Tandem repeat cancer targeting peptides for molecular conjugation or modification and their use in cancer theranostics

Abstract

An isolated cancer targeting peptide comprising at least two copies of the amino acid sequence PFLP (SEQ ID NO: 1) or PELF (SEQ ID NO: 2). A pharmaceutical composition for treating cancer is also disclosed. The composition comprises the isolated cancer targeting peptide and an anticancer agent. A bispecific anticancer antibody is also disclosed, comprising the isolated cancer targeting peptide and an antigen binding peptide that stimulates T cell activity. A method for treating cancer by administering the pharmaceutical composition or the bispecific anticancer antibody is provided. A method for diagnosing cancer by administering a radionuclide-labeled cancer targeting peptide to an individual and imaging the location of the radionuclide is also provided.

Description

Background Art

[0001] Cancer recurrence is a major clinical challenge. Cancer stem cells, which exist as a subpopulation in tumors, are particularly resistant to chemotherapeutic drugs and radiation. See Lee et al., 2015, FASEB J. 29: Supplement 629.18. After conventional chemotherapy, the increased proportion of cancer stem cells in tumors is an important predictor of cancer recurrence. See Lee et al.

[0002] It is reported that the 78kDa glucose-regulated protein (GRP78), a member of the HSP70 protein family, is present on the surface of a variety of cancer cells, but not on normal cells. See Wang et al., 2016, Biomaterials 94:31-44 and Liu et al. 2013, Clin. Cancer Res. 19:6802-11. GRP78 is also associated with both cancer cell resistance and stem cell-like cell behavior, and is further shown to be a targetable cell surface receptor. See Bachelder, 2018. Therefore, GRP78 is an attractive target for anticancer therapy that should reduce damage to normal cells and reduce recurrence.

[0003] Cancer-targeting peptides (CTPs) that specifically bind to GRP78 have been previously identified. See Wang et al. CTPs were found to interact with the peptide-binding domain (PBD) of GRP78 in a linear peptide conformation. These CTPs were shown to enhance the anti-tumor efficacy of the chemotherapeutic drug doxorubicin when conjugated to this drug, and also target both cancer cells and cancer stem cells, thereby reducing the recurrence rate of cancer. See Liu et al.

[0004] There is a need to develop CTPs with higher affinity for cancer stem cells and to develop CTP-based anticancer therapeutic modalities. Summary of the invention

[0005] To meet this need, isolated cancer targeting peptides are disclosed that comprise at least two copies of the amino acid sequence PFLP (SEQ ID NO: 1) or PFLF (SEQ ID NO: 2).

[0006] The present invention also discloses a pharmaceutical composition for treating cancer, which comprises the isolated cancer targeting peptide and an anticancer agent.

[0007] In addition, a bispecific anti-cancer antibody is disclosed, which comprises the isolated cancer targeting peptide and an antigen-binding peptide that stimulates T cell activity.

[0008] Furthermore, methods for treating cancer by administering the pharmaceutical composition or the bispecific anti-cancer antibody are both within the scope of the present invention.

[0009] Another method for diagnosing cancer is disclosed. The method is performed by administering a radionuclide-labeled cancer targeting peptide to an individual and subjecting the individual to imaging techniques to determine the location and amount of the radionuclide-labeled cancer targeting peptide, wherein the cancer targeting peptide comprises at least two copies of the amino acid sequence PFLP (SEQ ID NO: 1) or PFLF (SEQ ID NO: 2).

[0010] Details of several embodiments of the invention are described below and in the accompanying drawings. Figure 2 All features, objects and advantages of the present invention will be apparent from this description and the accompanying drawings and the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The following description refers to the accompanying drawings, in which:

[0012] Figure 1 is a bar graph showing the percent injected dose / gram (%ID / g) determined by positron emission tomography (PET) analysis following injection into tumor-bearing mice for each of the four designated radiolabeled peptides. The peptide sequences are shown in Table 1 below.

[0013] Figure 2 is a bar graph showing the relative signal ratios in tumor tissue sections of tumor-bearing mice injected with radiolabeled peptides P6, F4P6, P13, or F4P13.

[0014] Figure 3 is a diagram of a bispecific anti-cancer antibody construct of the present invention. α-CD3 = anti-CD3 scFv, Fc(K) = heavy chain with knob dimerization sequence, Fc(H) = heavy chain with hole dimerization sequence, F4P6-TR-CTP = F4P6 tandem repeat cancer targeting peptide that binds to GRP-78.

[0015] Figure 4A is a bar graph showing the percentage of lysis (% Lysis) of TOV21G ovarian cancer cells after 40 hours of incubation with PBMC effector cells in the absence (PBMC) or presence (Ctrl-BsAb and (F4P6-BsAb) of the indicated concentrations of bispecific antibodies.

[0016] Figure 4B is a bar graph showing the percentage of lysis (% Lysis) of NCI-N87 gastric cancer cells after 90 hours of incubation with PBMC effector cells in the absence (PBMC) or presence (Ctrl-BsAb and (F4P6-BsAb) of the indicated concentrations of bispecific antibodies.

[0017] Figure 5 Anticancer micellar nanocomplex (MNC) is schematically shown. EGCG = epigallocatechin-3-O-gallate; oEGCG = oligomeric epigallocatechin-3-O-gallate; PEG = poly(ethylene glycol); TR-CTP = tandem repeat cancer targeting peptide.

[0018] Figure 6 is a bar graph showing the percent survival of BT474 human breast cancer cells incubated with vehicle, Herceptin, MNC-Herceptin, or F4P6-MNC-Herceptin as indicated.

[0019] Figure 7 is a graph of tumor volume versus days after injection of N87 tumor cells into mice. Mice were injected once a week for 4 weeks with the indicated treatments. *p=0.041. DETAILED DESCRIPTION

[0020] As described above, an isolated cancer targeting peptide is provided, which comprises at least two copies of the amino acid sequence PFLP (SEQ ID NO: 1).

[0021] The two copies of PFLP in the cancer targeting peptide may overlap each other. For example, the cancer targeting peptide may be RPFLPFLPY (SEQ ID NO: 5) and RPFLPFLPYRPFLPFLPY (SEQ ID NO: 6).

[0022] Another isolated cancer targeting peptide comprises at least two copies of PFLF (SEQ ID NO: 2). Some examples of such peptides include RPFLFPFLFY (SEQ ID NO: 7) and RPFLFPFLFYRPFLFPFLFY (SEQ ID NO: 8).

[0023] The cancer targeting peptides described above can specifically bind to GRP-78 on cancer cells. In other words, no additional amino acids are required. Therefore, the present invention encompasses any of the cancer targeting peptides described above that do not contain the sequence RLLDT (SEQ ID NO: 15).

[0024] Also within the scope of the present invention is a pharmaceutical composition for treating cancer comprising any of the above-described isolated cancer targeting peptides and an anti-cancer agent.

[0025] In certain compositions, the anti-cancer agent is a monoclonal antibody, such as anti-HER2 / neu, anti-PD-1, anti-PD-L1, or anti-CTLA4.

[0026] In other compositions, the anticancer agent is a chemotherapeutic agent, such as doxorubicin, vincristine, vinorelbine, paclitaxel, or irinotecan.

[0027] In addition, the anticancer agent in the pharmaceutical composition may include a radioactive isotope, e.g. 90 Y. 125 I. 188 Re, 68 Ga, 111 In or 131 I. In one embodiment, the radioisotope is chelated by a chelator conjugated to a cancer targeting peptide.

[0028] Referring again to the pharmaceutical composition for treating cancer, one specific example includes micellar nanocomplexes (MNCs) having a core encapsulating an anticancer agent and a shell comprising an isolated cancer targeting peptide.

[0029] The core can be, for example, a monoclonal antibody complexed with oligomeric epigallocatechin-3-O-gallate (EGCG). In a particular composition, the monoclonal antibody is an anti-HER2 / neu antibody, such as trastuzumab.

[0030] The shell can be formed by a conjugate of EGCG and polyethylene glycol (PEG) linked to a cancer targeting peptide, for example, the cancer targeting peptide is linked to PEG. The shell can also comprise a PEG / EGCG conjugate without a cancer targeting peptide.

[0031] Alternatively, the shell can be a liposome formed from distearoylphosphatidylcholine, cholesterol and PEG-distearoylphosphoethanolamine.Polymers such as poly(lactic-co-glycolic acid) and polyvinyl chloride can also be used as components of the shell.

[0032] The scope of the present invention also includes bispecific anticancer antibodies, which include any of the above-mentioned separated cancer targeting peptides and antigen binding peptides that stimulate T cell activity. Exemplary antigen binding peptides specifically bind to: CD3, PD-1, CTLA-4, LAG-3, TIM-3, TIGIT, VISTA, B7-H3, OX40, GITR, ICOS or 41BB. The antigen binding peptide can be, for example, a single-chain Fv (scFv) or a single-domain antibody. In an exemplary bispecific anticancer antibody, the antigen binding peptide is an anti-CD3 scFv. A specific example of a bispecific anticancer antibody is a heterodimer consisting of SEQ ID NO: 12 and SEQ ID NO: 14.

[0033] Methods for treating cancer using the cancer targeting properties of the above pharmaceutical compositions and bispecific anti-cancer antibodies are provided.

[0034] For example, a method of treating cancer is by administering to a cancer patient a pharmaceutical composition comprising a cancer targeting peptide and an anticancer agent as described above. In a specific method, cancer is treated by administering MNCs having a core of oligomeric EGCG complexed with trastuzumab and a shell comprising (i) a cancer targeting peptide linked to a PEG-EGCG conjugate and (ii) a PEG-EGCG conjugate lacking the peptide.

[0035] A different method of treating cancer is achieved by administering the above-mentioned bispecific anti-cancer antibody to a cancer patient. In one example, the bispecific anti-cancer antibody is a heterodimer consisting of SEQ ID NO:12 and SEQ ID NO:14.

[0036] Cancers that may be treated by the above methods include, but are not limited to, breast cancer, hepatocellular carcinoma, prostate cancer, lung cancer, ovarian cancer, kidney cancer, uterine cancer, cervical cancer, melanoma, embryonal carcinoma, leukemia, and osteosarcoma.

[0037] The above mentioned method of diagnosing cancer using a radionuclide-labeled cancer targeting peptide comprising at least two copies of the amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 2. The radionuclide-labeled cancer targeting peptide may have an amino acid sequence of SEQ ID NO: 5, 6, 7 or 8.

[0038] In order to realize the method, a radionuclide-labeled cancer targeting peptide is applied to, for example, injected into, an individual suspected of having cancer. Then, the individual is subjected to imaging techniques (e.g., positron emission tomography) to quantify the amount of the radionuclide-labeled cancer targeting peptide accumulated in each body tissue. If the amount of the radionuclide-labeled cancer targeting peptide accumulated in the local area of ​​the tissue is greater than the background level in the adjacent area of ​​the tissue, cancer is diagnosed.

[0039] In one specific method, a cancer targeting peptide having an amino acid sequence of SEQ ID NO: 6 is treated with a radionuclide 68 Ga is labeled and administered to an individual.

[0040] Although not described in further detail, it is believed that those skilled in the art will be able to maximize the use of the present disclosure based on the disclosure herein. Therefore, the following specific examples should be interpreted as merely illustrative and not limiting the remainder of the present disclosure in any way. All publications and patent documents cited herein are incorporated by reference in their entirety.

[0041] Example

[0042] Example 1: Tandem Repeat Cancer Targeting Peptide (TR-CTP)

[0043] TR-CTP was designed to contain at least one tandem repeat of the binding motif, PFLX1 (where X1 is P or F) present in CTPs previously described in U.S. Pat. No. 8,846,623. The peptide sequences are shown in Table 1 below, with the repeats underlined and in bold. Note that in F4P6-TR-CTP and F5P6-TR-CTP, the repeats overlap by one amino acid.

[0044] Without being bound by theory, it is believed that the increased length of TR-CTP compared to CTP will prevent steric hindrance when incorporated into a bispecific antibody. In addition, the repetition of the binding motif should increase binding affinity. Furthermore, the novel TR-CTP with a repetitive sequence described herein should be suitable for conjugation at its N-terminus or its C-terminus.

[0045] Table 1. Amino acid sequences of TR-CTP and CTP

[0046]

[0047] Example 2: Binding affinity of N-terminally extended TR-CTP

[0048] Biotin-labeled TR-CTP and CTP (Biotools Co., Ltd, Taiwan, China) were synthesized to evaluate the kinetics of binding to GRP78 by surface plasmon resonance. TR-CTP and CTP were extended with five amino acids (GGGGS; SEQ ID NO: 9) at their N-termini. Each TR-CTP / CTP was labeled with a single biotin molecule at the N-terminus via an aminohexanoic acid bond.

[0049] Two negative control peptides were also biotinylated, where the L-Leu residues in the F4P6-TR-CTP and F4P13-TR-CTP sequences were replaced with D-Leu residues (dF4P6 and dF4P13, respectively).

[0050] Streptavidin was immobilized to a sensor chip (CM5; GE Healthcare) with a matrix of carboxymethylated dextran covalently attached to a gold surface using standard amine coupling methods according to established procedures (GE Healthcare). TR-CTP labeled with biotin was immobilized on the chip by flowing it through the chip at a flow rate of 5 μl / min using HBS-P+ (GE Healthcare) as the running buffer.

[0051] Then use BIACORE TM The binding affinity of the peptide binding domain of GRP78 (GRP78-PBD, amino acids 421 to 639) bound to the chip was analyzed by a T200 instrument (GM Healthcare) at various concentrations (approximately 0.1 to 15 μM) to TR-CTP and CTP. The sensor chip was regenerated by washing the chip surface with glycine buffer (10 mM; pH 11.5) for 30 seconds. The chip was reused after two chip regeneration cycles and subsequent washing with running buffer for 120 seconds. The results are shown in Table 2 below.

[0052] Table 2. Binding affinity of GRP78-PBD to N-conjugated CTP and TR-CTP

[0053]

[0054] a = No binding signal detected

[0055] The dissociation constants (K) of the P6 series peptides (repeating sequence PFLP): P6-CTP (one copy), F4P6-TR-CTP (two copies), and F5P6-TR-CTP (four copies) D ) are 1.9×10 -6 M, 1.2×10 -6 M and 1.0×10 -6M. Improvement of binding affinity (i.e., K D The decrease in ) is associated with an increase in the copy number of the binding sequence present in the TR-CTP peptide.

[0056] Similar results were found for the P13 series peptides (repeating sequence PFLF). The K values ​​of P13-CTP (one copy), F4P13-TR-CTP (two copies), and F5P13-TR-CTP (four copies) were D The values ​​are 2.7×10 -6 M, 2.1×10 -6 M and 6.1×10 -7 M. Among all the peptides tested, F5P13-TR-CTP exhibited the highest binding affinity to the peptide-binding domain of GRP78.

[0057] Dissociation rate (K off ) reflects the binding stability of the TR-CTP / GRP78-PBD complex. K off The smaller the value, the slower the dissociation rate, that is, the higher the stability of the complex. As shown in Table 2, the K values ​​of P6-CTP, F4P6-TR-CTP and F5P6-TR-CTP off The values ​​are 3.0×10 -3 S -1 , 2.8×10 -3 S -1 and 9.1×10 -4 S -1 K of P13-CTP, F4P13-TR-CTP and F5P13-TR-CTP off The values ​​are 3.4×10 -3 S -1 , 1.7×10 -3 S -1 and 1.2×10 -3 S -1 There is a general correlation between the number of repeat sequences in TR-CTP and the dissociation rate.

[0058] As expected, the two D-Leu-substituted negative control peptides (i.e., dF4P6 and dF4P13) showed negligible binding to GRP78-PBD; their K D Value cannot be determined. See last two rows of Table 2.

[0059] Example 3: Binding affinity of C-terminally extended TR-CTP

[0060] The P13 series peptides described in Example 1 above were also biotin labeled after their C-terminus was extended with the sequence GGGGSK (SEQ ID NO: 10). Biotin was conjugated to the C-terminal lysine residue using standard techniques. The binding affinity of GRP78-PBD to the sensor chip bound peptides was determined as described in Example 2 above. The results are shown in Table 3 below.

[0061] Table 3. Kinetics of GRP78-PBD binding to C-conjugated CTP and TR-CTP

[0062]

[0063] The binding affinity of GRP78-PBD to the C-terminally biotinylated peptide was similar to that to its N-terminally tagged counterpart. In addition, the binding affinities of F4P13-TR-CTP and MF4P13-TR-CTP-c-biotin, both of which have two copies of PFLF, with C-terminal biotin (K D ) and dissociation rate (K off ) are superior to P13-CTP-c-biotin with only one copy. The peptide MF4P13-TR-CTP has the same amino acid sequence as F4P13-TR-CTP with methionine added to the N-terminus.

[0064] Example 4. In vivo tumor targeting of TR-CTP

[0065] The ability of TR-CTP to target tumor cells in vivo was tested in N87 tumor-bearing mice (a HER2-positive gastric cancer xenograft model). N87 tumors were established in NOD SCIDγ (NSG) mice using standard protocols. Briefly, NSG mice were injected subcutaneously with 3 × 10 6 Tumors were established using N87 cells / animal. Tumors were allowed to grow to 100 to 200 mm before the following treatments were performed. 3 volume.

[0066] Using standard techniques 68 Ga labeled four peptides, namely F4P13-TR-CTP, P13-CTP, F4P6-TR-CTP and P6-CTP. Briefly, dodecanetetraacetic acid polyethylene glycol (DOTA-PEG3350) was conjugated to each peptide to form DOTA-CTP-PEG3350 (Mission Biotech, Taipei, Taiwan). 68 GaCl3 (itG, Germany) was mixed in 0.1 M sodium acetate buffer (pH 5.5) and incubated at 95°C for 10 min. 68Ga-DOTA-CTP-PEG3350 was used directly without further purification.

[0067] More specifically, the above four 68 A dose of 250 μCi of each of the Ga-labeled peptides was injected intravenously into four NSG mice. Fifteen minutes after injection, each mouse was scanned for 15 minutes using nanoScan PET / CT (Mediso Pacific), and static microPET images of the tumor were obtained. Quantification of the images can be found in Figure 1 The results showed that the percentage injected dose / gram (%ID / g) in the tumor was significantly higher after administration of F4P13-TR-CTP and F4P6-TR-CTP compared to P13-CTP and P6-CTP, respectively. Apparently, the presence of tandem repeats of the binding sequences (i.e., PFLP and PFLP) improved tumor targeting of CTP.

[0068] The PET studies were confirmed by autoradiographic studies of frozen sections of tumor tissue excised from injected mice. 68 The distribution pattern of Ga-conjugated peptides was heterogeneous throughout the tumor. Quantification of the radiographic signal in tumor sections is shown in Figure 2 In tumor tissue, compared with injection 68 Ga-P6-CTP and 68 Radiographic signal of Ga-P13-CTP mice, injected 68 Ga-F4P6-TR-CTP and 68 The radiographic signals in mice with Ga-F4P13-TR-CTP were higher, respectively, indicating that the tandem repeat peptide (i.e., TR-CTP) can better target cancer compared to CTP. 68 Radiographic signal of Ga-P6-CTP, 68 Ga-P13-CTP, 68 Ga-F4P6-TR-CTP and 68 The radiographic signals of Ga-F4P13-TR-CTP are 1.8, 2.3, and 11, respectively.

[0069] Example 5. Bispecific Antibodies

[0070] The bispecific antibody-like protein was modified using the so-called "knob-hole" technology to efficiently form heterodimers to evaluate the suitability of TR-CTP for immunotherapy. See, for example, U.S. Patent 8,961,971. Briefly, F4P6-TR-CTP was fused to the C-terminus of Fc-hole peptide and the C-terminus of Fc-knob peptide, respectively. Anti-CD3 scFv was fused to the N-terminus of Fc-hole peptide. The bispecific antibody-like construct named F4P6-BsAb was used in Figure 3 In this example, the F4P6-TR-CTP Fc-hole fusion has the amino acid sequence of SEQ ID NO: 12, and the F4P6-TR-CTP Fc-knob fusion has the amino acid sequence of SEQ ID NO: 14. The control antibody (i.e., Ctrl-BsAb) lacks the F4P6-TR-CTP sequence.

[0071] After expression and purification of F4P6-BsAb, the kinetics of GRP78-PDB binding to it was evaluated by surface plasmon resonance as described in Example 1 above, except that F4P6-BsAb was directly coupled to the CM5 sensor chip via amine coupling. The results showed that the K of GRP78-PBD to F4P6-BsAb was D The value is 1.1×10 -6 M and K off The value is 9.4×10 -4 S -1 , these values ​​are close to those of GRP78-PBD for the isolated peptide F4P6-TR-CTP (see Table 2).

[0072] Example 6. Cancer cell killing induced by F4P6-BsAb

[0073] The ability of the bispecific antibody-like construct F4P6-BsAb to induce cell killing by peripheral blood mononuclear cells (PBMCs) was tested on TOV21G ovarian cancer target cells and N87 gastric cancer target cells.

[0074] 2×10 cells / well were plated in 96-well electronic microtiter plates (“E-plates”; ACEA Biosciences, Inc.). 4 Target cells were inoculated with 10 cells and allowed to adhere for 2 hours. PBMC effector cells, PBMC effector cells + F4P6-BsAb, and PBMC effector cells + Ctrl-BsAb lacking the P4P6-TC-CTP sequence were added to achieve a final effector to target ratio of 10:1. The concentration of F4P6-BsAb and Ctrl-BsAb was 12.5 nM. Data were collected and quantified using the xCELLigence Real-Time Cell Analysis System ("RTCA"; ACEA Biosciences, Inc.) according to the manufacturer's instructions. The results are shown in Figure 4A and 4B middle.

[0075] In the presence of F4P6-BsAb, PBMCs mediated TOV21G cells ( Figure 4A ) and N87 cells ( Figure 4B) showed 22% and 15% lysis, while no cell lysis was detectable in the absence of F4P6-BsAb or in the presence of Ctrl-BsAb.

[0076] Example 7. TR-CTP micellar nanocomplex

[0077] It is known that the following micellar nanocomplexes (MNCs) have better tumor selectivity, greater cancer cell growth inhibitory activity and longer blood half-life compared to free Herceptin, the micellar nanocomplexes (MNCs) having: (i) a combination of the anticancer monoclonal antibody trastuzumab ( The invention relates to a novel nanostructured ...

[0078] MNCs were prepared to test the ability of TR-CTP to improve the effectiveness of MNCs. F4P6-TR-CTP was conjugated to PEG-EGCG as follows to produce F4P6-TR-CTP-PEG-EGCG. F4P6-TR-CTP was PEGylated using CHO-PEG-NHS in dimethylformamide with the addition of N,N-diisopropylethylamine. F4P6-TR-CTP-PEG-EGCG was synthesized by the Baeyer reaction between the aldehyde (CHO) group of the PEGylated F4P6-TR-CTP and the nucleophilic ring of EGCG. The resulting product was dialyzed (MWCO=3500) and lyophilized to give F4P6-TR-CTP-PEG-EGCG. See Chung et al.

[0079] The Herceptin / oEGCG core was prepared as previously described. See Chung et al. MNCs were prepared by mixing the Herceptin / oEGCG core with PEG-EGCG to form MNC-Herceptin or with F4P6-TR-CTP-PEG-EGCG to form F4P6-MNC-Herceptin. Figure 5 It is shown diagrammatically in FIG.

[0080] Example 8. In vitro cancer cell killing by TR-CTP-MNC

[0081] The ability of F4P6-TR-CTP-MNC-Herceptin to kill cancer cells was compared with that of MNC-Herceptin in HER-2 / neu. Briefly, BT-474 human breast cancer cells overexpressing HER2 / neu were cultured at 1×10 cells / well.4 Cells were seeded in 96-well E plates and cultured for one day. The wells were treated with vehicle, Herceptin alone, MNC-Herceptin, or F4P6-TR-CTP-MNC-Herceptin in amounts corresponding to 0.5 mg / ml and 1.25 mg / ml Herceptin. Cell viability was assessed after 3 days using RTCA as described above in Example 6. The results, expressed as percent survival, are shown in Figure 6 middle.

[0082] At the concentrations tested, Herceptin alone did not inhibit the survival of BT-474 cells compared to vehicle controls. MNC-Herceptin reduced cell viability to 30% to 40% at both concentrations tested, compared to 100% viability of vehicle-treated cells.

[0083] F4P6-MNC-Herceptin was more effective in killing BT-474 breast cancer cells than either Herceptin or MNC-Herceptin alone. In fact, treating the cells with F4P6-MNC-Herceptin at a concentration corresponding to 1.25 mg / ml Herceptin unexpectedly reduced their viability to only 1% of the vehicle control. Clearly, TR-CTP enhanced the in vitro cell-killing activity of MNC-Herceptin.

[0084] Example 9. In vivo cancer cell killing by TR-CTP-MNC

[0085] Because Herceptin inhibits tumor growth, we sought to determine whether TR-CTP-MNC-Herceptin enhances the anticancer activity of Herceptin in vivo. N87 tumors were established as described above in Example 4. Tumors carrying 100 to 200 mm were injected intravenously with 125 μg of free Herceptin, MNC-Herceptin, or F4P6-MNC-Herceptin weekly. 3 The results are shown in Figure 7 middle.

[0086] F4P6-MNC-Herceptin inhibited tumor growth to a greater extent than free Herceptin. In fact, 61 days after injection of tumor cells, the tumor size in mice treated with F4P6-MNC-Herceptin was significantly smaller than those in the Herceptin-treated group. Figure 7 The tumor growth inhibition mediated by F4P6-MNC-Herceptin was slightly greater than that mediated by MNC-Herceptin.

[0087] Other Implementations

[0088] All features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature for the same, equivalent or similar purpose. Therefore, unless otherwise expressly stated, each feature disclosed is only an example of a general series of equivalent or similar features.

[0089] Through the above description, those skilled in the art can easily determine the essential characteristics of the present invention, and without departing from the spirit and scope of the present invention, various changes and modifications can be made to the present invention to adapt it to various uses and conditions. Therefore, other embodiments are also within the scope of the appended claims. Sequence Listing <110> UCT <120> Tandem repeat cancer targeting peptides for molecular conjugation or modification and their use in cancer theranostics <130> 218514-0001PCT <150> US 62 / 976,811 <151> 2020-02-14 <160> 15 <170> PatentIn version 3.5 <210> 1 <211> 4 <212> PRT <213> Artificial sequence <220> <223> CTP repeat 1 <400> 1 Pro Phe Leu Pro 1 <210> 2 <211> 4 <212> PRT <213> Artificial sequence <220> <223> CTP repeat 2 <400> 2 Pro Phe Leu Phe 1 <210> 3 <211> 12 <212> PRT <213> Artificial sequence <220> <223> P6-CTP <400> 3 Arg Leu Leu Asp Thr Asn Arg Pro Phe Leu Pro Tyr 1 5 10 <210> 4 <211> 12 <212> PRT <213> Artificial sequence <220> <223> P13-CTP <400> 4 Arg Leu Leu Asp Thr Asn Arg Pro Phe Leu Phe Tyr 1 5 10 <210> 5 <211> 9 <212> PRT <213> Artificial sequence <220> <223> F4P6-TR-CTP <400> 5 Arg Pro Phe Leu Pro Phe Leu Pro Tyr 1 5 <210> 6 <211> 18 <212> PRT <213> Artificial sequence <220> <223> F5P6-TR-CTP <400> 6 Arg Pro Phe Leu Pro Phe Leu Pro Tyr Arg Pro Phe Leu Pro Phe Leu 1 5 10 15 Pro Tyr <210> 7 <211> 10 <212> PRT <213> Artificial sequence <220> <223> F4P13-TR-CTP <400> 7 Arg Pro Phe Leu Phe Pro Phe Leu Phe Tyr 1 5 10 <210> 8 <211> 20 <212> PRT <213> Artificial sequence <220> <223> F5P13-TR-CTP <400> 8 Arg Pro Phe Leu Phe Pro Phe Leu Phe Tyr Arg Pro Phe Leu Phe Pro 1 5 10 15 Phe Le Phe Tyr 20 <210> 9 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic connector <400> 9 Gly Gly Gly Gly Ser 1 5 <210> 10 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Synthetic connector <400> 10 Gly Gly Gly Gly Ser Lys 1 5 <210> 11 <211> 1677 <212> DNA <213> Artificial sequence <220> <223> Mortar arm F4P6-BsAb <220> <221> CDS <222> (1)..(1677) <400> 11 atg ggc ggt agg cgt gta cgg tgg gag gtc tat ata agc aga gct ggg 48 Met Gly Gly Arg Arg Val Arg Trp Glu Val Tyr Ile Ser Arg Ala Gly 1 5 10 15 tac gtg aac cgt cag atc gcc tgg aga cgc cat cac aga tct gcc acc 96 Tyr Val Asn Arg Gln Ile Ala Trp Arg Arg His His Arg Ser Ala Thr 20 25 30 atg ggt tgg agc ctc atc ttg ctc ttc ctt gtc gct gtt gct acg cgt 144 Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg 35 40 45 gtc ctg tcc cag gtg cag ctg gtg cag agc ggc gct gaa gtg aag aaa 192 Val Leu Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 50 55 60 cct ggc gcc tcc gtg aag gtg tcc tgc aag gct tct ggc tac acc ttt 240 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 65 70 75 80 acc cgg tac acc atg cat tgg gtg cga cag gct cca ggc cag ggg ctg 288 Thr Arg Tyr Thr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu 85 90 95 gaa tgg att ggc tac atc aac ccc agc cgg ggc tac acc aac tac aat 336 Glu Trp Ile Gly Tyr Ile Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn 100 105 110 cag aag ttc aag gat aag gcc acc ctg acc acc gac aag tcc atc tcc 384 Gln Lys Phe Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ile Ser 115 120 125 acc gcc tac atg gaa ctg tcc cgg ctg aga tcc gac gat acc gct gtg 432 Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val 130 135 140 tac tac tgc gcc cgg tac tac gac gac cac tac acc ctg gac tac tgg 480 Tyr Tyr Cys Ala Arg Tyr Tyr Asp Asp His Tyr Thr Leu Asp Tyr Trp 145 150 155 160 gga cag ggt act ctc gtg act gtg tca agt ggc ggc gga gga tct ggc 528 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly 165 170 175 gga ggt gga agt ggc gga ggc ggt tct gaa atc gtg ctg aca cag agc 576 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser 180 185 190 ccc gcc acc ctg tca ctg tct cca ggc gag aga gct acc ctg agc tgc 624 Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys 195 200 205 tct gcc tcc tcc tcc gtg tct tac atg aac tgg tat cag cag aag ccc 672 Ser Ala Ser Ser Ser Val Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro 210 215 220 ggc cag gcc ccc aga cgg tgg atc tac gat acc tcc aag ctg gcc tcc 720 Gly Gln Ala Pro Arg Arg Trp Ile Tyr Asp Thr Ser Lys Leu Ala Ser 225 230 235 240 ggc atc cct gcc aga ttc tcc ggc tct ggc tcc ggc acc tcc tat acc 768 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Thr 245 250 255 ctg aca atc tcc agc ctg gaa ccc gag gac ttt gcc gtg tat tac tgc 816 Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 260 265 270 cag cag tgg tcc tcc aac ccc ttc acc ttc gga cag ggc aca aag gtg 864 Gln Gln Trp Ser Ser Asn Pro Phe Thr Phe Gly Gln Gly Thr Lys Val 275 280 285 gaa atc aag cgc tcc gga ggc gga gga agc gga ggc gga ggt tca ggt 912 Glu Ile Lys Arg Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 290 295 300 ggc gga gga tca acc ggt acc cac acc tgt cct cca tgc cct gcc cct 960 Gly Gly Gly Ser Thr Gly Thr His Thr Cys Pro Pro Cys Pro Ala Pro 305 310 315 320 gag ctg gcc ggc gcc ccc tcc gtg ttc ctg ttc cct cca aag cct aag 1008 Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 325 330 335 gac acc ctg atg atc tcc cgg acc cct gaa gtg acc tgc gtg gtg gtg 1056 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 340 345 350 gac gtg tcc cac gag gac cct gaa gtg aag ttc aat tgg tac gtg gac 1104 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 355 360 365 ggc gtg gaa gtg cac aac gcc aag acc aag ccc aga gag gaa cag tac 1152 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 370 375 380 aac tcc acc tac cgg gtg gtg tcc gtg ctg acc gtg ctg cac cag gat 1200 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 385 390 395 400 tgg ctg aac ggc aaa gag tac aag tgc aag gtg tcc aac aag gcc ctg 1248 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 405 410 415 cct gcc ccc atc gaa aag acc atc tcc aag gcc aag ggc cag cct cgg 1296 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 420 425 430 gaa cct caa gtg tgc acc ctg ccc cct agc cgg gaa gag atg acc aag 1344 Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 435 440 445 aac cag gtg tcc ctg tcc tgc gcc gtg aag ggc ttc tac ccc tcc gac 1392 Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp 450 455 460 att gcc gtg gaa tgg gag tcc aac ggc cag cct gag aac aac tac aag 1440 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 465 470 475 480 acc acc ccc cct gtg ctg gac tcc gac ggc tca ttc ttc ctg gtg tcc 1488 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser 485 490 495 aag ctg aca gtg gac aag tcc cgg tgg cag cag ggc aac gtg ttc tcc 1536 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 500 505 510 tgc tcc gtg atg cac gag gcc ctg cac aac cac tac acc cag aag tcc 1584 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 515 520 525 ctg agc ctg tcc ccc ggg aag ggc ggt ggt ggt tca ggc ggt ggc ggc 1632 Leu Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly 530 535 540 agc ggc gga ggc gga tcc aga cca ttt ttg ccc ttt ctg ccc tac 1677 Ser Gly Gly Gly Gly Ser Arg Pro Phe Leu Pro Phe Leu Pro Tyr 545 550 555 <210> 12 <211> 559 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 12 Met Gly Gly Arg Arg Val Arg Trp Glu Val Tyr Ile Ser Arg Ala Gly 1 5 10 15 Tyr Val Asn Arg Gln Ile Ala Trp Arg Arg His His Arg Ser Ala Thr 20 25 30 Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg 35 40 45 Val Leu Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 50 55 60 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 65 70 75 80 Thr Arg Tyr Thr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu 85 90 95 Glu Trp Ile Gly Tyr Ile Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn 100 105 110 Gln Lys Phe Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ile Ser 115 120 125 Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val 130 135 140 Tyr Tyr Cys Ala Arg Tyr Tyr Asp Asp His Tyr Thr Leu Asp Tyr Trp 145 150 155 160 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly 165 170 175 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser 180 185 190 Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys 195 200 205 Ser Ala Ser Ser Ser Val Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro 210 215 220 Gly Gln Ala Pro Arg Arg Trp Ile Tyr Asp Thr Ser Lys Leu Ala Ser 225 230 235 240 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Thr 245 250 255 Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 260 265 270 Gln Gln Trp Ser Ser Asn Pro Phe Thr Phe Gly Gln Gly Thr Lys Val 275 280 285 Glu Ile Lys Arg Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 290 295 300 Gly Gly Gly Ser Thr Gly Thr His Thr Cys Pro Pro Cys Pro Ala Pro 305 310 315 320 Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 325 330 335 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 340 345 350 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 355 360 365 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 370 375 380 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 385 390 395 400 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 405 410 415 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 420 425 430 Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 435 440 445 Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp 450 455 460 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 465 470 475 480 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser 485 490 495 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 500 505 510 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 515 520 525 Leu Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly 530 535 540 Ser Gly Gly Gly Gly Ser Arg Pro Phe Leu Pro Phe Leu Pro Tyr 545 550 555 <210> 13 <211> 906 <212> DNA <213> Artificial Sequence <220> <223> Stalk Arm F4P6-BsAb <220> <221> CDS <222> (1)..(906) <400> 13 atg ggc ggt agg cgt gta cgg tgg gag gtc tat ata agc aga gct ggg 48 Met Gly Gly Arg Arg Val Arg Trp Glu Val Tyr Ile Ser Arg Ala Gly 1 5 10 15 tac gtg aac cgt cag atc gcc tgg aga cgc cat cac aga tct gcc acc 96 Tyr Val Asn Arg Gln Ile Ala Trp Arg Arg His His Arg Ser Ala Thr 20 25 30 atg ggt tgg agc ctc atc ttg ctc ttc ctt gtc gct gtt gct acg cgt 144 Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg 35 40 45 gtc ctg tcc acc ggt acc cac acc tgt cct cca tgc cct gcc cct gag 192 Val Leu Ser Thr Gly Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 50 55 60 ctg gcc ggc gcc ccc tcc gtg ttc ctg ttc cct cca aag cct aag gac 240 Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 65 70 75 80 acc ctg atg atc tcc cgg acc cct gaa gtg acc tgc gtg gtg gtg gac 288 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 85 90 95 gtg tcc cac gag gac cct gaa gtg aag ttc aat tgg tac gtg gac ggc 336 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 100 105 110 gtg gaa gtg cac aac gcc aag acc aag ccc aga gag gaa cag tac aac 384 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 115 120 125 tcc acc tac cgg gtg gtg tcc gtg ctg acc gtg ctg cac cag gat tgg 432 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 130 135 140 ctg aac ggc aaa gag tac aag tgc aag gtg tcc aac aag gcc ctg cct 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 145 150 155 160 gcc ccc atc gaaag acc atc tcc aag gcc aag ggc cag ccc cgc gaa 528 Ala Pro Ile Glu Lys Thr Ser Ile Lys Ala Lys Gly Gln Pro Arg Glu 165 170 175 ccc cag gtg tac aca ctg ccc cct tgc cgg gaa gag atg acc aag aac 576 Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn 180 185 190 cag gtg tcc ctg tgg tgc ctc gtg aag ggc ttc tac ccc tcc gac att 624 Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 195 200 205 gcc gtg gaa tgg gag tcc aac ggc cag cct gag aac aac tac aag acc 672 Only Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Tyr Lys Thr 210 215 220 acc ccc cct gtg ctg gac tcc gac ggc tc ttc ctg tac tcc aag 720 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 225 230 235 240 ctg aca gtg gac aag tcc cgg tgg cag cag ggc aac gtg ttc tcc tgc 768 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 245 250 255 tcc gtg atg cac gag gcc ctg cac aac cac tac acc cag aag tcc ctg 816 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 260 265 270 agc ctg tcc ccc ggg aag ggc ggt ggt ggt tca ggc ggt ggc ggc agc 864 Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 275 280 285 ggc gga ggc gga tcc aga cca ttt ttg ccc ttt ctg ccc tac 906 Gly Gly Gly Gly Ser Arg Pro Phe Leu Pro Phe Leu Pro Tyr 290 295 300 <210> 14 <211> 302 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 14 Met Gly Gly Arg Arg Val Arg Trp Glu Val Tyr Ile Ser Arg Ala Gly 1 5 10 15 Tyr Val Asn Arg Gln Ile Ala Trp Arg Arg His His Arg Ser Ala Thr 20 25 30 Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg 35 40 45 Val Leu Ser Thr Gly Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 50 55 60 Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 65 70 75 80 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 85 90 95 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 100 105 110 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 115 120 125 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 130 135 140 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 145 150 155 160 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 165 170 175 Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn 180 185 190 Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 195 200 205 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 210 215 220 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 225 230 235 240 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 245 250 255 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 260 265 270 Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 275 280 285 Gly Gly Gly Gly Ser Arg Pro Phe Leu Pro Phe Leu Pro Tyr 290 295 300 <210> 15 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Polypeptide <400> 15 Arg Leu Leu Asp Thr 1 5

Claims

1. An isolated cancer targeting peptide consisting of the amino acid sequence of RPFLFPFLFY (SEQ ID NO: 7) or RPFLFPFLFYRPFLFPFLFY (SEQ ID NO: 8).

2. A pharmaceutical composition for treating cancer, comprising the isolated cancer targeting peptide according to claim 1 and an anticancer agent. 3 . The pharmaceutical composition of claim 2 , further comprising a micellar nanocomplex having a core encapsulating the anticancer agent and a shell comprising the isolated cancer targeting peptide.

4. The pharmaceutical composition of claim 3, wherein the isolated cancer targeting peptide is conjugated to polyethylene glycol.

5. The pharmaceutical composition of claim 4, wherein the anticancer agent is a therapeutic monoclonal antibody selected from anti-HER2 / neu, anti-PD-1, anti-PD-L1 and anti-CTLA4.

6. The pharmaceutical composition of claim 5, further comprising epigallocatechin-3-O-gallate, wherein the anticancer agent is an anti-HER2 / neu monoclonal antibody.

7. The pharmaceutical composition of claim 4, wherein the anticancer agent is doxorubicin, vincristine, vinorelbine, paclitaxel or irinotecan.

8. The pharmaceutical composition of claim 2, wherein the anticancer agent comprises a radioactive isotope.

9. The pharmaceutical composition of claim 8, wherein the radioisotope is 90 Y. 125 I. 68 Ga, 188 Re, 111 In or 131 I.

10. Use of the isolated cancer targeting peptide and anticancer agent of claim 1 in the preparation of a medicament for treating cancer, wherein the cancer is ovarian cancer, gastric cancer or breast cancer.

11. The use of claim 10, wherein the anticancer agent comprises a 90 Y. 125 I. 68 Ga, 188 Re, 111 In or 131 Radioactive isotopes of I.

12. Use of the isolated cancer targeting peptide, anti-Her2 / neu monoclonal antibody and epigallocatechin-3-O-gallate of claim 1 in the preparation of a medicament for treating cancer, wherein the cancer is ovarian cancer, gastric cancer or breast cancer.

13. Use of a cancer targeting peptide labeled with a radionuclide in the preparation of an agent for diagnosing cancer using imaging techniques, The amino acid sequence of the radionuclide-labeled cancer targeting peptide is RPFLFPFLFY (SEQ ID NO: 7) or RPFLFPFLFYRPFLFPFLFY (SEQ ID NO: 8), and the cancer is ovarian cancer, gastric cancer or breast cancer.

14. The use of claim 13, wherein the radionuclide is 68 Ga, and the imaging technique is positron enhanced tomography.

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