Preclinical study of LMP1-RNAi-based anti-tumor therapy in EBV-positive nasopharyngeal carcinoma

RNA interference (RNAi) treatment has been proven to be an important therapeutic approach in cancer based on downregulation of target-oncogenes, but its clinical efficacy still needs further investigation. LMP1 is usually presented by Epstein-Barr virus (EBV)-positive tumor cells like EBV-associated nasopharyngeal carcinoma (NPC) and acts as an oncogene in tumorigenesis. However, the mechanism of LMP1 as a proto-oncogene in nasopharyngeal carcinoma is still unclear. Two sequence-specific shRNAs 1 and 2 were designed to target the different nucleotide loci of EBV latent antigen LMP1 gene and a series of in vivo and in vitro experiments were performed to investigate the therapeutic effect of sequence-specific shRNAs targeting LMP1 and its related molecular mechanisms in EBV-positive NPC. LMP1-shRNA2 generated a truncated LMP1 mRNA and protein, whereas LMP1-shRNA1 completely blocked LMP1 mRNA and protein expression. Both LMP1-shRNAs inhibited the proliferation and migration of NPC cells overexpressing LMP1 (NPC-LMP1) as well as the NPC-associated myeloid-derived suppressor cell (MDSC) expansion in vitro. However, LMP1-shRNA2 maintained the immunogenicity of NPC-LMP1 cells, which provoked MHC-class I-dependent T cell recognition. LMP1-shRNAs inhibited tumor growth in nude mice but did not reach statistical significance compared to control groups, while the LDH nanoparticle loaded LMP1-shRNAs and the antigen-specific T cells induced by NPC-LMP1 cells treated with LMP1-shRNA2 significantly reduced tumor growth in vivo. LMP1-RNAi-based anti-tumor therapy could be a new hope for the clinical efficacy of RNAi treatment of tumors like NPC.

Progress in preclinical study of combination of PARP inhibitors against tumor / 中国药房

Poly（ADP-ribose）polymerase（PARP）is a kind of DNA damage repair enzyme. PARP inhibitors include Olaparib （AZD2281），Niraparib（MK-4827），Rucaparib，Veliparib（ABT-888），Fluzoparib and Talazoparib （BMN-673），etc. This article reviews the preclinical research on the combined application of PARP inhibitors against tumor by searching the relevant literatures. Through the synthetic lethal mode ，PARP inhibitors have a strong killing effect on tumor cells with homologous recombination repair defects. However ，for tumor cells with intact DNA damage repair function ，PARP inhibitors often need to be combined with radiotherapy or other drugs to play a role. Combined application drugs include antiangiogenic drugs ，heat shock protein 90 inhibitors，cyclin-dependent kinase 12 inhibitors，immune checkpoint inhibitors ，histone deacetylase inhibitors ，etc. The combined application of PARP inhibitors is expected to enhance the efficacy of anti-tumor drugs and achieve the goals of sensitization ， synergism and reversal of drug resistance ，which is worthy of further in-depth research and exploration of new combined treatment schemes.

Safety and Biodistribution of Human Bone Marrow-Derived Mesenchymal Stromal Cells Injected Intrathecally in Non-Obese Diabetic Severe Combined Immunodefi ciency Mice: Preclinical Study

BACKGROUND: Mesenchymal stromal cells (MSCs) have potent immunomodulatory and neuroprotective properties, and have been tested in neurodegenerative diseases resulting in meaningful clinical improvements. Regulatory guidelines specify the need to perform preclinical studies prior any clinical trial, including biodistribution assays and tumourigenesis exclusion. We conducted a preclinical study of human bone marrow MSCs (hBM-MSCs) injected by intrathecal route in Non-Obese Diabetic Severe Combined Immunodeficiency mice, to explore cellular biodistribution and toxicity as a privileged administration method for cell therapy in Friedreich's Ataxia. METHODS: For this purpose, 3 × 10⁵ cells were injected by intrathecal route in 12 animals (experimental group) and the same volume of culture media in 6 animals (control group). Blood samples were collected at 24 h (n = 9) or 4 months (n = 9) to assess toxicity, and nine organs were harvested for histology and safety studies. Genomic DNA was isolated from all tissues, and mouse GAPDH and human β2M and β-actin genes were amplified by qPCR to analyze hBM-MSCs biodistribution. RESULTS: There were no deaths nor acute or chronic toxicity. Hematology, biochemistry and body weight were in the range of normal values in all groups. At 24 h hBM-MSCs were detected in 4/6 spinal cords and 1/6 hearts, and at 4 months in 3/6 hearts and 1/6 brains of transplanted mice. No tumours were found. CONCLUSION: This study demonstrated that intrathecal injection of hBM-MSCs is safe, non toxic and do not produce tumors. These results provide further evidence that hBM-MSCs might be used in a clinical trial in patients with FRDA.

Considerations on the dose determination for first-in-human clinical trials with novel biopharmaceuticals / 药物评价研究

In the development process from the preclinical stage to the subsequent clinical phase,one critical risk controlling step is the determination of the first-in-human (FIH) dose.There is difference in the mechanisms of action and toxic risks between biopharmaceuticals and small molecule drugs,therefore different considerations will be involved in the determination of the FIH dose for biopharmaceuticals.This paper presents the overall review of the preclinical studies supporting the determination of FIH dose and the approaches based on the NOAEL,MABLE,and PK/PD model.The experience-based views that the MABLE-based FIH dose for immune activating products would be reasonably safe,although the NOAEL approach remains conservative and widely used for a majority of biopharmaceuticals are also presented.It is suggested that sponsors should determine the appropriate and safe FIH dose by diverse approaches according to the characteristics of product.Early communication between sponsors and regulators is always beneficial.

Considerations on the dose determination for first-in-human clinical trials with novel biopharmaceuticals / 药物评价研究

In the development process from the preclinical stage to the subsequent clinical phase,one critical risk controlling step is the determination of the first-in-human (FIH) dose.There is difference in the mechanisms of action and toxic risks between biopharmaceuticals and small molecule drugs,therefore different considerations will be involved in the determination of the FIH dose for biopharmaceuticals.This paper presents the overall review of the preclinical studies supporting the determination of FIH dose and the approaches based on the NOAEL,MABLE,and PK/PD model.The experience-based views that the MABLE-based FIH dose for immune activating products would be reasonably safe,although the NOAEL approach remains conservative and widely used for a majority of biopharmaceuticals are also presented.It is suggested that sponsors should determine the appropriate and safe FIH dose by diverse approaches according to the characteristics of product.Early communication between sponsors and regulators is always beneficial.

Construction of innovative pharmacy experiments based on new drug research and development / 中华医学教育探索杂志

In order to improve the comprehensive quality and innovation ability of undergraduates, we integrate the pharmacognosy, natural medicine chemistry, pharmaceutical chemistry, pharmaceutical analysis, pharmacy, pharmacology experiment module in accordance with the drug development program, to construct multidisciplinary innovative pharmaceutical experiment course. Besides, we carry out teaching with scientific research mode, scientific evaluation to assess the effects and making analysis to expose the prob-lems, which lay a solid foundation for establishment of the innovative pharmacy experiment in the future.

Technical considerations on the study and evaluation of the biosimilars' immunogenicity / 中国药学杂志

OBJECTIVE: To evaluate the immunogenicity of biosimilars. METHODS: Non-clinical and clinical studies carried out in accordance with the guidelines in immunological characteristics and immunogenicity of biosimilars should focus on the production of anti-drug antibodies and detection of drug-resistant properties, immune complexes and their potential effects on pharmacodynamics/pharmacokinetics. Assays for immunogenicity of biological agents mainly include enzyme linked immunosorbent assay (ELISA), radioimmunoassay, surface plasm on resonance method, enzyme linked immunospot assay, immuno-PCR method, cell proliferation assay, etc. RESULTS AND CONCLUSION: Biosimilars immunogenicity should be comprehensively interpreted based on both the preclinical/clinical evaluation results and the prevalence of clinical adverse events.

Adoptive Cell Therapy of Melanoma with Cytokine-induced Killer Cells

Melanoma is the most aggressive skin cancer and its incidence is gradually increasing worldwide. Patients with metastatic melanoma have a very poor prognosis (estimated 5-year survival rate of <16%). In the last few years, several drugs have been approved for malignant melanoma, such as tyrosine kinase inhibitors and immune checkpoint blockades. Although new therapeutic agents have improved progression-free and overall survival, their use is limited by drug resistance and drug-related toxicity. At the same time, adoptive cell therapy of metastatic melanoma with tumor-infiltrating lymphocytes has shown promising results in preclinical and clinical studies. In this review, we summarize the currently available drugs for treatment of malignant melanoma. In addition, we suggest cytokine-induced killer (CIK) cells as another candidate approach for adoptive cell therapy of melanoma. Our preclinical study and several previous studies have shown that CIK cells have potent anti-tumor activity against melanomas in vitro and in an in vivo human tumor xenograft model without any toxicity.

Estudio preclínico del efecto de las esencias florales de Bach en la inflamación aguda / Preclinical effect of the Bach flower essence in acute inflammation

Introducción: los estudios preclínicos que de forma habitual preceden al uso de un medicamento convencional pudieran convertirse en la solución a la comprobación de las acciones terapéuticas de medicinas naturales como las flores de Bach. La inflamación aguda y los modelos experimentales que la simulan son herramientas útiles en este camino. Objetivo: evaluar la actividad antiinflamatoria de las esencias florales de Bach en un modelo de inflamación aguda en ratas. Métodos: se realizó un estudio preclínico experimental piloto controlado y aleatorizado a ciegas en la UTEX de la Universidad de Ciencias Médicas de Villa Clara en el mes de Diciembre del 2010. El modelo experimental consistió en la inducción de edema plantar con dextrán. Se conformaron en 5 grupos de 6 animales (en total 30 ratas). Se trataron con las esencias Beech, Vervain y Rescue Remedy, respectivamente; mientras que los 2 restantes fueron control (negativo con placebo y positivo con Difenhidramina). Resultados: se observaron diferentes efectos en los tratamientos utilizados. Beech y Vervain mostraron propiedades antiinflamatorias con diferencias significativas al placebo, cuestión que avala su utilización en el tratamiento de patologías donde la inflamación es un denominador común. Estas acciones fueron selectivas ya que Beech tuvo efecto inmediato y Vervain mostró actividad sobre la inflamación en tercera hora. Otro preparado, el Rescue Remedy, no mostró esta actividad diferente al placebo

Background: the preclinical studies that in their habitual form precede the use of a conventional drug, could become a solution to testing therapeutic actions of natural medications such as the Bach flowers. The acute inflammation and the experimental models that simulate it are very useful tools in this direction. Objective: to evaluate the antinflammatory activity of the Bach flower essence in an acute inflammation model in rats. Methods: a pilot experimental preclinical study, controlled and blindly randomized, was carried out at the UTEX of the Medical Sciences University of Villa Clara on December 2010. The experimental model consisted on the induction of plantar oedema with Dextran. 5 groups of 6 animals were formed (30 rats in total). They were treated with Beech and Vervain essences and Rescue Remedy respectively; while the remaining 2 formed the control group (negative with placebo and positive with Diphenhydramine). Results: different effects in the treatments were observed. Beech and Vervain showed antinflammatory properties with significant differences form placebo, condition that endorse their use in the treatment of pathologies where inflammation is a common factor. These actions were selective because Beech had an immediate effect and Vervain showed its activity on the inflammation in the third hour. Other preparation, the Rescue Remedy, showed that its activity was not different from placebo