Use of small molecule polypeptide TP-7 in preparation of medicines for treating chronic kidney diseases

Abstract

The present invention relates to a small molecule polypeptide TP-7 having a function of inhibiting a renal fibrosis-promoting function of extracellular matrix tenascin C (TNC) and particularly to a use of the small molecule polypeptide TP-7 in preparation of medicines for treating chronic kidney diseases (CKD). An amino acid sequence of the polypeptide TP-7 is shown in SEQ ID NO.1. In multiple mouse kidney fibrosis models, the TP-7 has a significant effect, can obviously restore renal functions, inhibits progress of renal fibrosis, is also free of obvious toxic and side effects, and thus can be prepared into pharmaceutical preparations for treating the chronic kidney diseases.

Description

technical field

[0001] The present invention relates to the use of a small molecule polypeptide, the polypeptide is a partial peptide segment of Tenascin C (TNC), named TP-7, and specifically relates to the use of the small molecule polypeptide TP-7 in the preparation of chronic kidney disease drugs the use of. Background technique

[0002] Chronic Kidney Disease (CKD) refers to the disorder of chronic kidney structure and function caused by various reasons. In recent years, its incidence and prevalence have increased sharply, and it is another disease that seriously endangers human health after cardiovascular and cerebrovascular diseases, diabetes and malignant tumors. Globally, the number of people with chronic kidney disease progressing to end-stage renal disease (ESRD) is increasing at a rate of 5-6% per year. Patients need dialysis treatment to maintain their lives, which costs a lot and brings great harm to families and society. come a heavy economic burden. How to ...

Examples

Embodiment 1

[0019] Example 1: The source of the small molecule polypeptide TP-7

[0020] According to the partial amino acid composition of the FNⅢ4 domain of tendin C (TNC), a small molecular polypeptide composed of 30 amino acids was artificially synthesized by Hangzhou Dangang Biotechnology Co., Ltd., named TP-7.

[0021] The amino acid sequence of the small molecule polypeptide is as follows:

[0022] SEQ ID NO. 1: EWRNGKAAIDSYRIKYAPISGGDHAEVDVP.

[0023] After sequencing verification, the small molecule polypeptide TP-7 synthesized in this example was used in the experiments of the following examples.

Embodiment 2

[0024] Example 2: TP-7 inhibits the activation and proliferation of fibroblasts caused by tenascin C (TNC) in vitro

[0025] 1. Experimental materials

[0026] Cells: Rat kidney fibroblasts (NRK-49F).

[0027] Medium: DMEM / F12 (1:1) medium containing 10% FBS.

[0028] Culture conditions: 37°C with 5% CO 2 incubator.

[0029] 2. Experimental Treatment

[0030] Rat kidney fibroblasts (NRK-49F) were treated with 1.5×10 6 Sow in a 6cm culture dish, culture for 1 day, culture in serum-free DMEM / F12 (1:1) medium for 12 hours, add tenascin C recombinant protein (50ng / ml) to stimulate the cells, and add small molecule polypeptide TP-7 at the same time (50ng / ml) co-incubated for 48 hours to collect the protein for Western blot experiment.

[0031] 3. Experimental results

[0032] Experimental results such as Figure 1A to Figure 1E As shown, the TP-7 small molecule polypeptide can effectively inhibit the activation and proliferation of fibroblasts caused by tenascin C (TNC).

Embodiment 3

[0033] Example 3: The renoprotective effect of the small molecule polypeptide TP-7 on the mouse kidney unilateral ischemia / reperfusion model

[0034] 1. Experimental animals

[0035] BABL / c mice, male, 8 weeks old, weighing 20-22g, SPF grade.

[0036] First weigh the animals, put earrings numbered, select 18 healthy mice with a weight of 20-22g, and randomly divide them into 3 groups with 6 mice in each group, which are sham operation group and unilateral renal ischemia / reperfusion The model group and the TP-7 group were injected after renal unilateral ischemia / reperfusion modeling.

[0037] 2. Experimental grouping

[0038] 1) Sham-operated group: After anesthetizing mice with 1% pentobarbital sodium at 1ml / kg body weight at 26 degrees Celsius, choose the midline abdominal incision; after local disinfection, cut the skin, subcutaneous, muscle layer and peritoneum layer by layer, and find The lateral renal pedicles were then sutured layer by layer. After local disinfection...

technical field

[0001] The present invention relates to the use of a small molecule polypeptide, the polypeptide is a partial peptide segment of Tenascin C (TNC), named TP-7, and specifically relates to the use of the small molecule polypeptide TP-7 in the preparation of chronic kidney disease drugs the use of. Background technique

[0002] Chronic Kidney Disease (CKD) refers to the disorder of chronic kidney structure and function caused by various reasons. In recent years, its incidence and prevalence have increased sharply, and it is another disease that seriously endangers human health after cardiovascular and cerebrovascular diseases, diabetes and malignant tumors. Globally, the number of people with chronic kidney disease progressing to end-stage renal disease (ESRD) is increasing at a rate of 5-6% per year. Patients need dialysis treatment to maintain their lives, which costs a lot and brings great harm to families and society. come a heavy economic burden. How to ...

Examples

Embodiment 1

[0019] Example 1: The source of the small molecule polypeptide TP-7

[0020] According to the partial amino acid composition of the FNⅢ4 domain of tendin C (TNC), a small molecular polypeptide composed of 30 amino acids was artificially synthesized by Hangzhou Dangang Biotechnology Co., Ltd., named TP-7.

[0021] The amino acid sequence of the small molecule polypeptide is as follows:

[0022] SEQ ID NO. 1: EWRNGKAAIDSYRIKYAPISGGDHAEVDVP.

[0023] After sequencing verification, the small molecule polypeptide TP-7 synthesized in this example was used in the experiments of the following examples.

Embodiment 2

[0024] Example 2: TP-7 inhibits the activation and proliferation of fibroblasts caused by tenascin C (TNC) in vitro

[0025] 1. Experimental materials

[0026] Cells: Rat kidney fibroblasts (NRK-49F).

[0027] Medium: DMEM / F12 (1:1) medium containing 10% FBS.

[0028] Culture conditions: 37°C with 5% CO 2 incubator.

[0029] 2. Experimental Treatment

[0030] Rat kidney fibroblasts (NRK-49F) were treated with 1.5×10 6 Sow in a 6cm culture dish, culture for 1 day, culture in serum-free DMEM / F12 (1:1) medium for 12 hours, add tenascin C recombinant protein (50ng / ml) to stimulate the cells, and add small molecule polypeptide TP-7 at the same time (50ng / ml) co-incubated for 48 hours to collect the protein for Western blot experiment.

[0031] 3. Experimental results

[0032] Experimental results such as Figure 1A to Figure 1E As shown, the TP-7 small molecule polypeptide can effectively inhibit the activation and proliferation of fibroblasts caused by tenascin C (TNC).

Embodiment 3

[0033] Example 3: The renoprotective effect of the small molecule polypeptide TP-7 on the mouse kidney unilateral ischemia / reperfusion model

[0034] 1. Experimental animals

[0035] BABL / c mice, male, 8 weeks old, weighing 20-22g, SPF grade.

[0036] First weigh the animals, put earrings numbered, select 18 healthy mice with a weight of 20-22g, and randomly divide them into 3 groups with 6 mice in each group, which are sham operation group and unilateral renal ischemia / reperfusion The model group and the TP-7 group were injected after renal unilateral ischemia / reperfusion modeling.

[0037] 2. Experimental grouping

[0038] 1) Sham-operated group: After anesthetizing mice with 1% pentobarbital sodium at 1ml / kg body weight at 26 degrees Celsius, choose the midline abdominal incision; after local disinfection, cut the skin, subcutaneous, muscle layer and peritoneum layer by layer, and find The lateral renal pedicles were then sutured layer by layer. After local disinfection...

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