ABSTRACT
The generation of acylated impurities has represented an important hurdle in the development of long acting injectables for therapeutic peptides using biocompatible polymers with a polyester moiety. We investigated here an in situ forming depot (ISFD) technology that uses polyethylene glycol - polyester copolymers and a solvent exchange mechanism to promote depot formation. This technology has shown promise in formulating small molecules as well as therapeutic proteins. In the present work, using the well-known somatostatin analog octreotide acetate (OctAc) as a model molecule, we evaluated this delivery platform to release therapeutic peptides. Peptide acylation was found to be pronounced in the formulation, while it was very limited once the depot was formed and during the release process. The octreotide acylation pattern was fully characterized by LC-MS/MS. Moreover, it was demonstrated that exchanging the acetate anion with more hydrophobic counterions like pamoate or lauryl sulfate allowed to greatly improve the peptide stability profile, as well as the formulation release performance. Finally, the in vivo evaluation through pharmacokinetics studies in rat of these new octreotide salts in ISFD formulations showed that octreotide was quantifiable up to four weeks post-administration with a high bioavailability and an acceptable initial burst.
Subject(s)
Octreotide , Tandem Mass Spectrometry , Animals , Chromatography, Liquid , Kinetics , Polylactic Acid-Polyglycolic Acid Copolymer , Rats , TechnologyABSTRACT
Small therapeutic proteins represent a promising novel approach to treat cancer. Nevertheless, their clinical application is often adversely impacted by their short plasma half-life. Controlled long-term delivery of small biologicals has become a challenge because of their hydrophilic properties and in some cases their limited stability. Here, an in situ forming depot-injectable polymeric system was used to deliver BiJ591, a bispecific T-cell engager (BiTE) targeting both prostate-specific membrane antigen (PSMA) and the CD3 T-cell receptor in prostate cancer. BiJ591 induced T-cell activation, prostate cancer-directed cell lysis, and tumor growth inhibition. The use of diblock (DB) and triblock (TB) biodegradable polyethylene glycol-poly(lactic acid; PEG-PLA) copolymers solubilized in tripropionin, a small-chain triglyceride, allowed maintenance of BiJ591 stability and functionality in the formed depot and controlled its release. In mice, after a single subcutaneous injection, one of the polymeric candidates, TB1/DB4, provided the most sustained release of BiJ591 for up to 21 days. Moreover, the use of BiJ591-TB1/DB4 formulation in prostate cancer xenograft models showed significant therapeutic activity in both low and high PSMA-expressing tumors, whereas daily intravenous administration of BiJ591 was less efficient. Collectively, these data provide new insights into the development of controlled delivery of small therapeutic proteins in cancer. Mol Cancer Ther; 17(9); 1927-40. ©2018 AACR.
Subject(s)
Antibodies, Bispecific/pharmacology , Antineoplastic Agents/pharmacology , Drug Delivery Systems/methods , Polymers/chemistry , Prostatic Neoplasms/drug therapy , Xenograft Model Antitumor Assays/methods , Animals , Antibodies, Bispecific/administration & dosage , Antibodies, Bispecific/immunology , Antigens, Surface/immunology , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/pharmacokinetics , CD3 Complex/immunology , Cell Line, Tumor , Drug Liberation , Glutamate Carboxypeptidase II/immunology , Humans , Male , Mice, SCID , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/pathology , Survival Analysis , Tumor Burden/drug effectsABSTRACT
BACKGROUND: The aim of this paper was to validate an inductively coupled plasma-mass spectrometry (ICP-MS) method to quantify tungsten in human plasma and to study its percentage binding to plasma proteins. METHODS: This method was validated with respect to accuracy, precision, selectivity and limits of quantification and of detection according to Good Laboratory Practice Guidelines. Calibration curves were obtained in the range 10-500 ng/ml. The extent of plasma protein binding was determined by ultrafiltration in the range 40-2000 ng/ml. RESULTS: A significant matrix effect was observed. The linearity of this method was statistically proven. Precision ranged from 0.76% to 6.49%, and accuracy from 97% to 102%. The lower limit of quantification (LLOQ) was 10 ng/ml. The mean percentage of unbound fraction was 89%. CONCLUSIONS: The results obtained indicate that the method described fulfills the accuracy and precision requirements necessary to carry out pharmacokinetic studies in man.