Formulation development and manufacturing of a gastrin/CCK-2 receptor targeting peptide as an intermediate drug product for a clinical imaging study

Sosabowski, Jane; Lee, Myrto; Dekker, Bronwen A; Simmons, Ben P.; Singh, Surjeet; Beresford, Hayden; Hagan, Susan A.; McKenzie, Andrew J.; Mather, Stephen J.; Watson, Susan A.

doi:10.1016/j.ejps.2007.02.007

Cited by 12 publications

(8 citation statements)

References 19 publications

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“…The conjugation of gastrin-10 to CPNPs via the PEGϪmaleimide coupling strategy permits targeting of the gastrin receptor as previously described by Sosabowski and colleagues. 35 A 9 mL aliquot of Citrate-CPNPs was chemically conjugated with maleimide polyethylene glycol amine (PEGϪmaleimide; JenKem Technology Inc.), through the ethyl-N-(3-dimethylaminopropyl)-N=-hydrochloride carbodiimide reaction (EDCI, Fluka BioChemika Ն99.0% (AT); Sigma-Aldrich Co.). The sample was first stirred at 550 rpm on a combined magnetic stir/hot plate set to 50 °C.…”

Section: Preparation Of Nanoparticlesmentioning

confidence: 99%

Bioconjugation of Calcium Phosphosilicate Composite Nanoparticles for Selective Targeting of Human Breast and Pancreatic Cancers In Vivo

et al. 2010

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The early diagnosis of cancer is the critical element in successful treatment and long term favorable patient prognoses. The high rate of mortality is mainly attributed to the tendency for late diagnoses as symptoms may not occur until the disease has metastasized, as well as the lack of effective systemic therapies. Late diagnosis is often associated with the lack of timely sensitive imaging modalities. The promise of nanotechnology is presently limited by the inability to simultaneously seek, treat and image cancerous lesions. This study describes the design and synthesis of fluorescent calcium phosphosilicate nanocomposite particles (CPNPs) that can be systemically targeted to breast and pancreatic cancer lesions. The CPNPs are a ~20nm diameter composite composed of an amorphous calcium phosphate matrix doped with silicate in which a near infra-red imaging agent indocyanine green (ICG) is embedded. In the present studies, we describe and validate CPNP bioconjugation of human holotransferrin, anti-CD71 antibody, and short gastrin peptides via an avidin-biotin-or a novel PEG-maleimide-coupling strategy. The conjugation of biotinylated human holotransferrin (diferric transferrin) and biotinylated anti-CD71 antibody (anti-transferrin receptor antibody) to avidin conjugated CPNPs (Avidin-CPNPs) permits targeting of transferrin receptors, which are highly expressed on breast cancer cells. Similarly, the conjugation of biotinylated pentagastrin to AvidinCPNPs and decagastrin (gastrin-10) to PEG-CPNPs via PEG-maleimide coupling permits targeting of gastrin receptors, which are over-expressed in pancreatic cancer lesions. These bioconjugated CPNPs have the potential to perform as a theranostic modality, simultaneously enhancing drug delivery, targeting and imaging of breast and pancreatic cancer tumors. Keywords bioconjugation; transferrin receptor; gastrin receptor; breast cancer; pancreatic cancer; calcium phosphate; whole animal imaging Calcium phosphate nanoparticles (CPNPs) have been engineered to be a non-toxic vehicle for the delivery of a diverse range of therapeutic and imaging agents in biological systems. [1][2][3][4] Previous studies have shown that encapsulation within CPNPs improved the lifetime and RESULTS AND DISCUSSION Physical Characterization of CPNPsCitrate functionalized CPNPs were utilized as a platform for functionalization, which allowed the characterization of bioconjugation via zeta potential analysis (Figure 1). Figure 1 shows the zeta potential distribution of Citrate-CPNPs prior to bioconjugation (blue line), and the zeta (violet). Prior to bioconjugation, the Citrate-CPNPs display a negative mean zeta potential value of −16 ± 1.3 mV, which is consistent with previous reports. 1 However, after bioconjugation, the Avidin-CPNPs displayed a relatively high positive mean zeta potential value of +29 ± 8.7 mV. The isoelectric point for avidin is pH 10. Thus, the shift from a negative zeta potential to a positive zeta potential distribution is strong evidence of avidin bioconjugation on...

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Section: Preparation Of Nanoparticlesmentioning

confidence: 99%

Bioconjugation of Calcium Phosphosilicate Composite Nanoparticles for Selective Targeting of Human Breast and Pancreatic Cancers In Vivo

et al. 2010

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“…Trityl deprotection ( Figure 1 B ) and subsequent conjugation of Ga-10 polymer were confirmed by 1 H-NMR, which showed complete removal of the trityl group and more than 70% conjugation ( Figure 1 C ), rendering the NPs target-specific to the CCK-B receptor. 27 , 31 The conjugation of Ga-10 to the PEG-PLL polymer backbone was confirmed further by gel permeation chromatography using Ga-10–associated tryptophan fluorescence (e x /e m , 280/350) from the Ga-PEG-PLL polymer ( Figure 1 D1 ). Untargeted polymer also was evaluated under identical conditions and no fluorescent peak was detected, indicating the absence of any peptide-associated fluorescent moiety in the polymer Figure 1 D2 ).…”

Section: Resultsmentioning

confidence: 83%

“… 25 Currently, radiopeptide imaging with 111 In-minigastrin labeling to detect CCK-B receptors is established and in clinical use for medullary cancer (another cancer that expresses CCK-B receptors). 26 By using a similar maleimide coupling technique to target the CCK-B receptor as previously described, 27 we developed a polyplex nanoparticle (NP) that selectively targets the CCK-B receptor and serves as a target-specific vehicle to deliver gene therapy to inhibit growth and metastasis of PDAC. Biodegradable nontoxic nanoparticles that serve as vehicles to carry siRNA without off-target toxicity, such as the polyplex NP described in this work, have the potential to revolutionize cancer therapeutics.…”

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confidence: 99%

Cholecystokinin Receptor-Targeted Polyplex Nanoparticle Inhibits Growth and Metastasis of Pancreatic Cancer

Burks

Nadella

Mahmud

et al. 2018

Cellular and Molecular Gastroenterology and Hepatology

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Background & AimsPancreatic ductal adenocarcinoma (PDAC) remains the most aggressive malignancy with the lowest 5-year survival rate of all cancers in part owing to the lack of tumor-specific therapy and the rapid metastatic nature of this cancer. The gastrointestinal peptide gastrin is a trophic peptide that stimulates growth of PDAC in an autocrine fashion by interaction with the cholecystokinin receptor that is overexpressed in this malignancy.MethodsWe developed a therapeutic novel polyplex nanoparticle (NP) that selectively targets the cholecystokinin receptor on PDAC. The NP was characterized in vitro and stability testing was performed in human blood. The effects of the target-specific NP loaded with gastrin small interfering RNA (siRNA) was compared with an untargeted NP and with an NP loaded with a scrambled siRNA in vitro and in 2 orthotopic models of PDAC. A polymerase chain reaction metastasis array examined differentially expressed genes from control tumors compared with tumors of mice treated with the targeted polyplex NP.ResultsThe polyplex NP forms a micelle that safely delivers specific gastrin siRNA to the tumor without off-target toxicity. Consistent with these findings, cellular uptake was confirmed only with the targeted fluorescently labeled NP by confocal microscopy in vitro and by IVIS fluorescent based imaging in mice bearing orthotopic pancreatic cancers but not found with untargeted NPs. Tumor uptake and release of the gastrin siRNA NP was verified by decreased cellular gastrin gene expression by quantitative reverse-transcription polymerase chain reaction and peptide expression by immunohistochemistry. Growth of PDAC was inhibited in a dose-related fashion in cell culture and in vivo. The targeted NP therapy completely blocked tumor metastasis and altered tumor-specific genes.ConclusionsOur polyplex nanoparticle platform establishes both a strong foundation for the development of receptor-targeted therapeutics and a unique approach for the delivery of siRNA in vivo, thus warranting further exploration of this approach in other types of cancers.

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“…Several different types of antioxidants (Met, Cys, Glut, MTG, and EDTA) were selected and tested based on the presence in marketed protein formulations and their mechanism of action. [60][61][62][63][64][65] Results from our stability studies demonstrate that incorporation of Met and EDTA as antioxidants in the PS80 formulation substantially improved the stability of this mAb and provided protection from oxidation without altering the protein structure and solution properties. These data also suggested that oxidation occurs not only through a peroxide-mediated pathway, but also through a metal-catalyzed reaction and highlighted the value of a combination approach toward ensuring a stable formulation.…”

Section: Discussionmentioning

confidence: 99%

Development of a stable low-dose aglycosylated antibody formulation to minimize protein loss during intravenous administration

Morar-Mitrica

Puri

Sassi

et al. 2015

mAbs

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The physical and chemical integrity of a biopharmaceutical must be maintained not only during long-term storage but also during administration. Specifically for the intravenous (i.v.) delivery of a protein drug, loss of stability can occur when the protein formulation is compounded with i.v. bag diluents, thus modifying the original composition of the drug product. Here we present the challenges associated with the delivery of a low-dose, highly potent monoclonal antibody (mAb) via the i.v. route. Through parallel in-use stability studies and conventional formulation development, a drug product was developed in which adsorptive losses and critical oxidative degradation pathways were effectively controlled. This development approach enabled the i.v. administration of clinical doses in the range of 0.1 to 0.5 mg total protein, while ensuring liquid drug product storage stability under refrigerated conditions.

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Formulation development and manufacturing of a gastrin/CCK-2 receptor targeting peptide as an intermediate drug product for a clinical imaging study

Cited by 12 publications

References 19 publications

Bioconjugation of Calcium Phosphosilicate Composite Nanoparticles for Selective Targeting of Human Breast and Pancreatic Cancers In Vivo

Bioconjugation of Calcium Phosphosilicate Composite Nanoparticles for Selective Targeting of Human Breast and Pancreatic Cancers In Vivo

Cholecystokinin Receptor-Targeted Polyplex Nanoparticle Inhibits Growth and Metastasis of Pancreatic Cancer

Development of a stable low-dose aglycosylated antibody formulation to minimize protein loss during intravenous administration

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