阅读说明：本技术 一种多肽及其在制备治疗和预防肿瘤的药物中的应用 (Polypeptide and application thereof in preparing medicine for treating and preventing tumors ) 是由 胡卓伟 崔冰 于金梅 王振贺 于 2019-03-24 设计创作，主要内容包括：本发明公开了一种靶向TRIB3/AKT相互作用的多肽或所述多肽的衍生物及其在制备治疗肿瘤的药物中的应用。所述多肽的氨基酸序列表SEQ ID No.1所示。所述多肽或多肽衍生物能够特异性地与TRIB3结合,从而阻断TRIB3/AKT1相互作用,促进FOXO1蛋白降解,抑制SOX2表达,因此应用于治疗和预防肿瘤的药物制备中。所制备的药物在治疗肿瘤疾病中,具有疗效显著,毒副作用少,使用安全的优点。(The invention discloses a polypeptide of a targeted TRIB3/AKT interaction or a derivative of the polypeptide and application thereof in preparing a medicament for treating tumors. The amino acid sequence table of the polypeptide is shown in SEQ ID No. 1. The polypeptide or the polypeptide derivative can be specifically combined with TRIB3, so that the interaction of TRIB3/AKT1 is blocked, FOXO1 protein degradation is promoted, and SOX2 expression is inhibited, therefore, the polypeptide or the polypeptide derivative is applied to preparation of medicines for treating and preventing tumors. The prepared medicine has the advantages of obvious curative effect, less toxic and side effect and safe use in treating tumor diseases.)

1. The polypeptide or the derivative thereof is characterized in that the amino acid sequence of the polypeptide is shown as SEQ ID No. 2.

2. The polypeptide or derivative thereof of claim 1, wherein the polypeptide or derivative thereof is modified conventionally.

3. The polypeptide or derivative thereof of claim 2, wherein the conventional modification comprises acetylation, amidation, cyclization, glycosylation, phosphorylation, alkylation, biotinylation, fluorophore modification, polyethylene glycol (PEG) modification, and immobilization modification.

4. The polypeptide or derivative thereof of claim 2, wherein the derivative comprises a chimeric peptide of the polypeptide and a cell-penetrating peptide, a fusion peptide of the polypeptide and a virus, and the methylated polypeptide.

5. The polypeptide or derivative thereof according to claim 1, wherein the polypeptide targets interaction with TRIB3/AKT, promotes FOXO1 protein degradation, inhibits SOX2 expression, and is used for targeted therapy of diseases related to TRIB 3/AKT.

6. Use of the polypeptide or derivative thereof according to claim 1 for the preparation of a medicament for the treatment and/or prevention of tumors.

7. The use according to claim 6, wherein the tumor is breast cancer or intestinal cancer.

8. The use of claim 7, wherein the breast cancer is a Luminal type breast cancer, HER2+ breast cancer, or Basal-like breast cancer; the intestinal cancer is colon cancer or rectal cancer.

9. A pharmaceutical composition comprising the polypeptide or derivative thereof of any one of claims 1 to 4 and a biologically or pharmaceutically acceptable carrier or excipient.

10. The pharmaceutical composition according to claim 9, which comprises the targeted TRIB3/AKT interaction according to any one of claims 1 to 4, a polypeptide that promotes FOXO1 protein degradation, inhibits SOX2 expression, or a derivative of the polypeptide as an active ingredient; or, it contains the targeted TRIB3/AKT interaction as described in any one of claims 1-4, promotes FOXO1 protein degradation, inhibits SOX2 expression polypeptide or its derivative and other compounds with anti-tumor activity as active ingredients.

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a polypeptide and application thereof in preparing a medicament for treating and preventing tumors.

Background

The tumor stem cells are the source of tumorigenesis and are the key factors of tumor recurrence, metastasis and drug resistance. At present, the treatment strategies of tumors mainly comprise operations and chemotherapy, and although the treatment strategies are successful, the survival rate of patients is seriously reduced due to drug resistance, relapse and metastasis phenomena generated in the treatment process. Therefore, targeting tumor stem cells is an important idea for solving the above problems. The tumor microenvironment is soil for maintaining tumor stem cells, can stimulate the tumor cells to highly express stress protein TRIB3, TRIB3/AKT1 interaction, inhibit the formation of p-AKT, further inhibit the generation of p-FOXO1, block the FOXO1 protein degradation pathway, accumulate FOXO1 in cell nuclei in a large amount, promote the expression level of stem cell transcription factor SOX2, and thus maintain the functions of various tumor stem cells.

Thus, as a key mechanism for the tumor microenvironment to maintain tumor stem cell function, the TRIB3/AKT1 interaction is a potential target for targeting tumor stem cells. The targeting TRIB3/AKT1 interaction promotes the generation of p-AKT, promotes the degradation of FOXO1, inhibits the expression of SOX2, and plays an important role in treating tumor recurrence and metastasis caused by various tumor stem cells and a drug resistance strategy.

Protein/protein interactions play important roles in a variety of biological functions and are also potential therapeutic targets for a variety of human diseases. The current drugs that block protein/protein interactions are a hot area of new drug development. Protein polypeptides and analogs thereof can act as leads to interference with protein/protein interactions.

The compound or polypeptide drug targeting TRIB3/AKT interaction can activate p-AKT, promote FOXO1 degradation, and inhibit the activity of SOX2 downstream target genes. Has strong targeting property, small side effect and good prospect of patent medicine for inhibiting the generation and development of tumors.

Disclosure of Invention

The invention aims to solve the technical problems that TRIB3/AKT causes high drug resistance rate of drugs and lacks of drugs for directly targeting tumor stem cells, and provides a polypeptide or a derivative thereof for recovering AKT phosphorylation, promoting FOXO1 protein degradation and inhibiting SOX2 expression and application thereof in preparing drugs for treating tumors.

The inventor of the invention discovers through intensive research and repeated experiments that the polypeptide Ae (amino acid sequence is shown in a sequence table SEQ ID No.2) capable of targeted inhibition of TRIB3/AKT interaction is obtained, and the polypeptide Ae can inhibit multiple tumor cells from balling in vitro and forming tumors in vivo, so that the polypeptide Ae can be applied to preparation of a medicine for treating tumors. Based on the research work of the inventor, the invention provides the following technical scheme.

One of the technical schemes provided by the invention is as follows: a polypeptide or a derivative of the polypeptide which targets TRIB3/AKT interaction, promotes FOXO1 protein degradation and inhibits SOX2 expression, wherein the amino acid sequence of the polypeptide is shown as the amino acid sequence in a sequence table SEQ ID No. 2.

The derivatives of the polypeptide are conventional derivatives in the field, and preferably comprise chimeric peptide formed by connecting the polypeptide and cell-penetrating peptide, fusion peptide formed by the polypeptide and virus, and conventional modifications of the polypeptide or the derivatives thereof, including acetylation, amidation, cyclization, glycosylation, phosphorylation, alkylation, biotinylation, fluorescent group modification, polyethylene glycol (PEG) modification and immobilization modification.

Among them, the cell-penetrating peptide of the present invention is a cell-penetrating peptide that is conventional in the art as long as it can assist in delivering the polypeptide into a cell to function. Generally, the cell-penetrating peptide is a short peptide molecule consisting of 10-30 amino acids.

Wherein, appropriate amino acid substitution, deletion or addition can be carried out in the amino acid sequence shown in SEQ ID No.2, as long as the modified amino acid sequence can still be specifically combined with TRIB3 and the activity before modification is kept.

The second technical scheme provided by the invention is as follows: an application of a polypeptide for targeting and promoting the interaction of TRIB3/AKT, promoting the degradation of FOXO1 protein and inhibiting the expression of SOX2 or a derivative of the polypeptide in preparing a medicament for treating and/or preventing tumors.

The tumor is a tumor which is conventional in the art. Preferably breast cancer and intestinal cancer. Wherein the breast cancer is Luminal type breast cancer, HER2+ breast cancer or Basal-like breast cancer; the intestinal cancer is colon cancer or rectal cancer.

Such prevention is conventional in the art and preferably means preventing or reducing the development of a tumor after use in the presence of a potential tumor agent. The treatment is conventional in the art and preferably means reducing the extent of the tumor, or curing the tumor to normalize it, or slowing the progression of the tumor.

The third technical scheme provided by the invention is as follows: an anti-tumor pharmaceutical composition, which contains the polypeptide or the derivative of the polypeptide for targeting and promoting TRIB3/AKT, promoting FOXO1 protein degradation and inhibiting SOX2 expression.

The active component is a compound with the function of preventing or treating tumors. In the pharmaceutical composition, the targeted TRIB3/AKT interaction promotes FOXO1 protein degradation, and the polypeptide inhibiting SOX2 expression or the polypeptide can be used as an active ingredient alone or together with other compounds with antitumor activity.

The administration route of the pharmaceutical composition of the present invention is preferably injection administration or oral administration. The injection administration preferably comprises intravenous injection, intramuscular injection, intraperitoneal injection, intradermal injection or subcutaneous injection and the like. The pharmaceutical composition is various dosage forms which are conventional in the field, preferably in the form of solid, semisolid or liquid, and can be aqueous solution, non-aqueous solution or suspension, and more preferably tablet, capsule, granule, injection or infusion, etc.

Preferably, the pharmaceutical composition of the present invention further comprises one or more pharmaceutically acceptable carriers. The medicinal carrier is a conventional medicinal carrier in the field, and can be any suitable physiologically or pharmaceutically acceptable medicinal auxiliary material. The pharmaceutical adjuvant is conventional in the field, and preferably comprises pharmaceutically acceptable excipient, filler or diluent and the like. More preferably, the pharmaceutical composition comprises 0.01-99.99% of the protein and 0.01-99.99% of a pharmaceutical carrier, wherein the percentage is the mass percentage of the pharmaceutical composition.

Preferably, the pharmaceutical composition is administered in an effective amount, which is an amount that alleviates or delays the progression of the disease, degenerative or damaging condition. The effective amount can be determined on an individual basis and will be based in part on the consideration of the condition to be treated and the result sought.

On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows: the polypeptide or the polypeptide derivative can target the interaction of TRIB3/AKT, promote the degradation of FOXO1 protein, and inhibit the activation of a FOXO1 downstream signal channel, thereby being applied to the preparation of antitumor drugs. The prepared medicine has the advantages of obvious curative effect, less toxic and side effect and safe use in treating tumor diseases.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

Unless otherwise stated, the PBS solution described in the examples refers to a phosphate buffer solution with a concentration of 0.1M, pH and a value of 7.2.