A phase II dose-escalation trial of perioperative desmopressin (1-desamino-8-d-arginine vasopressin) in breast cancer patients.

Desmopressin (dDAVP) is a well-known peptide analog of the antidiuretic hormone vasopressin, used to prevent excessive bleeding during surgical procedures. dDAVP increases hemostatic mediators, such as the von Willebrand factor (vWF), recently considered a key element in resistance to metastasis. Studies in mouse models and veterinary trials in dogs with locally-advanced mammary tumors demonstrated that high doses of perioperative dDAVP inhibited lymph node and early blood-borne metastasis and significantly prolonged survival. We conducted a phase II dose-escalation trial in patients with breast cancer, administering a lyophilized formulation of dDAVP by intravenous infusion in saline, 30-60 min before and 24 h after surgical resection. Primary endpoints were safety and tolerability, as well as selection of the best dose for cancer surgery. Secondary endpoints included surgical bleeding, plasma levels of vWF, and circulating tumor cells (CTCs) as measured by quantitative PCR of cytokeratin-19 transcripts. Only 2 of a total of 20 patients experienced reversible adverse events, including hyponatremia (grade 4) and hypersensitivity reaction (grade 2). Reactions were adequately managed by slowing the infusion rate. A reduced intraoperative bleeding was noted with increasing doses of dDAVP. Treatment was associated with higher vWF plasma levels and a postoperative drop in CTC counts. At the highest dose level evaluated (2 µg/kg) dDAVP appeared safe when administered in two slow infusions of 1 µg/kg, before and after surgery. Clinical trials to establish the effectiveness of adjunctive perioperative dDAVP therapy are warranted. This trial is registered on www.clinicaltrials.gov (NCT01606072).

Anti-idiotype antibodies in cancer treatment: the pharmaceutical industry perspective.

Active immunotherapy is an interesting field from the industry's perspective and in the last years, regulatory agencies and the medical community have showed renewed expectations and interest in cancer vaccines. The development of new immune therapies offers many challenges, and this is reflected in the small number of phase III trials showing clear benefits. Traditional concepts applied in clinical trials for the development of chemotherapeutic agents may be inadequate for immunotherapies and a new paradigm is emerging. It is possible that organized efforts and funding will accelerate the development of therapeutically effective cancer vaccines. This article reviews the attributes of cancer vaccines which make them attractive from the industry's perspective, and focuses especially in the characteristics of Racotumomab, an anti-idiotype antibody vaccine.

Racotumomab: an anti-idiotype vaccine related to N-glycolyl-containing gangliosides - preclinical and clinical data.

Neu-glycolyl (NeuGc)-containing gangliosides are attractive targets for immunotherapy with anti-idiotype mAbs, because these glycolipids are not normal components of the cytoplasmic membrane in humans, but their expression has been demonstrated in several human malignant tumors. Racotumomab is an anti-idiotype mAb specific to P3 mAb, an antibody which reacts to NeuGc-containing gangliosides, sulfatides, and other antigens expressed in tumors. Preparations containing racotumomab were able to induce a strong anti-metastatic effect in tumor-bearing mice. Different Phase I clinical trials have been conducted in patients with advanced melanoma, breast cancer, and lung cancer. The results of these clinical trials demonstrated the low toxicity and the high immunogenicity of this vaccine. The induced antibodies recognized and directly killed tumor cells expressing NeuGcGM3. A Phase II/III multicenter, controlled, randomized, double blind clinical trial was conducted to evaluate the effect of aluminum hydroxide-precipitated racotumomab vaccine in overall survival in patients with advanced non-small cell lung cancer. The clinical results of this study showed a significant clinical benefit in the patients who were treated with the anti-idiotype vaccine.

Sensitivity of tumor cells towards CIGB-300 anticancer peptide relies on its nucleolar localization.

CIGB-300 is a novel anticancer peptide that impairs the casein kinase 2-mediated phosphorylation by direct binding to the conserved phosphoacceptor site on their substrates. Previous findings indicated that CIGB-300 inhibits tumor cell proliferation in vitro and induces tumor growth delay in vivo in cancer animal models. Interestingly, we had previously demonstrated that the putative oncogene B23/nucleophosmin (NPM) is the major intracellular target for CIGB-300 in a sensitive human lung cancer cell line. However, the ability of this peptide to target B23/NPM in cancer cells with differential CIGB-300 response phenotype remained to be determined. Interestingly, in this work, we evidenced that CIGB-300's antiproliferative activity on tumor cells strongly correlates with its nucleolar localization, the main subcellular localization of the previously identified B23/NPM target. Likewise, using CIGB-300 equipotent doses (concentration that inhibits 50% of proliferation), we demonstrated that this peptide interacts and inhibits B23/NPM phosphorylation in different cancer cell lines as evidenced by in vivo pull-down and metabolic labeling experiments. Moreover, such inhibition was followed by a fast apoptosis on CIGB-300-treated cells and also an impairment of cell cycle progression mainly after 5 h of treatment. Altogether, our data not only validates B23/NPM as a main target for CIGB-300 in cancer cells but also provides the first experimental clues to explain their differential antiproliferative response. Importantly, our findings suggest that further improvements to this cell penetrating peptide-based drug should entail its more efficient intracellular delivery at such subcellular localization.

NGcGM3 ganglioside: a privileged target for cancer vaccines.

Active specific immunotherapy is a promising field in cancer research. N-glycolyl (NGc) gangliosides, and particularly NGcGM3, have received attention as a privileged target for cancer therapy. Many clinical trials have been performed with the anti-NGc-containing gangliosides anti-idiotype monoclonal antibody racotumomab (formerly known as 1E10) and the conjugated NGcGM3/VSSP vaccine for immunotherapy of melanoma, breast, and lung cancer. The present paper examines the role of NGc-gangliosides in tumor biology as well as the available preclinical and clinical data on these vaccine products. A brief discussion on the relevance of prioritization of cancer antigens in vaccine development is also included.

Anticancer peptide CIGB-300 binds to nucleophosmin/B23, impairs its CK2-mediated phosphorylation, and leads to apoptosis through its nucleolar disassembly activity.

CIGB-300, formerly known as P15-tat, is a proapoptotic peptide with established antiproliferative activity in vitro and antitumoral activity in vivo. This hypothesis-driven peptide was initially selected for its ability to impair the in vitro CK2-mediated phosphorylation in one of its substrates through direct binding to the conserved acidic phosphoaceptor domain. However, the actual in vivo target(s) on human cancer cells among the hundreds of CK2 substrates as well as the subsequent events that lead to apoptosis on tumor cells remains to be determined. In this work, we identified the multifunctional oncoprotein nucleophosmin/B23 as a major target for CIGB-300. In vivo, the CIGB-300-B23 interaction was shown by pull-down experiments and confirmed by the early in situ colocalization of both molecules in the cell nucleolus. Moreover, CIGB-300 inhibits the CK2-mediated phosphorylation of B23 in a dose-dependent fashion both in vitro and in vivo as shown using the recombinant GST fusion protein and the metabolic labeling approach, respectively. Such phosphorylation impairment was correlated with the ability of CIGB-300 to induce nucleolar disassembly as documented by the use of established markers for nucleolar structure. Finally, we showed that such a sequence of events leads to the rapid and massive onset of apoptosis both at the molecular and cellular levels. Collectively, these findings provide important clues by which the CIGB-300 peptide exerts its proapoptotic effect on tumor cells and highlights the suitability of the B23/CK2 pathway for cancer-targeted therapy.

Molecular detection of circulating tyrosinase mRNA: optimization in a preclinical xenograft mouse melanoma model and further evaluation in samples from advanced melanoma patients.

We designed high-affinity primers for the mRNA sequence of human tyrosinase to test the value of molecular detection of circulating melanoma cells by reverse transcription-polymerase chain reaction (RT-PCR). The optimization process included in vitro settings and in vivo studies in a xenograft mouse model. We detected tyrosinase expression with at least 40 pg and 1.5 pg of total RNA extracted from cultured SKmel human melanoma cells, using a first round of PCR amplification and nested PCR, respectively. Human tyrosinase expression was found in the blood of nude mice bearing subcutaneous SKmel tumors, and the expression bands were stronger after manipulation of the tumor mass. We also examined the fate of circulating melanoma cells in the present melanoma model. Tyrosinase expression declined in blood 6 h after a direct intravenous injection of SKmel cells. A preliminary study in human blood samples demonstrated a baseline positive tyrosinase determination in 64% (16/25) of advanced melanoma patients using the RT-PCR nested assay. Baseline tyrosinase expression was significantly associated with disease progression after 12 months, and sequential determination during follow-up of the remaining disease-free patients showed a progressive increase of negative results.

Systemic administration of a peptide that impairs the protein kinase (CK2) phosphorylation reduces solid tumor growth in mice.

The antitumor efficacy of the CK2 inhibitors so far described has not been extensively evaluated in cancer animal models. We have previously demonstrated that a proapoptotic cyclic peptide termed P15 delivered into the cells by the Tat Cell Penetrating Peptide was able to abrogate the CK2-mediated phosphorylation and induce tumor regression when injected directly into solid tumors in mice. Here we explored the antitumor effect by systemic administration of P15-Tat in a consecutive 5-day schedule through either intraperitoneal or intravenous route. Importantly, significant delay of tumor growth was observed at 2 mg/kg (p < 0.05), 10 mg/kg (p < 0.01) or 40 mg/kg (p < 0.001) after P15-Tat administration both in syngeneic murine tumors and human tumors xenografted in nude mice. In line with this, the systemic administration of P15-Tat induced apoptosis in the tumor as evidenced by in situ DNA fragmentation. Furthermore, we evidenced that 99mTc-labeled P15-Tat peptide was certainly accumulated on the tumors after administration by both routes. This report becomes the first describing the antitumor effect induced by systemic administration of a peptide that targets the acidic phosphorylation domain for CK2 substrates. Also, our data reinforces the perspectives of P15-Tat for the cancer targeted therapy.

1E10 anti-idiotype vaccine in non-small cell lung cancer: experience in stage IIIb/IV patients.

Conventional treatment of non-small cell lung cancer (NSCLC) has apparently reached a plateau of effectiveness in improving the survival of the patients. For that reason the search for new therapeutic strategies in this type of tumor is justified. 1E10 is an anti-idiotype murine monoclonal antibody (Ab2 MAb) specific to P3 Ab1 MAb, which reacts with NeuGc-containing gangliosides, sulfatides and with antigens expressed in some tumors, including those from the lung. We report the treatment with aluminum hydroxide-precipitated 1E10 MAb of 34 stage IIIb and 37 stage IV NSCLC patients. These patients were treated with the anti-idiotype vaccine, after received standard chemotherapy and radiotherapy, in a compassionate-use basis study. Patients received five bi-weekly injections of 1 mg of 1E10/Alum, other 10 doses at 28-day intervals and later the patients who maintained a good performance status continued to be immunized at this same time interval. No evidence of unexpected or serious adverse effects was reported. The median survival time of the 56 patients who entered the study with partial response or disease stabilization and with a PS 1 after the first line of chemo/radiotherapy, was 11.50 months from starting vaccination. In contrast, the median survival time calculated for patients who started vaccination with progressive disease and/or a PS2 was 6.50 months.

Suprapontine ablation in normal and Doc-salt or one-kidney, one-clip hypertensive rats

Se estududió, en ratas no anestesiadas, el papel de las áreas supraprotuberanciales en el mantenimiento agudo de la hipertensión producida por tratamiento con Doca-sal (4 semanas) o en el modelo 1 riñon, 1-clip (1R-1C) (4 semanas post-cirugia). Se midió la presión arterial media (PA) y la fecuencia cardíaca (FC) 15 minutos antes y 2 horas después de aspirar todos los tejidos rostrales a la protuberancia. Este procedimiento no modificó la PA de las ratas normales o 1R-1C. En las ratas tratadas con Doca-sal la PA cayo (154 ñ 4 vs. 110 ñ 5 mmHg; p < 0.001). La FC aumentó en las ratas normales (351 ñ 10 a 446, ñ20 1pm; p < 0.01) y en las 1R-1C. (350 ñ 10 a (351 ñ 12 1 pm; p < 0.001). Las clonidinas inyectada en la cisterna magna de las ratas 1R-1C luego de la aspiración de las áreas supraprotuberanciales baja la PA (146 ñ6 a 118 ñ 11 mmHg; p < 0.05) y la FC (515 ñ17 a 400 ñ 33; p < 0.05). El pentolinium, luego de la aspiración supraprotuberancial, baja la PA de las ratas normales (116 ñ 4 a 63 ñ 5 mmHg; p < 0.001), Doca-sal (111 ñ 5; p < 0.001) y 1R-1C (163 ñ 8 a 59 ñ 6 mmHg; P < 0.001). Estos resultados sugieren que las estructuras supraprotuberanciales tienen un papel importante en el mantenimiento agudo de la hipertensión por Doca-sal. En las ratas 1R-1C el mantenimiento agudo de la hipertensión depende de una actividad simpática originada en áeas caudales a la ablación (AU)

Suprapontine ablation in normal and Doc-salt or one-kidney, one-clip hypertensive rats

Se estududió, en ratas no anestesiadas, el papel de las áreas supraprotuberanciales en el mantenimiento agudo de la hipertensión producida por tratamiento con Doca-sal (4 semanas) o en el modelo 1 riñon, 1-clip (1R-1C) (4 semanas post-cirugia). Se midió la presión arterial media (PA) y la fecuencia cardíaca (FC) 15 minutos antes y 2 horas después de aspirar todos los tejidos rostrales a la protuberancia. Este procedimiento no modificó la PA de las ratas normales o 1R-1C. En las ratas tratadas con Doca-sal la PA cayo (154 ñ 4 vs. 110 ñ 5 mmHg; p < 0.001). La FC aumentó en las ratas normales (351 ñ 10 a 446, ñ20 1pm; p < 0.01) y en las 1R-1C. (350 ñ 10 a (351 ñ 12 1 pm; p < 0.001). Las clonidinas inyectada en la cisterna magna de las ratas 1R-1C luego de la aspiración de las áreas supraprotuberanciales baja la PA (146 ñ6 a 118 ñ 11 mmHg; p < 0.05) y la FC (515 ñ17 a 400 ñ 33; p < 0.05). El pentolinium, luego de la aspiración supraprotuberancial, baja la PA de las ratas normales (116 ñ 4 a 63 ñ 5 mmHg; p < 0.001), Doca-sal (111 ñ 5; p < 0.001) y 1R-1C (163 ñ 8 a 59 ñ 6 mmHg; P < 0.001). Estos resultados sugieren que las estructuras supraprotuberanciales tienen un papel importante en el mantenimiento agudo de la hipertensión por Doca-sal. En las ratas 1R-1C el mantenimiento agudo de la hipertensión depende de una actividad simpática originada en áeas caudales a la ablación