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Treatment of glioblastoma with thymosin-alpha 1

A glioblastoma, thymosin technology, applied in antitumor drugs, peptide/protein components, medical preparations containing active ingredients, etc.

Inactive Publication Date: 2005-05-11
RHODE ISLAND HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For many years chloroethylnitrosoureas have been used only as monochemotherapy agents for primary brain tumors; however, historical data indicate that the use of these compounds as single agents in the treatment of brain tumors is not particularly effective (e.g., Aquafedda, et al.)

Method used

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  • Treatment of glioblastoma with thymosin-alpha 1
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  • Treatment of glioblastoma with thymosin-alpha 1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Treatment of 9L and 293 Tumor Cell Lines with Thymosin α1

[0028] Tumor cell line: Add 5% FCS, 2mM L-glutamine, and 100μM non-essential amino acid (Gibco-BRL, Grand Island, NY) Dulbecco's modified Eagle's medium (DMEM) to rat 9L glioblastoma Tumor cells and 293 human kidney cells were cultured. All cell lines were placed in 5% CO 2 in a humid atmosphere of 37°C. All cell lines were purchased from the American Type Culture Center and certified free of pathogens.

[0029] Thymosin a1 treatment: for acute treatment of thymus fasin, the cells were planted in a 96-well culture plate with a cell density of 20,000 cells / well, and 10 -5 M's thymus was treated with fasin for 24 hours. Cells chronically treated with thymusfasin were inoculated in culture flasks, and during the treatment period (3 days) every 24 hours, 10 -5 M's thymus fasin (added to fresh medium) was treated. In addition, to determine the dose-response curve of 9L cells to thymofasin, cells wer...

Embodiment 2

[0037] Example 2: Study on telomerase-induced apoptosis

[0038] The above MICE detection study confirmed that thymofasin induced the expression of TNF-R1, FasL and FasR in 9L glioblastoma. In order to determine the difference in the sensitivity of 9L glioblastoma cells treated (or untreated) with thymus method to apoptosis induced by cytotoxic T-lymphocytes (CTL), the following experiments were performed: 9L cells were treated with Thymofasin was treated acutely for 24 hours and chronically treated for 72 hours, then the cells were collected and re-seeded in 96-well black culture plates (7.5×10 5 cells / 75 μl / well), and then contacted with 20,000 units / ml streptolysin O (SLO) plus 100 ng recombinant telomerase B (reaction volume 100 μl) at 37° C. for 1 or 3 hours. Cells were permeabilized with SLO instead of perforin and assays were normalized using recombinant telomerase B. Control studies included parallel reactions with no SLO, no telomerase B, or both. Using ATPlite for...

Embodiment 3

[0040] Example 3: Effect of Thymalfasin on Cell Viability of Primary Neuronal Cell Cultures

[0041] Primary cortical neuronal cell cultures were studied to determine doses of thymofasin that were not toxic to non-neoplastic brain cells. Since the previous studies have shown that as the cell density increases from 1×10 per well 4 up to 5×10 5 The absorbance of CV and MTT increases linearly with the change (de la Monte, 2001 & 2000). Therefore, crystal violet (CV) and MTT assays are used to measure cell viability and mitochondrial function.

[0042] Use serially diluted thymus fasin (final concentration at 3.3×10 -5 M to 1×10 -9 M) treated primary neuronal cortical cells in 96-well culture plates and found no decrease in cell viability at the experimental doses used. It can be found by MTT assay that the highest concentration of thymus fasin (3.3×10 -7 M) did reduce viability by 30%. However, this dose is higher than the experimental and clinical doses used.

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Abstract

Thymosin-alpha1 is used as an adjuvant in combination with carmustine (BCNU) as an effective treatment for malignant glioblastoma.

Description

[0001] Related application materials [0002] This application claims priority to provisional application 60 / 337,149, filed December 10,2001. Background of the invention [0003] Glioblastoma is the most common primary CNS malignancy in adults, accounting for nearly 75% of primary CNS malignancies. Although the treatment of glioblastoma has steadily improved due to advances in neuroimaging, microsurgery, and radiation therapy, the disease remains incurable (McDonald, 2001; Burton, 2000; Prados, 2000). The average life expectancy after diagnosis is less than 1 year, and the five-year survival rate is less than 10% after urgent treatment, including resection of visible tumors (Burton, 2000; Nieder, 2000; Napolitano, 1999; Dazzi, 2000). Glioblastoma causes death by rapid, aggressive and infiltrating growth in the brain. The invasive growth property is the reason why these tumors cannot be resected. [0004] Glioblastoma is also relatively resistant to radiotherapy and chemoth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/00A61K31/17A61K31/175A61K38/22A61K45/06A61P35/00
CPCA61K31/17A61K31/175A61K45/06A61K38/2292A61P35/00A61K2300/00A61K38/16A61K31/64
Inventor 杰克·R·万兹苏珊·德拉·蒙特
Owner RHODE ISLAND HOSPITAL
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