COMPOSITION FOR PREVENTING OR TREATING EXTRAHEPATIC BILE DUCT CANCER

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating extrahepatic bile duct cancer containing, as active ingredients, natural killer (NK) cells and an anti-PD-1 antibody.

Claims

1-10. (canceled)

11. A method for preventing or treating extrahepatic bile duct cancer comprising a step of administering effective amounts of natural killer (NK) cells and an anti-PD-1 antibody to a subject in need thereof.

12. The method of claim 11, wherein the method comprises a step of obtaining natural killer cells by culturing peripheral blood mononuclear cells (PBMCs) in a T75 flask containing a cytokine-containing culture medium for 5 to 7 days, and then transferring the cultured cells to a T175 flask, followed by further culturing for 7 to 14 days.

13. The method of claim 12, wherein the peripheral blood mononuclear cells (PBMCs) are those from which CD3.sup.+ T cells have been removed.

14. The method of claim 12, wherein the cytokine is contained in the culture medium in an amount of 500 to 1,500 IU/ml.

15. The method of claim 11, wherein the natural killer cells are administered in an amount of 110.sup.4 cells/kg to 110.sup.10 cells/kg.

16. The method of claim 11, wherein the anti-PD-1 antibody is pembrolizumab.

17. The method of claim 11, wherein the anti-PD-1 antibody is administered in an amount of 10 to 1,000 mg.

18. The method of claim 11, wherein the extrahepatic bile duct cancer is upper (proximal), middle or lower (distal) bile duct cancer.

19. The method of claim 11, wherein a combined positive score (CPS) of PD-L1 (programmed cell death-ligand 1) in the extrahepatic bile duct cancer is 5 to 50%.

20. The method of claim 11, wherein the extrahepatic bile duct cancer comprises a cancer resistant to at least one of gemcitabine and cisplatin drugs, or a cancer that has recurred or metastasized after treatment with the drug.

21. The method of claim 11, wherein the step comprises a first administration of the natural killer cells and the anti-PD-1 antibody to the subject, followed by a second administration of the natural killer cells to the subject.

22. The method of claim 21, wherein the second administration is performed after a period of at least 1 day.

23. The method of claim 21, wherein the cycle of the first and the second administrations in the step of administering is repeated at least 3 times.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0061] FIG. 1 shows a method of administering natural killer cells and pembrolizumab to a patient with extrahepatic bile duct cancer according to an embodiment of the present invention.

BEST MODE

[0062] An embodiment of the present invention is directed to a pharmaceutical composition for preventing or treating extrahepatic bile duct cancer containing, as active ingredients, natural killer (NK) cells and an anti-PD-1 antibody.

[0063] In the present invention, the anti-PD-1 antibody may be pembrolizumab, nivolumab, pidilizumab, AMP 514 (Amplimmune), PDR-001 (Novartis), MEDI-0690 (also known as AMP-514; MedImmune LLC), SHR-1210 (Incyte Corp.), REGN-2810 (Regeneron Pharmaceuticals, Inc.), PF-06801591 (Pfizer), TSR-042 (also known as ANB011; Tesaro, Inc.), BGB-A317 (BeiGene, Ltd.), or JS001 (Shanghai Junshi Bioscience Co., Ltd.). Preferably, the anti-PD-1 antibody may be pembrolizumab.

[0064] In the present invention, the extrahepatic bile duct cancer may be one in which the expression level of PD-L1 (programmed cell death-ligand 1) protein or a gene encoding the protein, preferably the combined positive score (CPS), is 5 to 50%.

MODE FOR INVENTION

[0065] Hereinafter, the present invention will be described in more detail with reference to examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention according to the subject matter of the present invention is not limited by these examples.

Examples

1. Preparation of Natural Killer Cell Therapy Product (SMT-NK Line)

[0066] 60 ml of blood was collected from the subject's family or a healthy blood donor and then centrifuged using Ficoll-Paque PLUS at 3,000 rpm for 30 minutes, and the peripheral blood mononuclear cell (PBMC) layer was collected. Thereafter, CD3.sup.+ T cells were removed from the collected PBMCs using MACSxpress (Milteyi Biotec, Germany), and the PBMCs from which the CD3.sup.+ T cells have been removed were washed twice with Dulbecco's phosphate-buffered saline (DPBS). Then, the cells were cultured in a T75 flask containing 20 ml of NK expansion medium (ALyS505NK-IL2 1000 IU/ml, Cell Science & Technology Inst). Fresh medium was added to the IL-2-activated natural killer cells every 3 days, and after 5 to 7 days of culture, the cells were transferred to a T175 flask. Natural killer cell expansion was further performed for 7 to 14 days until the desired cell number was reached, while fresh medium was continuously added. The viability and number of the expanded natural killer cells were measured using a trypan blue counting method and an automatic cell counter.

2. Selection of Patients to be Treated

[0067] 15 patients, who have been diagnosed with extrahepatic bile duct cancer and intrahepatic bile duct cancer, were selected and subjected to an experiment. However, criteria for patient selection included males and females between the ages of 19 and 80 years old, and patients with a combined positive score (CPS) (corresponding to the level of tumor PD-L1 expression) of 1% or more as determined by immunohistochemical staining (IHC 22C3 pharmDx test), or patients with a MSI-high positive tumor as determined by PCR, or patients with a dMMR** positive tumor as determined by immunohistochemical staining. Table 1 below summarizes the cancer type and combined positive score (CPS) of tumor PD-L1 in the selected patients, and drugs to which the patients were resistant.

TABLE-US-00001 TABLE 1 Patient Name of Drugs to serial PD-L1 diagnosed which patients No. score disease were resistant 1 1 Extrahepatic bile Gemcitabine, cisplatin, duct cancer 5-FU 2 95 Extrahepatic bile Gemcitabine, cisplatin, duct cancer radiotherapy 3 5 Intrahepatic bile Gemcitabine, cisplatin duct cancer 5-FU, levofolic 4 2 Extrahepatic bile Gemcitabine, cisplatin, duct cancer 5-FU 5 3 Extrahepatic bile Gemcitabine, cisplatin duct cancer 6 1 Intrahepatic bile Gemcitabine, cisplatin duct cancer 7 2 Intrahepatic bile Gemcitabine, cisplatin duct cancer 8 2 Intrahepatic bile Gemcitabine, cisplatin duct cancer 9 30 Extrahepatic bile Gemcitabine, cisplatin duct cancer 10 3 Extrahepatic bile Gemcitabine, cisplatin, duct cancer 5-FU 11 50 Extrahepatic bile Gemcitabine, cisplatin duct cancer 12 5 Extrahepatic bile Gemcitabine, cisplatin duct cancer 13 1 Extrahepatic bile Gemcitabine, cisplatin duct cancer 14 35 Intrahepatic bile Gemcitabine, cisplatin duct cancer 15 5 Intrahepatic bile Gemcitabine, cisplatin, duct cancer 5-FU

3. Determination of Dosage and Administration Method

[0068] To the patients selected as described above, the prepared SMT-NK line (allogeneic blood-derived natural killer cells) and pembrolizumab (Keytruda) were administered by the method shown in FIG. 1. Specifically, the SMT-NK line (blue arrow) was administered intravenously to the patients at a dose of 310.sup.6 (20%) cells/kg, once a week a total of 48 times, with a drug-free period of one week after two administrations, and pembrolizumab (Keytruda) (red arrow) was administered to the patients by intravenous instillation at a dose of 200 mg, once every 3 weeks for a total of 24 times.

4. Evaluation of Treatment Results

[0069] While the SMT-NK line and pembrolizumab (Keytruda) were administered to each patient as described above, computed tomography (CT) was performed every 3 cycles to evaluate the drug efficacy. After a total of 9 cycles (administration of the SMT-NK line for a total of 48 times, and administration of pembrolizumab for a total of 24 times), it was evaluated whether the cancer was in complete response (CR), partial response (PR), stable disease (SD), or progressive disease (PD). The results are shown in Table 2 below.

TABLE-US-00002 TABLE 2 Name of Drugs to Maximum Final Serial PD-L1 diagnosed which patients tumor tumor No. score disease were resistant response response 1 1 Extrahepatic Gemcitabine, cisplatin, SD PD bile duct cancer 5-FU 2 95 Extrahepatic Gemcitabine, cisplatin, SD PD bile duct cancer radiotherapy 3 5 Intrahepatic Gemcitabine, cisplatin PD PD bile duct cancer 5-FU, levofolic 4 2 Extrahepatic Gemcitabine, cisplatin, PD PD bile duct cancer 5-FU 5 3 Extrahepatic Gemcitabine, cisplatin PD PD bile duct cancer 6 1 Intrahepatic Gemcitabine, cisplatin PD PD bile duct cancer 7 2 Intrahepatic Gemcitabine, cisplatin PD PD bile duct cancer 8 2 Intrahepatic Gemcitabine, cisplatin SD PD bile duct cancer 9 30 Extrahepatic Gemcitabine, cisplatin PR CR bile duct cancer 10 3 Extrahepatic Gemcitabine, cisplatin, PR PD bile duct cancer 5-FU 11 50 Extrahepatic Gemcitabine, cisplatin PR CR bile duct cancer 12 5 Extrahepatic Gemcitabine, cisplatin SD SD bile duct cancer 13 1 Extrahepatic Gemcitabine, cisplatin SD PD bile duct cancer 14 35 Intrahepatic Gemcitabine, cisplatin SD PD bile duct cancer 15 5 Intrahepatic Gemcitabine, cisplatin, SD CR bile duct cancer 5-FU

[0070] As shown in Table 2 above, as a result of administering the natural killer cells and pembrolizumab to 9 patients with extrahepatic bile duct cancer and 6 patients with intrahepatic bile duct cancer, it was confirmed that, in terms of the maximum tumor response, the therapeutic effect was shown in 7 (77.78%) out of the 9 patients with extrahepatic bile duct cancer, whereas only 3 out (50%) of the 6 patients with intrahepatic bile duct cancer showed stable disease (SD). In addition, in terms of the final tumor response, the effective therapeutic effect was shown in 3 (33.33%) out of the 9 patients with extrahepatic bile duct cancer, and 2 out of the 9 patients showed complete response (CR). However, in the case of the patients with intrahepatic bile duct cancer, the therapeutic effect was shown in only one (16.67%) out of the 6 patients, and most (five patients; 83.33%) of the 6 patients showed advanced bile duct cancer. In addition, among all the patients, 3 (75%) out of 4 patients (3 CR and 1 SD) who showed the therapeutic effect corresponded to extrahepatic bile duct cancer, and 2 (66.67%) out of 3 patients who showed complete remission (CR) corresponded to extrahepatic bile duct cancer.

[0071] Furthermore, it could be seen found that, among the patients with extrahepatic bile duct cancer, in particular, patients with a PD-L1 combined positive score (CPS) of 5 to 50% showed better therapeutic effects of natural killer cells and pembrolizumab than those having a PD-L1 combined positive score (CPS) out of the above range.

[0072] Thereby, it could be confirmed that the therapeutic effects of natural killer cells and pembrolizumab were much better in the patients with extrahepatic bile duct cancer than in the patients with intrahepatic bile duct cancer, and it could be seen that the therapeutic effects of natural killer cells and pembrolizumab were better in patients with a PD-L1 combined positive score (CPS) of 5 to 50%, among the patients with extrahepatic bile duct cancer.

[0073] Although the present invention has been described in detail with reference to the specific features, it will be apparent to those skilled in the art that this detailed description is only of a preferred embodiment thereof, and does not limit the scope of the present invention. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereto.

INDUSTRIAL APPLICABILITY

[0074] The present invention relates to a composition capable of effectively preventing or treating bile duct cancer, particularly extrahepatic bile duct cancer, and a method of preventing or treating extrahepatic bile duct cancer using the same.