Yuping Xu

First Experience of 18F-Alfatide in Lung Cancer Patients Using a New Lyophilized Kit for Rapid Radiofluorination

18F-FPPRGD2, which was approved for clinical study recently, has favorable properties for integrin targeting and showed potential for antiangiogenic therapy and early response monitoring. However, the time-consuming multiple-step synthesis may limit its widespread applications in the clinic. In this study, we developed a simple lyophilized kit for labeling PRGD2 peptide (18F-AlF-NOTA-PRGD2, denoted as 18F-alfatide) using a fluoride–aluminum complex that significantly simplified the labeling procedure. Methods: Nine patients with a primary diagnosis of lung cancer were examined by both static and dynamic PET imaging with 18F-alfatide, and 1 tuberculosis patient was investigated using both 18F-alfatide and 18F-FDG imaging. Standardized uptake values were measured in tumors and other main organs at 30 min and 1 h after injection. Kinetic parameters were calculated by Logan graphical analysis. Immunohistochemistry and staining intensity quantification were performed to confirm the expression of integrin αvβ3. Results: Under the optimal conditions, the whole radiosynthesis including purification was accomplished within 20 min with a decay-corrected yield of 42.1% ± 2.0% and radiochemical purity of more than 95%. 18F-alfatide PET imaging identified all tumors, with mean standardized uptake values of 2.90 ± 0.10. Tumor-to-muscle and tumor-to-blood ratios were 5.87 ± 2.02 and 2.71 ± 0.92, respectively. Conclusion: 18F-alfatide can be produced with excellent radiochemical yield and purity via a simple, 1-step, lyophilized kit. PET scanning with 18F-alfatide allows specific imaging of αvβ3 expression with good contrast in lung cancer patients. This technique might be used for the assessment of angiogenesis and for planning and response evaluation of cancer therapies that would affect angiogenesis status and integrin expression levels.

Unexpected fluorescence from polymers containing dithio/amino-succinimides

Fluorescent polymers without conventional fluorescent units are attracting extensive interest because of their excellent biocompatibility. Succinimides are generally known as common fluorescence quenchers of proteins due to some degree of electron transfer between the excited state and the quencher. In this work, we observed unexpected fluorescence from succinimide-containing polymers. We synthesized a series of succinimides and found that 2,3-dithio-succinimide and 2-amino-succinimide were fluorescent, presenting solvent-dependent emission. This study is the first to report that nonfluorescent succinimides become fluorescent only by thiol and amine group substitutions, without employing any fluorescent units. The unusual fluorescence of dithio/amino-succinimides resulted from the spatial separation of HOMO and LUMO based on density functional theory calculations. Compared with monomers, polymers containing 2-amino-succinimide exhibited substantial fluorescence enhancement (maximum of ∼200-fold) because of the isolation effect, in which nonradiative decay among succinimide fluorophores was significantly suppressed.

Bioevaluation of a novel [32P]-CP-PLLA microparticle for pancreatic cancer treatment

A novel [32P]‐CP‐PLLA ([32P]‐chromic phosphate‐poly‐L‐lactic acid) microparticle was designed and evaluated for the treatment of pancreatic carcinoma. The microparticle was prepared from [32P] chromic phosphate and poly‐L‐lactic acid. Bioelimination, biodistribution, SPECT image, and therapeutic effect were studied in mice bearing human pancreatic tumor xenografts. [32P]chromic phosphate ([32P]‐CP) colloid at the same radioactivity dose was compared as the control. High radioactivity (>95%ID) of [32P]‐CP‐PLLA was retained at the tumor, and almost no radioactivity excretion (<1%ID) was observed in urine and feces for 14 days, while radioactivity of [32P]‐CP colloid, was distributed to the liver, spleen, and lung (varied individual), and the excretion increased over 5%ID. Compared with controls, reduced tumor volumes were seen in the [32P]‐CP‐PLLA microparticle treatment group (P<0.01). Dose dependence was seen histologically. [32P]‐CP‐PLLA microparticle retained high activity and long‐term residence in tumor. With minimal distribution to normal organs [32P]‐CP‐PLLA microparticle is superior to [32P]‐CP colloid and is more suitable for brachytherapy in solid tumor. Drug Dev Res 71:364–370, 2010.

Thiolactone-maleimide: a functional monomer to synthesize fluorescent aliphatic poly(amide-imide) with excellent solubility via in situ PEGylation

Fluorescent poly(amide-imide)s are promising materials for use in photonics due to their controllable optical properties and excellent flexibility. Although unconventional fluorescent polymers have been flourishing recently, the fluorescence of aliphatic poly(amide-imide)s has never been realized until now. Their poor solubility and intolerance to reactive groups during synthesis prohibit access for the modification of poly(amide-imide)s. Here, we synthesize a functional thiolactone-maleimide monomer via copper(I)-catalyzed azide alkyne cycloaddition and employ a strategy combining aminolysis of thiolactones and amine-maleimide Michael addition for the synthesis of fluorescent aliphatic poly(amide-imide)s with 2-aminosuccinimide fluorophores. Moreover, in situ generated thiols enable the poly(amide-imide)s to undergo facile PEGylation via a thiol–methacrylate Michael addition reaction to accomplish excellent solubility in organic solvents and in water.

Prostate cancer imaging of FSHR antagonist modified with a hydrophilic linker

Follicle-stimulating hormone receptor (FSHR) is selectively expressed in endothelial cells of prostate cancer (PCa) and becomes a potential target for tumor diagnosis and therapy. (18)F-Al-NOTA-MAL-FSH1 is a promising PET imaging probe for targeting FSHR; however, the unfavorable abdominal backgrounds may hamper clinical translation. GGGRDN is a new hydrophilic linker, which can improve the imaging quality of radiolabeled peptides. In this study, GGGRDN-FSH1 (denoted as FSH2) was designed and conjugated with NOTA-MAL for (18)F-Al radiolabeling. NOTA-MAL-FSH2 was obtained with about 50% yield and labeled using (18)F-Al in a one-step method within 20 min with a yield of 41.46 ± 10.36% (non-decay-corrected). The radiochemical purity was more than 95% and the specific activity was more than 50 GBq/µmol. The in vitro stability studies were determined in PBS and human serum. (18)F-Al-NOTA-MAL-FSH2 remained stable in PBS and human serum. Balb/c nude mice bearing PC-3 human PCa were used for in vivo study. PC-3 tumors were clearly visualized with good contrast to background through microPET. ROI analysis showed the tumor uptake values were 2.68 ± 0.52 and 1.97 ± 0.61%ID/g at 30 and 60 min post injection (p.i.), respectively. Biodistribution studies showed that the accumulations of (18)F-Al-NOTA-MAL-FSH2 in liver and intestine were 0.47 ± 0.11 and 0.12 ± 0.03%ID/g respectively at 60 min p.i. FSHR-binding specificity was also demonstrated by reduced tumor uptake after coinjection of excessive unlabeled FSH2. In conclusion, (18)F-Al-NOTA-MAL-FSH2 was successfully prepared in a one-step method and showed better pharmacokinetics than (18) F-Al-NOTA-MAL-FSH1. Favorable preclinical study revealed that (18)F-Al-NOTA-MAL-FSH2 appears to be a promising candidate for FSHR-positive tumor imaging.

Preliminary evaluation of [18F]AlF-NOTA-MAL-Cys39-exendin-4 in insulinoma with PET.

Abstract Background: High expression of glucagon-like peptide-1 receptor (GLP-1R) in insulinoma supplies a potential drug target for tumor imaging. Exendin-4 can specifically bind to GLP-1R as an agonist and its analogs are extensively used in receptor imaging studies. Purpose: A new GLP-1R imaging agent, [18F]AlF-NOTA-MAL-Cys39-exendin-4, was designed and prepared for insulinoma imaging. Methods: Cys39-exendin-4 was conjugated with NOTA-MAL, then the compound was radiolabeled with [18F]AlF complex to obtained [18F]AlF-NOTA-MAL-Cys39-exendin-4. The tumor-targeting characters of the tracer were evaluated in INS-1 cells and BALB/c nude mice models. Results: [18F]AlF-NOTA-MAL-Cys39-exendin-4 can be efficiently produced with a yield of 17.5 ± 3.2% (non-decay corrected) and radiochemical purity of >95%. The IC50 value of displacement [18F]AlF-NOTA-MAL-Cys39-exendin-4 with Cys39-exendin-4 was 13.52 ± 1.36 nM. PET images showed excellent tumor visualization with high uptake (9.15 ± 1.6%ID/g at 30 min and 7.74 ± 0.87%ID/g at 60 min). The tumor to muscle, pancreas and liver ratios were 63.25, 3.85 and 7.29 at 60 min after injection. GLP-1R binding specificity was demonstrated by co-injection with an excess of unlabeled Cys39-exendin-4 and the tumor uptake was found to be reduced significantly. Conclusion: [18F]AlF-NOTA-MAL-Cys39-exendin-4 shows favorable characteristics for insulinoma imaging and may be translated to clinical studies.

Development of a Novel PET Tracer [18F]AlF-NOTA-C6 Targeting MMP2 for Tumor Imaging

Background and Objective The overexpression of gelatinases, that is, matrix metalloproteinase MMP2 and MMP9, has been associated with tumor progression, invasion, and metastasis. To image MMP2 in tumors, we developed a novel ligand termed [18F]AlF-NOTA-C6, with consideration that: c(KAHWGFTLD)NH2 (herein, C6) is a selective gelatinase inhibitor; Cy5.5-C6 has been visualized in many in vivo tumor models; positron emission tomography (PET) has a higher detection sensitivity and a wider field of view than optical imaging; fluorine-18 (18F) is the optimal PET radioisotope, and the creation of a [18F]AlF-peptide complex is a simple procedure. Methods C6 was conjugated to the bifunctional chelator NOTA (1, 4, 7-triazacyclononanetriacetic acid) for radiolabeling [18F]AlF conjugation. The MMP2-binding characteristics and tumor-targeting efficacy of [18F]AlF-NOTA-C6 were tested in vitro and in vivo. Results The non-decay corrected yield of [18F]AlF-NOTA-C6 was 46.2–64.2%, and the radiochemical purity exceeded 95%. [18F]AlF-NOTA-C6 was favorably retained in SKOV3 and PC3 cells, determined by cell uptake. Using NOTA-C6 as a competitive ligand, the uptake of [18F]AlF-NOTA-C6 in SKOV3 cells decreased in a dose-dependent manner. In biodistribution and PET imaging studies, higher radioactivity concentrations were observed in tumors. Pre-injection of C6 caused a marked reduction in tumor tissue uptake. Immunohistochemistry showed MMP2 in tumor tissues. Conclusions [18F]AlF-NOTA-C6 was easy to synthesize and has substantial potential as an imaging agent that targets MMP2 in tumors.

Targeting of MMP2 activity in malignant tumors with a 68Ga-labeled gelatinase inhibitor cyclic peptide

INTRODUCTION Elevated levels of gelatinases (matrix metalloproteinases 2/9, i.e., MMP2 and MMP9) are associated with tumor progression, invasion and metastasis, so these enzymes are potential targets for tumor imaging. The peptide c(KAHWGFTLD)NH2 (herein, C6) is a selective gelatinase inhibitor. Cy5.5-C6 has been visualized in many tumor models in vivo. However, the sensitivity and penetrance of optical imaging are poor. It is well known that positron emission tomography (PET) has a high detection sensitivity and Gallium-68 ((68)Ga) is an optimal PET radioisotope. Thus, in the present study, we developed a novel ligand, (68)Ga-NOTA-C6, to image MMP2 activity in tumors. METHODS C6 was conjugated with the bifunctional chelator NOTA (1,4,7-triazacyclononanetriacetic acid) and labeled with (68)Ga. In vitro uptake and binding analyses were performed by using SKOV3 cell lines, coincubating with or without the MMP inhibitor doxycycline. The biodistribution and PET imaging were conducted on SKOV3 ovarian tumor models. MMP2 expression in tumors was analyzed by immunohistochemistry (IHC). RESULTS The non-decay corrected yield of (68)Ga-NOTA-C6 was 61.8%-63.3%. (68)Ga-NOTA-C6 was stable in both physiological saline and human serum. The uptake of (68)Ga-NOTA-C6 in SKOV3 cells increased with time, and could be blocked by doxycycline in a dose dependent manner. The results of biodistribution and PET imaging showed that high radioactivity concentrations of (68)Ga-NOTA-C6 occurred in tumors. The ratios of tumor to blood, muscle and ovary and oviduct at 30, 60 and 120min p.i. were 2.78±0.54, 3.86±0.65, 0.48±0.14, and 1.73±0.36, 10.31±3.12, 1.22±0.10, and 2.50±0.78, 7.03±1.85, 0.97±0.25, respectively. The tracer was excreted mainly through the renal system, as evidenced by high levels of radioactivity in the kidneys. These data support the possibility of using (68)Ga-NOTA-C6 in PET to visualize tumors that overexpress MMP2. CONCLUSIONS (68)Ga-NOTA-C6 is a potential radiopharmaceutical for the imaging of in vivo MMP2 activity in tumors.

Polyphenol–Poloxamer Self‐Assembled Supramolecular Nanoparticles for Tumor NIRF/PET Imaging

Self-assembled supramolecular nanoparticles have remarkable benefits in bioimaging and drug delivery. Here it is first reported that polyphenol and poloxamer self-assemble supramolecular nanoparticles (PPNPs). PPNPs are fabricated by multivalent hydrogen bonding between tannic acid and Pluronic F-127 together with hydrophobic interactions of poly(propylene oxide) chains, to be applied in tumor near-infrared fluorescence (NIRF) imaging and positron emission tomography (PET) imaging. With near-infrared fluorescent dyes such as IR780 encapsulated via hydrophobic interactions, PPNPs are used in NIRF imaging. PPNPs with excess phenolic hydroxyl groups chelating positron emitting radionuclide 89 Zr function as a PET contrast agent. The in vivo results show surprisingly higher fluorescence intensity in tumors than in other tissues. In addition, PPNPs exhibit good biocompatibility in various cell lines and do not induce hemolysis in vitro. In this study, it is demonstrated that biodegradable and biocompatible PPNPs are an excellent bimodal contrast agent for in vivo tumor imaging.

PET of HER2 Expression with a Novel 18FAl Labeled Affibody

Background: Human epidermal growth factor receptor type 2 (HER2) is abundant in a wide variety of tumors and associated with the poor prognosis. Radiolabeled affibodies are potential candidates for detecting HER2-positive lesions. However, laborious multiple-step synthetic procedure and high abdomen background may hinder the widespread use. Herein, cysteinylated ZHER2:342 modified with a new hydrophilic linker (denoted as MZHER2:342) was designed and labeled using 18FAl-NOTA strategies. The biologic efficacy of the novel tracer and its feasibilities for in vivo monitoring HER2 levels were also investigated in xenograft models with different HER2 expressions. Method: MZHER2:342 was conjugated with MAL-NOTA under standard reaction conditions. The affibody molecule was then radiolabeled with 18FAl complex. The binding specificity of the tracer, 18FAl-NOTA-MAL-MZHER2:342, with HER2 was primarily characterized via in vitro studies. MicroPET imaging were performed in nude mice bearing tumors (SKOV-3, JIMT-1 and MCF-7) after injection. The HER2 levels of xenografts were determined using Western blotting analysis. Results: 18FAl-NOTA-MAL-MZHER2:342 can be efficiently produced within 30 min with a non-decaycorrected yield of about 10% and a radiochemical purity of more than 95%. In vitro experiments revealed that the modified affibody retained the specific affinity to HER2. PET imaging showed that SKOV-3 and JIMT-1 xenografts were clearly visualized with excellent contrast and low abdomen backgrounds. On the contrary, the signals of MCF-7 tumor were difficult to visualize. The ROI values ranged from16.54±2.69% ID/g for SKOV-3 to 8.42±1.20 %ID/g for JIMT-1 tumors at 1h postinjection respectively. Poor uptake was observed from MCF-7 tumors with 1.71±0.34% ID/g at the same time point. Besides, a significant linear correlation between % ID/g values and relative HER2 expression levels was also found. Conclusions: 18FAl-NOTA-MAL-MZHER2:342 is a promising tracer for in vivo detecting HER2 status with the advantages of facile synthesis and favorable pharmacokinetics. It may be useful in differential diagnosis, molecularly targeted therapy and prognosis of the cancers.