FGF4 and application thereof

A protein and polynucleotide technology, applied in the field of protein drugs, can solve the problems of FGF4 and other problems without evidence, and achieve the effects of improving glucose tolerance, facilitating drug production, and good adaptability

Active Publication Date: 2020-11-17
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although two endocrine FGFs (i.e., FGF21 and FGF19) and one paracrine FGF (FGF1) have been reported to exert multiple beneficial effects in obesity and T2D in an insulin-dependent manner, including: potent hypoglycemic activity, promoting energy consumption and lipid metabolism, however, there is no evidence that FGF4 has the same properties

Method used

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  • FGF4 and application thereof
  • FGF4 and application thereof
  • FGF4 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 Experimental material and method

[0034] 1. Expression and Purification of Recombinant FGF4 (rFGF4)

[0035] The cDNA fragment (Ala67-Leu206) encoding FGF4 was amplified by PCR, and subcloned into the bacterial expression vector pET-15b via NcoI and XhoI sites. After sequencing, the amino acid sequence of the encoded protein is shown in SEQ ID NO: 1 Show( figure 1 ). Competent E. coli BL-21(DE3) cells transformed with FGF4 expression constructs were cultured in an incubator shaker at 37 °C and 200 rpm. The medium contains 2% glucose and 30 mg / mL kanamycin (LB medium). at lambda 600Recombinant protein expression was induced by adding isopropyl-L-thio-β-D-galactopyranoside (IPTG) to 1 mM at an optical density of 0.8 to 1.0 and further grown at 37 °C for 4 h . Cells were harvested and lysed in 25 mM Na / K phosphate buffer (pH 7.5) containing 300 mM NaCl using an Emulsiflex-C3 (Avestin, Inc., Ottawa, Ontario, Canada) high capacity homogenizer. Recombinan...

Embodiment 2

[0055] Embodiment 2 Experimental results and conclusions

[0056] 1. rFGF4 can induce significant hypoglycemic effect

[0057] hyperglycemia db / db A single dose (i.p., 0.25 mg / kg, 0.5 mg / kg or 1 mg / kg body weight) of FGF4 was injected intraperitoneally in mice to examine its therapeutic potential and a dose-dependent reduction in blood glucose levels was found ( figure 2 A). At a dose of 1 mg / kg body weight, rFGF4 elicited a significant hypoglycemic effect 3 hours after injection and reached its maximum effect at 6 hours. This effect lasts up to 36 hours ( figure 2 B). In diet-induced obese (DIO) mice ( figure 2 C) and the second diabetic mouse model ( ob / ob ) A similar hypoglycemic effect was also observed in parallel experiments ( figure 2 D). More importantly, subcutaneous (s.c.) or intravenous (i.v.) injection of rFGF4 also lowered blood glucose levels in a similar manner ( figure 2 e). Therefore, we concluded that the hypoglycemic effect of rFGF4 has noth...

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Abstract

The invention belongs to the field of protein drugs, particularly relates to a fibroblast growth factor FGF4 and application thereof, and provides FGF4 protein as a novel powerful hypoglycemic and insulin sensitizer, and application of the FGF4 protein in treatment of diabetes and other diseases. The protein can quickly and durably regulate and control blood sugar, enhance the insulin sensitivityof a whole body, improve the glucose tolerance and reduce liver fatty degeneration and adipose tissue inflammation, and does not cause hypoglycemia or other obvious negative effects; the adaptabilityadministration routes of the protein is good; the protein is convenient for patent medicine; and the prospect of effectively treating diseases, such as diabetes, is widened.

Description

technical field [0001] The invention belongs to the field of protein medicine, in particular, the invention relates to fibroblast growth factor 4 and its variants, and their application in treating diseases such as diabetes. Background technique [0002] The incidence of type 2 diabetes (T2D) is increasing at an alarming rate globally and poses a huge health and economic burden. Genetic susceptibility and chronic obesity are the main risk factors for the development of T2D. In healthy individuals, insulin maintains glucose and lipid homeostasis by stimulating glucose uptake into various tissues including muscle, liver, and fat, and by inhibiting adipose tissue lipolysis. However, in T2D patients, the diminished action of insulin (known as insulin resistance) leads to the development of various metabolic disturbances, including hyperglycemia and dyslipidemia. This further disrupts insulin secretion and its function, leading to severe organ damage and eventually organ failur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/50C12N15/12A61P3/10A61P3/04A61P5/50
CPCC07K14/50A61P3/10A61P3/04A61P5/50A61K38/00
Inventor 黄志锋应磊王璐瑶侯煜姝周洁金辉李校堃穆萨·穆罕穆迪
Owner WENZHOU MEDICAL UNIV
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