n an era of increasing health care costs and qualitycontrol mandates, compounding pharmacies have been under intense scrutiny. The health care system, including ophthalmologists, relies on compounded drug formulations particularly intravitreal bevacizumab (Avastin; Genentech/Roche) for the treatment of a variety of choroidal and retinal vascular diseases. Clinical trials, such as the Comparison of AMD Treatments Trials 1 and the Inhibition of Vascular Endothelial Growth Factor (VEGF) in Age-related choroidal Neovascularization trial, 2 support the effectiveness of intravitreal bevacizumab for the treatment of age-related macular degeneration. While both bevacizumab and ranibizumab (Lucentis; Genentech/Roche) have been found to have similar safety and efficacy, 1,2 substantial cost savings may be derived from using bevacizumab, which is reimbursed approximately $50 from Medicare in comparison with nearly $2000 for ranibizumab. 3 Indeed, the researchers of the Inhibition of Vascular Endothelial Growth Factor (VEGF) in Age-related choroidal Neovascularization trial estimated that the National Health Service in the United Kingdom could potentially save an estimated £84.5 million (US $140.9 million) per year by switching from ranibizumab to bevacizumab and administering medication on an as-needed basis.Compounding pharmacies prepare intravitreal bevacizumab for clinical use but the quality, consistency, and safety IMPORTANCE Bevacizumab acquired from compounding pharmacies for intravitreal injection may cause infectious and noninfectious inflammation. In addition to safety issues, the drug itself may have variable efficacy associated with product aliquoting, handling, and distribution.OBJECTIVE To conduct surveillance cultures, evaluate endotoxin levels, and assess protein concentrations of bevacizumab obtained from compounding pharmacies in the United States.DESIGN AND SETTING Prospective in vitro study of syringes containing intravitreal preparations of bevacizumab from compounding pharmacies. This study was conducted at a university-based, good manufacturing practice facility and academic ophthalmology practice.MAIN OUTCOMES AND MEASURES Microbial culture growth, endotoxin levels, and quantity and binding affinity of protein in each sample.RESULTS There were no microbial contaminants or endotoxin detected in any of the samples. Of the 21 compounded samples of bevacizumab obtained from 11 pharmacies, 17 (81%) had lower protein concentrations (mean [SD], 22.2 [4.9] mg/mL; range, 19.2-24.5 mg/mL) compared with bevacizumab acquired directly from Genentech (25 mg/mL; P < .05). In 3 of 10 compounding pharmacies where more than 1 sample was available, there were statistically significant differences in the protein concentration between samples from the same compounding pharmacy. CONCLUSIONS AND RELEVANCETest results from intravitreal preparations of bevacizumab acquired from compounding pharmacies were negative for microbial contaminants and endotoxin. However, there were significant variations in protein concentration th...

Coronary artery disease (CAD), a leading cause of mortality, is a chronic disease in which blood flow to the myocardium is obstructed, leading to ischemia. Although coronary artery stenting and surgical bypass are successful with localized coronary lesions, patients with diffuse CAD have only pharmacologic options. In a mouse ischemic hind-limb model, AdVEGF-All6A+, an Ad5 vector expressing the cDNA/genomic hybrid of the vascular endothelial growth factor (VEGF) gene (expressing isoforms, 121, 165, and 189) mediated recovery of blood flow at a dose of two logs less than that required with a single isoform. The objective of the current study was to ascertain the safety profile of good manufacturing practice (GMP)-grade AdVEGF-All6A+ in the adult rat ischemic heart model in support of a clinical study to treat humans with diffuse CAD. AdVEGF-All6A+ (10(5), 10(6), or 10(7) particle units), a control vector (AdNull, 10(7) particle units) with no translatable expression cassette and a vehicle sham control (phosphate buffered saline [PBS]) were administered separately to the left ventricle of rats immediately following acute coronary artery ligation to initiate myocardial infarction (MI), designed to evoke an extreme ischemic myocardium in cohorts (n=5 males; n=5 females), with sacrifice at 5, 14, or 30 days. Six cohorts received no ligation but were administered AdVEGF-All6A+ vector or PBS with sacrifice at 30 or 365 days. Although there were surgical-related abnormalities among the groups, blinded evaluation of gross and histopathology, complete blood count, and serum chemistry found no significant differences between control- and vector-treated groups and no adverse effects could be attributed to AdVEGF-All6A+. No changes in serum troponin-I levels persisted beyond those associated with the MI. Gross pathology and histopathology of all major organs showed no AdVEGF-All6A+-related changes. Overall, this safety profile suggests that AdVEGF-All6A+ or AdNull administration to the myocardium meets the criteria to proceed to clinical trial.

BackgroundDespite clinical success with T cell engagers (TCEs) targeting hematological malignancies, achieving a safe and efficacious dose in patients with solid tumors remains challenging. Due to potency, low levels of target antigen expression on normal tissues may not be tolerated. To overcome this, we engineered a novel conditionally active TCE design called COBRA (Conditional Bispecific Redirected Activation). Administered as prodrugs, COBRAs bind to cell surface antigens on both normal and tumor tissues but are preferentially activated within the tumor microenvironment.MethodsA COBRA was engineered to target EGFR, TAK-186. The potency of precleaved TAK-186 relative to a non-cleavable control was assessed in vitro. Mice bearing established solid tumors expressing a range of EGFR levels were administered a single bolus of human T cells, and concurrently treated with TAK-186 and associated controls intravenously. We assessed the plasma and tumor exposure of intact and cleaved TAK-186.ResultsTAK-186 shows potent redirected T cell killing of antigen expressing tumor cells. In vivo efficacy studies demonstrate regressions of established solid tumors, dependent on intratumoral COBRA cleavage. Pharmacokinetic studies reveal TAK-186 is stable in circulation, but once activated is rapidly cleared due to loss of its albumin-binding half-life extension domain.ConclusionsThe studies shown support the advancement of TAK-186, and the pursuit of additional COBRA TCEs for the treatment of solid tumors.

ImmunologIcal aspects of gene therapy I 75% (range 38-94%) of HIV-infected cells over 2-3 days, and reduced HIV DNA by approximately one log over 5 days. Beyond 72 hours of culture the anti-HIV CAR+ lymphocytes with CCR5 disruption resulted in much greater reduction in HIV than CAR+ lymphocytes without CCR5 disruption.Conclusions: It is feasible to construct anti-HIV CAR+ T-cells that are protected from HIV infection. This strategy warrants further study using in vivo models of HIV latency.

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