CN119039397A

CN119039397A - Polypeptide targeting fibroblast activation protein and application thereof

Info

Publication number: CN119039397A
Application number: CN202411451074.7A
Authority: CN
Inventors: 苗林; 梁旭博; 邹升; 王登; 王新波; 李向群
Original assignee: Hunan Zhongsheng Whole Peptide Biotechnology Co ltd
Current assignee: Hunan Zhongsheng Whole Peptide Biotechnology Co ltd
Priority date: 2024-10-17
Filing date: 2024-10-17
Publication date: 2024-11-29

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a polypeptide of a targeted fibroblast activation protein and application thereof, and in particular relates to the polypeptide of the targeted fibroblast activation protein, wherein the amino acid sequence of the polypeptide is shown as any one of SEQ ID No. 1-SEQ ID No. 6. Experimental results prove that the polypeptide of the targeting fibroblast activation protein has excellent binding specificity and affinity, has good endocytosis aiming at cells with high expression of FAP, can be used as candidate targeting molecules of antitumor drugs, and is used for treating or diagnosing cancers related to abnormal activation of the fibroblast activation protein.

Description

Polypeptide targeting fibroblast activation protein and application thereof

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to a polypeptide targeting fibroblast activation protein and application thereof.

Background

Fibroblast activation protein (FAP, also known as fibroblast activation protein alpha, fapα) is highly overexpressed on cancer-associated fibroblasts (CAFs) in solid tumors, but is not generally expressed in normal tissues and benign tumors. Tumor stroma CAFs can promote the growth and infiltration of tumor cells, and has become an important target for tumor intervention. The over-expression of the tumor biomarker molecule FAP is a remarkable characteristic of CAFs, and FAP is a potential target in the aspect of CAF targeting for tumor diagnosis and treatment.

FAP, a type II transmembrane serine protease, belongs to the dipeptidyl peptidase 4 (DPP 4) family, has dipeptidyl peptidase (DPPs) and proline endopeptidase (PREPs) activities, whose role involves in the remodeling and fibrosis of the extracellular matrix (ECM). CAF surface-specific FAP can enhance tumor cell invasive capacity along fiber orientation by promoting matrix reconstruction, participating in VEGF/AKT/ERK signal transduction pathway, participating in tumor angiogenesis and the like to form tumor biological barrier and inhibit the function of effector T cells, thereby promoting tumor progression. Inducible overexpression of FAP in the tumor stroma is also dependent on malignant transformation of tumor tissue. The high expression level of FAP is positively correlated with poor prognosis of tumors. Thus, FAP has become a marker of active fibroblasts in tumors, granulation tissue, and fibrotic lesions.

At present, the clinical success rate of FAP-targeted therapy is not high, but the overexpression of FAP in many diseases suggests that FAP is a potential molecular diagnostic biomarker. Although FAP inhibitor molecules do not have good direct inhibition of tumors, some of them have good specificity and affinity for FAP, and these molecules can be used as structural frameworks for new drugs. The polypeptide FAP targeting inhibitor subjected to structural modification and optimization screening has great advantages and development value for diagnosis and treatment of cancers.

Disclosure of Invention

The invention aims to provide a polypeptide of targeting fibroblast activation protein and application thereof, and the polypeptide is targeted to be combined with the fibroblast activation protein overexpressed in tumor cells, and has important application value in tumor molecular diagnosis and targeted therapy.

In order to achieve the above object, according to one aspect of the present invention, there is provided a fibroblast activation protein-targeted polypeptide or a pharmaceutically acceptable salt thereof, wherein the amino acid sequence of the polypeptide is one of the amino acid sequences shown in SEQ ID No.1 to SEQ ID No.6, GCFRQCQTAWPAWDCFHHCG (SEQ ID No. 1),

GCVERCTTDFPQGAAACQAWCAG(SEQ ID No.2)、

GCHSLCVKYYQEAFCHSHCG(SEQ ID No.3)、

GCYRKCRANFNDLWCYKHCG(SEQ ID No.4)、

GCYRRCVTQYAAKWCLAHCG(SEQ ID No.5)、

GCYHSCSREWHPDTCRGWCG(SEQ ID No.6)。

In a second aspect, the present invention provides a polypeptide derivative which is a modified product of a polypeptide as described in the first aspect or a variant obtained by addition and/or substitution of one or more amino acids.

Preferably, the polypeptide derivative is a variant obtained by adding and/or replacing one, two or three amino acids of any one of the polypeptides shown in SEQ ID No. 1-SEQ ID No. 6.

Further preferably, the polypeptide derivative is a variant of the polypeptide shown in SEQ ID No.1, SEQ ID No.2, SEQ ID No.3 or SEQ ID No.4 obtained by adding and/or replacing an amino acid.

Preferably, the polypeptide derivative is an N-terminal or C-terminal modification product of the polypeptide shown in SEQ ID No.1, SEQ ID No.2, SEQ ID No.3 or SEQ ID No.4, wherein the modification group is selected from one or more of acetyl, amino, alkyl, aryl, fatty acid group, glycosyl, phosphate group, sulfate group, polyethylene glycol (PEG) group or peptide linking group.

Further preferably, the modification is selected from the group consisting of C-terminal amidation blocking and/or N-terminal modified acetyl.

In another aspect, the invention provides a polynucleotide encoding any of the FAP-targeted polypeptides or polypeptide derivatives described above.

In another aspect, the present invention provides a pharmaceutical formulation comprising as an active ingredient the FAP-targeted polypeptide described above or a pharmaceutically acceptable salt or polypeptide derivative or polynucleotide thereof and a pharmaceutical carrier.

In another aspect, the present invention provides the use of the FAP-targeted polypeptide described above, or a pharmaceutically acceptable salt, polypeptide derivative, polynucleotide or pharmaceutical formulation thereof, in the manufacture of a medicament for the prevention, diagnosis and/or treatment of cancer associated with abnormal activation of FAP.

Preferably, the cancer is selected from the group consisting of prostate cancer, breast cancer, pancreatic cancer, liver cancer, lung cancer, sarcoma, colorectal cancer, cholangiocellular carcinoma, chordoma, small intestine cancer, pheochromocytoma, gastric cancer, renal cancer, ovarian cancer, bladder cancer, esophageal cancer, head and neck cancer, thymus cancer, cervical cancer, endometrial cancer, neuroendocrine tumor, thyroid cancer, intestinal cancer, and solid tumors such as bone metastasis. Further, the cancer is selected from pancreatic cancer, thyroid cancer, liver cancer.

The beneficial effects are that:

1. The invention develops a series of novel high-affinity polypeptides of fibroblast activation protein, which can be used for targeting the fibroblast activation protein. The FAP receptor is utilized to be highly expressed in tumors, and can be used for preventing, diagnosing and/or treating FAP high-expression tumors.

2. The polypeptides are all low molecular weight polypeptides, the synthesis cost is low, and the endocytosis of the polypeptides in HEK293-FAP cells is obvious.

As used herein, "amino acid" refers to both natural and unnatural amino acids. The three-letter code containing the prefix "L-" or "D-" (except for achiral glycine) indicates the three-letter configuration of an amino acid, e.g., L-form amino acid such as alanine ("L-Ala" or "A"), arginine ("L-Arg" or "R"), asparagine ("L-Asn" or "N"), aspartic acid ("L-Asp" or "D"), cysteine ("L-Cys" or "C"), glutamine ("L-Gln" or "Q"), glutamic acid ("L-Glu" or "E"), glycine ("Gly" or "G"), histidine ("L-His" or "H"), isoleucine ("L-Ile" or "I"), leucine ("L-Leu" or "L"), lysine ("L-Lys" or "K"), methionine ("L-Met" or "M"), phenylalanine ("L-Phe" or "F"), proline ("L-Pro" or "P"), serine ("L-Asp" or "S"), threonine ("L-Thr" or "T"), glycine ("L-P" or "Tyr" or "L-Tyr" and Val "L-Tyr" or "L-Val". L-norleucine and L-norvaline can be represented as (NLeu) and (NVal), respectively. Nineteenth naturally occurring chiral amino acids have the corresponding D-isomer indicated by the three letter code containing the prefix "D-", alanine ("D-Ala" or "a"), arginine ("D-Arg" or "r"), asparagine ("D-Asn" or "a"), aspartic acid ("D-Asp" or "D"), cysteine ("D-Cys" or "c"), glutamine ("D-Gln" or "q"), glutamic acid ("D-Glu" or "e"), histidine ("D-His" or "h"), isoleucine ("D-Ile" or "i"), leucine ("D-Leu" or "l"), lysine ("D-Lys" or "k"), methionine ("D-Met" or "m"), phenylalanine ("D-Phe" or "f"), proline ("D-Pro" or "p"), serine ("D-Ser" or "s"), threonine ("D-Thr" or "t"), tryptophan ("D-Trp" or "w"), tyrosine ("D-Tyr" or "y"), and ("D-Val" v ").

"Unnatural amino acid" refers to any derivative of a natural amino acid, including alpha-and beta-amino acid derivatives. It should be noted that certain amino acids (e.g., hydroxyproline) that may be categorized as unnatural amino acids in the present invention may also be found in certain biological tissues or in certain proteins in nature. Amino acids having many different protecting groups suitable for direct use in solid phase peptide synthesis are commercially available. In addition to the twenty most common natural amino acids, exemplary unnatural amino acids and amino acid derivatives (common abbreviations in brackets) may be used in accordance with the invention, 2-aminoadipic acid (Aad), 3-aminoadipic acid (β -Aad), 2-aminobutyric acid (2-Abu), α, β -dehydro-2-aminobutyric acid (8-AU), 1-aminocyclopropane-1-carboxylic Acid (ACPC), aminoisobutyric acid (Aib), 3-aminoisobutyric acid (β -Aib), 2-aminothiazoline-4-carboxylic acid, 5-aminopentanoic acid (5-Ava), 6-aminocaproic acid (6-Ahx), 2-aminoheptanoic acid (Ahe), 8-aminocaprylic acid (8-Aoc), 11-aminoundecanoic acid (11-Aun), 12-aminododecanoic acid (12-Ado), 2-aminobenzoic acid (2-Abz), 3-aminobenzoic acid (3-Abz), 4-aminobenzoic acid (4-Abz), 4-amino-3-hydroxy-6-methylheptanoic acid (6-Ahx), 2-aminoheptanoic acid (2-hydroxy-37-5-oxolanic acid), tetralin, para-aminophenylalanine (4-NH 2-Phe), 2-aminopimelic acid (Apm), biphenylalanine (Bip), para-bromophenylalanine (4-Br-Phe), ortho-chlorophenylalanine (2-Cl-Phe), meta-chlorophenylalanine (3-Cl-Phe), para-chlorophenylalanine (4-Cl-Phe), meta-chlorotyrosine (3-Cl-Tyr), para-benzoylphenylalanine (Bpa), t-butylglycine (TLG), cyclohexylalanine (Cha), cyclohexylglycine (Chg), desmin (Des), 2-diaminopimelic acid (Dpm), 2, 3-diaminopropionic acid (Dpr), 2, 4-diaminobutyric acid (Dbu), 3, 4-dichlorophenylalanine (3, 4-Cl 2-Phe), 3, 4-difluorophenylalanine (3, 4-F2-Phe), 3, 5-diiodo (3, 5-I2-Tyr), N-ethylglycine (EtGly), N-ethyl glycine (EtAsn), asparagine (HfF) and homofluorophenylalanine (3-F-Phe), homotyrosine (Htyr), hydroxylysine (Hyl), isohydroxylysine (aHyl), 5-hydroxytryptophan (5-OH-Trp), 3-or 4-hydroxyproline (3-or 4-Hyp), p-iodophenylalanine (4-I-Phe), 3-iodotyrosine (3-I-Tyr), indoline-2-carboxylic acid (Idc), iso Ai Dumei (Ide), isoleucine (alpha-Ile), isopiperidinic acid (Inp), N-methylisoleucine (Melle), N-methyllysine (MeLys), m-methyltyrosine (3-Me-Tyr), N-methylvaline (MeVal), 1-naphthylalanine (1-Nal), 2-naphthylalanine (2-Nal), p-nitrophenylalanine (4-NO 2-Phe), 3-nitrotyrosine (3-NO 2-Tyr), norleucine (Nle), norvaline (Nva), ornithine (Orn), ortho-phosphotyrosine (H2 PO 3-Tyr), octahydroindole-2-carboxylic acid (Otic acid (Ph), penta-phenylalanine (Ph) (35 a), penta-phenylalanine (Ph-5-phenylalanine (Ph), sarcosine (Sar), tetrahydroisoquinoline-3-carboxylic acid (Tic), thienyl alanine and thiazolidine-4-carboxylic acid (thioproline, th).

As used herein, the meaning of "peptide" or "polypeptide" is well known to those skilled in the art. Typically, a peptide or polypeptide is one in which two or more amino acids are linked by an amide bond, which is formed by the amino group of one amino acid and the carboxyl group of an adjacent amino acid. The polypeptides described herein may comprise naturally occurring amino acids or non-naturally occurring amino acids. May be modified to include at least two amino acids such as analogs, derivatives, functional mimics, pseudopeptides, and the like.

As used herein, "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, or of drug conjugates, which are safe and effective for use in a mammal, non-limiting examples of pharmaceutically acceptable salts include hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, pear, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, mesylate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate.

Drawings

FIG. 1 shows the endocytosis of a polypeptide shown in SEQ ID No. 1;

FIG. 2 shows the endocytosis of the polypeptide shown in SEQ ID No. 2;

FIG. 3 shows the endocytosis of the polypeptide shown in SEQ ID No. 3;

FIG. 4 shows the endocytosis of the polypeptide shown in SEQ ID No. 4;

FIG. 5 shows the endocytosis of the polypeptide shown in SEQ ID No. 5;

FIG. 6 shows the endocytosis of the polypeptide shown in SEQ ID No. 6.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

The present invention will be described in further detail with reference to specific examples, but embodiments of the present invention are not limited thereto. Various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

The polypeptides of the invention may be prepared using methods well known to those skilled in the art, including well known methods of chemical synthesis. The polypeptide may be expressed in an organism and purified by well known purification techniques.

EXAMPLE 1 solid phase Synthesis of Polypeptides

The polypeptide compound and its derivative are synthesized into straight-chain precursor through solid phase synthesis and oxidized with DMSO to form two pairs of disulfide bonds inside molecule. The synthetic carrier is Fmoc-Gly-WANG RESIN resin. In the synthesis process, fmoc-Gly-WANG RESIN resin is fully swelled in N, N-Dimethylformamide (DMF), then the solid-phase carrier and the activated amino acid derivative are repeatedly condensed, washed, deprotected Fmoc, washed and subjected to the next round of amino acid condensation so as to reach the length of the polypeptide chain to be synthesized, and finally trifluoroacetic acid is used for: the mixed solution of water and triisopropylsilane and phenyl sulfide (90:2.5:2.5:5, v: v) reacts with resin to crack the polypeptide from the solid carrier, and then the solid crude product of the linear precursor is obtained after the solid crude product is settled by freezing methyl tertiary butyl ether. And performing disulfide bond oxidation on the cut linear precursor crude product in a weak alkaline solution to obtain a target polypeptide crude product. Purifying and separating the crude polypeptide in acetonitrile/water system of 0.1% trifluoroacetic acid by C-18 reversed phase preparative chromatographic column to obtain pure product of polypeptide and its derivative.

Experimental reagent

(1) Solid phase synthesis of the polypeptide shown in SEQ ID No.1

SEQ ID No.1:GCFRQCQTAWPAWDCFHHCG

Step 1, coupling Fmoc-Gly-OH of first amino acid

0.2Mmol of the 2-Chlorotrityl chloride resin was fully swollen in DCM for 1h. Fmoc-Gly-OH (0.16 mmol) and diisopropylethylamine (DIEA, 0.64 mmol) were weighed into 8mL of DCM and added to the resin and reacted at room temperature for 2h. After the reaction was completed, blocking solution (10 mL) DCM: methanol: DIEA (85:10:5, v:v: v) was added for 10min at room temperature. The blocked resin was washed 5 times with DCM and 5 times with DMF.

Step 2, synthesis of straight-chain peptide chain

The resin obtained in step 1 was fully swollen in DMF for 1h. The following linear precursor sequences were then synthesized in the order from the second amino acid at the carboxy terminus to the amino terminus. Each coupling cycle proceeds as follows:

fmoc-deprotection was performed twice for 8min each with 20% piperidine/DMF (20% v/v,10 mL).

DMF washing resin 6-8 times until neutral pH.

0.5Mmol of Fmoc-AA, 0.5mmol of 6-chlorobenzotriazole-1, 3-tetramethylurea Hexafluorophosphate (HCTU) and 1mmol of 4-methylmorpholine (NMM) were dissolved in DMF and the resin was added to react for 1h at room temperature.

The resin was washed 4-6 times with DMF before the next amino acid coupling.

The linear polypeptide was synthesized and the resin was washed 5 times with DMF and 5 times with DCM. The resin was dried in vacuo.

Step 3 cleavage of the Linear precursor peptide chain

Freshly prepared cut cocktail (10 mL) of trifluoroacetic acid in water in triisopropylsilane in phenylsulfide (90:2.5:2.5:5, v: v) was added to the resin from step 1 and reacted for 2 hours with shaking at room temperature. After the reaction was completed, the reaction solution was filtered, and the resin was washed with trifluoroacetic acid, combined with the reaction solution, and precipitated with 4-fold volume of cold MTBE to obtain a crude product. The crude product was washed 3 times with MTBE and placed in vacuum for drying.

Step 4 purification preparation of the polypeptide

After filtration through a 0.45um membrane, separation was performed using a reverse phase high performance liquid chromatography system with buffers A (0.1 wt% trifluoroacetic acid, aqueous solution) and B (0.1 wt% trifluoroacetic acid, acetonitrile). And collecting the relevant fractions of the product, combining the fractions with the purity of more than 95% after the purity is identified by HPLC, and freeze-drying to obtain the polypeptide pure product.

(2) Solid phase synthesis of the polypeptide shown in SEQ ID No.3

SEQ ID No.3:GCHSLCVKYYQEAFCHSHCG

Step 1, coupling Fmoc-Gly-OH of first amino acid

Step 2, synthesis of straight-chain peptide chain

DMF washing resin 6-8 times until neutral pH.

0.5Mmol Fmoc-AA,0.5mmol 6-chlorobenzotriazol-1, 3-tetramethyluronium Hexafluorophosphate (HCTU) and 1mmol 4-methylmorpholine (NMM) were dissolved in DMF and the resin was added and reacted for 1h at room temperature.

The resin was washed 4-6 times with DMF before the next amino acid coupling.

Step 3 cleavage of the Linear precursor peptide chain

Step 4 purification preparation of the polypeptide

The rest of the polypeptides of the invention can be prepared by referring to the Fmoc solid-phase synthesis method.

EXAMPLE 2 affinity of polypeptide samples with FAP protein

1. Experimental materials:

2. The experimental steps are as follows:

Polypeptide affinity assays with FAP were performed with Biacore T200. FAP protein was captured at 25 ℃ using a protein a chip at about 2000RU, 1 x HBS, pH 7.4 as runningbuffer, binding experiments were performed at 25 ℃, polypeptide analyte flow rates were 30 μl/min, association120s, association 600s, single-cycle or multi-concentration cycle kinetic/Affinity detection of binding of polypeptide samples to protein was selected, gly-HCl ph= 1.5,30 μl/min, chip regeneration was performed for 30s, and data were fitted using a 1:1 binding model.

3. Experimental results

Table 1 affinity results of polypeptide samples with FAP protein

The results show that the affinity of the polypeptide disclosed by the invention and FAP protein reaches the nm level, and the affinity example 3 shows that the endocytosis experiment is carried out

Experimental materials:

Positive cells were HEK293-FAP and negative cells were HEK293.

(1) Main reagent

Reagent name	Manufacturer' s	Goods number
			FBS	Excell	FSP500
Puromycin	Invivogen	ant-pr-1
			4% Paraformaldehyde fixing solution	Beyotime	P0099
DAPI solution (10 ug/ml, ready-to-use)	Solarbio	C0065
			Streptavidin-Cy5(SA-Cy5)	ApexBio	K1080
Streptomycin sulfate	Alatine	S105491
			Penicillin G sodium salt	Alatine	P105489)

(2) Main consumable

Name of the name	Production brands	Goods number	Storage environment
				96WellTC-TreatedBlackMicroplates	Agilent	204626-100	RT

(3) Cell lines

Cell lines	Culture medium
		HEK293 (negative cell)	F12K+10%FBS
HEK293-FAP (Positive cell)	F12K+10%FBS+1%P/S+4μg/mlpuromycin

The experimental steps are as follows:

1. SA-Cy5 dilutions were prepared at a dilution ratio of 1:50, and the dilutions were cell growth medium.

2. Sample preparation, namely directly diluting the mother solution to the required concentration (0.1-1 mu M), wherein the diluted solution is SA-Cy5 diluted solution prepared in the step 1. Premix for 1h at room temperature in dark.

3. And (3) sample adding, namely sucking out the culture medium in the hole, replacing the culture medium with a sample with the corresponding concentration (0.1-1 mu M), and incubating for 2h at the temperature of 37 ℃.

4. Fixation DPBS cells were washed 3 times, 60. Mu.L/well fixative was added, 4℃and protected from light for 30min.

5. The nuclear dyeing step, in which DPBS is used for washing cells for 3 times, 40-50 mu L/hole DAPI nuclear dyeing liquid is added, and the temperature is 37 ℃ and the light is prevented from being used for 20min.

6. Experimental results.

The experimental results are shown in figures 1-5, the polypeptide of the invention can enter cells through endocytosis in HEK293 (HEK 293-FAP) cells with high FAP expression, and the polypeptide corresponding to all sequences has endocytosis at 0.3 mu M.

In conclusion, the results prove that the polypeptide provided by the invention has targeting of FAP, has better penetrating capacity of high-expression FAP cells, and can realize high-sensitivity living imaging of micro tumors, so that in practical application, the polypeptide provided by the invention can be used as homing peptide to be combined with anticancer drugs or imaging agents for targeted treatment and imaging of tumors.

It should be noted that the above-mentioned embodiments are merely for illustrating the technical solution of the present invention, and not for limiting the same, and although the present invention has been described in detail with reference to the above-mentioned embodiments, it should be understood by those skilled in the art that the technical solution described in the above-mentioned embodiments may be modified or some technical features may be equivalently replaced, and these modifications or substitutions do not make the essence of the corresponding technical solution deviate from the spirit and scope of the technical solution of the embodiments of the present invention.

Claims

1. A polypeptide targeting fibroblast activation protein or a pharmaceutically acceptable salt thereof, wherein the amino acid sequence of the polypeptide is any one of the amino acid sequences shown in SEQ ID No. 1 to SEQ ID No. 6,

GCFRQCQTAWPAWDCFHHCG(SEQ ID No.1)、

GCVERCTTDFPQGAAACQAWCAG(SEQ ID No.2)、

GCHSLCVKYYQEAFCHSHCG(SEQ ID No.3)、

GCYRKCRANFNDLWCYKHCG(SEQ ID No.4)、

GCYRRCVTQYAAKWCLAHCG(SEQ ID No.5)、

GCYHSCSREWHPDTCRGWCG (SEQ ID No. 6).

2. A polypeptide derivative, characterized in that the polypeptide derivative is a modified product of the polypeptide according to claim 1 or a pharmaceutically acceptable salt thereof, or a variant obtained by adding and/or replacing one or more amino acids to the polypeptide according to claim 1 or a pharmaceutically acceptable salt thereof.

3. The polypeptide derivative according to claim 2 is characterized in that the polypeptide derivative is a modified product of the N-terminus or C-terminus of the polypeptide according to claim 1, and the modified group is selected from one or more of an acetyl group, an amino group, an alkyl group, an aromatic group, a fatty acid group, a sugar group, a phosphate group, a sulfate group, a PEG group or a peptide linking group; preferably, the modification is selected from: C-terminal amidation blocking and/or N-terminal modification of an acetyl group.

4. The polypeptide derivative according to claim 2, characterized in that the polypeptide derivative is a variant obtained by adding and/or replacing one, two or three amino acids of the polypeptide according to claim 1.

5. A polynucleotide, characterized in that the polynucleotide encodes the polypeptide according to claim 1 or the polypeptide derivative according to any one of claims 2 to 4.

6. A pharmaceutical preparation, characterized in that the pharmaceutical preparation comprises as an active ingredient the polypeptide or a pharmaceutically acceptable salt thereof according to claim 1, the polypeptide derivative according to any one of claims 2 to 4, or the polynucleotide according to claim 5.

7. Use of the polypeptide or pharmaceutically acceptable salt thereof according to claim 1, the polypeptide derivative according to any one of claims 2 to 4, the polynucleotide according to claim 5, or the pharmaceutical preparation according to claim 6 in the preparation of a medicament for preventing, diagnosing and/or treating cancer associated with abnormal activation of FAP.

8. The method of claim 7, wherein the cancer is selected from the group consisting of prostate cancer, breast cancer, pancreatic cancer, liver cancer, lung cancer, sarcoma, colorectal cancer, cholangiocarcinoma, chordoma, small intestine cancer, pheochromocytoma, gastric cancer, kidney cancer, ovarian cancer, bladder cancer, esophageal cancer, head and neck cancer, thymic cancer, cervical cancer, endometrial cancer, neuroendocrine tumors, thyroid cancer, intestinal cancer, and solid tumors such as bone metastasis.