CN114605517B

CN114605517B - Polypeptide LXP-7 with broad-spectrum anticancer effect and application thereof

Info

Publication number: CN114605517B
Application number: CN202210511432.3A
Authority: CN
Inventors: 梁海鹰; 熊继先; 申铖皓
Original assignee: Shenzhen University; Guangdong Ocean University
Current assignee: Shenzhen University; Guangdong Ocean University
Priority date: 2022-05-12
Filing date: 2022-05-12
Publication date: 2022-09-27
Anticipated expiration: 2042-05-12
Also published as: CN114605517A

Abstract

The invention provides a polypeptide LXP-7 with broad-spectrum anticancer effect and application thereof, belonging to the technical field of medicines. The amino acid sequence of the polypeptide LXP-7 with broad-spectrum anticancer effect is shown as SEQ ID NO. 1. The polypeptide LXP-7 of the invention can effectively kill various tumor cells, including lung cancer, colorectal cancer, liver cancer, stomach cancer, breast cancer, esophageal cancer and prostate cancer, has anticancer effect on various tumors, and can be used for preparing broad-spectrum tumor treatment medicines. Meanwhile, the LXP-7 has a remarkable killing effect on paclitaxel-resistant prostate cancer cells (PC 3/Tax), and can be used for preparing paclitaxel-resistant prostate cancer treatment medicines.

Description

Polypeptide LXP-7 with broad-spectrum anticancer effect and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a polypeptide LXP-7 with a broad-spectrum anticancer effect and application thereof.

Background

The current cancer treatment principle is comprehensive treatment mainly based on surgical treatment, including radiotherapy, chemotherapy, targeted therapy, immunotherapy and the like. Current cancer drugs are poorly effective and expensive, and most patients die due to drug resistance and recurrent metastasis of the tumor.

The peptide substance is a peptide mixture which naturally exists in organisms such as animals, plants and microorganisms, or exists in the modes of enzymolysis, artificial chemical synthesis, biological engineering and the like of animal and plant proteins, and has extremely strong activity and various biological functions. Anticancer polypeptides (ACP) have the characteristics of targeting property, safety, specificity, easiness in synthesis and customization, low cost and the like, are concerned in the development of antitumor drugs, and open up a new prospect for cancer treatment. ACP drugs have been studied extensively for many years in vitro, in vivo, and at various stages of the clinic, but only a few have been ultimately used for clinical treatment. Therefore, the discovery and identification of new anti-cancer polypeptides would provide new anti-tumor drug approaches.

Disclosure of Invention

In view of the above, the present invention aims to provide a polypeptide LXP-7 with broad-spectrum anticancer effect and still better killing effect on drug-resistant tumors.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a polypeptide LXP-7 with broad-spectrum anticancer effect, and the amino acid sequence of the polypeptide LXP-7 is shown as SEQ ID NO. 1.

The invention also provides an application of the polypeptide in preparing an anti-tumor product.

Preferably, the product comprises a pharmaceutical product.

Preferably, the tumor includes, but is not limited to: lung cancer, colorectal cancer, liver cancer, stomach cancer, breast cancer, esophageal cancer, and prostate cancer.

Preferably, the prostate cancer comprises paclitaxel resistant prostate cancer.

The invention also provides a medicine with anticancer effect, which comprises the polypeptide LXP-7.

Preferably, the administration of the pharmaceutical product includes, but is not limited to: oral administration and injection.

The invention has the beneficial effects that:

the polypeptide LXP-7 provided by the invention can effectively kill various tumor cells including lung cancer, colorectal cancer, liver cancer, gastric cancer, breast cancer, esophageal cancer and prostate cancer, shows that LXP-7 has an anticancer effect on various tumors, and can be used for preparing broad-spectrum tumor treatment medicines. In addition, the polypeptide LXP-7 has a remarkable killing effect on paclitaxel-resistant prostate cancer cells, can be used for preparing paclitaxel-resistant prostate cancer treatment medicines, and the effect of the LXP-7 on tumor cells is time-dependent and concentration-dependent.

The invention provides a new medicine and a scheme for clinical treatment of tumors, and has important significance for improving the clinical treatment effect of tumors and improving the life quality of patients.

Drawings

FIG. 1 shows the results of HPLC purification;

FIG. 2 shows the MS identification result of mass spectrum;

FIG. 3 shows the killing effect of LXP-7 on different tumor cells;

FIG. 4 shows the killing effect of LXP-7 at different concentrations on different cancer cells;

FIG. 5 shows the results of LXP-7 at different concentrations for different times on colorectal cancer cells (HCT 8).

Detailed Description

The polypeptide LXP-7 with broad-spectrum anticancer effect is designed according to a marjoram's pinctada histone PmH2A sequence, wherein the histone PmH2A sequence is as follows: MSGRGKGGKTKGKAKSRSSRAGLQFPVGRIHRLLRKGNYAERVGAGAPVYLAAVLEYLAAEVLELAGNAARDNKKSRIIPRHLQLAIRNDEELNKLLSGVTIAQGGVLPNIQAVLLPKKTQKPAK (SEQ ID No. 2), wherein the bold part is a reference sequence designed for a polypeptide of the invention. The polypeptide LXP-7 is obtained by adding a KLLK overlapping sequence on the basis of truncation PmH2A, and consists of 21 amino acid residues, wherein the amino acid sequence is as follows: RAGLQFPVGKLLKKLLKKLLK (SEQ ID No. 1). The preparation method of the polypeptide LXP-7 is not particularly limited, and any method for synthesizing the polypeptide which is conventional in the field can be adopted.

The present invention is not particularly limited with respect to the specific type of product, and preferably includes a pharmaceutical product. In the present invention, the tumor preferably includes, but is not limited to: lung cancer, colorectal cancer, liver cancer, stomach cancer, breast cancer, esophageal cancer and prostate cancer, preferably including paclitaxel-resistant prostate cancer.

In the invention, the medicine contains LXP-7 as a main active ingredient and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is not particularly limited, and any pharmaceutically acceptable carrier that is conventional in the art may be used. In the present invention, the administration mode of the pharmaceutical product preferably includes, but is not limited to: oral administration and injection, the injection preferably includes intratumoral injection, intradermal injection, subcutaneous injection, intramuscular injection and intravenous injection.

The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.

Example 1

The polypeptide LXP-7 is synthesized by adopting a conventional polypeptide synthesis method in the field, and is purified and identified by High Performance Liquid Chromatography (HPLC) and mass spectrum MS. The results are shown in fig. 1 and fig. 2, respectively. After High Performance Liquid Chromatography (HPLC) purification, the purity of the detected sample is more than 92.7%, mass spectrum MS identification is correct, and the molecular weight is 2392.13.

Example 2

Taking lung cancer cells (PC 9 and A549), colorectal cancer cells (HCT 116 and HCT 8), liver cancer cells (Huh 7), gastric cancer cells (AGS, HGC-27, MGC-803 and BGC-823), breast cancer cells (MDA-MB-231), esophageal cancer cells (KYSE 520), prostate cancer cells (PC 3) and paclitaxel-resistant prostate cancer cells (PC 3/Tax), wherein the concentration of the cancer cells is 1 × 10 ⁵ cells/mL, 100. mu.L each was inoculated into a 96-well plate, 3 wells per group, and each cell line was treated with LXP-7 (20. mu.M) purified in example 1 for 72 hours, with PBS as a control. CCK-8 was added to measure absorbance (A450) at a wavelength of 450nm, and cell growth curves were plotted for each group. The killing effect of LXP-7 on various tumor cells was compared. The results are shown in FIG. 3. As can be seen from figure 3, LXP-7 can effectively kill various tumor cells, has a significant killing effect on paclitaxel-resistant prostate cancer cells (PC 3/Tax), and has a significant effect on paclitaxel-resistant prostate cancer patients.

Example 3

Collecting lung cancer cells (PC 9 and A549), colorectal cancer cells (HCT 116 and HCT 8), gastric cancer cells (HGC-27), and breast cancer cells (MDA-MB-231), wherein the concentration of the above cancer cells is 1 × 10 ⁵ cells/mL, 100. mu.L each was inoculated into a 96-well plate, 3 wells per group, and each cell line was treated with LXP-7 (0, 0.5, 1, 2.5, 5, 10, 20. mu.M) purified according to example 1 at different concentrations for 72 h, with PBS as a control. Adding CCK-8 to measure the absorbance value (A4) with the wavelength of 450nm50) Drawing the growth curve of each group of cells, and calculating the IC ₅₀ The value is obtained. The results are shown in FIG. 4. As can be seen from FIG. 4, LXP-7 of the present invention is concentration dependent on the killing of different cancer cells and has IC's for different cancer cells ₅₀ The values were different as shown in table 1.

TABLE 1 IC of LXP-7 on different cancer cells ₅₀ Value of

Example 4

Collecting colorectal cancer cell (HCT 8) (1 × 10) ⁵ cells/mL, 100 μ L) were plated in 96-well plates, 3 wells per group, and cells were treated with different concentrations of LXP-7 (0, 0.5, 1, 2.5, 5, 10, 20 μ M) for 24, 48, 72 h, PBS as control. CCK-8 was added to measure absorbance (A450) at a wavelength of 450nm, and cell growth curves were plotted for each group. The results are shown in FIG. 5, which shows that LXP-7 is time-dependent in killing cancer cells.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Sequence listing

<110> Guangdong ocean university

SHENZHEN University

<120> polypeptide LXP-7 with broad-spectrum anticancer effect and application thereof

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 21

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 1

Arg Ala Gly Leu Gln Phe Pro Val Gly Lys Leu Leu Lys Lys Leu Leu

1 5 10 15

Lys Lys Leu Leu Lys

20

<210> 2

<211> 125

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 2

Met Ser Gly Arg Gly Lys Gly Gly Lys Thr Lys Gly Lys Ala Lys Ser

1 5 10 15

Arg Ser Ser Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Ile His Arg

20 25 30

Leu Leu Arg Lys Gly Asn Tyr Ala Glu Arg Val Gly Ala Gly Ala Pro

35 40 45

Val Tyr Leu Ala Ala Val Leu Glu Tyr Leu Ala Ala Glu Val Leu Glu

50 55 60

Leu Ala Gly Asn Ala Ala Arg Asp Asn Lys Lys Ser Arg Ile Ile Pro

65 70 75 80

Arg His Leu Gln Leu Ala Ile Arg Asn Asp Glu Glu Leu Asn Lys Leu

85 90 95

Leu Ser Gly Val Thr Ile Ala Gln Gly Gly Val Leu Pro Asn Ile Gln

100 105 110

Ala Val Leu Leu Pro Lys Lys Thr Gln Lys Pro Ala Lys

115 120 125

Claims

1. An application of polypeptide LXP-7 with broad-spectrum anticancer effect in the preparation of antitumor products is characterized in that the amino acid sequence of the polypeptide LXP-7 is shown as SEQ ID NO. 1; the tumor is paclitaxel-resistant prostate cancer.

2. The use of claim 1, wherein the product comprises a pharmaceutical product.