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Application of recombined ganoderma lucidum immnoregulation protein (rLZ-8) in preparation of medicine for treating melanin tumor

An immunoregulatory protein and melanoma technology, applied in the field of biomedicine, can solve the problems of rLZ-8 anti-tumor level research reports, etc., and achieve the effect of no drug resistance, strong toxic side effects, and good safety

Inactive Publication Date: 2013-12-04
张喜田 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Ganoderma lucidum immunoregulatory protein (LZ-8) is a member of the fungal immunomodulatory protein family. Studies have shown that LZ-8 (recombinant LZ-8, rLZ-8) obtained through genetic engineering recombination can effectively kill human Gastric adenocarcinoma tumor cells (SGC-7901) and human lung cancer tumors (A549), it is not believed that the immunoregulatory protein rLZ-8 can directly kill tumor cells in vivo, so there is still no research report on the anti-tumor level of rLZ-8 in vivo

Method used

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  • Application of recombined ganoderma lucidum immnoregulation protein (rLZ-8) in preparation of medicine for treating melanin tumor
  • Application of recombined ganoderma lucidum immnoregulation protein (rLZ-8) in preparation of medicine for treating melanin tumor
  • Application of recombined ganoderma lucidum immnoregulation protein (rLZ-8) in preparation of medicine for treating melanin tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: rLZ-8 kills orthotopic melanoma in mice and maintains the number of white blood cells in the body and improves the living conditions of tumor-bearing mice

[0018] experimental method

[0019] (1) Experimental materials and reagents

[0020] Kunming male mice aged 6-8 weeks, weighing 18-22 g, were purchased from the Experimental Animal Center of Bethune Medical University and raised under SPF conditions of Northeast Normal University. During the experiment, the temperature was controlled at (20 ± 2) °C, the humidity was 48%, and the lighting was alternated for 12 hours. Mouse melanoma B16-F10 cell line.

[0021] DMEM, fetal bovine serum, PBS, trypsin-EDTA, DMSO, 0.9% NaCl solution, pH7.6 Tris-HCl buffer solution for washing, 0.05% trypsin, rLZ-8, dacarbazine (positive control drug, etc.).

[0022] (2) Instruments, equipment and utensils

[0023] CO 2 Constant temperature incubator, inverted microscope, pipette, tweezers, ultra-clean bench, blood cell cou...

Embodiment 2

[0048] Example 2: rLZ-8 inhibits the growth of melanoma lung metastases and the life extension rate of melanoma lung metastases mice

[0049] experimental method

[0050] (1) Experimental materials and reagents

[0051] Kunming male mice aged 6-8 weeks, weighing 18-22 g, were purchased from the Experimental Animal Center of Bethune Medical University and raised under SPF conditions of Northeast Normal University. During the experiment, the temperature was controlled at (20 ± 2) °C, the humidity was 48%, and the lighting was alternated for 12 hours. Mouse melanoma B16-F10 cell line. DMEM, fetal bovine serum, PBS, trypsin-EDTA, DMSO, pH7.6 Tris-HCl buffer washing, 0.05% trypsin, rLZ-8, dacarbazine, etc.

[0052] (2) Instruments, equipment and utensils

[0053] Same as Example 1

[0054] (3) Experimental grouping and administration methods

[0055] Mouse melanoma cells B16-F10 were cultured in DMEM medium containing 10% fetal bovine serum at 37 °C, CO 2 cultured in an incu...

Embodiment 3

[0065] Example 3: Production of antibodies and neutralizing antibodies after repeated administration of rLZ-8 in cynomolgus monkeys

[0066] experimental method:

[0067] As rLZ-8 is a recombinant genetic engineering drug derived from fungi, tracking its antibody production is an important part of preclinical evaluation. The serum of the normal cynomolgus monkey group was selected to detect rLZ-8 antibody by ELISA method, and the neutralizing activity of anti-rLZ-8 antibody was detected by biological activity method.

[0068] Experimental results:

[0069] From the 9th to the 11th day after the administration, the three cynomolgus monkeys all developed low-titer (1:5) anti-rLZ-8 antibodies. The scale is 1:25 to 1:125. No anti-rLZ-8 antibody was detected in the cynomolgus monkeys in the control group. The effect of anti-rLZ-8 antibody positive (1:125) monkey serum (diluted 10 times) and monkey serum-free culture medium on the secretion and expression of IFN-γ stimulated by ...

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Abstract

The invention relates to application of recombined ganoderma lucidum immnoregulation protein (rLZ-8) in preparation of medicine for treating melanin tumor. In-situ tumor and metastatic tumor experiment animal models are used to research anti-tumor effect of rLZ-8, and the results show that the rLZ-8 can evidently inhibit in-situ tumor growing and metastasis forming of the melanin tumor.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of recombinant Ganoderma lucidum immunoregulatory protein (rLZ-8) in inhibiting the growth and metastasis of melanoma. Background technique [0002] Malignant melanoma (Malignant melanoma), also known as melanoma (Melanoma) or melanosar-coma (Melanosar-coma), is a highly malignant melanocyte tumor that mostly occurs in the skin and also in the mucous membranes close to the skin ( Such as conjunctiva, oral cavity, nasal cavity, anal canal, rectum, cervix, vagina, penis, glans, etc.). The European "Update of the Guideline on the Diagnosis and Treatment of Melanoma 2012" states that the incidence of melanoma is increasing and the incidence in Europe is 10-20 / 10 5 , 20-30 / 10 in the US 5 , with an incidence rate of 50-60 / 10 in Australia 5 . The incidence of melanoma in yellow-skinned Chinese is <1 / 10 5 . According to the statistics of the World Health Organization (WHO...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61P35/00
CPCA61K38/16A61P35/00A61P35/04A61K36/06
Inventor 孙非梁重阳张喜田
Owner 张喜田
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