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Compounds for therapy and diagnosis for lung cancer and method for their use

A lung cancer and enhancer technology, applied in the field of peptides, can solve the problem of difficult treatment of lung cancer

Inactive Publication Date: 2001-04-18
CORIXA CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite considerable research into lung cancer treatment, lung cancer remains difficult to treat

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1 Preparation of Lung Tumor-Specific cDNA Sequences Using Differential RT-PCR

[0090] This example illustrates the preparation of cDNA molecules encoding lung tumor-specific polypeptides using differential screening.

[0091] Tissue samples were prepared from breast tumor tissues and normal tissues of lung cancer patients, and the determination of lung cancer was proved by pathological examination of tissue samples from the patients. Normal RNA and tumor RNA were extracted from samples, mRNA was isolated, and analyzed by using a (dT) 12AG (SEQ ID NO: 47) anchored 3' primer was converted into cDNA. Differential PCR was performed by using a randomly selected primer (SEQ ID NO: 48). Amplification conditions: containing 1.5mM MgCl 2 , 20 pmol primer, 500 pmol dNTP and 1 unit Taq DNA polymerase (Perkin-Elmer, Branchburg, NJ) standard buffer. Denaturation at 94°C for 30 seconds, annealing at 42°C for 1 minute, and extension at 72°C for 30 seconds completed 40 amp...

Embodiment 2

[0094] Identification of DNA sequences encoding lung tumor antigens using patient serum

[0095] This example exemplifies the isolation of cDNA sequences encoding lung tumor antigens by screening lung tumor samples by expression using autologous patient serum.

[0096] A human lung tumor-targeted cDNA library was constructed using the Lambda ZAP Express Expression System (Stratagene, La Jolla, CA). All RNA used for the library was obtained from human squamous cell lung cancer passaged from a dead SCID mouse, and poly A+ RNA was isolated by using the INFO labeling kit (Gibco BRL, Gaithersburg, MD). The resulting library was screened using autologous patient sera absorbed from E. coli as described in Sambrook et al., (Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1989), and the secondary antibody was alkaline phosphatase-conjugated Goat anti-human IgG-A-M (H+L), developed with NBT / BCIP (Gibco BRL). Positive plaques express...

Embodiment 3

[0100] Identification of DNA sequences encoding lung tumor antigens using mouse antisera

[0101] This example illustrates the isolation of cDNA sequences encoding lung tumor antigens by screening a lung tumor cDNA library with mouse antitumor serum.

[0102] A directed cDNA lung tumor expression library was prepared as described in Example 2. Sera were obtained from SCID mice containing post-passage human squamous epithelial and adenocarcinoma tumors. These sera were pooled and injected into normal mice to produce anti-tumor serum. Approximately 200,000 PFU were screened from the unamplified library using this antiserum. Using goat anti-mouse IgG-A-M (H+L) alkaline phosphatase secondary antibody, NBT / BCIP (BRL Labs) was used for color development, and about 40 positive phage plaques were identified. The plaques were purified, and the phagemids were cut into 9 clones, which were inserted into the pBK-CMV vector for expression in prokaryotic cells or eukaryotic cells.

[01...

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PUM

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Abstract

Compounds and methods for treating lung cancer are disclosed. The inventive compounds include polypeptides containing at least a portion of a lung tumor protein. Vaccines and pharmaceutical compositions for immunotherapy of lung cancer comprising such polypeptides, or polynucleotides encoding such polypeptides, are also disclosed, together with polynucleotides for preparing the inventive polypeptides.

Description

technical field [0001] The present invention generally relates to compositions and methods for treating lung cancer. The present invention particularly relates to nucleotide sequences predominantly expressed in lung cancer tissues, and polypeptides encoded by these nucleotide sequences. The inventive nucleotide sequence and polypeptide can be used in vaccines and pharmaceutical compositions for treating lung cancer. Background of the invention [0002] Lung cancer is the leading cause of cancer death in both men and women in the United States, with an estimated 172,000 new cases reported in 1994. Excluding the diagnosis period of the disease, the 5-year survival rate among all lung cancer patients is only 13%. This was in contrast to the five-year survival rate of 46 percent for cases whose tumors had not metastasized at the time of testing. However, only 16 percent of lung cancers are detected before the disease has spread. [0003] Early detection is difficult because ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61KA61K35/14A61K38/00A61K39/00A61K39/39G01N33/53A61K39/395A61P11/00A61P35/00C07HC07KC07K14/47C07K16/18C07K19/00C12NC12N1/19C12N1/21C12N5/10C12N15/09C12N15/12C12P21/08C12Q1/68C12R1/19G01NG01N33/574G01N33/577
CPCC12Q1/6883C12Q2600/158A61K38/00C07K14/47A61P11/00A61P35/00C12N15/117
Inventor S·G·里德M·J·洛迪斯T·N·弗鲁达奇斯R·莫哈玛斯
Owner CORIXA CORP
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