Philip S. Low

PurposeWe recently developed an optical instrument to non-invasively detect fluorescently labeled circulating tumor cells (CTCs) in mice called ‘Diffuse in vivo Flow Cytometry’(DiFC). OTL38 is a folate receptor (FR) targeted near-infrared (NIR) contrast agent that is FDA approved for use in fluorescence guided surgery of ovarian and lung cancer. In this work, we investigated the use OTL38 for in vivo labeling and detection of FR+ CTCs with DiFC.

PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-]. C= 12C= CC (=[N+](CC) CC) C= C2OC2= CC (N (CC) CC)= CC= C2C= 1C1= CC= CC= C1C (O)= O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims description 7YMZMTOFQCVHHFB-UHFFFAOYSA-N 5-carboxytetramethylrhodamine Chemical compound C= 12C= CC (N (C) C)= CC2=[O+] C2= CC (N (C) C)= CC= C2C= 1C1= CC= C (C (O)= O) C= C1C ([O-])= O YMZMTOFQCVHHFB-UHFFFAOYSA-N 0.000 claims description 6

MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C (= O) C2= CC (N= C= S)= CC= C2C21C1= CC= C (O) C= C1OC1= CC (O)= CC= C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 claims description 31

Cancer immunotherapy has achieved notable clinical success, yet its broader application is impeded primarily by immune evasion mechanisms, predominantly stemming from factors such as tumor antigen loss, intrinsic low antigen presentation, and the inherent impermeability of current macromolecules, including cells, antibodies, and proteins. Consequently, there exists a pressing demand for more permeable agents that target intracellular components as viable alternatives. Among the various molecular players involved in immune evasion, PTPN1 and PTPN2, two protein tyrosine phosphatases (PTPs), play a non-redundant role in regulating tumor antigen presentation. They achieve this by modulating the JAK/STAT signaling pathway within tumor cells, with their deletion having been demonstrated to enhance MHC-I presentation across various cell types. Furthermore, PTPN1 and PTPN2 also negatively …

RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1= CC (OC)= CC= C1C [C@ H](N) C (= O) N [C@ H] 1 [C@@ H](O)[C@ H](N2C3= NC= NC (= C3N= C2) N (C) C) O [C@@ H] 1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 4UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN [C@ H] 1 [C@ H](O)[C@@ H](O)[C@ H](CO) O [C@ H] 1O [C@@ H] 1 [C@](C= O)(O)[C@ H](C) O [C@ H] 1O [C@@ H] 1 [C@@ H](NC (N)= N)[C@ H](O)[C@@ H](NC (N)= N)[C@ H](O)[C@ H] 1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4

2023-07-06 Assigned to PURDUE RESEARCH FOUNDATION reassignment PURDUE RESEARCH FOUNDATION ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: GARDEEN, Spencer S., SHI, RIYI, HERR, Seth, LOW, PHILIP

Our team recently developed “Diffuse in vivo Flow Cytometry” (DiFC) for detection and enumeration rare circulating tumor cells (CTCs) in mice with highly-scattered fluorescent light. We have used DiFC to study dissemination of CTCs in a number of mouse models of metastasis with fluorescent protein expressing cells. Because DiFC uses diffuse light and interrogates large blood vessels in relatively deep tissue, in principle it could be translated to larger limbs, species, and even humans clinically. In this perspective, we discuss the technical challenges of human translation of DiFC in the context of the current state of the technology, as well as potential strategies for labeling of CTCs with targeted fluorescent molecular probes. We also discuss potential advantages and disadvantages of DiFC as a clinical or research tool. In principle, DiFC could represent a powerful complementary technique (to liquid biopsy blood draws) for accurate and sensitive measurement of changes in CTC numbers over time.

Folate receptors can perform folate transport, cell adhesion, and/or transcription factor functions. The beta isoform of the folate receptor (FRβ) has attracted considerable attention as a biomarker for immunosuppressive macrophages and myeloid-derived suppressor cells, however, its role in immunosuppression remains uncharacterized. We demonstrate here that FRβ cannot bind folate on healthy tissue macrophages, but does bind folate after macrophage incubation in anti-inflammatory cytokines or cancer cell-conditioned media. We further show that FRβ becomes functionally active following macrophage infiltration into solid tumors, and we exploit this tumor-induced activation to target a toll-like receptor 7 agonist specifically to immunosuppressive myeloid cells in solid tumors without altering myeloid cells in healthy tissues. We then use single-cell RNA-seq to characterize the changes in gene expression induced by the targeted repolarization of tumor-associated macrophages and finally show that their repolarization not only changes their own phenotype, but also induces a proinflammatory shift in all other immune cells of the same tumor mass, leading to potent suppression of tumor growth. Because this selective reprogramming of tumor myeloid cells is accompanied by no systemic toxicity, we propose that it should constitute a safe method to reprogram the tumor microenvironment.

Spinal fusions are performed to treat congenital skeletal malformations, spondylosis, degenerative disk diseases, and other pathologies of the vertebrae that can be resolved by reducing motion between neighboring vertebrae. Unfortunately, up to 100,000 fusion procedures fail per year in the United States, suggesting that efforts to develop new approaches to improve spinal fusions are justified. We have explored whether the use of an osteotropic oligopeptide to target an attached bone anabolic agent to the fusion site might be exploited to both accelerate the mineralization process and improve the overall success rate of spinal fusions. The data presented below demonstrate that subcutaneous administration of a modified abaloparatide conjugated to 20 mer of D-glutamic acid not only localizes at the spinal fusion site but also outperforms the standard of care (topically applied BMP2) in both speed of mineralization (p < 0.05) and overall fusion success rate (p < 0.05) in a posterior lateral spinal fusion model in male and female rats, with no accompanying ectopic mineralization. Because the bone-localizing conjugate can be administered ad libitum post-surgery, and since the procedure appears to improve on standard of care, we conclude that administration of a bone-homing anabolic agent for improvement of spinal fusion surgeries warrants further exploration.

Excessive deposition of extracellular matrix is a hallmark of Idiopathic pulmonary fibrosis (IPF), it is advantageous to target the cells and the mechanisms associated with this process. By targeting myofibroblasts (specialized contractile fibroblasts) that are key for the development of IPF with drugs conjugated with fibroblast activation protein (FAP), this technology helps minimize the production of extracellular matrix in the lungs and provides a new treatment option for patients diagnosed with IPF.

The present disclosure relates to methods of treating a patient with a cancer by administering to the patient a composition comprising CAR T cells wherein the CAR T cells comprise a CAR and the CAR comprises an E2 anti-fluorescein antibody fragment, and administering to the patient a small molecule linked to a targeting moiety by a linker. The disclosure also relates to compositions for use in such methods.

The present disclosure generally relates to methods of treating hemolytic diseases such as sickle cell anemia using kinase inhibitors, for example, compounds that inhibit the spleen tyrosine kinase (SYK). In some embodiments a method is presented to prevent thrombosis in blood vessels of patients with a hemolytic disease comprising the step of: administrating to a patient having at least one hemolytic disease a therapeutically effective amount of at least one SYK inhibitor. In some embodiments the method includes repeatedly administering the SYK kinase inhibitor to a patient.

BackgroundSmall animal models remain invaluable for the preclinical study of emerging molecular imaging agents. However, the data obtained in this setting are generated in genetically homogenous animals that do not mimic human pathophysiology. The purpose of this study was to prospectively validate precision-cut lung slices (PCLSs) obtained from patients with lung cancer as a translational tool for the development of targeted fluorophores.MethodsThe lung tissue was gently inflated with 2% Low-Melt Agarose (Fisher, 16520050) to avoid lung damage and minimize inflation pressure. The slices were then loaded into specialized cylindrical cartridges and inserted into a compressotome, and slices 150 to 350 μm thick were cut. Samples were incubated with fluorophore conjugates for ex vivo validation and immunohistochemical staining for receptor expression.ResultsA total of 184 unique 3-dimensional …

BACKGROUNDIntraoperative molecular imaging(IMI) utilizes a cancer-targeted fluorescent agent injected into patients to localize tumor nodules. Pafolacianine is a folate receptor(FR)-targeted near-infrared fluorescent probe. Almost 10% of patients have false negative fluorescence intraoperatively. We hypothesized that the tumor histology explains why a lung cancer may not fluoresce.METHODSAdenocarcinoma(AC) (A549, LKR) and squamous cell carcinoma(SCC) (H127, H1264) cell lines were stained with pafolacianine. NIR fluorescent microscopy was used to quantify mean fluorescence intensity. Tissue microarrays slides of patients with AC and SCC were evaluated by immunohistochemistry for FR alpha(FRα) and beta(FRβ) expression. Finally, we retrospectively analyzed IMI data from clinical trials of patients with AC and SCC receiving pafolacianine.RESULTSAC(intesnity 30.31) cell lines have a higher …

The disclosure relates to methods, compositions, and kits for treatment of parasite-mediated disease. In one embodiment, the disclosure relates to compounds, compositions, methods and kits for the treatment of malaria. In still another embodiment, the disclosure relates to a method for treating malaria comprising the use of a Syk kinase inhibitor.

Altered by defects in p53, epigenetic silencing, and genomic loss, the microRNA miR-34a represents one of the most clinically relevant tumor-suppressive microRNAs. Without question, a striking number of patients with cancer would benefit from miR-34a replacement, if poor miR-34a stability, non-specific delivery, and delivery-associated toxicity could be overcome. Here, we highlight a fully modified version of miR-34a (FM-miR-34a) that overcomes these hurdles when conjugated to a synthetically simplistic ligand. FM-miR-34a is orders of magnitude more stable than a partially modified version, without compromising its activity, leading to stronger repression of a greater number of miR-34a targets. FM-miR-34a potently inhibited proliferation and invasion, and induced sustained downregulation of endogenous target genes for >120 h following in vivo delivery. In vivo targeting was achieved through conjugating FM …

A compound of the formula (II); a pharmaceutical composition comprising same; and methods for treating a fibrotic disease in a subject.

In vivo imaging using shortwave infrared (SWIR) light (1,000–2,000 nm) benefits from deeper penetration and higher resolution compared with using visible and near-infrared wavelengths. However, the development of biocompatible SWIR contrast agents remains challenging. Despite recent advancements, small-molecule SWIR fluorophores are often hindered by their significant hydrophobicity. We report a platform for generating a panel of soluble and functional dyes for SWIR imaging by late-stage functionalization of a versatile fluorophore intermediate, affording water-soluble dyes with bright SWIR fluorescence in serum. Specifically, a tetra-sulfonate derivative enables clear video-rate imaging of vasculature with only 0.05 nmol dye, and a tetra-ammonium dye shows strong cellular retention for tracking of tumor growth. Additionally, incorporation of phosphonate functionality enables imaging of bones in awake …

Purpose: Joint damage caused by trauma can proceed to post-traumatic osteoarthritis (PTOA) due to the limited tissue repair capability. Currently, there is no cure available to prevent progressive joint degeneration, therefore there is an immediate need to develop treatments to prevent PTOA. Systemic pharmacological treatments can cause serious side effects and there is a need in the field to develop joint local therapies. Synovial macrophages play a role in OA onset, progression, and pain. In previous studies, we monitored macrophage infiltration in two different PTOA mouse models, anterior cruciate ligament transection (ACLT) and the destabilization of the medial meniscus (DMM), using a macrophage targeted nanoemulsion that contains a near-infrared fluorescent marker (provided by Dr. Janjic’s group). We established that macrophage infiltration increases in injured knees and is maintained during PTOA …

The goal of this research is to develop a method to label, detect, and count circulating tumor cells directly in vivo with an injectable folate-receptor targeted fluorescent molecular probe and near infrared (NIR) light.

Introduction Non-invasive imaging techniques such as positron emission tomography (PET) are extremely important for cancer detection and characterization especially for difficult to biopsy or extremely delicate organs such as the brain. The folate analogue 1,4,7-triazacylononane-1,4,7-triacetic acid-conjugated folate radiolabeled with aluminum fluoride-18 ([18F]FOL) has been previously shown to accumulate preferentially in tumor cells with an overexpression of folate receptors (FRs) and here was investigated for its ability to detect orthotopic gliomas in a rat model. In addition, we studied the expression of FRs in human glioblastoma samples to investigate if an analogous relationship may exist. Methods Nine BDIX rats were injected with BT4C rat glioma cells into the right hemisphere of the brain. Animals were imaged with gadolinium-enhanced magnetic resonance imaging at on days prior to PET/computed tomography (CT) imaging. Animals were divided into two groups, and were PET/CT imaged with either [18F]FOL or 2-deoxy-2-18F-fluoro-D-glucose ([18F]FDG) on 19 and 32-days post glioma grafting. Two subjects were also PET/CT imaged with [18F]FOL on day 16. Biodistribution was studied and brains were cryosectioned for autoradiography, immunofluorescence, and histological studies. Patient-derived paraffin-embedded glioblastomas were sectioned and stained with similar methods. Results PET imaging showed an increase of [18F]FOL tumor-to-brain uptake ratio (TBR) over the study duration from day 16/19 (3.3 ± 0.9) increasing to 5.7 ± 1.0 by day 32. [18F]FDG PET-imaged rats had a consistent TBR of 1.6 ± 0.1 throughout the …

Background Nearly 10% of world’s population is affected by influenza virus infections and more than 26 million Americans were infected during the 2022-2023 flu season, with 290,000-640,000 patients requiring hospitalization. While flu vaccines are widely available, they were only 36% effective during the 2021-2022 season. Standard-of-care (SOC) drugs such as Tamiflu and Xofluza are only effective when taken in the early stages of infection requiring novel flu drugs for treating later stages of infection. Schematics of the mechanism of action of EV21 Methods Herein we report a targeted therapy (EV21) with a dual mechanism of action that elicits a host immune response to infections. Because neuraminidase is expressed on both the viral envelope and infected host cell surface, we repurposed the neuraminidase inhibitor zanamivir for use as the targeting ligand. We …

The targeted delivery of growth factors, vasoactive peptides and other representative anabolic peptide drugs from different signaling cascades to bone fracture for accelerated healing is disclosed herein.

T-cell protein tyrosine phosphatase (TC-PTP), encoded by PTPN2, has emerged as a promising target for cancer immunotherapy. TC-PTP deletion in B16 melanoma cells promotes tumor cell antigen presentation, while loss of TC-PTP in T-cells enhances T-cell receptor (TCR) signaling and stimulates cell proliferation and activation. Therefore, there is keen interest in developing TC-PTP inhibitors as novel immunotherapeutic agents. Through rational design and systematic screening, we discovered the first highly potent and selective TC-PTP PROTAC degrader, TP1L, which induces degradation of TC-PTP in multiple cell lines with low nanomolar DC50s and >110-fold selectivity over the closely related PTP1B. TP1L elevates the phosphorylation level of TC-PTP substrates including pSTAT1 and pJAK1, while pJAK2, the substrate of PTP1B, is unaffected by the TC-PTP degrader. TP1L also intensifies interferon …

Myeloid derived suppressor cells (MDSCs) and tumor associated macrophages (TAMs) are two of the more immunosuppressive cell types in a tumor microenvironment (TME). Although TAMs and MDSCs can be converted from tumor-supporting to tumor-suppressing phenotypes by treatment with proinflammatory immune stimulants (e.g. TLR7 agonists) in vitro, such strategies have proven to be too toxic in vivo because of their systemic activation of immune cells. Therefore, for such immune stimulants to be used to reprogram the immune system in solid tumors, they must be targeted specifically to the immune cells in the TME. One strategy for targeting TAMs/MDSCs is to exploit a receptor that is expressed solely on these cell types. Because folate receptor beta (FRβ) is expressed exclusively on myeloid cells and since FRβ acquires its ability to bind folic acid (FA) only in inflamed tissues, we have previously …

BackgroundTargeted real-time imaging during robot-assisted radical prostatectomy provides information on the localisation and extent of prostate cancer. We assessed the safety and feasibility of the prostate-specific membrane antigen (PSMA)-targeted fluorescent tracer OTL78 in patients with prostate cancer.MethodsIn this single-arm, phase 2a, feasibility trial with an adaptive design was carried out in The Netherlands Cancer Institute, Netherlands. Male patients aged 18 years or older, with PSMA PET-avid prostate cancer with an International Society of Urological Pathology (ISUP) grade group of 2 or more, who were scheduled to undergo robot-assisted radical prostatectomy with or without extended pelvic lymph node dissection were eligible. All patients had a robot-assisted radical prostatectomy using OTL78. Based on timing and dose, patients received a single intravenous infusion of OTL78 (0·06 mg/kg 1–2 …

Described herein are prostate specific membrane antigen (PSMA) binding conjugates that are useful for delivering therapeutic, diagnostic and imaging agents. Also described herein are pharmaceutical composition containing them and methods of using the conjugates and compositions. Also described are processes for manufacture of the conjugates and the compositions containing them.

Adoptive chimeric antigen receptor (CAR)-engineered natural killer (NK) cells have shown promise in treating various cancers. However, limited immunological memory and access to sufficient numbers of allogenic donor cells have hindered their broader preclinical and clinical applications. Here, we first assess eight different CAR constructs that use an anti-PD-L1 nanobody and/or universal anti-fluorescein (FITC) single-chain variable fragment (scFv) to enhance antigen-specific proliferation and anti-tumor cytotoxicity of NK-92 cells against heterogenous solid tumors. We next genetically engineer human pluripotent stem cells (hPSCs) with optimized CARs and differentiate them into functional dual CAR-NK cells. The tumor microenvironment responsive anti-PD-L1 CAR effectively promoted hPSC-NK cell proliferation and cytotoxicity through antigen-dependent activation of phosphorylated STAT3 (pSTAT3) and …

The present disclosure relates to methods of treating a patient with a cancer by administering to the patient a composition comprising CAR T cells and a small molecule linked to a targeting moiety by a linker. The disclosure also relates to compositions for use in such methods.

Aspects of the present disclosure generally relate to compounds for targeting and healing bone fractures. Some of these compounds include a negatively charged oligopeptide comprising an acidic oligopeptide, a linker, which may be hydrolyzable or may be a substrate for the protease cathepsin K, and at least one molecule that promotes bone healing. In some compounds the molecule that promotes bone healing is an anabolic compound that inhibits GSK3β, in some compounds the molecule that promotes the healing of bone fracture is a pro-inflammatory agent such as PGE1. Other embodiments include methods of treating a bone fracture comprising administering a therapeutic amount of any one of the compounds disclosed herein.

Fibroblast activation protein (FAP) has received increasing attention as an oncologic target because of its prominent expression in solid tumors but virtual absence from healthy tissues. Most radioligand therapies (RLTs) targeting FAP, however, suffer from inadequate tumor retention or clearance from healthy tissues. Herein we report a FAP-targeted RLT comprising an FAP6 ligand conjugated to DOTA and an albumin binder (4-p-iodophenylbutyric acid, or IP) for enhanced pharmacokinetics. We evaluated the performance of the resulting FAP6-IP-DOTA conjugate in 4 tumor models, 3 of which express FAP only on cancer-associated fibroblasts, that is, analogously to human tumors. Methods Single-cell RNA-sequencing data were analyzed from 34 human breast, ovarian, colorectal, and lung cancers to quantify FAP-overexpressing cells. FAP6-DOTA conjugates were synthesized with or without an albumin binder …

Folate receptor delta (FRδ) has been used as a biomarker for regulatory T cells (Tregs), because its expression is limited to Tregs and ovum. Although FRδ is unable to bind folate, we have used molecular docking software to identify a folate congener that binds FRδ with high affinity and have exploited this FRδ-specific ligand to target attached drugs (imaging agents, immune activators, and immune suppressors) specifically to Tregs in murine tumor xenografts. Analysis of treated tumors demonstrates that targeting of a Toll-like receptor 7 agonist inhibits Treg expression of FOXP3, PD-1, CTLA4, and HELIOS, resulting in 40-80% reduction in tumor growth and repolarization of other tumor-infiltrating immune cells to more inflammatory phenotypes. Targeting of the immunosuppressive drug dexamethasone, in contrast, promotes enhanced tumor growth and shifts the tumor-infiltrating immune cells to more anti-inflammatory phenotypes. Since Tregs comprise <1% of cells in the tumor masses examined, and since the targeted drugs are not internalized by cancer cells, these data demonstrate that Tregs exert a disproportionately large effect on tumor growth. Because the targeted drug did not bind to Tregs or other immune cells in healthy tissues, the data demonstrate that the immunosuppressive properties of Tregs in tumors can be manipulated without causing systemic toxicities associated with global reprogramming of the immune system.

Compounds, including Toll-like receptor (TLR) agonists, eg, TLR7 and TLR7/8 agonists and their folic acid or pteroyl amino acid conjugates, and use thereof to treat a cancer or a fibrotic disease or disorder; and methods of making conjugates comprising targeting ligands of folic acid receptor and TLR7 and TLR7/8 agonists.

A conjugate comprising FL-L-IA, wherein FL is a radical of a small molecule ligand that specifically binds with fibroblast activation protein (FAP), L is a linker, which binds an FL to IA, and IA is a radical of a magnetic resonance imaging (MRI) agent, or a pharmaceutically acceptable salt thereof; a composition comprising same; and a method of using the conjugate or composition to image cells, a tissue, or an organ that express (es) FAP with magnetic resonance imaging.

CAR T cell therapies have demonstrated considerable potency in treating hematologic cancers, but only limited efficacy in eliminating solid tumors. One reason for this discrepancy may derive from the immunologic and physical barriers created by infiltration of cancer associated fibroblasts (CAFs) into solid tumors. These CAFs, which can comprise 15-85% of the stromal cells in a tumor mass, are correlated with poor patient survival and can reduce CAR T cell efficacies by secreting immunosuppressive cytokines, stimulating tumor cell proliferation, and depositing fibrotic barriers to CAR T cell penetration. Here, we are pursuing a method to suppress or eliminate CAF activities in the tumor microenvironment (TME). A potential CAR T cell target that is uniquely expressed on CAF surfaces is fibroblast activation protein (FAP). In this study, we have employed a novel, highly specific FAP targeting small molecule to direct …

Accurate delineation of gross tumor volumes remains a barrier to radiotherapy dose escalation and boost dosing in the treatment of solid tumors, such as prostate cancer. Magnetic resonance imaging (MRI) of tumor targets has the power to enable focal dose boosting, particularly when combined with technological advances such as MRI‐linear accelerator. Fibroblast activation protein (FAP) is overexpressed in stromal components of >90% of epithelial carcinomas. Herein, the authors compare targeted MRI of prostate specific membrane antigen (PSMA) with FAP in the delineation of orthotopic prostate tumors. Control, FAP, and PSMA‐targeting iron oxide nanoparticles were prepared with modification of a lymphotropic MRI agent (FerroTrace, Ferronova). Mice with orthotopic LNCaP tumors underwent MRI 24 h after intravenous injection of nanoparticles. FAP and PSMA nanoparticles produced contrast …

A compound of the formula (I): G 1-LG 2, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof, wherein G 1 is a folate radical, an antifolate radical, or a folate analog radical; L is a linker; and G 2 is a radical of a steroid; compositions comprising such compounds; and the use of such compounds and compositions to treat, for example, inflammation associated with a disease or disorder.

Myocardial infarction (MI) is the leading cause of death worldwide. However, current therapies are unable to restore the function of the injured myocardium. Advanced approaches, such as stimulation of cardiomyocyte (CM) proliferation are promising, but suffer from poor pharmacokinetics and possible systemic adverse effects. Nanomedicines can be a solution to the above‐mentioned drawbacks. However, targeting the cardiac tissue still represents a challenge. Herein, a MI‐selective precision nanosystem is developed, that relies on the heart targeting properties of atrial natriuretic peptide (ANP) and lin‐TT1 peptide‐mediated hitchhiking on M2‐like macrophages. The system based on pH‐responsive putrescine‐modified acetalated dextran (Putre‐AcDEX) nanoparticles, shows biocompatibility with cultured cardiac cells, and ANP receptor‐dependent interaction with CMs. Moreover, treatment with nanoparticles …

2023-06-13 Assigned to PURDUE RESEARCH FOUNDATION reassignment PURDUE RESEARCH FOUNDATION ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: LOW, PHILIP STEWART, LOW, Stewart A., NIELSON, JEFFREY JAY HOWARD

Recent success in the use of chimeric antigen receptor (CAR) T cell therapies to treat hematologic cancers has encouraged the exploration of CAR T cell technologies to treat solid tumors. For establishment of a productive immunologic synapse between the CAR T cell and cancer cell, multiple CARs on the T cell surface must engage multiple tumor antigens on the cancer cell surface. Whereas these binding interactions are mediated in classical CAR T cells by direct recognition of the tumor antigen by the scFv on the CAR, in the case of universal CAR T cells this recognition must be created by a bispecific adapter that can bridge between the tumor antigen and the CAR. While a diversity of bispecific adapters have been explored in the literature, the bispecific adapter used here is comprised of a low molecular weight ligand that binds avidly to the desired tumor antigen, the yellow dye fluorescein (FL) that binds …

ObjectiveFolate receptor beta (FR-β) has been used as a clinical marker and target in multiple inflammatory diseases, including osteoarthritis (OA) and rheumatoid arthritis (RA). However, the conditions under which FR-β+ macrophages arise remain unclear and could be affected by corticosteroids. Therefore, we studied FR-β expression in vitro in macrophage subtypes and determined their response to triamcinolone acetonide (TA), a clinically often-used corticosteroid.DesignHuman monocyte–derived macrophages were differentiated to the known M0, M1, or M2 macrophage phenotypes. The phenotype and FR-β expression and plasticity of the macrophage subtypes were determined using flow cytometry, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA).ResultsFR-β expression was low in granulocyte-macrophage colony-stimulating factor …

0* c1ccc2c (c1) C (C)(C) C (/C= C/C1= C (Oc3ccc (C (C) C) cc3)/C (= C/C= C3/N (CCC [N+](C)(C) C) c4ccc (*) cc4C3 (C) C) CCC1)=[N+] 2CCC [N+](C)(C) C.* c1ccc2c (c1) C (C)(C) C (/C= C/C1= C (c3ccc (C (C) C) cc3)/C (= C/C= C3/N (CCCCS (= O)(= O) O) c4ccc (*) cc4C3 (C) C) CCC1)=[N+] 2CCCCC.* c1ccc2c (c1) C (C)(C) C (/C= C/C1= C (c3ccc (C (C) C) cc3)/C (= C/C= C3/N (CCC [N+](C)(C) C) c4ccc (*) cc4C3 (C) C) CCC1)=[N+] 2CCC [N+](C)(C) C Chemical compound* c1ccc2c (c1) C (C)(C) C (/C= C/C1= C (Oc3ccc (C (C) C) cc3)/C (= C/C= C3/N (CCC [N+](C)(C) C) c4ccc (*) cc4C3 (C) C) CCC1)=[N+] 2CCC [N+](C)(C) C.* c1ccc2c (c1) C (C)(C) C (/C= C/C1= C (c3ccc (C (C) C) cc3)/C (= C/C= C3/N (CCCCS (= O)(= O) O) c4ccc (*) cc4C3 (C) C) CCC1)=[N+] 2CCCCC.* c1ccc2c (c1) C (C)(C) C (/C= C/C1= C (c3ccc (C (C) C) cc3)/C (= C/C= C3/N (CCC [N+](C)(C) C) c4ccc (*) cc4C3 (C) C) CCC1)=[N+] 2CCC …

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MicroRNAs (miRNAs) are small noncoding RNAs that regulate a wide array of genes. Because miRNA downregulate translation proteins associated with cancer progression, miRNA-based therapeutics have great potential as anticancer drugs. Nonetheless, the therapeutic potential of miRNA replacement is limited due to lack of safe and efficient delivery vehicles, and inability to target miRNAs to the intended cells/tissues. Thus, identifying molecules that are able to deliver therapeutic miRNA is of critical need. This could be achieved by targeting receptors that are overexpressed on target tumor cells. For example, the folate receptor (FR) is overexpressed on various tumor types including breast, lung, ovarian and brain tumors. Our laboratory has previously developed a unique strategy that directly links a miRNA mimic to the high affinity FR ligand, folate (FolamiRs). In the present work, to further improve the potency …

Soft-tissue sarcomas (STS) are a group of lethal mesenchymal tumors that are a major cause of cancer related morbidity for those under the age of 20.PI3K inhibitors have been found to be effective against subset of STS but are associated with significant toxicities limiting their use.We have thought to test a selective fibroblast activation protein (FAP) targeted delivery of PI3K/AKT/mTOR inhibitor omipalisib to STS in pre-clinical setting as FAP is overwhelmingly expressed agressive soft tissue malignancies. Experimental Design: Patient derived fibrosarcomas (aSMA-,FAP+,pAKT+) were isolated by flow cytometry.FAP(-)HT1080 fibrosarcoma cell lines were used as a negative control whereas methylcoanthrene induced mouse fibrosarcoma transfected with human FAP cDNA were used as positive control.Cell expression of FAP,AKT,mTOR was compared using SDS Page,IHC,and flow cytometry.FAP ligand coupled …

Background: The scientific rationale for mitapivat (AG-348) as an anti-sickling therapy is that it enhances red cell pyruvate kinase (PKR) activity and glycolytic flux, leading to a reduction of intracellular 2, 3-diphosphoglycerate (2, 3-DPG) and a concomitant increase in adenosine triphosphate (ATP). Reducing 2, 3-DPG destabilizes hemoglobin S (HbS) fibers, while increasing ATP improves red blood cell (RBC) integrity and cellular hydration. Mitapivat increased ATP and decreased 2, 3-DPG levels in a dose-dependent manner in this Phase 1 study (Xu et al, 2022). The Hb increase was sustained after stopping mitapivat until end of study (EOS) at 4 weeks, suggesting an improvement in RBC survival. However, pathways modulated by mitapivat that may contribute to sickle RBC survival remain largely unknown. Mitapivat has been shown to improve oxidative stress by increasing GSH/GSSG ratio (Matte et al 2021). In …

Related US Application Data (62) Division of application No. 16/894,287, filed on Jun. 5, 2020, now Pat. No. 11,185,505, which is a division of application No. 13/700,358, filed on Nov. 27, 2012, now abandoned, filed as application No. PCT/US2011/038437 on May 27, 2011.

Although chimeric antigen receptor (CAR) T cells have demonstrated significant promise in suppressing hematopoietic cancers, their applications in treating solid tumors have been limited by onset of CAR T cell exhaustion that accompanies continuous CAR T cell exposure to tumor antigen. To address this limitation, we have exploited the abilities of recently designed universal CARs to bind fluorescein and internalize a fluorescein‐TLR7 agonist conjugate by CAR‐mediated endocytosis. We demonstrate here that anti‐fluorescein CAR‐mediated uptake of a fluorescein‐TLR7‐3 conjugate can reactivate exhausted CAR T cells, leading to dramatic reduction in T cell exhaustion markers (PD‐1+Tim‐3+) and shrinkage of otherwise resistant tumors without inducing systemic activation of the immune system. We conclude that CAR T cell exhaustion can be reversed by administration of a CAR‐targeted TLR7 agonist …

Chimeric antigen receptor (CAR) T-cell therapies have proven to be effective in treating hematologic malignancies but demonstrate only marginal efficacy in eradicating solid tumors. Although several mechanisms can account for these differences, a major cause is thought to derive from CAR T-cell exhaustion, where chronic exposure to tumor antigen can activate feedback pathways that suppress CAR T-cell cytotoxicity. We describe here a strategy to reverse this CAR T-cell exhaustion using a universal anti-fluorescein CAR that concurrently serves as (i) a cancer recognition receptor that enables engagement of multiple cancer cell clones upon addition of a cocktail of bispecific fluorescein-linked tumor-targeting ligands, and (ii) a drug-internalizing receptor that mediates uptake of a CAR T-cell activator comprised of fluorescein linked to an immune stimulant. By attaching a Toll-like receptor 7 …

Methods for treating cancers using one or more compounds comprising a folate receptor binding ligand attached to a drug via a linker are described. Methods for treating cancers using one or more compounds comprising a folate receptor binding ligand attached to a drug via a linker to target tumor associated macrophages are described. Specification includes a Sequence Listing.

The disclosure relates to lipid-fluorescein conjugates, compositions comprising same, and methods of synthesis and use, such as in the treatment of cancer.

A microfluidic detection system for micrometer-sized entities, such as biological cells, includes a detector component incorporating a plate with a plurality of opening, the plate separating two chambers, one in communication with a fluid source containing target cells bound to magnetic beads. The openings are sized to always permit passage of the magnetic beads therethrough into a lower one of the chambers and are further sized to always prevent passage of the target cells from the upper one of the chambers. The detector component further includes a magnet positioned to pull unbound magnetic beads through the openings and to capture target cells bound to magnetic beads on the surface of the plate. The microfluidic detection system includes a pump flowing the fluid through the detector component at high flow rates of milliliters per minute for high throughput detection of target cells.

Although CAR T cell therapies have proven to be effective in treating hematopoietic cancers, their abilities to regress solid tumors have been less encouraging. Mechanisms to explain these disparities have focused primarily on differences in cancer cell heterogeneity, barriers to CAR T cell penetration of solid tumors, and immunosuppressive microenvironments. To evaluate the contributions of immunosuppressive tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) on CAR T cell efficacies, we have exploited the ability of a folate-targeted Toll-like receptor 7 agonist (FA-TLR7-1A) to specifically reactivate TAMs and MDSCs from an immunosuppressive to pro-inflammatory phenotype without altering the properties of other immune cells. We report here that FA-TLR7-1A significantly augments standard CAR T cell therapies of 4T1 solid tumors in immune competent mice. We further show that co-administration of the FA-TLR7-1A with the CAR T cell therapy not only repolarizes TAMs and MDSCs from an M2-like anti-inflammatory to M1-like pro-inflammatory phenotype, but also enhances both CAR T cell and endogenous T cell accumulation in solid tumors while concurrently increasing their states of activation. Because analogous myeloid cells in healthy tissues ar not altered by administration of FA-TLR7-1A, no systemic activation of the immune system nor accompanying weight loss is observed. These data argue that immunosuppressive myeloid cells contribute prominently to the failure of CAR T cells to eradicate solid tumors and suggest that methods to reprogram tumor associated myeloid cells to a more …

Pulmonary squamous cell carcinoma is the second most common lung cancer subtype and has a low 5-year survival rate at 17.6%. Complete resection with negative margins can be curative, but a high number of patients suffer early postoperative recurrence due to inadequate disease clearance at the index operation. Intraoperative molecular imaging (IMI) with tumor-targeted optical contrast agents is effective in improving resection completeness for other tumor types, but there are no IMI tracers targeted to pulmonary squamous cell carcinoma. In this report, we describe the use of a novel prostate-specific membrane antigen (PSMA)-targeted near-infrared conjugate (OTL78) to identify pulmonary squamous cell carcinoma. We identified PSMA as a viable target by examining its expression in human lung tumor specimens from a surgical cohort. Ninety-four percent of tumors expressed PSMA in either the …

Although both protein tyrosine phosphatases and kinases are constitutively active in healthy human red blood cells (RBCs), the preponderance of phosphatase activities maintains the membrane proteins in a predominantly unphosphorylated state. We report here that unlike healthy RBCs, proteins in sickle cells are heavily tyrosine phosphorylated, raising the question regarding the mechanism underpinning this tyrosine phosphorylation. Upon investigating possible causes, we observe that protein tyrosine phosphatase 1B (PTP1B), the major erythrocyte tyrosine phosphatase, is largely digested to a lower molecular weight fragment in sickle cells. We further find that the resulting truncated form of PTP1B is significantly less active than its intact counterpart, probably accounting for the intense tyrosine phosphorylation of Band 3 in sickle erythrocytes. Because this tyrosine phosphorylation of Band 3 promotes …

Oxidative stress has been shown to play a critical pathogenic role in functional loss after spinal cord injury (SCI). As a direct result of oxidative stress, lipid peroxidation-derived aldehydes have emerged as key culprits that sustain secondary injury and contribute significantly to pathological outcomes. Acrolein, a neurotoxin, has been shown to be elevated in SCI and can result in post-SCI neurological deficits. Reducing acrolein has therefore emerged as a novel and effective therapeutic strategy in SCI. Previous studies have revealed that hydralazine, an FDA approved blood pressure lowering medication, when administered after SCI shows strong acrolein scavenging capabilities and significantly improves cellular and behavioral outcomes. However, while effective at scavenging acrolein, hydralazine's blood pressure lowering activity can have a detrimental impact on neurotrauma patients. Here, our goal was to …

Conjugates are described herein where CCK2R targeting ligands are attached to an active moiety, such as therapeutic agent or an imaging agent, through a linker. The conjugates can be used in the detection, diagnosis, imaging and treat ment of cancer.Related US Application Data (63) Continuation of application No. 16/942,537, filed on Jul. 29, 2020, now Pat. No. 11,344,623, which is a continuation of application No. 16/105,734, filed on Aug. 20, 2018, now Pat. No. 10,765,756, which is a continuation of application No. 14/380,273, filed on Aug. 21, 2014, now Pat. No. 10,080,805, filed as application No. PCT/US2013/027463 on Feb. 22,

CIHQJFPNVHQIDJ-YYMSGCCCSA-N Cc1cc2c ([nH] c (= O) n2Cc2ccc (NCCN3CCCC (COCCOCCNC (= O) CC [C@ H](NC (= O) c4ccc (NCc5cnc6nc (N)[nH] c (= O) c6n5) cc4) C (= O) O) C3) nc2) c (N) n1 Chemical compound Cc1cc2c ([nH] c (= O) n2Cc2ccc (NCCN3CCCC (COCCOCCNC (= O) CC [C@ H](NC (= O) c4ccc (NCc5cnc6nc (N)[nH] c (= O) c6n5) cc4) C (= O) O) C3) nc2) c (N) n1 CIHQJFPNVHQIDJ-YYMSGCCCSA-N 0.000 description 8PVGDMICIFYXCSC-UHFFFAOYSA-N CC (C) NC (CSSCCOC (= O) C (C) C) C (= O) O Chemical compound CC (C) NC (CSSCCOC (= O) C (C) C) C (= O) O PVGDMICIFYXCSC-UHFFFAOYSA-N 0.000 description 5

The present disclosure relates to compositions and methods of carbonic anhydrase IX inhibitors. The present disclosure also relates to targeting conjugates of carbonic anhydrase IX inhibitors. The present disclosure also relates to the use of targeting conjugates of carbonic anhydrase IX inhibitors in methods of treating disease and for imaging of disease.

Background:In recent decades, the use of near-infrared light and fluorescence-guidance during open and laparoscopic surgery has exponentially expanded across various clinical settings. However, tremendous variability exists in how it is performed.Objective:In this first published survey of international experts on fluorescence-guided surgery, we sought to identify areas of consensus and nonconsensus across 4 areas of practice: fundamentals; patient selection/preparation; technical aspects; and effectiveness and safety.Methods:A Delphi survey was conducted among 19 international experts in fluorescence-guided surgery attending a 1-day consensus meeting in Frankfurt, Germany on September 8 th, 2019. Using mobile phones, experts were asked to anonymously vote over 2 rounds of voting, with 70% and 80% set as a priori thresholds for consensus and vote robustness, respectively.Results:Experts from 5 …

Non-invasive imaging modalities constitute an increasingly important tool in diagnostic and therapy response monitoring of patients with autoimmune diseases, including rheumatoid arthritis (RA). In particular, macrophage imaging with positron emission tomography (PET) using novel radiotracers based on differential expression of plasma membrane proteins and functioning of cellular processes may be suited for this. Over the past decade, selective expression of folate receptor β (FRβ), a glycosylphosphatidylinositol-anchored plasma membrane protein, on myeloid cells has emerged as an attractive target for macrophage imaging by exploiting the high binding affinity of folate-based PET tracers. This work discusses molecular, biochemical and functional properties of FRβ, describes the preclinical development of a folate-PET tracer and the evaluation of this tracer in a translational model of arthritis for diagnostics and therapy-response monitoring, and finally the first clinical application of the folate-PET tracer in RA patients with active disease. Consequently, folate-based PET tracers hold great promise for macrophage imaging in a variety of (chronic) inflammatory (autoimmune) diseases beyond RA.

Retrieval of circulating tumor cells (CTC) has proven valuable for assessing a patient's cancer burden, evaluating response to therapy, and analyzing which drug might treat a cancer best. Although most isolation methods retrieve CTCs based on size, shape, or capture by tumor-specific antibodies, we explore here the use of small molecule tumor-specific ligands linked to magnetic beads for CTC capture. We have designed folic acid-biotin conjugates with different linkers for the capture of folate receptor (FR) + tumor cells spiked into whole blood, and application of the same technology to isolate FR + CTCs from the peripheral blood of both tumor-bearing mice and non-small cell lung patients. We demonstrate that folic acid linked via a rigid linker to a flexible PEG spacer that is in turn tethered to a magnetic bead enables optimal CTC retrieval, reaching nearly 100% capture when 100 cancer cells are spiked into …

Related US Application Data (63) Continuation of application No. 16/157,024, filed on Oct. 10, 2018, now abandoned, which is a continu ation of application No. 15/035,936, filed on May 11, 2016, now abandoned, filed as application No. PCT/US2014/065467 on Nov. 13, 2014.Described herein are compounds, compositions, and meth ods for diagnosing and/or monitoring pathogenic disease using positron emission tomography. Also described are conjugates of the formula B-L-P, wherein B is a radical of a targeting agent selected from vitamin receptor binding ligands (such as folate), PSMA binding ligands, or PSMA inhibitors; L is a divalent linker comprising aspartic acid, lysine, or arginine, and P is a radical of an imaging agent or radiotherapy agent, such as a radionuclide or radionuclide containing group, or a radical of a compound capable of

PYWVYCXTNDRMGF-UHFFFAOYSA-N Rhodamine B Chemical class [Cl-]. C= 12C= CC (=[N+](CC) CC) C= C2OC2= CC (N (CC) CC)= CC= C2C= 1C1= CC= CC= C1C (O)= O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 24

Targeted drug delivery, often referred to as “smart” drug delivery, is a process whereby a therapeutic drug is delivered to specific parts of the body in a manner that increases its concentration at the desired sites relative to others. This approach is poised to revolutionize medicine as exemplified by the recent FDA approval of Cytalux (FDA approves pioneering drug for ovarian cancer surgery - Purdue University News) which is a folate-receptor targeted intraoperative near infrared (NIR) imaging agent that was developed in our laboratories. Fracture-associated morbidities and mortality affect a significant portion of world population. United states, Canada and Europe alone spent $48 billion in treating osteoporosis related fractures although this number doesn't count the economic burden due to loss in productivity. It is estimated that by 2050 ca 21 million hip fractures would occur globally which will be leading cause of …

Inhibitors of erythrocyte band 3 tyrosine phosphorylation, compositions comprising same, and methods of using the inhibitors and compositions to treat sickle cell diseases.

(57) ABSTRACT Related US Application Data (63) Continuation of application No. 15/296,666, filed on Oct. 18, 2016, now abandoned, which is a continu ation of application No. 14/654,227, filed on Jun. 19, 2015, now abandoned, filed as application No. PCT/US2013/076986 on Dec. 20, 2013.(60) Provisional application No. 61/740,384, filed on Dec.

PYWVYCXTNDRMGF-UHFFFAOYSA-N Rhodamine B Chemical compound [Cl-]. C= 12C= CC (=[N+](CC) CC) C= C2OC2= CC (N (CC) CC)= CC= C2C= 1C1= CC= CC= C1C (O)= O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims description 20

We developed a near-infrared instrument for non-invasive and continuous enumeration of circulating tumor cells in mice with diffuse light. The instrument is designed for compatibility with clinical cancer-targeted fluorescence contrast agents.

Suspicious nodules detected by radiography are often investigated by biopsy, but the diagnostic yield of biopsies of small nodules is poor. Here we report a method—NIR-nCLE—to detect cancer at the cellular level in real-time during biopsy. This technology integrates a cancer-targeted near-infrared (NIR) tracer with a needle-based confocal laser endomicroscopy (nCLE) system modified to detect NIR signal. We develop and test NIR-nCLE in preclinical models of pulmonary nodule biopsy including human specimens. We find that the technology has the resolution to identify a single cancer cell among normal fibroblast cells when co-cultured at a ratio of 1:1000, and can detect cancer cells in human tumors less than 2 cm in diameter. The NIR-nCLE technology rapidly delivers images that permit accurate discrimination between tumor and normal tissue by non-experts. This proof-of-concept study analyzes …

MicroRNAs (miRNAs) inhibit the expression of genes through imperfect base pairing with target messenger RNAs (mRNAs), which allows a single miRNA the ability to regulate the expression of multiple genes, potentially acting as a multi-drug cocktail. In principle, tumor-suppressive miRNAs, such as miRNA-34a (miR-34a) are excellent anti-cancer agents; however, due to toxicity associated with currently used delivery vehicles and poor in vivo miRNA stability, the clinical application of miRNAs is thwarted. To overcome these challenges, we engineered two modifications to the miRNA. The first entails conjugating the miRNA to a ligand that promotes delivery specifically to cancer cells while the second involves using a fully chemically modified miRNA (FM-miR) to enhance miRNA stability. The ligand chosen for delivery, 2- [3-(1,3-dicarboxy propyl) ureido] pentanedioic acid (DUPA), is a high-affinity binding partner …

BackgroundThe diagnostic yield of biopsies of solitary pulmonary nodules (SPNs) is low, particularly in sub-solid lesions. We developed a method (NIR-nCLE) to achieve cellular level cancer detection during biopsy by integrating (i) near-infrared (NIR) imaging using a cancer-targeted tracer (pafolacianine), and (ii) a flexible NIR confocal laser endomicroscopy (CLE) system that can fit within a biopsy needle. Our goal was to assess the diagnostic accuracy of NIR-nCLE ex vivo in SPNs.MethodsTwenty patients with SPNs were preoperatively infused with pafolacianine. Following resection, specimens were inspected to identify the lesion of interest. NIR-nCLE imaging followed by tissue biopsy was performed within the lesion and in normal lung tissue. All imaging sequences (n = 115) were scored by 5 blinded raters on the presence of fluorescent cancer cells and compared to diagnoses by a thoracic pathologist …

BackgroundPulmonary ground glass opacities (GGOs) are early-stage adenocarcinoma spectrum lesions that are not easily palpable. Challenges in localizing GGOs during intraoperative pathology can lead to imprecise diagnoses and additional time under anesthesia. To improve localization of GGOs during frozen section diagnosis, we evaluated a novel technique, 3-dimensional near-infrared specimen mapping (3D-NSM).MethodsFifty-five patients with a cT1 GGO were enrolled and received a fluorescent tracer preoperatively. After resection, specimens were inspected to identify lesions. Palpable and nonpalpable nodules underwent 3D-NSM and the area of highest fluorescence was marked with a suture. Time for 3D-NSM, time for frozen section diagnosis, and number of tissue sections examined were recorded. To compare 3D-NSM with standard-of-care techniques, a control cohort of 20 subjects with …