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KR-20260067508-A - Peptides specifically binding to CCR8 and uses thereof

KR20260067508AKR 20260067508 AKR20260067508 AKR 20260067508AKR-20260067508-A

Abstract

The present invention relates to peptides, antibodies, or fragments having immunological activity that specifically bind to CCR8, and to the use thereof. The CCR8-binding peptides newly discovered in the present invention bind to the TM subpocket of CCR8 in a pattern similar to CCL1 and have been confirmed to specifically bind to cells that express CCR8 on their cell membranes. Therefore, these can be used as target substances for designing drugs targeting Tregs that express CCR8. By using these, cytotoxic drugs can be specifically delivered to the tumor microenvironment or cancer cells without affecting other effector T cells or peripheral Tregs, thereby having the effect of reducing the side effects of existing immunotherapies.

Inventors

  • 배현수
  • 양주원
  • 박선영
  • 양혜진

Assignees

  • 경희대학교 산학협력단

Dates

Publication Date
20260513
Application Date
20241105

Claims (20)

  1. A peptide, antibody, or fragment having the same immunological activity that specifically binds to an epitope comprising an amino acid at position 20 to 50, an amino acid at position 80 to 120, an amino acid at position 160 to 210, or an amino acid at position 250 to 290 of CCR8 (C-C chemokine receptor 8).
  2. In claim 1, a peptide, antibody, or fragment having the same immunological activity comprising any one selected from the group consisting of the amino acid sequences of SEQ ID NOs 1 to 3.
  3. In claim 1, a peptide, antibody, or fragment having the immunological activity thereof comprising the amino acid sequence of SEQ ID NO. 5.
  4. In claim 1, a peptide, antibody, or fragment having the immunological activity thereof comprising the amino acid sequence of SEQ ID NO. 6.
  5. In claim 1, human CCR8 is a peptide, antibody, or fragment having the same’s immunological activity, comprising the amino acid sequence of SEQ ID NO. 7.
  6. A composition for detecting tumor-infiltrating regulatory T cells (Ti-Treg), comprising the peptide of claim 1, an antibody, or a fragment having the immunological activity thereof.
  7. A composition for detecting tumor-infiltration-regulating T cells, further comprising a label in claim 6.
  8. A composition for detecting tumor-infiltration-regulating T cells, wherein the label is a chromogenic enzyme, a radioisotope, a chromopore, a luminescent substance, a fluorescent substance, a probe, or a tag.
  9. A drug delivery system comprising the peptide of claim 1, an antibody or a fragment having the immunological activity thereof, and a drug combined therewith.
  10. In paragraph 9, the drug is a drug delivery vehicle that is a compound, RNA, DNA, antibody, effector, prodrug, toxin, peptide, or radionuclide.
  11. In claim 9, the drug is a drug delivery vehicle that is an immunogenic apoptosis inducer, a pro-apoptotic peptide, a microtubulin structure formation inhibitor, a meiosis inhibitor, a topoisomerase inhibitor, a DNA intercalator, a toxin, or an anticancer agent.
  12. In claim 11, the pre-apoptotic peptide is a drug delivery vehicle selected from the group consisting of KLA, alpha-defensin-1, BMAP-28, Brevenin-2R, Buforin IIb, cecropin A-Magainin 2 (CA-MA-2), cecropin A, cecropin B, chrysophsin-1, D-K6L9, Gomesin, Lactoferricin B, LLL27, LTX-315, Magainin 2, Magainin II-bombesin conjugate (MG2B), Pardaxin, and combinations thereof.
  13. In claim 11, the immunogenic apoptosis inducer is a drug delivery vehicle selected from the group consisting of anthracycline anticancer agents, taxane anticancer agents, anti-EGFR antibodies, BK channel agonists, bortezomib, cardiac glycosides, cyclophosphamide anticancer agents, GADD34/PP1 inhibitors, LV-tSMAC, Measles virus, bleomycin, mitoxantrone, oxaliplatin, and combinations thereof.
  14. In paragraph 11, the anticancer agents are SN-38 (7-ethyl-10-hydroxy-camptothecin), daunorubicin, doxorubicin, epirubicin, idarubicin, pixantrone, sabarubicin, valrubicin, paclitaxel, docetaxel, mechloethamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), Streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin (actinomycin D), plicamycin, mitomycin C, vincristine, vinblastine, teniposide, topotecan, iridotecan, uramustine, melphalan, bendamustine, dacarbazine, temozolomide, altretamine, duocarmycin, nedaplatin, oxaliplatin, satraplatin, Triplatin tetranitrate, 5-fluorouracil, 6-mercaptopurine, capecitabine, cladribine, clofarabine, cystarbine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, pemetrexed, pentostatin, thioguanine, etoposide, mitoxantrone, izabepilone, vindesine, vinorelbine, estramustine, A drug delivery system selected from the group consisting of maytansine, DM1 (mertansine), DM4, dolastatin, auristatin E, auristatin F, monomethyl auristatin E (MMAE), monomethyl auristatin F, and derivatives thereof.
  15. In claim 9, a drug delivery system that specifically delivers a drug to the tumor microenvironment or cancer cells.
  16. In claim 9, a drug delivery system that specifically reduces tumor-infiltration regulatory T cells.
  17. A pharmaceutical composition for the prevention or treatment of cancer comprising the drug delivery system of claim 9 as an active ingredient.
  18. In paragraph 17, cancer is any one or more selected from the group consisting of brain tumor, melanoma, multiple myeloma, non-small cell lung cancer, oral cancer, liver cancer, gastric cancer, colon cancer, breast cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cervical cancer, ovarian cancer, colorectal cancer, small intestine cancer, rectal cancer, fallopian tube carcinoma, proanal cancer, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, lymphoma, bladder cancer, gallbladder cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, renal or ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma, of Pharmaceutical composition for prevention or treatment.
  19. Clause 9. An anticancer adjuvant comprising a drug delivery system as an active ingredient.
  20. In paragraph 19, an anticancer adjuvant administered in combination with an immunotherapy agent, simultaneously, separately, or sequentially.

Description

Peptides specifically binding to CCR8 and uses thereof The present invention relates to the use of a peptide, antibody, or fragment having the same’s immunological activity that specifically binds to CCR8. Cancer cells occur periodically in the human body, but immune cells such as NK cells and cytotoxic T cells kill them, preventing them from developing into cancer. However, if cancer cells evade immunity by inducing immunosuppressive effects through immune checkpoints such as PD-1 and CTLA-4, or by inducing regulatory T cells (Treg), the normal immune response breaks down and cancer develops (Van Damme et al., 2021). Conventional anticancer treatments have been researched in the direction of directly attacking cancer cells or enhancing the activity of the body's immune cells that attack cancer cells. However, these anticancer drugs also attack normal cells other than cancer cells, leading to numerous side effects such as hair loss, nausea, and vomiting, as well as causing adverse reactions due to the excessive proliferation of immune cells. Anticancer immunotherapy, which can minimize side effects compared to conventional chemotherapy or radiation therapy, is a method of treating cancer by utilizing the body's immune system. Among these anticancer immunotherapy techniques, active research is being conducted on cell therapy methods, which involve activating therapeutic immune cells such as T cells (including CAR-T), dendritic cells, and natural killer cells outside the body and injecting them directly into the body, and anticancer vaccine methods, which enhance anticancer efficacy by directly activating existing immune cells through the injection of cancer antigens and immune-activating substances. However, these cell therapies and cancer vaccines are primarily used for blood cancers, and they have the disadvantage of generally having very low therapeutic efficacy in solid tumors. One of the reasons for this is attributed to microenvironmental factors that suppress immune function around solid tumors. In fact, cells that impair immune function (MDSCs: myeoloid-derived stromal cells, Treg: regulatory T cells, TAM: tumor-assisted macrophages), as well as immunosuppressive cytokines and metabolites, act actively within the tumor microenvironment, thereby drastically reducing the activity of immune-activating substances and therapeutic immune cells. Therefore, it is becoming increasingly important to develop therapeutic agents that possess anticancer effects by regulating only the tumor microenvironment without directly affecting tumor cells or immune cells, thereby blocking nutrient supply to tumor cells and angiogenesis around them. The tumor microenvironment is considered a major therapeutic target as it contributes to the proliferation and survival of malignant cells, angiogenesis, metastasis, abnormal adaptive immunity, and reduced response to hormones and chemotherapy agents. Accordingly, efforts are being made to develop immunotherapies that activate the immune response of T effector cells by reducing regulatory T cells (Tregs) in the tumor microenvironment. Most strategies to reduce Tregs target peripheral Tregs; while the death of Tregs can reduce cancer cells, it can destroy resistance to inflammation, thereby causing serious autoimmune risks. Therefore, there is a need to develop new markers to target and destroy only tumor-infiltrating regulatory T cells (Ti-Tregs) without affecting other effector T cells or peripheral Tregs (Hatzioannou et al., 2021; Moser, 2022). Figure 1 is a diagram showing the process of selecting peptides that bind to CCR8 through M13 phage display biopanning and NGS. Figure 2 is a figure confirming the selective increase of peptides attached to the target according to the biopanning round: A: Tittering results of eluted phage and amplified phage; and B: Number of eluted and amplified phages (log 10 ). Figure 3 is a figure showing the number of NGS reads in each round: A: Total sorted reads; and B to E: Number of peptides according to the number of NGS reads for each lead. Figure 4 is a heatmap showing clustered 336 peptides with more than 10 leads in 4 rounds. Figure 5 is a figure showing the derivation of the common sequence of cluster A. Figure 6 is a figure showing the derivation of the common sequence of cluster B. Figure 7 is a figure showing the derivation of the common sequence of cluster C. Figure 8 is a figure confirming the actual read number change of three candidate peptides. Figure 9 shows the secondary and tertiary structures of human CCR8 predicted by the in silico method: TMs: transmembranes; ECL: extracellular loop; A: Secondary structure of human CCR8 predicted by PSIPRED 4.0; B: Predicted human CCR8 structure & region, and amino acid sequence number; and C: 3D structure of human CCR8 predicted by trRosetta server: Purple: Spiral; Yellow: strand; and Other: Coil structure. FIG. 10 is a figure showing the simulation results predicting the binding of three typ