Application of interleukin 35 to preparation of drugs used for treating autoimmune dermatoses, therapeutic drug, and IL-35-Fc fusion protein

An autoimmune, fusion protein technology, applied in the field of biochemical drugs, can solve the problems of trouble, strong toxicity, easy recurrence, etc., and achieve the effects of inhibiting local infiltration, less toxic and side effects, and easy treatment

Inactive Publication Date: 2016-11-09
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the disadvantages of traditional methods for treating psoriasis are urgently needed to be solved in this field, such as troublesome treatment (laser therapy), strong toxic reaction (chemotherapy methotrexate) and easy recurrence (hormonal drug dexamethasone), etc., to develop new Effective psoriasis treatments

Method used

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  • Application of interleukin 35 to preparation of drugs used for treating autoimmune dermatoses, therapeutic drug, and IL-35-Fc fusion protein
  • Application of interleukin 35 to preparation of drugs used for treating autoimmune dermatoses, therapeutic drug, and IL-35-Fc fusion protein
  • Application of interleukin 35 to preparation of drugs used for treating autoimmune dermatoses, therapeutic drug, and IL-35-Fc fusion protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation of IL-35-Fc plasmid expression vector and expression and purification of IL-35-Fc protein

[0042] Construction of plasmid expression vector: pcDNA3.1 containing IL-35-Fc target gene sequence (coding sequence with signal peptide, its nucleotide sequence is shown in SEQ ID NO.6) synthesized by Nanjing GenScript -IL-35-Fc plasmid, using PCR to amplify the full-length sequence of the IL-35-Fc gene from the pcDNA3.1-IL-35-Fc plasmid, the amplified full-length sequence of the IL-35 gene is the same as that of pcDNA 3.4 TA cloning vectors were connected and transformed into competent bacteria. Colony PCR identification was performed on the grown clones, and then the positive clones identified by PCR were sequenced and analyzed. The correct comparison was the successful construction of recombinant IL-35- Fc expression plasmid pcDNA 3.4-IL35-Fc.

[0043] The expression plasmid pcDNA3.4-IL-35-Fc was transfected into suspension-cultured Expi293F cells (pur...

Embodiment 2

[0045] Example 2 Results of animal experiment evaluation of psoriasis mice treated with IL-35-Fc fusion protein of the present invention

[0046] Seven K14-VEGF transgenic mice (purchased from Jackson Lab (Bar Harbor, ME, USA)) (10 weeks old) in each group were divided into 2 groups, according to the expression kinetics of IL-35-Fc fusion protein in mice , we treated the mice once every other day, 10 times in total, 5 μg / mouse / time (pre-experimental injection doses were 2.5 μg / mouse / time, 5 μg / mouse / time and 10 μg / mouse / time, and finally Determine the optimal therapeutic dose is 5μg / only / time). After the treatment, the ear thickness of the mice was measured, and the ears of the mice were taken to weigh the ears, and the ear tissues of the mice were scored according to the PISA scoring system, including redness, scales, and erythema. The ear tissues were taken for H&E staining, and the ear tissues were scored according to the Baker scoring standard. This treatment experiment ...

Embodiment 3

[0050] Example 3 The animal experiment evaluation results of the IL-35-Fc fusion protein of the present invention in the treatment of imiquimod IMQ-induced psoriasis mice.

[0051] The purchased Babl / c mice were divided into two groups for treatment: IgG group, 8 mice; IL-35-Fc fusion protein (5 μg), 8 mice (IL-35 group shown in the figure); after a total of seven treatments Do model building. The back hair of the mice was shaved clean with a razor to expose the back skin, and then the back skin was smeared with 5% imiquimod cream (IMQ) (62.5 mg / mouse / time). Apply it continuously for 6 days, and observe the morbidity of mice of various races every day. During the modeling process, the above-mentioned treatment plan was continued for three times, and the skin tissue of the mice was collected on the second day after the last treatment. Skin tissue was used for histopathological examination, and H&E staining was used to evaluate the severity of psoriasis lesions. The results s...

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Abstract

The invention specifically relates to application of interleukin 35 to preparation of drugs used for treating autoimmune dermatoses, a therapeutic drug, and an IL-35-Fc fusion protein, belonging to the field of biochemical drugs. The objective of the invention is to provide a novel effective approach for treatment of autoimmune dermatoses. According to a technical scheme in the invention, application of interleukin 35 to preparation of drugs for treating psoriasis is provided. Results show that IL-35 has good treatment effect, small toxic and side effect and simple operation in treatment of psoriasis, so a novel option is provided for treatment of autoimmune dermatoses like psoriasis, chorionitis and pemphigus.

Description

technical field [0001] The invention belongs to the field of biochemical medicines, and in particular relates to the use of interleukin-35 in the preparation of medicines for treating autoimmune skin diseases, the medicine and a new IL-35-Fc fusion protein. Background technique [0002] Autoimmune skin diseases refer to diseases caused by the body's excessive immune response to self-antigens, resulting in severe damage to its own skin tissue. Autoimmune skin diseases mainly include psoriasis, scleroderma, and pemphigus. Psoriasis is a representative chronic autoimmune skin disease, which is greatly influenced by genetic factors. Clinically, psoriasis presents as diverse, prominent, and easily identifiable symptoms with prominent redness and erythema. The disease is mainly caused by increased vascular permeability of the dermis and up-regulated expression of vascular endothelial growth factor (VEGF). In recent years, new progress has been made on the complex pathogenesis an...

Claims

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Application Information

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IPC IPC(8): A61K38/20A61P37/06A61P17/06A61P17/00A61P17/02C07K19/00C12N15/62
CPCA61K38/208C07K14/54C07K2319/30
Inventor 邓洪新张俊凤黎一鸣魏于全
Owner SICHUAN UNIV
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