ADP-ribose-binding peptide for preventing or treating cancer, and combination therapy of ADP-ribose

Abstract

The present invention pertains to: an adenosine diphosphate (ADP)-ribose-binding peptide; and an anticancer use of ADP-ribose. The pharmaceutical composition of the present invention causes ADP-ribose to accumulate in cancer cells, thereby destroying the cell balance and leading to the death of the cancer cells, and in particular, has excellent anticancer effects even with respect to cancers resistant to conventional PRAP inhibitors.

Description

mL / min, and detection was performed at 220 nm. Mobile phase A was 0.1% trifluoroacetic acid (TFA) in water and mobile phase B was 0.1% TFA in acetonitrile. The gradient was 35 %B for 2 minutes, 30 %B for 10 minutes, 10 %B for 1 minute, and 77 %B for 4 minutes. The results are shown in Table 4 below, where the LC- MS [M+H]+ value was found to be 5363.43. [Table 4] Detector,A -1 (220nm) Pk# Retention.Tiine Area Area % Height Height % X 5.217 935'0 0.401 1640' 0.581 2 5375 2235 0,096 479' 0,170 3 s 4758: 0.204 1.002 0,355 4 6.325 2279816 •97.658: 273458 9003 P 7242 7468 0.320' 1206 0.427 6 '7.567 1W01 0.814 3026 1.072 •7 13.475. 11857 0,508 '1317 0A!H 5 I Otdh 23^1485 100.000 282198 iMwsw ; Experimental Example 1. Changes in cancer cell viability by combination treatment of peptides according to Examples and ADP-ribose Experimental Example 1-1. Changes in Caki-1 cell 10 viability by combination treatment of peptides according to Examples and ADP-ribose 2.5 X 103 Kidney cancer (Caki-1) cells were cultured in a 96-well plate for 24 hours under 37°C and 5% CO2 conditions, and then divided into the following groups and treated with 15 each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADPR at 1 mM concentration alone 3) Group treated with peptide of Example 1 at 25 pM concentration alone 4) Group treated with each CPP-peptide of Example 15 at 10 pM concentration alone 5) Group treated in combination with ADPR at 1 mM concentration and peptide of Example 1 at 25 pM concentration (Combination 1) 6) Group treated in combination with ADPR at 1 mM concentration and CPP-peptide of Example 15 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37°C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, it was found that Caki-1 cells were growing rapidly in the peptide untreated group, while the viability was reduced in all the groups treated with peptides of Examples of the present disclosure and ADPR. In particular, in the group treated in combination with the peptides of Examples and ADPR, Caki-1 cells were significantly inhibited in growth, with observable signs of cell death (FIG. 2). Experimental Example 1-2. Changes in HCC1937 cell viability by combination treatment of peptides according to Examples and ADP-ribose 2.5 X 103 Breast cancer (HCC1937) cells were cultured in a 96-well plate for 24 hours under  37°C and 5% CO2 conditions, and then divided into the following groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADPR at 1 mM concentration alone 3) Group treated with peptide of Example 2 at 25 pM concentration alone 4) Group treated with each CPP-peptide of Example 16 at 10 pM concentration alone 5) Group treated in combination with ADPR at 1 mM concentration and peptide of Example 2 at 25 pM concentration (Combination 1) 6) Group treated in combination with ADPR at 1 mM concentration and CPP-peptide of Example 16 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37°C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium       bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, it was found that HCC1937 cells were growing rapidly in the peptide untreated group, while the viability was reduced in all the groups treated with peptides of Examples of the present disclosure and ADPR. In particular, in the group treated in combination with the peptides of Examples and ADPR, HCC1937 cells were significantly inhibited in growth, with observable signs of cell death (FIG. 3). Experimental Example 1-3. Changes in AsPC-1 cell viability by combination treatment of peptides according to Examples and ADP-ribose 2.5 X 103 Pancreatic cancer (AsPC-1) cells were cultured in a 96-well plate for 24 hours under 37°C and 5% CO2 conditions, and then divided into the following groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADPR at 1 mM concentration alone 3) Group treated with peptide of Example 3 at 25 pM concentration alone 4) Group treated with each CPP-peptide of Example 17 at 10 pM concentration alone 5) Group treated in combination with ADPR at 1 mM concentration and peptide of Example 3 at 25 pM concentration (Combination 1) 6) Group treated in combination with ADPR at 1 mM concentration and CPP-peptide of Example 17 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, it was found that AsPC-1 cells were growing rapidly in the peptide untreated group, while the viability was reduced in all the groups treated with peptides of Examples of the present disclosure and ADPR. In particular, in the group treated with the combination of the peptides of Examples and ADPR, AsPC-1 cells were significantly inhibited in growth, with observable signs of cell death (FIG. 4). Experimental Example 1-4. Changes in HepG2 cell viability by combination treatment of peptides according to Examples and ADP-ribose 2.5 X 103 Liver cancer (HepG2) cells were cultured in a 96-well plate for 24 hours under 37C and 5% CO2 conditions, and then divided into the following groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADPR at 1 mM concentration alone 3) Group treated with peptide of Example 4 at 25 pM concentration alone 4) Group treated with each CPP-peptide of Example 18 at 10 pM concentration alone 5) Group treated in combination with ADPR at 1 mM concentration and peptide of Example 4 at 25 pM concentration (Combination 1) 6) Group treated in combination with ADPR at 1 mM concentration and CPP-peptide of Example 18 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium       bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, it was found that HepG2 cells were growing rapidly in the peptide untreated group, while the viability was reduced in all the groups treated with peptides of Examples of the present disclosure and ADPR. In particular, in the group treated in combination with the peptides of Examples and ADPR, HepG2 cells were significantly inhibited in growth, with observable signs of cell death (FIG. 5). Experimental Example 1-5. Changes in H1975 cell viability by combination treatment of peptides according to Examples and ADP-ribose 2.5 X 103 Lung cancer (H1975) cells were cultured in a 96-well plate for 24 hours under 37C and 5% CO2 conditions, and then divided into the following groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADPR at 1 mM concentration alone 3) Group treated with peptide of Example 5 at 25 pM concentration alone 4) Group treated with each CPP-peptide of Example 19 at 10 pM concentration alone 5) Group treated in combination with ADPR at 1 mM concentration and peptide of Example 5 at 25 pM concentration (Combination 1) 6) Group treated in combination with ADPR at 1 mM concentration and CPP-peptide of Example 19 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, it was found that H1975 cells were growing rapidly in the peptide untreated group, while the viability was reduced in all the groups treated with peptides of Examples of the present disclosure and ADPR. In particular, in the group treated in combination with the peptides of Examples and ADPR, H1975 cells were significantly inhibited in growth, with observable signs of cell death (FIG. 6). Experimental Example 1-6. Changes in HCT116 cell viability by combination treatment of peptides according to Examples and ADP-ribose 2.5 X 103 Colon cancer (HCT116) cells were cultured in a 96-well plate for 24 hours under 37OC and 5% CO2 conditions, and then divided into the following groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADPR at 1 mM concentration alone 3) Group treated with peptide of Example 6 at 25 pM concentration alone 4) Group treated with each CPP-peptide of Example 20 at 10 pM concentration alone 5) Group treated in combination with ADPR at 1 mM concentration and peptide of Example 6 at 25 pM concentration (Combination 1) 6) Group treated in combination with ADPR at 1 mM concentration and CPP-peptide of Example 20 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, it was found that HCT116 cells were growing rapidly in the peptide untreated group, while the viability was reduced in all the groups treated with peptides of Examples of the present disclosure and ADPR. In particular, in the group treated in combination with the peptides of Examples and ADPR, HCT116 cells were significantly inhibited in growth, with observable signs of cell death (FIG. 7). Experimental Example 2. Confirmation of normal cytotoxicity upon combination treatment of peptides according to Examples and ADP-ribose In a 96-well plate, 2.5 X 103 human-derived normal CCD-18-Co or MRC5 cells were cultured for 24 hours under 37C and 5% CO2 conditions, and then treated in combination with the peptides of Examples 1 to 15 and 17 to 29 and ADP-ribose. The cells were then cultured for another 96 hours under 37°C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium       bromide reagent was added to each well and reacted for 1 hour.  The MTT reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, it was found that normal CCD-18-Co cells survived by approximately 90% or more when treated in combination with each peptide of Examples and ADPR as compared to the peptide untreated group. Thus, it was found that combination treatment of each peptide of Examples and ADPR showed little toxicity to normal cells (FIG. 8). Experimental Example 3. Comparison and verification of anti-cancer efficacy by combination treatment of peptides of Examples and ADP-ribose using animal models Experimental Example 3-1. Changes in tumor volume by combination treatment of peptides according to Examples and ADP-ribose in AsPC-1 cells Pancreatic cancer (AsPC-1) cells (1 x 107) were inoculated into 5-week-old BALB / c nude mice on the back flank, and the mice were divided into the following treatment groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADP-ribose alone 3) Group treated with each peptide of Examples 1 to 6 alone 4) Groups treated in combination with ADPR and each peptide of Examples 1 to 6 (Combinations 1 to 6). When the tumor volume reached about 75 mm3, ADP-ribose and the peptides of Examples 1 to 6 were administered alone or in combination. Here, ADP-ribose at a concentration of 10 mg / kg and the peptides of Examples 1 to 6 at a concentration of 20 mg / kg were administered by subcutaneous route twice weekly for 4 weeks. Tumor size was then measured using a digital caliper and the results of the change in tumor volume were compared between groups. The results showed that tumor volume was significantly reduced in all treatment groups compared to the control group. In particular, the combination treatment group showed a significant reduction in tumor volume compared to the single treatment group (FIGS. 9 and 10). Experimental Example 3-2. Changes in tumor volume by combination treatment of CPP-peptides according to Examples and ADP-ribose in HCC1937 cells Breast cancer (HCC1937) cells (1 x 107) were inoculated into 5-week-old BALB / c nude mice on the back flank, and the mice were divided into the following treatment groups and treated with each CPP-peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with ADP-ribose alone 3) Group treated with each CPP-peptide of Examples 15 to 20 alone 4) Groups treated in combination with ADPR and each CPP-peptide of Examples 15 to 20 (Combinations A to F). When the tumor volume reached about 75 mm3, ADP-ribose and the CPP-peptides of Examples 15 to 20 were administered alone or in combination. Here, ADP-ribose at a concentration of 10 mg / kg and the CPP-peptides of Examples 15 to 20 at a concentration of 10 mg / kg were administered by subcutaneous route twice weekly for 4 weeks. Tumor size was then measured using a digital caliper and the results of the change in tumor volume were compared between groups. The results showed that tumor volume was significantly reduced in all treatment groups compared to the control group. In particular, the combination treatment group showed a significant reduction in tumor volume compared to the single treatment group (FIGS. 11 and 12). Experimental Example 4. Confirmation of anti-cancer synergistic effects of combination treatment of peptides according to Examples and ADP-ribose in PARP-resistant cell lines Next, the effect of combination treatment of the peptide of Examples with ADP-ribose was confirmed in a PARP inhibitor-resistant cancer cell line. Specifically, 2.5 X 103 PARP inhibitor-resistant breast cancer (HCC1937) cells were cultured in a 96-well plate for 24 hours under 37°C and 5% CO2 conditions. The cells were then divided into the following groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with PARP inhibitor (Olaparib) at 2 pM concentration 3) Group treated with ADPR at 1 mM concentration alone 4) Groups treated with each peptide of Examples 1 to 6 at 25 pM concentration alone 5) Groups treated with each CPP-peptide of Examples 15 to 20 at 10 pM concentration alone 6) Group treated in combination with ADPR at 1 mM concentration and each peptide of Examples 1 to 6 at 25 pM concentration (Combination 1) 6) Group treated in combination with ADPR at 1 mM concentration and CPP-peptide of Examples 15 to 20 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37°C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium       bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. The results showed that when HCC1937 cells were treated with Olaparib, a PARP inhibitor, alone, approximately 80% of the cancer cells were viable, but treatment with the peptides of Examples or ADP-ribose significantly reduced cell viability in all cases.    In particular, all cases of combination treatment with the peptides according to Examples and ADP-ribose showed a significant reduction in cancer cell viability compared to the single treatment group (FIG. 13). Experimental Example 5. Changes in cancer cell viability by combination treatment of peptides according to Examples and NAD+ Considering the possibility that NAD+, a precursor of ADP-ribose, could replace ADP-ribose, the present inventors confirmed its anti-cancer effects when administered with NAD+ alone and in combination with ADP-ribose binding peptides according to Examples. Specifically, 2.5 X 103 ovarian cancer (OVCAR-3) cells were cultured in a 96-well plate for 24 hours under 37OC and 5% CO2 conditions, and then divided into the following groups and treated with each peptide according to Examples at GI50 concentration alone or in combination. 1) Untreated group 2) Group treated with NAD+ at 1 mM concentration alone 3) Group treated with peptide of Example 2 at 25 pM concentration alone 4) Group treated with CPP-peptide of Example 16 at 10 pM concentration alone 5) Group treated in combination with NAD+ at 1 mM concentration and peptide of Example 2 at 25 pM concentration (Combination 1) 6) Group treated in combination with NAD+ at 1 mM concentration and CPP-peptide of Example 16 at 10 pM concentration (Combination 2) The cells were then cultured for another 96 hours under 37°C  and 5% CO2 conditions, and 10 pl of 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium       bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. The results showed that the peptide untreated group had dramatic growth of OVCAR-3 cells, and the NAD+ treatment alone group was not significantly different from the control group. However, the combination treatment of the peptides of Examples according to the present disclosure and NAD+ significantly reduced the viability of cancer cells compared to the treatment of the peptides of Examples alone, indicating a superior anti-cancer effect (FIG. 14). Experimental Example 6. Changes in cancer cell viability by combination treatment of peptides according to Examples, ADP-ribose, NAD+, and various anti-cancer drugs Each of 2.5 X 103 human-derived pancreatic cancer (Aspc-1), kidney cancer (Caki-1), breast cancer (HCC1937), liver cancer (HepG2), lung cancer (H1975), colon cancer (HCT116), ovarian cancer (OVCAR-3), brain cancer (U87) cells were cultured in a 96-well plate under 37C and 5% CO2 conditions for 24 hours, and then divided into the following groups and treated with each formulation at GI20 / IC20 concentrations alone or in combination. 1) Untreated group 2) Group in which each cancer cell was treated with peptides of Examples 1 to 14 at 25 pM concentration and Examples 15, 17 to 29 at 10 pM concentration, respectively, alone. 3) Group treated with ADPR at 2 mM concentration alone 4) Group treated with NAD+ at 2 mM concentration alone 5) Groups treated with various anti-cancer drugs alone (Cisplatin at 2 pM, Docetaxel at 1 pM, Dichloroacetate at 3 mM, Doxorubicin at 2 pM, Sorafenib at 2 pM, Osimertinib at 10 nM, Trastuzumab at 0.2 pM, Bevacizumab at 0.2 nM, Tamoxifen at 0.2 pM, or Olaparib at 1 pM) 6) Groups treated in combination with ADPR at 2 mM concentration and each peptide according to Examples (Examples 1 to 14 at 25 pM, and Examples 15, 17 to 29 at 10 pM). 7) Groups treated in combination with ADPR at 2 mM concentration, NAD+ at 2 mM concentration, and each peptide according to Examples (Examples 1 to 14 at 25 pM, and Examples 15, 17 to 29 at 10 pM). 8) Groups treated in combination with ADPR at 2 mM concentration, peptides according to Examples (Examples 1 to 14 at 25 pM, and Examples 15, 17 to 29 at 10 pM), and various anti-cancer drugs (same concentration as Group 5 above), respectively. The cells were then cultured for another 96 hours under 37C and 5% CO2 conditions, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reagent was added to each well and reacted for 1 hour. The MTT reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, in all cancer cells used in the experiment, compared to treatment with the peptides of Examples 1 to 15 and 17 to 29, ADPR, NAD+, and various anti-cancer drugs alone, the combination treatment of each peptide according to Examples and ADPR resulted in about 40% cancer cell viability, and the combination treatment of each peptide according to Examples, ADPR, and NAD+ resulted in about 20% cancer cell viability.    Further, the combination treatment of the peptides according to Examples 1 to 15 and 17 to 29, ADPR, and various anti-cancer drugs significantly reduced the viability of cancer cells compared to all other groups, resulting in a viability of about 10%. Thus, it was found that the combination treatment of the peptides according to Examples, ADPR, and various anti-cancer drugs resulted in excellent anti-cancer effects (FIGS. 15 to 28). Experimental Example 7. Changes in cancer cell viability upon treatment with sustained release derivative prepared using the peptide of Example 29, alone or in combination Each of 2.5 X 103 human-derived pancreatic cancer (Aspc-1), kidney cancer (Caki-1), breast cancer (HCC1937), liver cancer (HepG2), lung cancer (H1975), colon cancer (HCT116), ovarian cancer (OVCAR-3), brain cancer (U87) cells were cultured in a 96-well plate under 37°C and 5% CO2 conditions for 24 hours, and then treated with the sustained release derivative alone. In addition, MDA-MB-231 cancer cells were divided into the following groups and treated with each formulation at GI20 concentration alone or in combination. 1) Untreated group 2) Group treated with sustained-release derivative alone at 5 pM concentration 3) Group treated with ADPR at 2 mM concentration alone 4) Group treated with NAD+ at 2 mM concentration alone 5) Groups treated with various anti-cancer drugs alone (Cisplatin at 2 pM, Docetaxel at 1 pM, Dichloroacetate at 3 mM, Doxorubicin at 2 pM, Sorafenib at 2 pM, Osimertinib at 10 nM, Trastuzumab at 0.2 pM, Bevacizumab at 0.2 nM, Tamoxifen at 0.2 pM, or Olaparib at 1 pM) 6) Group treated in combination with ADPR at 2 mM concentration and a sustained-release derivative at 5 pM concentration 7) Group treated in combination with ADPR at 2 mM concentration, NAD+ at 2 mM concentration, and a sustained-release derivative at 5 pM concentration 8) Group treated in combination with ADPR at 2 mM concentration, a sustained-release derivative at 5 pM concentration, and various anti-cancer drugs (same concentration as Group 5 above). The cells were then cultured for another 96 hours at 37°C and 5% CO2, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reagent was added to each well and reacted for 1 hour. The reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. As a result, treatment with the sustained-release derivative alone resulted in viability of less than 10% for most cancer cell lines (FIG. 29, left). Meanwhile, MDA-MB-231, a triple-negative breast cancer cell line, showed drug resistance with viability of 80% when treated with the sustained-release derivative, ADPR, NAD+, or various anticancer drugs alone, but showed cytotoxicity when combined with sustained-release derivative and ADPR, and showed significantly superior cytotoxicity at the time of tripledrug combination treatment with NAD+ or various anti-cancer drugs (FIG. 29, right). Experimental Example 8. Confirmation of normal cytotoxicity upon treatment with sustained release derivative prepared using the peptide of Example 29, alone or in combination In a 96-well plate, 2.5 X 103 human-derived normal MRC5 cells were cultured for 24 hours under 37°C and 5% CO2 conditions, and then treated with the sustained-release derivative and ADPR alone or in combination. The cells were then cultured for another 96 hours at 37°C and 5% CO2, and 10 pl of 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide reagent was added to each well and reacted for 1 hour. The MTT reagent was removed at the end of the reaction, 200 pl of dimethyl sulfoxide was then added to each well, and the absorbance was measured to determine the cell viability. The results showed that compared to the untreated group, treatment with the sustained-release derivative alone or in combination with ADPR resulted in approximately 90% or more viability of normal MRC5 cells. Thus, it was found that treatment with the sustained release derivative exhibited little toxicity to normal cells, regardless of whether the sustained release derivative was used alone or in combination (FIG. 30). Experimental Example 9. Comparison and verification of anti-cancer efficacy of sustained-release derivative alone or in combination treatment with immunotherapeutic agent using animal models Colon cancer (MC38) cells (1 x 106 / 100 pL) were inoculated into 6-week-old male C57BL / 6-hPD1 (PD-1 humanized mice) on the right dorsal forelimb area of mice, and the treatment groups were divided as follows. 1) Untreated group 2) Group treated with sustained-release derivative alone at GI50 concentration by intravenous injection 3) Group treated with sustained-release derivative alone at GI50 concentration by subcutaneous injection 4) Group treated with immunotherapeutic agent Nivolumab alone at GI50 concentration by intraperitoneal injection 5) Group treated in combination with sustained-release derivative at GI50 concentration (intravenous injection) and Nivolumab at GI50 concentration (intraperitoneal injection) (Combination 1) 6) Group treated in combination with sustained-release derivative at GI50 concentration (subcutaneous injection) and Nivolumab at GI50 concentration (intraperitoneal injection) (Combination 2) When the tumor volume reached approximately 100 mm3, the sustained-release derivative or Nivolumab was administered alone or in combination. Here, the sustained-release derivative at a concentration of 5 mg / kg was mixed with albumin in a 3:1 ratio, and Nivolumab was used at a concentration of 5 mg / kg, each administered twice weekly for 4 weeks. Tumor size was then measured using a digital caliper and the results of the change in tumor volume were compared between groups. The results showed that tumor volume was significantly reduced in all treatment groups compared to the control group. In particular, the combination treatment group showed a significant reduction in tumor volume compared to the single treatment group (FIG. 31). Further, no change in body weight was observed in all treatment groups, confirming the absence of toxicity (FIG. 32). From the above description, those skilled in the art to 5 which the present disclosure pertains will understand that the present disclosure may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof.    In this regard, it should be understood that the embodiments described above are 10 illustrative in all respects and not restrictive. The scope of the present disclosure is indicated by the following claims rather than the detailed description, and should be construed as including all changes or modifications derived from the meaning and scope of the claims and equivalent 15 concepts within the scope of the present disclosure.

Claims

1. Use of a composition comprising:a) an adenosine diphosphate (ADP)-ribose binding peptide or a pharmaceutically acceptable salt thereof; andb) ADP-ribose, a precursor thereof, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the prevention or treatment of cancer,wherein the ADP-ribose binding peptide consists of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14; andthe precursor of the ADP-ribose is nicotinamide adenine dinucleotide (NAD+), cADPR, or poly-ADPR.

2. A method of treating or preventing cancer comprising administering to a subjectin need thereof a pharmaceutical composition comprising:a) an adenosine diphosphate (ADP)-ribose binding peptide or a pharmaceutically acceptable salt thereof; andb) ADP-ribose, a precursor thereof, or a pharmaceutically acceptable salt thereof, wherein the ADP-ribose binding peptide consists of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14; andthe precursor of the ADP-ribose is nicotinamide adenine dinucleotide (NAD+), cADPR, or poly-ADPR.

3. The use of claim 1, or the method of claim 2, wherein the peptide further comprises a cell-penetrating peptide at the N-terminus, C-terminus, or both termini.

4. The use or method of claim 3, wherein the cell-penetrating peptide is at least one selected from the group consisting of TAT, buforin, maurocalcine, penetratin, poly-argininederived peptides, Antennapedia, Transportan, VP22, Hph-1, poly-arginine R11(R9), Pep-1, HP4, LAH4, Vetofusin-1, signal sequence-based peptides, and amphipathic peptides.

5. The use or the method of claim 3, wherein the peptide comprises any one amino acid sequence selected from the group consisting of SEQ ID NOs: 15 to 29.

6. The use of any one of claims 1 or 3-5 or the method of any one of claims 2 or 3-2024227135   07 Jul 20265, wherein the peptide comprises one or more of its constituent amino acids that are acetylated, hydroxylated, methylated, amidated or PEGylated.

7. The use of any one of claims 1 or 3-6 or the method of any one of claims 2 or 36, wherein the peptide comprises a fatty acid, carbohydrate, lipid component, or cofactor bound to one or more of its constituent amino acids.

8. The use of any one of claims 1 or 3-7 or the method of any one of claims 2 or 37, wherein the cancer is at least one solid cancer selected from the group consisting of brain cancer, head and neck cancer, lung cancer, breast cancer, thymoma, esophageal cancer, colon cancer, liver cancer, stomach cancer, pancreatic cancer, biliary tract cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, germ cell tumor, ovarian cancer, cervical cancer, endometrial cancer, colorectal cancer, lymphoma, acute leukemia, chronic leukemia, multiple myeloma, sarcoma, malignant melanoma, and skin cancer.

9. The use of any one of claims 1 or 3-8, or the method of any one of claims 2 or 3-8, wherein the cancer is a poly(ADP-ribose) polymerase (PARP) inhibitor-resistant cancer.

10. The use of any one of claims 1 or 3-9 or the method of any one of claims 2 or 39, wherein the pharmaceutical composition is formulated as a sustained-release preparation.

11. The use of any one of claims 1 or claims 3-10 or the method of any one of claims 2 or 3-10, further comprising a third anti-cancer drug.

12. The use or the method of claim 11, wherein the third anti-cancer drug is a cytotoxic anti-cancer drug, a targeted anti-cancer drug, an immune anti-cancer drug, a metabolic anti-cancer drug, a synthetic lethality anti-cancer drug, or a combination thereof.

13. Use of a composition comprising an ADP-ribose binding peptide or a pharmaceutically acceptable salt thereof as an active ingredient, wherein the ADP-ribose binding peptide is derivatized by PEGylation of one or more of its constituent amino acids or binding of a fatty acid to one or more of its constituent amino acids to have sustained-release properties in the manufacture of a medicament for prevention or treatment of cancer, andwherein the ADP-ribose binding peptide consists of any one amino acid sequence2024227135   07 Jul 2026selected from the group consisting of SEQ ID NOs: 1 to 14.

14. A method of treating or preventing cancer in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising an ADP-ribose binding peptide or a pharmaceutically acceptable salt thereof as an active ingredient, wherein the ADP-ribose binding peptide is derivatized by PEGylation of one or more of its constituent amino acids or binding of a fatty acid to one or more of its constituent amino acids to have sustained-release properties, andwherein the ADP-ribose binding peptide consists of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14.

15. The use of claim 13 or the method of claim 14, wherein the cancer is at least one solid cancer selected from the group consisting of brain cancer, head and neck cancer, lung cancer, breast cancer, thymoma, esophageal cancer, colon cancer, liver cancer, stomach cancer, pancreatic cancer, biliary tract cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, germ cell tumor, ovarian cancer, cervical cancer, endometrial cancer, colorectal cancer, lymphoma, acute leukemia, chronic leukemia, multiple myeloma, sarcoma, malignant melanoma, and skin cancer.

16. The use of claim 13 or 15 or the method of claim 14 or 15, wherein the derivatized ADP-ribose binding peptide is represented by the following Chemical Formula 3:2024227135   07 Jul 2026

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