Agonist plus an Immunogenic Cell Death Stimulus Triggers Effective Pancreatic Cancer Chemo-immunotherapy.

doi: 10.1021/acsnano.2c06300

Figure Lengend Snippet: Figure 2. Development of a co-formulated silicasome drug carrier to deliver a combination of 3M-052, plus the ICD-chemotherapeutic agents IRIN. (A) Schematic to outline the synthesis steps for constructing the silicasome. Briefly, MSNP was soaked in TEA8SOS trapping agent at 65 °C (step 1), prior to adding to preheated (65 °C) pure ethanol, into which we dissolved a mixture of 3M-052 and lipids (DSPC/Chol/DSPE- PEG2000/3M-052, in the molar ratio of 55.5:38.5:2.7:3.3) (step 2). This mixture was sonicated to provide LB coating (step 3). For IRIN remote loading, irinotecan was dissolved in HEPES-buffered dextrose, before mixing and incubation of the purified TEA8SOS-loaded 3M- silicasome at 65 °C (step 4). After quenching in ice water, the 3M-silicasome-IR was purified (step 5). As a control, we also synthesized a silicasome that incorporates 3M-052 only, without irinotecan loading (3M-silicasome-IR). CryoEM visualization of the (B) 3M-silicasome and (C) 3M-silicasome-IR. The scale bar is 100 nm. (D) Physicochemical properties of both silicasomes, including hydrodynamic size, polydispersity index (PDI), zeta potential, and drug loading capacities.

Article Snippet: 3M-052 (aka Telratolimod) and Cell Counting Kit-8 (CCK-8) were purchased from MedChemExpress, USA.

Techniques: Sonication, Incubation, Purification, Control, Synthesized, Zeta Potential Analyzer

Journal: ACS nano

Article Title: Nanocarrier Co-formulation for Delivery of a TLR7 Agonist plus an Immunogenic Cell Death Stimulus Triggers Effective Pancreatic Cancer Chemo-immunotherapy.

doi: 10.1021/acsnano.2c06300

Figure Lengend Snippet: Figure 4. Therapeutic impact on tumor growth in a subcutaneous KPC model. (A) Experimental timeline to assess the therapeutic impact of single- and dual-delivery carriers in subcutaneous KPC tumor-bearing mice. (B) Averaged tumor growth kinetics (n = 6, 7) through assessment of tumor volume in animals treated with saline, free 3M-052, free IRIN, 3M-silicasome, and 3M-silicasome-IR at dose equivalents of 2 and 40 mg/kg for 3M-052 and IRIN, respectively, every 3 or 4 days, for a total of 4 IV injections. Please notice that the inhibition of tumor growth by free 3M-052, free IRIN, 3M-silicasome, and 3M-silicasome-IR is statistically significant compared to the saline control (p < 0.001) on day 20. Although limited animal availability during COVID-19 precluded the inclusion of a carrier delivering encapsulated IRIN only, we have previously shown the advantage of an IRIN silicasome over liposomal or free drug delivery in KPC tumors.7−9 (C) Photographic images of harvested tumors at the same magnification on day 21. (D) Spaghetti plots to depict the individual tumor growth kinetics over the course of the study. These plots further depict that inhibition of tumor growth by 3M-silicasome becomes statistically significant compared to free 3M-052 (p < 0.05) from day 18 onward. Moreover, inhibition of tumor growth by 3M-silicasome-IR becomes statistically significant from free IRIN and the 3M-silicasome (p < 0.01) from day 8 onward and from free 3M-052 (p < 0.01) from day 10 onward. Data represent mean ± SEM, #p < 0.05, ##p < 0.01, ###p < 0.001.

Article Snippet: 3M-052 (aka Telratolimod) and Cell Counting Kit-8 (CCK-8) were purchased from MedChemExpress, USA.

Techniques: Saline, Inhibition, Control

Journal: ACS nano

Article Title: Nanocarrier Co-formulation for Delivery of a TLR7 Agonist plus an Immunogenic Cell Death Stimulus Triggers Effective Pancreatic Cancer Chemo-immunotherapy.

doi: 10.1021/acsnano.2c06300

Figure Lengend Snippet: Figure 6. Pharmacokinetics and drug delivery by silicasomes versus free drugs. Healthy B6129SF1/J mice received one IV injection of free 3M- 052, free IRIN, and 3M-silicasome-IR at dose equivalents of 2 and 40 mg/kg for 3M-052 and IRIN, respectively. Blood collection was performed at 1, 5, 24, and 48 h postinjection (n = 3). Free 3M-052 and IRIN were extracted by methanol, and plasma drug concentrations were measured by HPLC, as described in the Methods section. While free IRIN (panel A) and free 3M-052 (panel B) rapidly disappeared from the blood (i.e., <60 min, prompting the inserted graphs display), the 3M-silicasome-IR significantly increased both plasma IRIN and 3M-052 concentrations.

Article Snippet: 3M-052 (aka Telratolimod) and Cell Counting Kit-8 (CCK-8) were purchased from MedChemExpress, USA.

Techniques: Drug discovery, IV Injection, Clinical Proteomics

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Millimeter Wave Treatment Inhibits Apoptosis of Chondrocytes via Regulation Dynamic Equilibrium of Intracellular Free Ca 2+

doi: 10.1155/2015/464161

Figure Lengend Snippet: Identification of third generation of chondrocytes (×200). (a) Chondrocytes were stained using toluidine blue. Immunohistochemical staining of type II collagen demonstrated that (b) the cytoplasm of the negative group of chondrocytes was unstained, whereas (c) the cytoplasm of chondrocytes was stained.

Article Snippet: Toluidine blue was obtained from Guoyao Group (Beijing, China), and a type II collagen immunostaining kit from Boster Inc. (Wuhan, China).

Techniques: Staining, Immunohistochemical staining