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EP-4739704-A2 - RECOMBINANT POLYPEPTIDES, RECOMBINANT NUCLEIC ACIDS ENCODING THE SAME, AND USES THEREOF IN TREATING CANCERS

EP4739704A2EP 4739704 A2EP4739704 A2EP 4739704A2EP-4739704-A2

Abstract

Disclosed herein are recombinant polypeptides and recombinant nucleic acids encoding the same. According to some embodiments of the present disclosure, the recombinant polypeptide comprises a first bi-functional domain, and a first single-chain fragment variable (scFv) or a peptide linked to the N-terminus of the first bi-functional domain. Optionally, the recombinant polypeptide further comprises a second scFv linked to the N-terminus of the first scFv or peptide. Also disclosed herein are methods of treating cancers by using the immune cells expressing the recombinant polypeptides.

Inventors

  • CHEN, MING-WEI

Assignees

  • Rephimmune Biotechnology Inc.

Dates

Publication Date
20260513
Application Date
20240624

Claims (20)

  1. WHAT IS CLAIMED IS: 1. A recombinant polypeptide comprising a first bi-functional domain, and a first single-chain fragment variable (scFv) or a peptide linked to the N-terminus of the first bi-functional domain, wherein the first bi-functional domain comprises, in sequence, an intracellular loop 1 (ICL1), an ICL2, an ICL3 and a C-terminal region of a first G protein-coupled receptor (GPCR), and is characterized in not having an extracellular domain nor a transmembrane domain of the first GPCR.
  2. 2. The recombinant polypeptide of claim 1, wherein the first bi-functional domain consists of the ICL1, ICL2, ICL3 and C-terminal region of the first GPCR.
  3. 3. The recombinant polypeptide of claim 1, wherein the first GPCR is a class A GPCR.
  4. 4. The recombinant polypeptide of claim 3, wherein the first GPCR is cannabinoid receptor 2 (CNR2), hydroxycarboxylic acid receptor 2 (HCAR2), G-protein coupled receptor 84 (GPR84), or P2Y purinoceptor 14 (P2Y14).
  5. 5. The recombinant polypeptide of claim 4, wherein the first GPCR is the CNR2, and the first bi- functional domain comprises the amino acid sequence of SEQ ID NO: 2.
  6. 6. The recombinant polypeptide of claim 4, wherein the first GPCR is the HCAR2, and the first bi-functional domain comprises the amino acid sequence of SEQ ID NO: 4.
  7. 7. The recombinant polypeptide of claim 4, wherein the first GPCR is the GPR84, and the first bi-functional domain comprises the amino acid sequence of SEQ ID NO: 6.
  8. 8. The recombinant polypeptide of claim 4, wherein the first GPCR is the P2Y14, and the first bi-functional domain comprises the amino acid sequence of SEQ ID NO: 8.
  9. 9. The recombinant polypeptide of claim 1, wherein the first scFv is specific to CD3 or NKp46.
  10. 10. The recombinant polypeptide of claim 1, further comprising a second scFv that is linked to the N-terminus of the first scFv or peptide, and is specific to a tumor-associated antigen (TAA).
  11. 11. The recombinant polypeptide of claim 1, further comprising a second bi-functional domain disposed at and connected to the C-terminus of the first bi-functional domain, wherein the second bi-functional domain comprises in sequence, an ICL1, an ICL2, an ICL3 and a C-terminal region of a second GPCR; the second bi-functional domain is characterized in not having an extracellular domain nor a transmembrane domain of the second GPCR; and the second GPCR is different from the first GPCR.
  12. 12. The recombinant polypeptide of claim 11, wherein the second bi-functional domain consists of the ICL1, ICL2, ICL3 and C-terminal region of the second GPCR.
  13. 13. The recombinant polypeptide of claim 11, wherein each of the first and second GPCRs is a class A GPCR.
  14. 14. The recombinant polypeptide of claim 13, wherein the first and second GPCRs are independently selected from the group consisting of CNR2, HCAR2, GPR84 and P2Y14.
  15. 15. The recombinant polypeptide of claim 14, wherein the first and second GPCRs are respectively the CNR2 and P2Y14; and the first and second bi-functional domains respectively comprise the amino acid sequences of SEQ ID NOs: 2 and 8.
  16. 16. The recombinant polypeptide of claim 14, wherein the first and second GPCRs are respectively the CNR2 and HCAR2; and the first and second bi-functional domains respectively comprise the amino acid sequences of SEQ ID NOs: 2 and 4.
  17. 17. The recombinant polypeptide of claim 11, further comprising a third bi-functional domain disposed at and connected to the C-terminus of the second bi-functional domain, wherein the third bi-functional domain comprises, in sequence, an ICL1, an ICL2, an ICL3 and a C- terminal region of a third GPCR; the third bi-functional domain is characterized in not having an extracellular domain nor a transmembrane domain of the third GPCR; and the first, second and third GPCRs are different from one another.
  18. 18. The recombinant polypeptide of claim 17, wherein the third bi-functional domain consists of the ICL1, ICL2, ICL3 and C-terminal region of the third GPCR.
  19. 19. The recombinant polypeptide of claim 17, wherein each of the first, second and third GPCRs is a class A GPCR.
  20. 20. The recombinant polypeptide of claim 17, wherein the first, second and third GPCRs are independently selected from the group consisting of CNR2, HCAR2, GPR84 and P2Y14.

Description

RECOMBINANT POLYPEPTIDES, RECOMBINANT NUCLEIC ACIDS ENCODING THE SAME, AND USES THEREOF IN TREATING CANCERS BACKGROUND OF THE INVENTION [0001] 1. FIELD OF THE INVENTION [0002] The present disclosure in general relates to the field of disease treatment. More particularly, the present disclosure relates to the treatment of cancers via novel cell therapy. [0003] 2. DESCRIPTION OF RELATED ART [0004] Cancer is a leading cause of death worldwide, accounting for nearly 10 million deaths in 2020, or nearly one in six deaths. One defining feature of cancer is the rapid growth of abnormal cells that develop uncontrollably and can invade or spread to any part of the body. The most common cancers include, breast, lung, colorectal, prostate, skin and gastric cancers. Signs and symptoms associated with cancer usually vary depending on what part of the body is affected. The general signs and symptoms of cancer include fatigue, lump, weight changes (e.g., unintended loss or gain), changes in bowel or bladder habits, difficulty swallowing, nausea, vomiting, fever, night sweats, bleeding, bruising, cough and/or trouble breathing. Symptoms of advanced cancer further include pain, weakened physical strength, and paralysis that affect the movement of cancer patients, resulting in depression, loss of independence and disruption of social relationships. In addition to the patients themselves, cancer also affects patients’ family and friends. Many caregivers experienced a physical, mental, and emotional struggle. According to some reports, a cancer diagnosis even has a negative effect on the caregivers’ health; for example, they may have higher rates of depression and weakened immune response, as well as cardiovascular diseases. [0005] While different types of therapeutic approaches are currently available, including chemotherapy, surgery, radiation therapy, hormone therapy, targeted therapy, and most recently, immunotherapy and cell-based therapy, none of these treatments provides a satisfactory result. Cancer still poses significant health risks, and increases the personal, family, social and economic burden. In view of the foregoing, there is a continuing interest in developing a novel agent and method for treating cancer. SUMMARY [0006] The following presents a simplified summary of the disclosure to provide a basic understanding to the reader. This summary is not an extensive overview of the disclosure, and it does not identify key/critical elements of the present invention or delineate the scope of the present invention. Its sole purpose is to present some concepts disclosed herein in a simplified form as a prelude to the more detailed description that is presented later. [0007] As embodied and broadly described herein, the first aspect of the present disclosure is directed to a recombinant polypeptide comprising a bi-functional domain, and a single-chain fragment variable (scFv) or peptide linked to the N-terminus of the bi-functional domain. According to the embodiments of the present disclosure, the bi-functional domain is capable of spanning cell membrane (i.e., serving as a transmembrane domain) and mediating signal transduction in cells (i.e., serving as an intracellular domain). [0008] According to some embodiments of the present disclosure, the bi-functional domain comprises, from its N-terminus to C-terminus, in sequence, an intracellular loop 1 (ICL1), an ICL2, an ICL3 and a C-terminal region of a G protein-coupled receptor (GPCR), without comprising an extracellular domain and a transmembrane domain of the GPCR. According to some preferred embodiments, the bi-functional domain consists of the ICL1, ICL2, ICL3 and C- terminal region of the GPCR. [0009] The GPCR is preferably a class A GPCR. According to some exemplary embodiments, the GPCR is cannabinoid receptor 2 (CNR2), hydroxycarboxylic acid receptor 2 (HCAR2), G-protein coupled receptor 84 (GPR84), or P2Y purinoceptor 14 (P2Y14). [0010] According to one embodiment, the bi-functional domain comprises the ICL1, ICL2, ICL3 and C-terminal region of CNR2. In the embodiment, the CNR2-derived bi-functional domain comprises the amino acid sequence of SEQ ID NO: 2. [0011] According to another embodiment, the bi-functional domain comprises the ICL1, ICL2, ICL3 and C-terminal region of HCAR2. In the embodiment, the HCAR2-derived bi- functional domain comprises the amino acid sequence of SEQ ID NO: 4. [0012] According to another embodiment, the bi-functional domain comprises the ICL1, ICL2, ICL3 and C-terminal region of GPR84. In the embodiment, the GPR84-derived bi- functional domain comprises the amino acid sequence of SEQ ID NO: 6. [0013] According to still another embodiment, the bi-functional domain comprises the ICL1, ICL2, ICL3 and C-terminal region of P2Y14. In the embodiment, the P2Y14-derived bi- functional domain comprises the amino acid sequence of SEQ ID NO: 8. [0014] According to certain embodiments of the present disclosure, the scFv is specific to CD3 or