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CN-121985957-A - Use of coronavirus 3CL protease for preventing or treating tumors

CN121985957ACN 121985957 ACN121985957 ACN 121985957ACN-121985957-A

Abstract

Use of a 3CL protease of a coronavirus or a nucleic acid molecule encoding said 3CL protease of a coronavirus for the preparation of a medicament for the prevention and/or treatment of a tumor.

Inventors

  • ZHONG WU
  • CAO RUIYUAN
  • TAO HUIMIN
  • YANG XIAOTONG
  • LI WEI
  • LI SONG

Assignees

  • 中国人民解放军军事科学院军事医学研究院

Dates

Publication Date
20260505
Application Date
20240904
Priority Date
20230907

Claims (13)

  1. Use of a 3CL protease of a coronavirus or a nucleic acid molecule encoding the same for the preparation of a medicament for the prevention and/or treatment of a tumor.
  2. Use according to claim 1, wherein the coronavirus is SARS-CoV, MERS-CoV or SARS-CoV-2, preferably SARS-CoV-2.
  3. The use of claim 1 or 2, wherein the 3CL protease has an amino acid sequence selected from the group consisting of: i) A sequence shown in SEQ ID NO.6, 10 or 13; ii) sequences having one or more amino acid substitutions, deletions or additions (e.g.1, 2,3,4 or 5 amino acid substitutions, deletions or additions) compared to the sequence shown in SEQ ID NO:6, 10 or 13, and Iii) A sequence having at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID No. 6, 10 or 13; preferably, the substitution described in ii) is a conservative substitution.
  4. The use of any one of claims 1-3, wherein the nucleic acid molecule is a DNA molecule or an RNA molecule; Preferably, the nucleic acid molecule is single-stranded or double-stranded.
  5. The use of claim 4, wherein the nucleic acid molecule is an RNA molecule, preferably an mRNA molecule; Preferably, the nucleic acid molecule comprises the coding sequence of the 3CL protease; Preferably, the nucleic acid molecule further comprises one or more selected from the group consisting of a 5'UTR, a Kozak sequence, a start codon, a stop codon, a 3' UTR, a poly-A tail; Preferably, the nucleic acid molecule comprises, in order from the 5 'end to the 3' end, a 5'UTR, a Kozak sequence, a start codon, a coding sequence for the 3CL protease, a stop codon, a 3' UTR, a poly-A tail; Preferably, the nucleic acid molecule consists of, in order from the 5 'end to the 3' end, a 5'UTR, a Kozak sequence, a start codon, a coding sequence for the 3CL protease, a stop codon, a 3' UTR, a poly-A tail.
  6. The use according to claim 5, which is provided with one or more features selected from the group consisting of: (1) The coding sequence of the 3CL protease is codon optimized or non-optimized according to the codon preference of the host cell (e.g., human cell); (2) The coding sequence of the 3CL protease has at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleotide sequence shown in SEQ ID NO:1, 8, or 11, preferably the coding sequence of the 3CL protease has a sequence as shown in SEQ ID NO:1, 8, or 11; (3) The 5 'end of the nucleic acid molecule is modified 5' (e.g., CAP0, CAP1, CAP 2); (4) The nucleic acid molecule comprises one or more N1-methyl pseudouridine (m1ψ) modifications, preferably the uracil nucleotides in the nucleic acid molecule are replaced in whole or in part by N1-methyl-pseudouracil nucleotides; (5) One or more stop codons are present at the 3' end of the coding sequence of the 3CL protease; (6) The 5'UTR has a nucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleotide sequence shown in SEQ ID NO. 2, preferably the 5' UTR has the nucleotide sequence shown in SEQ ID NO. 2; (7) The 3'UTR has a nucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleotide sequence shown in SEQ ID NO. 4, preferably the 3' UTR has the nucleotide sequence shown in SEQ ID NO. 4; (8) The Kozak sequence has a sequence shown in SEQ ID NO. 3; (9) The poly-A tail comprises one or more poly A sequences, each independently consisting of 20-120 consecutive adenylates, preferably the poly-A tail comprises a plurality of poly A sequences and the adjacent poly A sequences are linked by a spacer sequence comprising non-A, preferably the poly-A tail has a sequence as shown in SEQ ID NO 5; (10) The nucleic acid molecule has a nucleotide sequence having at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleotide sequence set forth in SEQ ID NO 7, 9, or 12, preferably the nucleic acid molecule has a sequence set forth in SEQ ID NO 7, 9, or 12.
  7. The use of any one of claims 1-6, wherein the tumor is a solid tumor or a hematological tumor (e.g., leukemia, lymphoma, myeloma); preferably, the tumor is a solid tumor; preferably, the tumor is selected from pancreatic cancer, gastric cancer, liver cancer, melanoma, intestinal cancer, renal cancer, cervical cancer, osteosarcoma, prostate cancer, glioma and lung cancer; Preferably, the pancreatic cancer classification includes, but is not limited to, pancreatic acinar cell carcinoma, adenosquamous carcinoma, squamous cell carcinoma, ring cell carcinoma, undifferentiated carcinoma, giant cell undifferentiated carcinoma, ampulla carcinoma, pancreatic neuroendocrine tumor, and the like; the gastric cancer type includes but is not limited to cardiac cancer, gastric body cancer and pyloric cancer; the liver cancer types include, but are not limited to, hepatocellular carcinoma, fibrolamellar liver carcinoma, intrahepatic duct carcinoma, angiosarcoma, hepatoblastoma, secondary liver cancer, benign liver tumor; such melanoma types include, but are not limited to, localized melanoma, localized area melanoma, distant metastatic melanoma, such intestinal cancer types include, but are not limited to, colon cancer and rectal cancer, including in particular adenocarcinoma, undifferentiated carcinoma, adenosquamous carcinoma, squamous cell carcinoma, small cell carcinoma, carcinoid, mucinous carcinoma, etc., such renal cancer types include, but are not limited to, clear cell carcinoma, papillary cell carcinoma, chromophobe carcinoma, low malignant potential multiple chamber renal cell carcinoma, manifold carcinoma, renal medullary carcinoma, such cervical cancer types include, but are not limited to, squamous carcinoma, adenocarcinoma, adenosquamous carcinoma, such osteosarcoma types include, but are not limited to, telangiectasia type osteosarcoma, small round cell type osteosarcoma, fibrohistiocyte type osteosarcoma, intramedullary hyperdifferentiated osteosarcoma, multicenter type osteosarcoma, intracortical osteosarcoma, paramyomatoid, dedifferentiated osteosarcoma, highly malignant surface osteosarcoma, such prostate cancer types include, but are not limited to, ductal carcinoma (acinar), ductal carcinoma, astrocytoma, anaplastic carcinoma, astrocytoma, etc., such as described in the patient's, the patient may be in need to be in need of the patient, or the patient, to be in need of the patient, or to be in the patient, or to be, anaplastic oligodendrocytomas, glioblastomas, and lung cancer types including, but not limited to, small cell lung cancer and non-small cell lung cancer (adenocarcinoma, squamous carcinoma, large cell carcinoma, adenosquamous carcinoma, carcinoid, etc.).
  8. An isolated nucleic acid molecule comprising a nucleotide sequence encoding a 3CL protease, wherein the 3CL protease is as defined in any one of claims 1-3; preferably, the isolated nucleic acid molecule is a nucleic acid molecule as defined in any one of claims 4 to 6, or the isolated nucleic acid molecule encodes a nucleic acid molecule as defined in any one of claims 4 to 6.
  9. A vector comprising the isolated nucleic acid molecule of claim 8.
  10. A delivery composition comprising a delivery vehicle, which comprises one or more selected from the group consisting of a 3CL protease, the isolated nucleic acid molecule of claim 8, the vector of claim 9, wherein the 3CL protease is as defined in any one of claims 1-3; Preferably, the delivery vehicle is for entrapment, carrying or delivery of the 3CL protease, isolated nucleic acid molecule or vector; Preferably, the delivery vector comprises a non-viral vector, a viral vector, and a Viroid (VLP) vector interposed between the viral vector and the non-viral vector; Preferably, the non-viral vectors include, but are not limited to, cationic liposomes, lipid nanoparticles, lipopolymers, artificial microspheres, micelles, lipid-polymer hybrid systems, extracellular vesicles, natural or engineered exosomes, etc., including, but not limited to, adeno-associated viruses, lentiviruses, adenoviruses, retroviruses, vaccinia viruses, measles viruses, herpes simplex viruses, alphaviruses, vesicular stomatitis viruses, influenza viruses, etc.; preferably, the delivery vehicle is a lipid nanoparticle.
  11. A pharmaceutical composition comprising a 3CL protease, an isolated nucleic acid molecule of claim 8, a vector of claim 9 and/or a delivery composition of claim 10, and a pharmaceutically acceptable carrier and/or excipient, wherein the 3CL protease is as defined in any one of claims 1-3; preferably, the pharmaceutical composition further comprises other antitumor drugs; Preferably, the other antitumor drug is selected from small molecule drugs, biological macromolecule antibodies or proteins, chimeric antigen receptor cell drugs, and antibody-coupled drugs; Preferably, the pharmaceutical composition is administered orally, by inhalation spray, rectally, nasally, bucally, vaginally, topically, parenterally, e.g., subcutaneously, intravenously, intramuscularly, intraperitoneally, intrathoracic, intrathecally, intraventricularly, intrasternally and intracranially by injection or infusion, intratracheal installation, surgical implantation, transdermal delivery, local injection, overdrive injection/bombardment, or by means of an explanted reservoir, Preferably, the medicament is administered orally, intraperitoneally, intrathoracic or intravenously.
  12. A method of inhibiting a tumor cell in vivo or in vitro comprising contacting the tumor cell with, or delivering into, a 3CL protease, the isolated nucleic acid molecule of claim 8, the vector of claim 9, the delivery composition of claim 10 or the pharmaceutical composition of claim 11, wherein the 3CL protease is as defined in any one of claims 1-3, Preferably, the tumor cell is a cell of a solid tumor or a hematological tumor (e.g., leukemia, lymphoma, myeloma); Preferably, the tumor cells are selected from the group consisting of pancreatic cancer, gastric cancer, liver cancer, melanoma, intestinal cancer, renal cancer, cervical cancer, osteosarcoma, prostate cancer, glioma, and lung cancer cells; Preferably, the pancreatic cancer classification includes, but is not limited to, pancreatic acinar cell carcinoma, adenosquamous carcinoma, squamous cell carcinoma, ring cell carcinoma, undifferentiated carcinoma, giant cell undifferentiated carcinoma, ampulla carcinoma, pancreatic neuroendocrine tumor, and the like; the gastric cancer type includes but is not limited to cardiac cancer, gastric body cancer and pyloric cancer; the liver cancer types include, but are not limited to, hepatocellular carcinoma, fibrolamellar liver carcinoma, intrahepatic duct carcinoma, angiosarcoma, hepatoblastoma, secondary liver cancer, benign liver tumor; such melanoma types include, but are not limited to, localized melanoma, localized area melanoma, distant metastatic melanoma, such intestinal cancer types include, but are not limited to, colon cancer and rectal cancer, including in particular adenocarcinoma, undifferentiated carcinoma, adenosquamous carcinoma, squamous cell carcinoma, small cell carcinoma, carcinoid, mucinous carcinoma, etc., such renal cancer types include, but are not limited to, clear cell carcinoma, papillary cell carcinoma, chromophobe carcinoma, low malignant potential multiple chamber renal cell carcinoma, manifold carcinoma, renal medullary carcinoma, such cervical cancer types include, but are not limited to, squamous carcinoma, adenocarcinoma, adenosquamous carcinoma, such osteosarcoma types include, but are not limited to, telangiectasia type osteosarcoma, small round cell type osteosarcoma, fibrohistiocyte type osteosarcoma, intramedullary hyperdifferentiated osteosarcoma, multicenter type osteosarcoma, intracortical osteosarcoma, paramyomatoid, dedifferentiated osteosarcoma, highly malignant surface osteosarcoma, such prostate cancer types include, but are not limited to, ductal carcinoma (acinar), ductal carcinoma, astrocytoma, anaplastic carcinoma, astrocytoma, etc., such as described in the patient's, the patient may be in need to be in need of the patient, or the patient, to be in need of the patient, or to be in the patient, or to be, anaplastic oligodendrocytomas, glioblastomas, and lung cancer types including, but not limited to, small cell lung cancer and non-small cell lung cancer (adenocarcinoma, squamous carcinoma, large cell carcinoma, adenosquamous carcinoma, carcinoid, etc.).
  13. Use of an isolated nucleic acid molecule according to claim 9, a vector according to claim 10, a delivery composition according to claim 11 or 12 or a pharmaceutical composition according to claim 13 for the preparation of a medicament for the prevention and/or treatment of tumors.

Description

Use of coronavirus 3CL protease for preventing or treating tumors The present application is based on and claims priority from CN application number 202311153380.8, day 2023, month 9, 7, the disclosure of which is incorporated herein by reference in its entirety. Technical Field The application belongs to the technical field of medical biology, and particularly relates to application of 3CL protease of coronavirus or a nucleic acid molecule encoding the same in preparation of medicines for preventing and/or treating tumors. Background Cancer is a disease characterized by uncontrolled growth of cells induced by mutation or abnormal regulation of genes, and exhibits strong spatiotemporal heterogeneity according to the site of occurrence. The application of the cancer therapeutic drugs not only needs to overcome the heterogeneity, but also achieves the aim of minimizing toxicity to normal cells and selectively killing cancer cells. In the aspect of anti-cancer drug development, how to realize the reduction of toxicity of clinical drugs to organisms at the present stage and to maintain the broad spectrum of anti-cancer spectrum of drugs has been a challenging problem. For different cancer types, including solid and non-solid tumors, current clinical treatment protocols typically include one or a combination of surgery, chemotherapy, radiation therapy, bone marrow/stem cell transplantation, hormonal therapy, targeted therapy and immunotherapy. The seven main types of mainstream anti-tumor therapies can achieve the purpose of eliminating tumors to a certain extent, but the patients can have drug resistance and recurrence risk when taking the drugs, and the patients feel pain or the body is irreversibly damaged in the process of repeatedly receiving the treatment. Development and application of novel effective anti-tumor therapeutic methods are urgently needed. Gene drug therapy and gene vaccination belong to anti-tumor strategies which develop faster in recent years, and provide specific and individual choices for the treatment and prevention of various tumors. Currently, there are only several gene drug therapies approved for the tumor field worldwide, such as GENDICINE for treating head and neck squamous cell carcinoma, imlygic for treating melanoma lesions that cannot be completely resected by surgery, kymriah for treating recurrent or refractory acute lymphoblastic leukemia of 3 to 25 years old, yescarta for treating recurrent or refractory diffuse large B cell lymphoma and primary mediastinal large B cell lymphoma systematically twice or more in adults, and gene drug therapies represented by these four therapeutic methods bring suicide genes, immunomodulating genes into cells by using viruses, engineered immune cells or other delivery vehicles, and insert them into existing genomes, thereby eliminating the influence of harmful mutations in genes or stimulating anti-tumor immune responses of patients to achieve anti-tumor purposes. In addition to this mode of introducing exogenous genes, delivery vehicles may also be used to mediate the entry of oligonucleotides into tumor cells, inhibit or interfere with endogenous genes for therapeutic purposes. In addition, researchers have developed a variety of DNA vaccines or RNA vaccines encoding tumor antigens for the prevention and treatment of tumors, belonging to the field of genetic vaccines. Related studies have shown that messenger RNA vaccines have become a more promising cancer immunotherapy strategy. Following vaccination with either naked or drug-loaded mRNA vaccines, tumor antigens will be expressed in Antigen Presenting Cells (APCs), thereby promoting APC activation and innate/adaptive immunity. The mRNA cancer vaccine has the advantages of high efficiency, safety, great development potential, low production cost and the like, and is advanced in front of other conventional vaccine platforms. Tumor vaccines are used as an immunotherapy means and highly depend on the immune system of the body, but the immune system of cancer patients is often inhibited and the defense function is low, so the application of the tumor vaccines has a certain limit. MRNA therapy in gene therapy has been explored as the safest and effective method of treating tumors. mRNA consists of four nucleotides, abbreviated as letters A, U, C and G. The sequence of the nucleotide arrangement is read by the cell, if the information encoded by the mRNA cannot be read, the mRNA cannot be translated into a protein with a specific structure and function, and the mRNA is finally metabolized and degraded by the cell and does not generate corresponding biological effects. If the opposite happens, the body will only make the protein it needs at the right time and place. Thus, mRNA drug therapy is considered to be a precise, safe anti-tumor strategy. Thus, there remains a great medical need for anti-tumor mRNA drugs to improve patient survival, bringing more therapeutic benefit to the patient himself