WO2021131944A1 - 癌遺伝子治療薬 - Google Patents

癌遺伝子治療薬 Download PDF

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WO2021131944A1
WO2021131944A1 PCT/JP2020/046891 JP2020046891W WO2021131944A1 WO 2021131944 A1 WO2021131944 A1 WO 2021131944A1 JP 2020046891 W JP2020046891 W JP 2020046891W WO 2021131944 A1 WO2021131944 A1 WO 2021131944A1
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nucleic acid
protein
base sequence
hif
sequence
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WO2021131944A8 (ja
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利朗 白川
尚人 國村
孝一 北川
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Kobe University NUC
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Kobe University NUC
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Priority to US17/789,447 priority Critical patent/US12459984B2/en
Priority to EP20908300.5A priority patent/EP4083213A4/en
Priority to CN202080090551.5A priority patent/CN115151644B/zh
Priority to JP2021567329A priority patent/JP7737708B2/ja
Publication of WO2021131944A1 publication Critical patent/WO2021131944A1/ja
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    • C12N15/62—DNA sequences coding for fusion proteins
    • C12N15/625—DNA sequences coding for fusion proteins containing a sequence coding for a signal sequence
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    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
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    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Definitions

  • the present disclosure relates to, for example, a novel oncogene therapeutic agent or method, an active ingredient thereof, and the like.
  • a novel oncogene therapeutic agent or method an active ingredient thereof, and the like.
  • gene therapy using a viral vector has been attracting attention as one of the new treatments for cancer.
  • This utilizes the characteristics of various viruses and uses the virus as a "carrier" of genes and a viral vector.
  • a recombinant viral vector incorporating a therapeutic gene can be directly administered into the body to treat a disease.
  • Oncogene therapy has a different mechanism of action from existing treatments such as chemotherapy, in which genes are introduced into cancer cells to directly suppress genes involved in cancer development and growth, or tumor suppressor genes are introduced into cells. Directly induce cell death. Therefore, it is expected to be an effective treatment method for cancers that are difficult to treat with existing chemotherapy and radiation therapy.
  • HIF-3alpha4 is a dominant-negative regulator of HIF-1 and is down-regulated in renal cell . FASEB J. 2005 Sep; 19 (11): 1396-406.
  • the present inventors conducted a study with the main purpose of providing a novel oncogene therapeutic agent or method.
  • a viral vector that carries a gene is important for expressing a gene introduced from the outside in cancer cells.
  • Viral vectors include, for example, retroviruses, lentiviruses, and Sendai viruses.
  • the present disclosure includes, for example, the subjects described in the following sections.
  • Item 1. Nucleic acid encoding a protein having a CD44 extracellular function (B) Nucleic acid encoding a protein having a Notch core region function (C) Nucleic acid encoding a protein having a HIF-3 ⁇ 4 function (A)-(B) -Nucleic acid having a structure linked in the order of (C).
  • Item 2. Nucleic acid (A), nucleic acid (B), and nucleic acid (C) are nucleic acids having a structure in which (A)-(B)-(C) are linked in this order.
  • Item 4. (D-1) Does it consist of the nucleotide sequence of SEQ ID NO: 4? (D-2) In the base sequence of SEQ ID NO: 4, a portion consisting of a base sequence in which one or more bases are deleted, substituted, or added and encoding a protein capable of binding to hyaluronic acid, a protease.
  • the portion encoding the protein that can be cleaved and the portion encoding the protein that can bind to HIF-1 ⁇ have a structure in which they are linked in this order, or (D-3) In the base sequence of SEQ ID NO: 4, it comprises a base sequence in which one or more bases are deleted, substituted, or added, and encodes a protein having an anticancer activity.
  • Item 8. Item 7. The vector according to Item 7, wherein the vector is an adenovirus vector.
  • Item 10. Item 9. The anti-cancer composition according to Item 9, which is an injection.
  • the anticancer composition according to Item 9 or 10 which is used for treating breast cancer, prostate cancer, gastric cancer, or pancreatic cancer.
  • Item 12. Item 8. The anti-cancer composition according to Item 9 or 10, which is used for treating triple-negative breast cancer.
  • a novel oncogene therapeutic agent having a very high anticancer effect can be provided.
  • the therapeutic agent may be effective against small cancer types that are not effective with existing cancer therapeutic agents, and may also be effective against, for example, triple-negative (estrogen receptor negative, progesterone receptor negative, HER2-negative) breast cancer and the like. It can be preferably used.
  • the sequence of the prepared CD44 / Notch / HIF-3 ⁇ 4 fusion gene is shown.
  • the base sequence and amino acid sequence of human CD44 are shown.
  • the underlined part is the part used for the production of the CD44 / Notch / HIF-3 ⁇ 4 fusion gene.
  • the base sequence of Notch is shown.
  • the base sequence of Notch (continued) is shown.
  • the underlined part is the part used for the production of the CD44 / Notch / HIF-3 ⁇ 4 fusion gene.
  • the base sequence of Notch (continued) is shown.
  • the amino acid sequence of Notch is shown.
  • the underlined part is the part used for the production of the CD44 / Notch / HIF-3 ⁇ 4 fusion gene.
  • the base sequence and amino acid sequence of human HIF-3 ⁇ 4 are shown.
  • the underlined part is the part used for the production of the CD44 / Notch / HIF-3 ⁇ 4 fusion gene.
  • the results of infecting cells in vitro with each recombinant adenovirus vector and evaluating whether each gene is normally introduced by a real-time PCR test are shown.
  • Infect cells in vitro with each recombinant adenovirus vector containing ADX730 (genetically modified adenovirus vector incorporating a CD44 / Notch / HIF-3 ⁇ 4 fusion gene), and decoy of the CD44 Decoy receptor of ADX730.
  • the results of a comparative study by a real-time PCR test are shown to see if the expression of the CD44 downstream genes Survivin and CCL2 genes is suppressed by function.
  • Cells are infected in vitro with each recombinant adenovirus vector containing ADX730, and the HIF-1 ⁇ target genes VEGF, CyclinG2, and Bcl-xL genes are affected by the HIF-1 ⁇ function-suppressing effect of HIF-3 ⁇ 4 of ADX730.
  • the results of a comparative study by a Real-time PCR test are shown to see if the expression is suppressed.
  • each recombinant adenovirus vector containing ADX730 to nude mice transplanted with MDA-MB-231 human triple-negative breast cancer cells and examination of its tumor growth inhibitory effect are shown.
  • a schematic diagram of the administration schedule is shown on the upper side.
  • the lower side is a graph showing how the tumor volume was changed by the administration of each recombinant adenovirus vector or PBS (phosphate buffered saline).
  • the present disclosure preferably includes, for example, a specific artificial nucleic acid, a viral vector incorporating the artificial nucleic acid, an anticancer composition containing the viral vector, a method for treating cancer using the composition, and the like. Not limited to these, the present disclosure includes everything disclosed herein and recognizable to those skilled in the art.
  • the artificial nucleic acids included in the present disclosure include (A) a nucleic acid encoding a protein having a CD44 extracellular function, (B) a nucleic acid encoding a protein having a Notch core region function, and (C) a nucleic acid encoding a HIF-3 ⁇ 4 function. It is preferable that the nucleic acid encoding the protein is a nucleic acid having a structure in which (A)-(B)-(C) are linked in this order.
  • the nucleic acid may be referred to as "nucleic acid of the present disclosure".
  • the nucleic acid may be DNA, RNA, PNA or the like, but DNA is particularly preferable.
  • the (A) side may be the 3'end or the 5'end, but the 5'end is preferable.
  • CD44 is one of the receptors for hyaluronic acid and the like, and by binding to a ligand (for example, hyaluronic acid), it clusters and transmits a signal. It is known that this causes intracellular phenomena such as activation of kinases involved in various cell proliferation and cell migration such as c-Src, FAK, and MAPK. It is also known that after the signal transduction, the intracellular domain is transferred to the nucleus by being cleaved by a protease, and the isolated extracellular domain is released as soluble CD44. Due to these properties, CD44 is highly expressed in many types of cancer cells such as colon cancer, breast cancer, gastric cancer, pancreatic cancer, and prostate cancer, and is also being studied as a marker for cancer stem cells.
  • a ligand for example, hyaluronic acid
  • the extracellular function of CD44 is a receptor function for a ligand, and the function preferably includes, for example, hyaluronic acid binding ability.
  • the protein having the extracellular function of CD44 may be, for example, the protein of the entire extracellular portion of CD44, or may include the cell membrane portion of CD44 as long as the receptor function for the ligand is not impaired. It may be a protein of a part of the extracellular component of CD44. Furthermore, one or more amino acids may be deleted, substituted, or added in such proteins as long as they have this function.
  • the nucleic acid (A) is not particularly limited as long as it is a nucleic acid encoding such a polypeptide.
  • examples of the preferred nucleic acid as the nucleic acid (A) include the following (a-1) or (a-2).
  • A-1) Nucleic acid consisting of the nucleotide sequence of SEQ ID NO: 1 (a-2): In the nucleotide sequence of (a-1), 1 or 2 or more (for example, 1 to 30, 1 to 20, 1 to 10, Or a nucleic acid that comprises a base sequence in which 1, 2, 3, 4, or 5) bases are deleted, substituted, or added, and encodes a protein that can bind to hyaluronic acid.
  • Whether the protein can bind to hyaluronic acid can be determined, for example, by obtaining these dissociation constants.
  • Notch is a receptor expressed on the surface of cells
  • the Notch signaling system is one of the main signaling systems responsible for intercellular signaling.
  • the mechanism of intercellular signal transduction is that the signal-sending cell produces and releases a soluble ligand, which binds to a receptor on the cell surface of the signal-receiving cell. ..
  • gene expression is regulated by activating intracellular signal transduction pathways such as the downstream phosphorylation cascade and changing the activity of specific transcription factors.
  • the Notch signaling system is characterized in that signal transduction is performed by direct interaction between adjacent cells.
  • Notch which acts as a receptor, binds to the ligand Delta or Serrate (Jagged in mammals) on the cell surface, and structural changes occur when physical force is applied, resulting in protein-cleaving enzymes such as ADAM protease or ⁇ -secretase.
  • ADAM protease or ⁇ -secretase.
  • the Notch core region is a region containing a site to be cleaved by the action of a protease, and the Notch core region function can be cleaved by a protease capable of cleaving the Notch core region (preferably limited separation). It is a function called.
  • the protein having the Notch core region function may be, for example, a protein containing only a portion necessary for protease cleavage in the Notch core region, and is necessary for protease cleavage as long as it is cleaved by the action of protease.
  • a protein containing one or more amino acids before and after a portion (for example, 1 to 30, 1 to 20, 1 to 10, or 1, 2, 3, 4, or 5) together with a portion required for protease cleavage. May be good.
  • one or more eg, 1-30, 1-20, 1-10, or 1,2,3,4, or 5
  • Amino acids may be deleted, substituted, or added.
  • the nucleic acid (B) is not particularly limited as long as it is a nucleic acid encoding such a protein.
  • examples of the preferred nucleic acid as the nucleic acid (B) include the following (b-1) or (b-2).
  • B-1) Nucleic acid consisting of the base sequence of SEQ ID NO: 2
  • b-2) From the base sequence in which one or more bases are deleted, substituted, or added in the base sequence of (b-1).
  • ADAM protease As the protease, as described above, ADAM protease (ADAM protease) and ⁇ -secretase ( ⁇ -secretase) are preferably mentioned, and a polypeptide that is cleaved by either or both of these proteases can be encoded. More preferred. ADAM protease is a proteolytic enzyme belonging to a group called A Disintegrin And Metalloprotease family.
  • protease Whether the protein can be cleaved by protease can be confirmed by treating the protein with protease and then electrophoresis (for example, SDS-PAGE).
  • (b-2) is composed of a base sequence in which one or more bases are deleted, substituted, or added in the base sequence of (b-2'): (b-1), and is described above.
  • a protein encoded by a nucleic acid having a structure linked in the order of (A)-(B)-(C) it encodes a protein that can be cleaved by an intracellular protease when a ligand binds to the protein encoded by the nucleic acid (A). More preferably, it is a nucleic acid.
  • HIF Hydrofluid Inducible Factor
  • hypoxia-inducing factor is a transcription factor that is activated when the inside of a cell falls into a hypoxic state, and is a heterodimer composed of HIF-1 ⁇ and HIF-1 ⁇ . It has been clarified that HIF-1 ⁇ is suppressed not only by decomposition by PHD under normal oxygen concentration but also by a transcription factor called IPAS (Inhibitory PAS domain protein) found in mice. IPAS was identified as a splicing variant of HIF-3 ⁇ , one of the HIFs. Although IPAS does not exhibit transcriptional activity by itself, it inhibits binding to DNA by interacting with HIF-1 ⁇ and suppresses the function of HIF-1 ⁇ . In humans, HIF-3 ⁇ 4, identified as a splicing variant of HIF-3 ⁇ , has been shown to perform a function similar to IPAS.
  • the HIF-3 ⁇ 4 function is a function capable of suppressing HIF-1 ⁇ , and more specifically, a function of interacting (binding) with HIF-1 ⁇ .
  • the protein having the HIF-3 ⁇ 4 function may be, for example, HIF-3 ⁇ 4 itself, or 1 or 2 or more (for example, 1 to 30, 1) in HIF-3 ⁇ 4 as long as HIF-1 ⁇ can be suppressed. It may be a protein to which amino acids of ⁇ 20, 1-10, or 1, 2, 3, 4, or 5) are further added. Furthermore, in such proteins, as long as HIF-1 ⁇ can be suppressed, one or two or more (for example, 1 to 30, 1 to 20, 1 to 10, or 1, 2, 3, 4, or 5) Amino acids may be deleted, substituted, or added.
  • the nucleic acid (C) is not particularly limited as long as it is a nucleic acid encoding such a protein.
  • examples of the preferred nucleic acid as the nucleic acid (C) include the following (c-1) or (c-2).
  • C-1) Nucleic acid consisting of the base sequence of SEQ ID NO: 3
  • c-2 From the base sequence in which one or more bases are deleted, substituted, or added in the base sequence of (c-1).
  • Whether or not the protein can bind to HIF-1 ⁇ can be examined by a co-immunoprecipitation method or the like using the target protein, HIF-1 ⁇ , and an antibody that recognizes these.
  • nucleic acid (A), nucleic acid (B), and nucleic acid (C) these nucleic acids may be directly linked, or each nucleic acid may be linked via a linker.
  • the linker is not particularly limited as long as the effect of the nucleic acid of the present disclosure is not impaired, but is preferably a nucleic acid consisting of, for example, one or two or more bases.
  • the base length of the nucleic acids of the present disclosure is preferably, for example, 8000 bp or less, and 7500, 7000, 6500, 6000, 5500, More preferably, it is 5000, 4500, 4000, 3500, or 3000 bp or less.
  • nucleic acid of the present disclosure a nucleic acid consisting of the base sequence of (d-1) SEQ ID NO: 4 can be mentioned.
  • the base sequence of SEQ ID NO: 4 comprises a base sequence in which one or more bases are deleted, substituted, or added, and Nucleic acids in which a portion encoding a protein that can bind to hyaluronic acid, a portion that encodes a protein that can be cleaved by a protease, and a portion that encodes a protein that can bind to HIF-1 ⁇ are linked in this order can also be mentioned. ..
  • nucleic acid comprising a base sequence in which one or more bases are deleted, substituted, or added, and which encodes a protein having an anticancer activity is also available. Can be mentioned. Whether or not the protein encoded by the nucleic acid has an anticancer effect can be examined by incorporating the nucleic acid into an adenovirus vector so that it can be expressed and administering it to cancer cells.
  • the number of bases, or the number of bases constituting the linker is preferably 1 to 100 (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, etc.).
  • the type of base in the present disclosure is not particularly limited, but is preferably A (adenitin), T (thymine), G (guanine), C (cytosine), or U (uracil).
  • the nucleic acid of the present disclosure can be produced by a known method or a method that can be easily conceived from a known method.
  • it can be produced by a genetic engineering method.
  • DNA or RNA encoding CD44, Notch, and HIF-3 ⁇ 4 may be extracted from a human-derived sample, artificially mutated as necessary, and then ligated to produce the product.
  • it may be produced by chemical synthesis.
  • Fusion proteins encoded by the nucleic acids of the present disclosure include (i) a protein moiety having CD44 extracellular function, (ii) a protein moiety having Notch core region function, and (iii) a protein moiety having HIF-3 ⁇ 4 function. Therefore, (i) can function as a Decoy receptor, and a signal can be transmitted to (iii) via (ii). This becomes more effective when (i) is fused upstream of (ii).
  • HIF-1 ⁇ activated in the tumor can be suppressed. It is considered that this makes it possible to suppress the gene to be inserted into the viral vector within an acceptable range and to exert an antitumor effect composed of a plurality of mechanisms of action.
  • the vector in which the nucleic acid of the present disclosure is incorporated in an expressible manner is useful for treating cancer or also for increasing the production of the nucleic acid of the present disclosure.
  • the vector include a plasmid vector, a cosmid vector, a phosmid vector, a virus vector and the like.
  • viral vectors are particularly suitable.
  • the viral vector include an adenoviral vector, a retroviral vector, a lentiviral vector, a Sendai viral vector and the like. Of these, the adenovirus vector is preferable.
  • both a proliferative viral vector and a non-proliferative viral vector can be used. In particular, proliferative or non-proliferative adenovirus vectors are preferred.
  • the nucleic acid of the present disclosure can be incorporated into a vector by a known method or a method that can be easily conceived from a known method.
  • the anti-cancer composition containing the vector into which the nucleic acid of the present disclosure is expressively incorporated exhibits a very excellent anti-cancer effect (particularly a cancer therapeutic effect).
  • the administration form of the anticancer composition is not particularly limited as long as the anticancer effect is exhibited.
  • intratumoral administration is generally preferred, but intravenous administration may be used depending on the vector.
  • the dosage form of the anticancer composition is not particularly limited as long as the anticancer effect is exhibited. For example, it is preferably an injection.
  • the anti-cancer composition can appropriately contain components other than the above vector as needed.
  • examples of such other components include a pharmaceutically acceptable carrier (for example, water), and an appropriate carrier can be selected and used according to the treatment site and the dosage form of the anticancer composition. ..
  • Such an anti-cancer composition can also be prepared based on a known method.
  • the cancer type to be treated with the anticancer composition is not particularly limited as long as the anticancer effect is exhibited.
  • examples thereof include solid tumors and hematological malignancies, and more specifically, but not limited to, brain tumors, maxillary cancers, nasopharyngeal cancers, lung cancers, esophageal cancers, rectal cancers, colon cancers, liver cancers, gastric cancers, bile sac cancers.
  • Pancreatic cancer skin cancer, breast cancer, uterine cancer, ovarian cancer, prostate cancer, kidney cancer, bladder cancer, thyroid cancer, multiple myeloma, lymphoma, acute myeloid leukemia, chronic myeloid leukemia, etc.
  • the anti-cancer effect of the anti-cancer composition is very high, it is preferable because it can be used for cancer types for which existing cancer therapeutic agents are ineffective or small. For example, it can be preferably used for triple negative (estrogen receptor negative, progesterone receptor negative, HER2-negative) breast cancer and the like. Further, the type of anticancer effect of the anticancer composition is not particularly limited. For example, those capable of suppressing tumor growth and / or infiltration are preferable.
  • CD44 / Notch / HIF-3 ⁇ 4 fusion gene was outsourced to GENEWIZ Solid science. Superior service. From each of these three types of human genes, restriction enzyme treatment with SwaI (TaKaRa, Siga, Japan), electrophoresis and purification were used as insert DNA.
  • Each domain of the fusion gene is described in publicly known documents (particularly the above non-patent documents 9, 10 and 12) and the transmembrane domain search tool TMHMM (http://www.cbs.dtu.dk/services/TMHMM/). It was designed with reference to the signal peptide sequence search tool SignalP (http://www.cbs.dtu.dk/services/SignalP/).
  • the sequence of the prepared CD44 / Notch / HIF-3 ⁇ 4 fusion gene is shown in FIG.
  • the nucleotide sequences of the three human genes used are shown in FIGS. 2a to 2c, respectively.
  • the pAxCAwtit2 cosmid vector attached to the Adenovirus Dual Expression Kit (TaKaRa, Siga, Japan) was used as the vector DNA for incorporating the insert DNA. Similar to the insert DNA, the cosmid vector was also purified by phenol-chloroform extraction after cleaving the SwaI sequence at the cloning site by restriction enzyme treatment.
  • coli HST08 Premium Competent Cells were seeded on 100 ⁇ g / ml ampicillin-added LB agar medium (Nacalai Tesque, Kyoto, Japan) and cultured overnight at 37 ° C. After culturing, insert check PCR was performed using KOD FX Neo (TOYOBO CO., LTD., Osaka, Japan) using the colonies grown on the LB agar medium as a template to check whether the transformation was normal or not. Confirmed by gel electrophoresis.
  • Lipofectamine LTX (invitrogen) using 10 ⁇ g of BspT104I digested cosmid was applied to HEK293 cells (National Institutes of Biomedical Innovation, Health and Nutrition) cultured in 60 mm cell culture Petri dish (TPP) to become confluent. , Waltham, MA) performed lipofection. Then, the cultured cells were collected and seeded on Biocoat Collagen I Cellware 96-Well Plate (CORNING, NY, USA).
  • D-MEM Dulbecco's modified Eagle's medium
  • FBS Fetal bovine serum
  • ADX730 The adenovirus vector in which the CD44 / Notch / HIF-3 ⁇ 4 fusion gene is integrated so that it can be expressed is referred to as ADX730.
  • HEK293 cells were cultured in Collagen Type I-Coated 25 cm 2 Flask (IWAKI) until they became 70 to 100% confluent, and 0.5 ml of 5% FBS-D-MEM and 15 ⁇ l of the secondary virus solution prepared above were added. Gently added to infect the virus. For infection, the plate was shaken several times in an incubator (37 ° C, 5% CO 2 ) and slowly shaken four times every 15 minutes. After 1 hour of infection, an additional 4.5 ml of 5% FBS-D-MEM was added to each well and cultured for 3 days. After culturing, it was confirmed that all the cells were denatured, and the cells were collected together with the culture medium.
  • IWAKI Collagen Type I-Coated 25 cm 2 Flask
  • HEK293 cells were cultured in Collagen Type I-Coated 75 cm 2 Flask (IWAKI) until 70-100% confluent, and 2 ml of 5% FBS-D-MEM and 50 ⁇ l of the tertiary virus solution prepared above were gently added. And infected the virus.
  • the plate was shaken several times in an incubator (37 ° C, 5% CO 2 ) and slowly shaken four times every 15 minutes. After 1 hour of infection, another 13 ml of 5% FBS-D-MEM was added to each well and cultured for 3 days. After culturing, it was confirmed that all the cells were denatured, and the cells were collected together with the culture medium. The collected cells were frozen in liquid nitrogen and thawed in a warm bath at 37 ° C 6 times in the same manner as the primary virus solution. After the final freeze-thaw, centrifugation (3,000 rpm, 10 minutes, 4 ° C) was performed, and the collected supernatant was stored as a quaternary virus solution. All of these processes were performed according to the manual of the Adenovirus Dual Expression Kit (TaKaRa, Siga, Japan).
  • the prepared cell pack was sufficiently stirred by vortex, 4 ⁇ l of 10% SDS was added, and the mixture was further stirred by vortex. After incubating at 50 ° C. for 1 hour, phenol-chloroform extraction and chloroform extraction were performed twice, and ethanol precipitation was performed. After ethanol precipitation, it was dissolved in 50 ⁇ l of TE Buffer containing RNase A, and 15 ⁇ l of it was used for restriction enzyme treatment with restriction enzyme XhoI. After the restriction enzyme treatment, the migration pattern of the obtained product was confirmed by agarose gel electrophoresis. All of these processes were performed according to the manual of Adenovirus Dual Expression Kit (TaKaRa, Siga, Japan).
  • Mass culture and purification of recombinant adenovirus vector In order to use the recombinant adenovirus vector prepared above for subsequent experiments, mass culture and purification were performed.
  • HEK293 cells were seeded to 70-100 % confluence in 5 Corning225cm 2 Flasks (CORNING, NY, USA).
  • the virus was infected by gently adding 15 ml of 5% FBS-D-MEM and 150 ⁇ l of the tertiary virus solution prepared above. For infection, the plate was shaken several times in an incubator (37 ° C, 5% CO 2 ) and slowly shaken four times every 15 minutes.
  • the filter was removed, the filter was attached to 5 ml syringe (TaKaRa, Siga, Japan) containing 3 ml of 1 ⁇ Elution Buffer (TaKaRa, Siga, Japan), and 1 ml of 1 ⁇ Elution Buffer was extruded and collected from the filter. After recovery, the filter was incubated once (room temperature, 5 minutes), and then the remaining 2 ml of 1 ⁇ Solution Buffer was extruded to elute the virus. All of these processes were performed according to the manual of the Adeno-X Maxi Purification Kit (TaKaRa, Siga, Japan).
  • HEK293 cells were seeded into 12-Well Cell Culture Plate Flat Bottom (CORNING, NY, USA), and virus solution diluted 10-fold from 100-fold dilution to 10-fold dilution was added dropwise to each well for 2 days. It was cultured. After culturing, all the medium was removed, the cells were slightly dried, and then 1 ml of chilled methanol (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) was added dropwise and incubated (-20 ° C, 10 minutes).
  • HEK293 cells used in cell and medium experiments were D-MEM containing 10% FBS and 1% 100U / ml penicillin and 100mg / ml streptomycin (P / S; Nacalai Tesque, Kyoto, Japan), and A549 cells were 10% FBS.
  • the cells were cultured using Ham's F-12K (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) containing 1% P / S at 37 ° C. under 5% CO 2 conditions.
  • MDA-MB-231 cells (The European Collection of Cell Cultures) were used at 37 ° C using Leibovitz's L-15 Medium (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) containing 15% FBS and 1% P / S. Culturing was performed under conditions without CO 2 equilibration.
  • ADX730 used in various recombinant virus experiments, recombinant adenovirus vector (rAd-SOCS3) incorporating SOCS3 gene, recombinant adenovirus vector (rAd-p53) incorporating p53 gene, and LacZ gene.
  • the engineered recombinant adenovirus vector (rAd-LacZ) is mass-cultured and purified, and then Glycerol (Nacalai Tesque) is used using Slide-A-Lyzer Dialysis Cassette (Extra Strength) (Thermo Fisher Scientific, Waltham, MA).
  • the buffer was exchanged with a dialysis buffer containing 10% and 1 mol / l-Tris-HCl Buffer Solution (Nacalai Tesque, Kyoto, Japan) 1%, and appropriate amounts of each were dispensed and stored at -80 ° C.
  • rAd-SOCS3 and rAd-p53 are adenovirus vectors that are known candidates for oncogene therapy.
  • Flow cytometry The cells were actually infected with ADX730 in vitro, and the comparison of the increase / decrease in the expression level of the CD44 region of the fusion gene incorporated into AD730 was evaluated using Flow cytemetry.
  • MDA-MB-231 cells were seeded at 1 ⁇ 10 6 cells / well against 6-Well Cell Culture Plate Flat Bottom (CORNING, NY, USA) overnight at 37 ° C. without CO 2 equilibrium. It was cultured. After culturing, ADX730, rAd-SOCS3, rAd-p53, and rAd-LacZ were each infected with 40 multiplicity of infection (MOI) and cultured for another 48 hours.
  • MOI multiplicity of infection
  • the cells were washed with PBS and collected, and blocked using Blocking One Histo (Nacalai Tesque, Kyoto, Japan) at room temperature for 10 minutes. After blocking, wash the cells again with PBS and dilute 200-fold FITC anti-mouse / human CD44 Clone: IM7 (BioLegend, San Diego, CA) or 100-fold diluted FITC Rat IgG2a, ⁇ Isotype Ctrl Clone: RTK2758 (BioLegend, San) Diego, CA) was added dropwise, and the mixture was allowed to react for 30 minutes under shading on ice.
  • IM7 BioLegend, San Diego, CA
  • RTK2758 BioLegend, San) Diego, CA
  • the cells were washed again with PBS, 100-fold diluted BD Pharmingen 7-AAD (BD Biosciences, San Diego, CA) was added dropwise, and the cells were reacted under shading on ice for 5 minutes. After the reaction, the cells were washed with PBS and measured using Guava easy Cyte (Merck Millipore, Burlington, MA). Data analysis was performed according to the attached software InCyte.
  • the real-time PCR method was used to confirm the gene transfer by each recombinant adenovirus vector containing ADX730 and the effect obtained by its function.
  • MDA-MB-231 cells were seeded at 1 ⁇ 10 6 cells / well against 6-Well Cell Culture Plate Flat Bottom, and Aneropack Kenki 5% (Mitsubishi Gas Chemical Company, Inc., Tokyo, Japan) and Sodium Hyaluronate. The cells were cultured overnight under the conditions of 37 ° C., 5% CO 2 , 1% ⁇ O 2, and 0.04 mg / ml by using (40 kDa to 80 kDa) (PG Research, Tokyo, Japan).
  • ADX730, rAd-SOCS3, rAd-p53, and rAd-LacZ were each infected with 40 MOI, and further cultured for 48 hours. The cells were then harvested and Total RNA was extracted using NucleoSpin RNA (TaKaRa, Siga, Japan).
  • cDNA was synthesized from the extracted RNA using PrimeScript RT reagent Kit with gDNA Eraser (TaKaRa, Siga, Japan), and the primers prepared using it as a template (Table 3), TB Green Premix Ex Taq II (TaKaRa, Siga) , Japan) and Thermal Cycler Dice Real Time System (TaKaRa, Siga, Japan) were used for PCR reaction and comparative C t method ( ⁇ C t method) analysis.
  • mice Collection of mice and specimens In vivo experiments were conducted using mice in order to examine and compare the antitumor effects of ADX730 with other oncogene therapy agents.
  • MDA-MB-231 cells 1 ⁇ 10 6 cells / 70 ⁇ l and Matrigel Martrix Basement Membrane HC (CORNING, NY, USA) for 6-week-old female BALB / cAJcl-nu / nu (CLEA Japan, Inc., Tokyo, Japan) )
  • ADX730 (1 ⁇ 10 9 PFU / 50 ⁇ l), rAd-SOCS3 (1 ⁇ 10 9 PFU / 50 ⁇ l), rAd-p53 (1 ⁇ 10 9 PFU / 50 ⁇ l), rAd -LacZ (1 ⁇ 10 9 PFU / 50 ⁇ l) or PBS 50 ⁇ l was administered intratumorally every other day for a total of eight times (Days 14, 16, 18, 20, 22, 24, 26, 28). Tumor diameter measurement was performed twice a week from the start of virus administration, a total of 5 times.
  • the tumor was collected and fixed at 4 ° C with 4% Paraformaldehyde Phosphate Buffer Solution (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan). Alternatively, it was stored at -80 ° C.
  • the volume of the tumor was calculated by measuring the major axis (L) and the minor axis (W) and using the formula (W 2 ⁇ L) / 2.
  • Each recombinant adenovirus vector containing ADX730 was infected with cells in vitro, and the normal introduction of each gene was evaluated by a real-time PCR test.
  • the TBP (TATA-Box binding protein) gene was used as the endogenous control gene (forward primer used for measuring the expression of the TBP gene: 5'-GCCAGCTTCGGAGAGTTCTGGGATT-3', reverse primer: 5'-CGGGCACGAAGTGCAATGGTCTTTA-3'. ).
  • the relative expression ratio with the control gene is shown in FIG. 3 as a result.
  • ADX730 shows a high cancer therapeutic effect even if it is a cancer that is difficult to treat with existing therapeutic agents such as triple-negative breast cancer. Furthermore, in the above-mentioned Real-time PCR study, MDA-MB-231 cells derived from breast cancer were used, and the cells used were DU-145 cells derived from prostate cancer, MKN45 cells derived from gastric cancer, or The cells were changed to PANC-1 cells derived from pancreatic cancer, and the gene transfer by each recombinant adenovirus vector containing ADX730 was confirmed, and the effect obtained by the function was confirmed by the real-time PCR method.
  • the number of seeded cells was 5 ⁇ 10 5 cells / well, and the introduction of the CD44 / Notch / HIF-3 ⁇ 4 fusion gene by ADX730 was confirmed and compared using a primer set targeting HIF-3 ⁇ 4 (Table 3). went.
  • the results are shown in FIGS. 8a (DU-145 cells), 8b (MKN45 cells), and 8c (PANC-1 cells). It was confirmed that the expression of the CD44 / Notch / HIF-3 ⁇ 4 fusion gene was significantly increased regardless of which cell was used.

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025110147A1 (ja) * 2023-11-20 2025-05-30 国立大学法人神戸大学 がんの遺伝子治療薬
US12459984B2 (en) 2019-12-27 2025-11-04 National University Corporation Kobe University Cancer gene therapy drug

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2017504601A (ja) * 2013-12-20 2017-02-09 セレクティスＣｅｌｌｅｃｔｉｓ 免疫療法のためにマルチインプットシグナル感受性ｔ細胞を操作する方法
JP2018506293A (ja) * 2015-02-24 2018-03-08 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 結合誘発型転写スイッチ及びその使用方法
JP2019528077A (ja) * 2016-08-23 2019-10-10 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア タンパク質分解により切断可能なキメラポリペプチド及びそれらの使用方法

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090118175A1 (en) * 2005-05-06 2009-05-07 Macina Roberto A Compositions and Methods for Detection, Prognosis and Treatment of Breast Cancer
EP2853595A1 (en) * 2013-09-30 2015-04-01 Soluventis GmbH NOTCH 1 specific siRNA molecules
US20160125127A1 (en) * 2014-10-29 2016-05-05 Council Of Scientific & Industrial Research Identification of minimal combinations of oncoproteins in notch pathway to suppress human glioblastoma
CN110437339B (zh) * 2018-05-04 2021-08-13 免疫靶向有限公司 一种以白介素15为活性成分的融合蛋白型药物前体
CN115151644B (zh) 2019-12-27 2024-06-21 国立大学法人神户大学 癌基因治疗药

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2017504601A (ja) * 2013-12-20 2017-02-09 セレクティスＣｅｌｌｅｃｔｉｓ 免疫療法のためにマルチインプットシグナル感受性ｔ細胞を操作する方法
JP2018506293A (ja) * 2015-02-24 2018-03-08 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 結合誘発型転写スイッチ及びその使用方法
JP2019528077A (ja) * 2016-08-23 2019-10-10 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア タンパク質分解により切断可能なキメラポリペプチド及びそれらの使用方法

Non-Patent Citations (14)

* Cited by examiner, † Cited by third party
Title
BAZIL VHOREJSI V.: "Shedding of the CD44 adhesion molecule from leukocytes induced by anti-CD44 monoclonal antibody simulating the effect of a natural receptor ligand", J IMMUNOL., vol. 149, no. 3, 1 August 1992 (1992-08-01), pages 747 - 53, XP008156689
BOURGUIGNON L.YZHU HSHAO L.CHEN Y.W.: "CD44 interaction with c-Src kinase promotes cortactin-mediated cytoskeleton function and hyaluronic acid-dependent ovarian tumor cell migration", J BIOL CHEM., vol. 276, no. 10, 17 November 2000 (2000-11-17), pages 7327 - 36
CANCER RES., vol. 59, no. 5, 1 March 1999 (1999-03-01), pages 1141 - 5
FUJITA Y.KITAGAWA M.NAKAMURA S.AZUMA K.ISHII G.HIGASHI M.KISHI H.HIWASA TKODA K.NAKAJIMA N.: "CD44 signaling through focal adhesion kinase and its anti-apoptotic effect", FEBS LETT, vol. 528, no. 1-3, 25 September 2002 (2002-09-25), pages 101 - 8, XP004383241, DOI: 10.1016/S0014-5793(02)03262-3
GAO N.SHEN LZHANG Z.LEONARD S.S.HE H.ZHANG X.G.SHI X.JIANG B.H.: "Arsenite induces HIF-lalpha and VEGF through PI3K, Akt and reactive oxygen species in DU145 human prostate carcinoma cells", MOL CELL BIOCHEM, vol. 255, no. 1-2, January 2004 (2004-01-01), pages 33 - 45
ISHIWATARI-HAYASAKA, H. ET AL.: "Requirements for signal delivery through CD 44: analysis using CD 44-Fas chimeric proteins", THE JOURNAL OF IMMUNOLOGY, vol. 163, 1999, pages 1258 - 1264, XP055838666 *
KOPAN R.ILAGAN MX: "The canonical Notch signaling pathway: unfolding the activation mechanism", CELL, vol. 137, no. 2, 17 April 2009 (2009-04-17), pages 216 - 33, XP055411700, DOI: 10.1016/j.cell.2009.03.045
MAKINO Y., CAO R., SVENSSON K., BERTILSSON G., ASMAN M. TANAKA H., CAO Y., BERKENSTAM A., POELLINGER L.: "Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression", NATURE, vol. 414, no. 6863, 29 November 2001 (2001-11-29), pages 550 - 4
MAYNARD M.A., EVANS A.J., HOSOMI T., HARA S., JEWETT M.A., OHH M.: "Human HIF-3alpha4 is a dominant-negative regulator of HIF-1 and is down-regulated in renal cell carcinoma", FASEB J, vol. 19, no. 11, September 2005 (2005-09-01), pages 1396 - 406
MORSUT L.ROYBAL K.T.XIONG X.GORDLEY R.MCOYLE S.M.THOMSON M.LIM W.A.: "Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors", CELL, vol. 164, no. 4, 2016, pages 780 - 91, XP029416809, DOI: 10.1016/j.cell.2016.01.012
MURAKAMI D.OKAMOTO I.NAGANO 0.KAWANO Y.TOMITA T.IWATSUBO TDE STROOPER B.YUMOTO ESAYA H.: "Presenilin-dependent gamma-secretase activity mediates the intramembranous cleavage of CD44", ONCOGENE, vol. 22, no. 10, 13 March 2003 (2003-03-13), pages 1511 - 6, XP037737092, DOI: 10.1038/sj.onc.1206298
See also references of EP4083213A4
SHONA, T.GRAEME, J.: "A Cancer Gene Therapy Approach that Targets Tumor-associated Hyaluronan", CANCER GROWTH AND METASTASIS 2009 (2, November 2009 (2009-11-01)
UEDA M.SAJI H.: "Radiolabeled Probes Targeting Hypoxia-Inducible Factor-1-Active Tumor Microenvironments", SCIENTIFIC WORLD JOURNAL, vol. 2014, 18 August 2014 (2014-08-18), pages 165461

Cited By (2)

* Cited by examiner, † Cited by third party
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US12459984B2 (en) 2019-12-27 2025-11-04 National University Corporation Kobe University Cancer gene therapy drug
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