Polypeptides regulating sugar metabolism and uses thereof

A technology of glucose metabolism and peptides, applied in the field of biomedicine, can solve problems such as deficiency and insulin resistance

Active Publication Date: 2021-12-21
深圳中科新进生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Type 2 diabetes, also known as non-insulin-dependent diabetes, usually develops after the age of 35-40. The ability to produce insulin in the body is not completely lost or even compensatoryly increased, but insulin resistance occurs in the body, resulting in a relative deficiency.
[0003] Currently, there is no cure for type 2 diabetes, and the drug must be accompanied for a lifetime and has other side effects. Therefore, the current treatment of diabetes still has great defects, and new drugs with better performance need to be developed.

Method used

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  • Polypeptides regulating sugar metabolism and uses thereof
  • Polypeptides regulating sugar metabolism and uses thereof
  • Polypeptides regulating sugar metabolism and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0082] DMEM medium was purchased from Sigma, wherein every 500ml medium supplemented with 10% FBS (fetal bovine serum), 1% non-essential amino acids, 1g glucose, 0.75g sodium bicarbonate, 0.1g bovine serum albumin and 1.5ml HEPES (4 -Hydroxyethylpiperazineethanesulfonic acid).

[0083] ISAPs 1 The sequence is Tyr-Leu-Gly-Ala-Ser-Val-Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro-Thr (SEQ ID NO. 22). ISAPs 2 The sequence is Tyr-Leu-Tyr-Gln-Trp-Leu-Gly-Ala-Ser-Val-Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro (SEQ ID NO.2). ISAPs 3 The sequence of is Tyr-Leu-Tyr-Gln-Trp-Leu-Gly-Ala-Ser-Val-Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro-Arg (SEQ ID NO.3). ISAPs 4 The sequence is Ser-Val-Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro (SEQ ID NO.15). ISAPs 5 The sequence is Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro (SEQ ID NO.16). ISAPs 6 The sequence is Pro-Asp-Pro-Leu-Glu-Pro (SEQ ID NO.14).

[0084] βTC-6 was purchased from the American Type Culture Collection (ATCC). This cell line is derived from mouse pancreatic cancer and has introd...

Embodiment 1

[0087] ISAPs 1 function of

[0088] 1. ISAPs 1 Effects on βTC-6 cells.

[0089] The experimental steps are as follows:

[0090] 1. Cell plating: Dilute the βTC-6 cell suspension at 1.6×10 5 / mL density plated in 24-well plates, 0.5 mL per well, at 37 °C, 5% CO 2 Incubate for 24 hours.

[0091] 2. Dilute ISAP with sugar-free DMEM 1 Dilute to 2nmol / L, 200pmol / L, 20pmol / L, 2pmol / L respectively and store in ice bath.

[0092] 3. Perform starvation pretreatment on βTC-6 cells, using KRBB buffer (129g NaCl, 5g NaHCO 3 ,4.8gKCl,1.2gKH 2 PO 4 ,1.2g MgSO 4 ,2.5g CaCl 2 , 10mM HEPES and 0.1% BSA at pH 7.4) for 4 hours.

[0093]4. The cell culture medium was discarded, and ISAP containing different concentrations were added to the experimental group and the positive control group respectively. 1 and glucose culture solution, the negative control group only added medium (blank), at 37 ° C, 5% CO 2 Neutralize for 1 hour.

[0094] 5. Wash the cells at the bottom of the plate ...

Embodiment 2

[0123] ISAPs 2 The function ISAP 2 Binding to human GPRC6A

[0124] 2.1 Overexpression of hGPRC6A in Hela cells 1) Cell plating: Hela cell suspension was mixed with 1.6×10 5 / mL density plated in 6-well plates, 2 mL per well of DMEM medium, at 37 °C, 5% CO 2Incubate for 24 hours.

[0125] 2) Using Lipofectamine2000 (Invitrogen), the hGPRC6 overexpression vector (pReceiver-M61) was transfected into Hela cells according to the manufacturer's instructions, and the normal medium was replaced 4 hours after transfection.

[0126] 3) 48 hours after transfection, discard the medium, wash the cells at the bottom of the plate twice with sterile PBS, add 300 μL of Trizol solution, extract cellular RNA according to the instructions of RNAiso Plus (TaKaRa), after DNase treatment, use SuperScript T (Invitrogen, Canada) kit was reverse transcribed into cDNA by DNAEngine. Using cDNA as a template, use SYBRGr ee The n method was used to monitor and analyze the fluorescent PCR products at...

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PUM

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Abstract

The invention relates to a polypeptide regulating sugar metabolism and its use in the preparation of medicines for treating diseases related to sugar metabolism. The polypeptide of the present invention can lower blood sugar and promote insulin secretion; it can be used to prepare medicine for treating diabetes, and can be administered in combination with existing treatment schemes. The polypeptide of the present invention has the remarkable advantages of low therapeutically effective dose, oral administration and the like.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a polypeptide regulating sugar metabolism and its use in preparing medicines for treating diseases related to abnormal sugar metabolism. Background technique [0002] Diabetes is a major and growing health problem worldwide. Diabetes mellitus is a chronic disease of carbohydrate, fat and protein metabolism disorder caused by relative or absolute deficiency of insulin or decreased sensitivity of target cells to insulin, or structural defects of insulin itself. Diabetes is clinically manifested as polydipsia, polyphagia, polyuria, and weight loss, accompanied by various complications such as ketoacidosis, limb gangrene, polyneuritis, blindness, and renal failure. According to the pathogenesis, diabetes can be divided into type 1 and type 2, and 95% of the patients are type 2. Type 2 diabetes, also known as non-insulin-dependent diabetes, usually occurs after the age of 35-40. The abili...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/12A61K38/17A61P3/08A61P3/10A61P5/50
CPCC07K14/4728A61K38/00
Inventor 任培根张键滕斌李健姚振宇
Owner 深圳中科新进生物科技有限公司
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