WO2012063894A1 - マイクロrnaのアンチセンスオリゴヌクレオチドを含む組成物 - Google Patents
マイクロrnaのアンチセンスオリゴヌクレオチドを含む組成物 Download PDFInfo
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- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
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- C12N2310/00—Structure or type of the nucleic acid
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- C12N2310/32—Chemical structure of the sugar
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Definitions
- the present invention relates to a composition comprising an antisense oligonucleotide of microRNA.
- Micro RNA is a small molecule RNA consisting of 18-25 bases, and inhibits translation into the protein by binding to the target mRNA.
- miRNA / miRNA is a small molecule RNA consisting of 18-25 bases, and inhibits translation into the protein by binding to the target mRNA.
- miRBase® http://www.miRbase.org/
- these miRNA expression or functional abnormalities are involved in various diseases.
- Patent Document 1 many miRNAs whose expression is enhanced or suppressed in cancer tissues or cancer cells have been identified by comprehensive expression analysis using miRNA microarrays.
- MiR-21, miR-155, miR-17-5p, miR-19, etc. are known as miRNAs whose expression is enhanced in cancer.
- miR-19 is necessary and sufficient for cell carcinogenesis, and its target One of them was shown to be a tumor suppressor gene PTEN.
- PTEN a tumor suppressor gene
- miRNAs have decreased expression due to cancer, and such miRNAs have been reported such as let-7, miR-15a, miR-34a, miR-143, miR-145 and the like.
- let-7 has been shown to correlate with decreased expression and prognosis in lung cancer, and its target is known to contain the oncogene Ras.
- MiRNA that exerts tumor suppressor-like functions targeting oncogenes is called Tumor suppressor miR.
- the present invention provides a composition containing an antisense oligonucleotide of microRNA that can inhibit the growth of cancer cells.
- the present invention includes an antisense oligonucleotide of microRNA, wherein the microRNA is from hsa-miR-133a, hsa-miR-133b, hsa-miR-346, and hsa-miR-361-3p.
- the present invention relates to a composition for suppressing the growth of human cancer cells, selected from the group consisting of:
- the present invention includes an antisense oligonucleotide of microRNA, and the microRNA is hsa-miR-92a, hsa-miR-133a, hsa-miR-133b, hsa-miR-139-5p, hsa-miR-197, hsa-miR-328, hsa-miR-346, hsa-miR-361-3p, hsa-miR-605, hsa-miR-766, hsa-miR-1228, hsa-miR-1252,
- the present invention relates to a composition for suppressing the growth of human head and neck cancer cells selected from the group consisting of hsa-miR-1260 and hsa-miR-1271.
- the present invention growth inhibition of cancer cells becomes possible. Therefore, according to the present invention, it can contribute to cancer treatment and prevention, recurrence prevention, and the like.
- FIG. 1 is an example of a fluorescent photograph showing the effect of a microRNA antisense oligonucleotide on the growth of human oral squamous cell carcinoma cell GFP-SAS.
- FIG. 2 is an example of a graph showing the effect of microRNA antisense oligonucleotides on the growth of various cancer cells.
- FIG. 3 is a graph showing an example of the cancer cell proliferation inhibitory effect of microRNA antisense oligonucleotides in vivo.
- FIG. 4 is a photograph showing an example of the cancer cell proliferation inhibitory effect of microRNA antisense oligonucleotides in vivo.
- the present invention includes the following aspects: [1] comprising an antisense oligonucleotide of microRNA, wherein the microRNA is selected from the group consisting of hsa-miR-133a, hsa-miR-133b, hsa-miR-346, and hsa-miR-361-3p A composition for inhibiting the growth of human cancer cells; [2] A pharmaceutical composition comprising an antisense oligonucleotide of microRNA, wherein the microRNA comprises hsa-miR-133a, hsa-miR-133b, hsa-miR-346, and hsa-miR-361-3p A pharmaceutical composition for the treatment or prevention of human cancer selected from the group consisting of: [3] In order to suppress the growth of non-human cancer cells, comprising an antisense oligonucleotide of microRNA, wherein the microRNA is selected from the group consisting of miR-133, miR-3
- An antisense oligonucleotide of microRNA wherein the microRNA is hsa-miR-92a, hsa-miR-133a, hsa-miR-133b, hsa-miR-139-5p, hsa-miR-197, hsa-miR-328, hsa-miR-346, hsa-miR-361-3p, hsa-miR-605, hsa-miR-766, hsa-miR-1228, hsa-miR-1252, hsa-miR-1260, And a composition for inhibiting the growth of human head and neck cancer cells selected from the group consisting of hsa-miR-1271; [6] A pharmaceutical composition comprising an antisense oligonucleotide of microRNA, wherein the microRNA comprises hsa-miR-92a, hs
- microRNA / miRNA refers to a kind of low-molecular non-coding RNA, and has a normal meaning used in this technical field.
- the microRNA referred to using an ID can refer to a database (for example, miRBase® (http://www.miRbase.org/)).
- microRNA refers to mature microRNA consisting of 18 to 25 bases.
- microRNA antisense oligonucleotide refers to an oligonucleotide having a base sequence complementary to a target microRNA, and a preferred form is a single-stranded oligonucleotide.
- the antisense oligonucleotide of the microRNA is an oligonucleotide comprising a base sequence complementary to all or part of the base sequence of the target mature microRNA, more preferably, the target An oligonucleotide consisting of a base sequence complementary to all or part of the base sequence of mature microRNA.
- the length of the “microRNA antisense oligonucleotide” in the present invention is, for example, 8-25 nucleotides, 10-24 nucleotides, 10-22 nucleotides, 12-22 nucleotides, 10-20 nucleotides, 12 -20 nucleotides, 10-19 nucleotides, 12-19 nucleotides, 10-18 nucleotides, 12-18 nucleotides, 10-17 nucleotides, 12-17 nucleotides, 10-16 nucleotides, 12-16 nucleotides. It is considered that nonspecific microRNA inhibition can be further suppressed by shortening the length of the oligonucleotide sequence.
- complementary means that two nucleotide sequences have the ability to correctly pair with each other. For example, if a certain oligonucleotide can hydrogen bond with a nucleotide at the corresponding position in the target microRNA at a certain position, the oligonucleotide and the microRNA are considered to be complementary to each other at that position. . When a sufficient number of nucleotides in the oligonucleotide can form hydrogen bonds with the corresponding nucleotides in the target microRNA and can form a stable complex, the oligonucleotide and the microRNA Are considered complementary to each other.
- the antisense oligonucleotide of the microRNA is preferably an oligonucleotide comprising or consisting of a base sequence that is 100% complementary to the target microRNA.
- nucleotide pairing refers to hydrogen bonding between bases.
- DNA it refers to the binding form of A and T and G and C.
- RNA A and U.
- G and C a bonding form of G and C.
- the antisense oligonucleotide of the microRNA may be an analog other than the above A, T (U), G, and C as long as it can pair with the nucleotide of the target microRNA.
- the “oligonucleotide” refers to a known nucleic acid analog in addition to DNA and RNA, and an oligonucleotide formed by mixing them.
- the oligonucleotide may include a modified oligonucleotide in which a known modification is performed.
- nucleic acid analogs include known nucleic acid analogs such as LNA (Locked Nucleic Acid) and PNA (Peptide Nucleic Acid).
- the antisense oligonucleotide in the present specification preferably contains a nucleic acid analog, more preferably contains an LNA, and more preferably contains a nucleic acid analog from the viewpoint of improving specificity to a target microRNA and improving cell growth suppression. More preferably, the rate exceeds 50%.
- microRNA antisense oligonucleotides containing LNA refer to, for example, JP2009-532392 (WO2007 / 000169).
- the base in the “microRNA antisense oligonucleotide” may be a non-DNA / RNA base other than the base of DNA and RNA, as long as the function of the target microRNA can be inhibited.
- the RNA may be substituted with fluorine or the like.
- the bond between nucleosides may be a bond other than a phosphate bond, and from the viewpoint of stability, a sulfur (S) -containing bond. (That is, it is a phosphorothioate type oligonucleotide).
- S sulfur
- “suppression of cell growth” includes inhibition of cell growth and cell death, and includes suppression of cell growth in in vivo, in vitro, and ex vivo.
- “cell” includes human cells and non-human cells. Non-human cells include organisms with microRNA mechanisms. In the present specification, “non-human” includes primates excluding humans, mammals excluding humans, and vertebrates excluding humans.
- the present invention includes an antisense oligonucleotide of microRNA, wherein the microRNA is from hsa-miR-133a, hsa-miR-133b, hsa-miR-346, and hsa-miR-361-3p.
- the present invention relates to a composition for suppressing the growth of human cancer cells (hereinafter also referred to as “first composition of the present invention”) selected from the group consisting of:
- the sequences of the microRNA (mature type) are described in SEQ ID NOs: 1 to 4 in Table 1 below, respectively.
- microRNA antisense oligonucleotide that can be contained in the first composition of the present invention is an oligonucleotide consisting of any one of the base sequences of SEQ ID NOS: 5 to 8 in Table 1 or a partial sequence thereof.
- the length of the oligonucleotide and the nucleic acid analog are as described above.
- the 1st composition of this invention can contain 1 type or multiple types of antisense oligonucleotide.
- one embodiment of the antisense oligonucleotide for the microRNA of hsa-miR-133a or hsa-miR-133b is an anti-sense that can inhibit both hsa-miR-133a and hsa-miR-133b. Includes sense oligonucleotides.
- the cancer of human cancer cells for which the first composition of the present invention suppresses growth is not particularly limited, and examples thereof include head and neck cancer, malignant melanoma, basal cell cancer, ovarian cancer, and breast cancer. , Non-small cell lung cancer, renal cell cancer, bladder cancer, recurrent superficial bladder cancer, stomach cancer, prostate cancer, biliary tract cancer, pancreatic cancer, lung cancer, cervical cancer, cervical dysplasia Including laryngeal papillomatosis, colon cancer, colorectal cancer, and carcinoid tumors.
- the cancer of human cancer cells for which the composition of this embodiment suppresses growth preferably includes head and neck cancer, lung cancer, biliary tract cancer, pancreatic cancer, and prostate cancer.
- head and neck cancer refers to cancer that can be formed from the neck excluding the brain and eyes, and generally includes oral cancer, nasal sinus cancer, lip cancer, and pharynx. Cancer, laryngeal cancer, cervical tumor, ear cancer.
- oral cancer generally refers to cancer that is caused by the mucous membrane of a part constituting the oral cavity, such as gingiva, tongue, cheek, palate, floor of mouth, and salivary gland.
- the first composition of the present invention may contain a reagent, a drug, and a medium that are suitable for bringing the target human cancer cell into contact with the subject and suppressing the growth of the cell.
- the first composition of the present invention may be in a lyophilized form.
- the above-described first composition of the present invention can be used as a pharmaceutical composition for treating or preventing the above-described cancer. Therefore, in another aspect, the present invention provides a pharmaceutical composition comprising an antisense oligonucleotide of microRNA, wherein the microRNA is hsa-miR-133a, hsa-miR-133b, hsa-miR-346, And a pharmaceutical composition for treating or preventing human cancer selected from the group consisting of hsa-miR-361-3p (hereinafter also referred to as “first pharmaceutical composition of the present invention”). As used herein, prevention of cancer includes prevention of recurrence of cancer.
- microRNA antisense oligonucleotide that can be included in the first pharmaceutical composition of the present invention is an oligonucleotide comprising any one of the base sequences of SEQ ID NOS: 5 to 8 in Table 1 above or a partial sequence thereof. .
- the length of the oligonucleotide and the nucleic acid analog are as described above.
- the 1st pharmaceutical composition of this invention may contain 1 type or multiple types of antisense oligonucleotide.
- the first pharmaceutical composition of the present invention may further contain a pharmaceutically acceptable carrier.
- the pharmaceutical carrier is not particularly limited, but, for example, a carrier capable of increasing the efficiency with which a antisense oligonucleotide of micro RNA enters a target site, tissue, cell, etc., for example, atelocollagen, liposome, cationic liposome Etc.
- the dosage form of the pharmaceutical composition of this embodiment is not particularly limited, and examples thereof include injections, creams, ointments, tablets, suspensions and the like.
- the administration method is not particularly limited, and examples thereof include injection, oral, topical, intranasal, rectal, intravenous, and intraarterial administration.
- the present invention provides, as a further aspect, the growth of a non-human cancer cell comprising an antisense oligonucleotide of microRNA, wherein the microRNA is selected from the group consisting of miR-133, miR-346, and miR-361. Relates to a composition for suppressing the above (hereinafter, also referred to as "second composition of the present invention").
- Antisense nucleotides of microRNAs selected from the group consisting of can be used, and antisense nucleotides for the corresponding microRNA sequences can be used for other organisms as well.
- the 2nd composition of this invention can contain 1 type or multiple types of antisense oligonucleotide.
- the cancer of the non-human cancer cell for which the second composition of the present invention suppresses growth is not particularly limited, and examples thereof include head and neck cancer, malignant melanoma, basal cell cancer, ovarian cancer, Breast cancer, non-small cell lung cancer, renal cell cancer, bladder cancer, recurrent superficial bladder cancer, gastric cancer, prostate cancer, biliary tract cancer, pancreatic cancer, lung cancer, cervical cancer, cervical formation Includes abnormalities, laryngeal papillomatosis, colon cancer, colorectal cancer, and carcinoid tumors.
- the cancer of the non-human cancer cell for which the composition of this embodiment suppresses growth preferably includes head and neck cancer, lung cancer, biliary tract cancer, pancreatic cancer, and prostate cancer.
- the 2nd composition of this invention may contain the reagent, chemical
- the second composition of the present invention may be in a lyophilized form.
- the present invention provides a non-human pharmaceutical composition comprising an antisense oligonucleotide of microRNA, wherein the microRNA comprises hsa-miR-133a, hsa-miR-133b, hsa-miR.
- second pharmaceutical composition of the present invention a pharmaceutical composition for treatment or prevention of non-human cancer selected from the group consisting of hsa-miR-361-3p (hereinafter also referred to as "second pharmaceutical composition of the present invention") .)
- the length of the antisense oligonucleotide and the nucleic acid analog of microRNA that can be contained in the second pharmaceutical composition of the present invention are as described above.
- the second pharmaceutical composition of the present invention may contain one or more types of antisense oligonucleotides.
- the second pharmaceutical composition of the present invention may further contain a pharmaceutically acceptable carrier.
- the pharmaceutical carrier is not particularly limited, but, for example, a carrier capable of increasing the efficiency with which a antisense oligonucleotide of micro RNA enters a target site, tissue, cell, etc., for example, atelocollagen, liposome, cationic liposome Etc.
- the dosage form of the pharmaceutical composition of this embodiment is not particularly limited, and examples thereof include injections, creams, ointments, tablets, suspensions and the like.
- the administration method is not particularly limited, and examples thereof include injection, oral, topical, intranasal, rectal, intravenous, and intraarterial administration.
- the present invention includes an antisense oligonucleotide of microRNA, and the microRNA is hsa-miR-92a, hsa-miR-133a, hsa-miR-133b, hsa-miR-139-5p, hsa-miR-197, hsa-miR-328, hsa-miR-346, hsa-miR-361-3p, hsa-miR-605, hsa-miR-766, hsa-miR-1228, hsa-miR-1252,
- a composition for suppressing the growth of human head and neck cancer cells selected from the group consisting of hsa-miR-1260 and hsa-miR-1271 (hereinafter also referred to as “third composition of the present invention”) .)
- the third composition of the present invention may contain one or more types of antisense oligonucleotide
- the cancer whose growth is to be suppressed is human oral squamous cell carcinoma, hsa-miR-92a, hsa-miR-133a, hsa-miR-133b, hsa-miR-139-5p, hsa-miR-197, hsa-miR-328, hsa-miR-346, hsa-miR-361-3p, hsa-miR-605, hsa-miR-766, hsa-miR- It includes an antisense nucleotide targeting a microRNA selected from the group consisting of 1228, hsa-miR-1252, hsa-miR-1260, and hsa-miR-1271.
- the sequence of the target microRNA is preferably SEQ ID NOS: 1-4 and 9-18 in Table 2 below.
- an antisense oligonucleotide of microRNA that can be included in the third composition of the present embodiment, from the nucleotide sequence of any of SEQ ID NOs: 5 to 8 and 19 to 28 in the following Table 2, or a partial sequence thereof The following oligonucleotide is mentioned.
- the length of the oligonucleotide and the nucleic acid analog are as described above.
- the cancer whose growth is to be suppressed is a human salivary gland cancer, hsa-miR-133a, hsa-miR-133b, and hsa-miR-361-
- An antisense nucleotide targeting a microRNA selected from the group consisting of 3p is included.
- the sequence of the target microRNA (mature type) is preferably SEQ ID Nos. 1, 2 and 4 in Table 3 below.
- a microRNA antisense oligonucleotide that can be included in the third composition of the present embodiment, an oligo consisting of any one of the base sequences of SEQ ID NOs: 5, 6, and 8 in Table 3 below or a partial sequence thereof Nucleotides are mentioned. The length of the oligonucleotide and the nucleic acid analog are as described above.
- the third composition of the present invention may contain a reagent, a drug, and a medium that are suitable for bringing the target human cancer cell into contact with the cell and suppressing the growth of the cell.
- the first composition of the present invention may be in a lyophilized form.
- the present invention provides a pharmaceutical composition comprising an antisense oligonucleotide of microRNA, wherein the microRNA is hsa-miR-92a, hsa-miR-133a, hsa-miR-133b, hsa-miR-139-5p, hsa-miR-197, hsa-miR-328, hsa-miR-346, hsa-miR-361-3p, hsa-miR-605, hsa-miR-766, hsa-miR- 1228, hsa-miR-1252, hsa-miR-1260, and a pharmaceutical composition for treating or preventing human head and neck cancer selected from the group consisting of hsa-miR-1271
- the cancer to be treated or prevented is human oral squamous cell carcinoma, hsa-miR-92a, hsa-miR-133a, hsa-miR-133b, hsa-miR-139-5p, hsa-miR-197, hsa-miR-328, hsa-miR-346, hsa-miR-361-3p, hsa-miR-605, hsa-miR-766, hsa-miR- It includes an antisense nucleotide targeting a microRNA selected from the group consisting of 1228, hsa-miR-1252, hsa-miR-1260, and hsa-miR-1271.
- the target microRNA (mature type) sequences are preferably SEQ ID NOs: 1 to 4 and 9 to 18 in Table 2 above.
- the oligonucleotide consists of any one of the base sequences of SEQ ID NOS: 5 to 8 and 19 to 28 in Table 2 or a partial sequence thereof. An oligonucleotide is mentioned. The length of the oligonucleotide and the nucleic acid analog are as described above.
- the cancer to be treated or prevented is human salivary gland cancer, hsa-miR-133a, hsa-miR-133b, and hsa-miR-361-
- An antisense nucleotide targeting a microRNA selected from the group consisting of 3p is included.
- the sequence of the target microRNA (mature type) is preferably SEQ ID Nos. 1, 2 and 4 in Table 3 above.
- an oligo consisting of the base sequence of any of SEQ ID NOs: 5, 6 and 8 in Table 3 above or a partial sequence thereof Nucleotides are mentioned.
- the length of the oligonucleotide and the nucleic acid analog are as described above.
- the third pharmaceutical composition of the present invention may further contain a pharmaceutically acceptable carrier.
- the pharmaceutical carrier is not particularly limited, but, for example, a carrier capable of increasing the efficiency with which a antisense oligonucleotide of micro RNA enters a target site, tissue, cell, etc., for example, atelocollagen, liposome, cationic liposome Etc.
- the dosage form of the pharmaceutical composition of this embodiment is not particularly limited, and examples thereof include injections, creams, ointments, tablets, suspensions and the like.
- the administration method is not particularly limited, and examples thereof include injection, oral, topical, intranasal, rectal, intravenous, and intraarterial administration.
- the present invention comprises an antisense oligonucleotide of microRNA, wherein the microRNA is miR-92, miR-133, miR-139, miR-197, miR-328, miR-346, miR-361. , MiR-605, miR-766, miR-1228, miR-1252, miR-1260, and a composition for suppressing the growth of non-human cancer cells selected from the group consisting of miR-1271 (hereinafter, Also referred to as “fourth composition of the present invention”.
- the 4th composition of this invention can contain 1 type or multiple types of antisense oligonucleotide. Oligonucleotide lengths and nucleic acid analogs that can be included are as described above.
- the fourth composition of the present invention may be in a lyophilized form.
- the cancer whose growth is to be suppressed is a non-human oral squamous cell carcinoma, miR-92, miR-133, miR-139, miR-197, miR -328, miR-346, miR-361, miR-605, miR-766, miR-1228, miR-1252, miR-1260, and antisense targeting a microRNA selected from the group consisting of miR-1271 Contains nucleotides.
- the target microRNA (mature type) sequence an antisense nucleotide for the corresponding microRNA sequence in the target non-human organism can be used.
- the cancer whose growth is to be suppressed is a non-human salivary gland cancer, and is from the group consisting of miR-133a, miR-133b, and miR-361-3p.
- antisense nucleotides that target the selected microRNA As the target microRNA (mature type) sequence, an antisense nucleotide for the corresponding microRNA sequence in the target non-human organism can be used.
- the fourth composition of the present invention described above can be used as a pharmaceutical composition for treating or preventing non-human head and neck cancer. Therefore, in another aspect, the present invention provides a non-human pharmaceutical composition comprising an antisense oligonucleotide of microRNA, wherein the microRNA comprises miR-92, miR-133, miR-139, miR-197. , MiR-328, miR-346, miR-361, miR-605, miR-766, miR-1228, miR-1252, miR-1260, and miR-1271 human head and neck cancer selected from the group
- the present invention relates to a pharmaceutical composition for treating or preventing (hereinafter also referred to as “the fourth pharmaceutical composition of the present invention”).
- the length of the antisense oligonucleotide and the nucleic acid analog of microRNA that can be contained in the fourth pharmaceutical composition of the present invention are as described above. Note that the fourth pharmaceutical composition of the present invention may contain one or more types of antisense oligonucleotides.
- the cancer to be treated or prevented is a non-human oral squamous cell carcinoma, miR-92, miR-133, miR-139, miR-197, Anti-targeting microRNAs selected from the group consisting of miR-328, miR-346, miR-361, miR-605, miR-766, miR-1228, miR-1252, miR-1260, and miR-1271 Contains sense nucleotides.
- the target microRNA (mature type) sequence an antisense nucleotide for the corresponding microRNA sequence in the target non-human organism can be used.
- the fourth pharmaceutical composition of the present invention is a group consisting of miR-133a, miR-133b, and miR-361-3p, wherein the cancer to be treated or prevented is a non-human salivary gland cancer.
- An antisense nucleotide targeting a microRNA selected from As the target microRNA (mature type) sequence an antisense nucleotide for the corresponding microRNA sequence in the target non-human organism can be used.
- the fourth pharmaceutical composition of the present invention may further contain a pharmaceutically acceptable carrier.
- the pharmaceutical carrier is not particularly limited, but, for example, a carrier capable of increasing the efficiency with which a antisense oligonucleotide of micro RNA enters a target site, tissue, cell, etc., for example, atelocollagen, liposome, cationic liposome Etc.
- the dosage form of the pharmaceutical composition of this embodiment is not particularly limited, and examples thereof include injections, creams, ointments, tablets, suspensions and the like.
- the administration method is not particularly limited, and examples thereof include injection, oral, topical, intranasal, rectal, intravenous, and intraarterial administration.
- compositions in this invention growth of the human cancer cell made into object can be suppressed. Therefore, this invention relates to the method of suppressing the growth of the cancer cell including making these compositions contact the human cancer cell of object as another aspect.
- the contact method is not particularly limited, and a method that can introduce the antisense oligonucleotide of microRNA in the composition into a cancer cell as a target can be employed.
- One embodiment of introducing an antisense oligonucleotide of microRNA into a cancer cell is the lipofection method.
- examples of the dose of the antisense oligonucleotide to cells during the lipofection method and other introduction methods include 10 to 25 nM.
- the dosage form of the composition to be administered relating to a method for suppressing the growth of cancer cells including administering these pharmaceutical compositions to a subject is not particularly limited. Injections, creams, ointments, tablets, suspensions and the like can be mentioned.
- the administration method is not particularly limited, and examples thereof include injection, oral, topical, intranasal, rectal, intravenous, and intraarterial administration. Administration will depend on the severity and responsiveness of the condition being treated and the course of treatment.
- Optimal dosing schedules can be calculated from measurements of drug accumulation in the subject's body.
- the optimal dose may vary depending on the relative effectiveness of the individual oligonucleotides. In general, it can be estimated based on the EC50 found to be effective in in vitro and in vivo animal models. Generally, the dose is 0.01 ⁇ g to 1 g / kg body weight, preferably 0.01 to 100 mg / kg body weight, more preferably 1 to 10 mg / kg body weight.
- the administration frequency may be one or more times per day, one week, one month or one year, or once every 2 to 10 years, or by continuous infusion for several hours to several months. The number of repeated administrations can be estimated based on the measured drug concentration in the body fluid or tissue and the residence time. After successful treatment, it may be desirable for the subject to receive maintenance therapy to prevent recurrence of the condition.
- the growth of the non-human cancer cell made into object can be suppressed. Therefore, this invention relates to the method of suppressing the growth of the cancer cell including making these compositions contact the nonhuman cancer cell of object as another aspect.
- the contact method is not particularly limited, and a method that can introduce the antisense oligonucleotide of microRNA in the composition into a cancer cell as a target can be employed.
- the dosage form of the composition to be administered relating to a method for inhibiting the growth of cancer cells including administering these pharmaceutical compositions to a non-human subject is not particularly limited, For example, injections, creams, ointments, tablets, suspensions and the like can be mentioned.
- the administration method is not particularly limited, and examples thereof include injection, oral, topical, intranasal, rectal, intravenous, and intraarterial administration. Administration will depend on the severity and responsiveness of the condition being treated and the course of treatment.
- Optimal dosing schedules can be calculated from measurements of drug accumulation in the subject's body.
- the optimal dose may vary depending on the relative effectiveness of the individual oligonucleotides. In general, it can be estimated based on the EC50 found to be effective in in vitro and in vivo animal models. Generally, the dose is 0.01 ⁇ g to 1 g / kg body weight, preferably 0.01 to 100 mg / kg body weight, more preferably 1 to 10 mg / kg body weight.
- the administration frequency may be one or more times per day, one week, one month or one year, or once every 2 to 10 years, or by continuous infusion for several hours to several months. The number of repeated administrations can be estimated based on the measured drug concentration in the body fluid or tissue and the residence time. After successful treatment, it may be desirable for the subject to receive maintenance therapy to prevent recurrence of the condition.
- the miRNA knockdown library was used to comprehensively analyze the effects on growth when microRNA was inhibited in the following two types of human head and neck cancer cell lines. Specifically, the following conditions were used. [Cell lines used] GFP stable expression strains GFP-SAS and GFP-ACCM, which were isolated and established by introducing a green fluorescent protein (GFP) gene into human oral squamous cell carcinoma cell line SAS and human salivary gland cancer cell line ACCM, were used.
- GFP green fluorescent protein
- Each well of a 96-well plastic plate (trade name: BD Falcon, manufactured by BD) has 160 ⁇ l of 10% FBS-containing DMEM medium containing 2 ⁇ 10 3 human oral cancer cell lines GFP-SAS or GFP-ACCM and 918 types of humans.
- Opti-MEM containing LNA (Locked Nucleic Acid) / DNA knockdown probe for microRNA (trade name: miRCURY LNA TM microRNA Knockdown Library-Human v12.0, manufactured by EXIQON) 5 pmol each, RNAiMAX (manufactured by Invitrogen) 0.4 ⁇ l 40 ⁇ l (Invitrogen) was mixed and added.
- OncomiR 14 types of human oral squamous cell carcinoma cell GFP-SAS (Table 4 below) and 2 types of human salivary gland cancer cell GFP-ACCM (Table 5 below) OncomiR was identified.
- the two types of OncomiR in human salivary gland cancer cells were OncomiR in human oral squamous cell carcinoma cells.
- IDs, accession numbers, and mature microRNA sequences in the identified OncomiR miRBase are shown.
- Antisense oligonucleotides for hsa-miR-133a / b, hsa-miR-346, and hsa-miR-361-3p are knockdown probes having the sequences shown in Table 6 below (trade name: miRCURY LNA microRNA Inhibitor, manufactured by EXIQON) Was used. As a negative control, a trade name: miRCURY Knockdown control probe (SEQ ID NO: 32, manufactured by EXIQON) was used. These oligonucleotides contain LNA (trade name: Locked Nucleic Acid) (including about 8 bases of LNA for about 20 bases of DNA).
- LNA Locked Nucleic Acid
- Opti-MEM containing 80 ⁇ l of complete medium containing 2 ⁇ 10 3 various human cancer cells in each well of a 96-well plastic plate, knockdown probe 2.5 pmol of the sequence shown in Table 3 below for miRNA, and 0.2 ⁇ l of RNAiMAX (Invitrogen) 20 ⁇ l (Invitrogen) was mixed and added. After culturing for 80 hours, the fluorescence of GFP emitted by viable cells in each well was observed with a fluorescence microscope. The result is shown in FIG.
- antisense oligonucleotides for hsa-miR-133a / b, hsa-miR-346, and hsa-miR-361-3p in Table 6 below are those of human oral squamous cell carcinoma cell GFP-SAS. Proliferation was completely suppressed.
- Antisense oligonucleotides to hsa-miR-133a / b, hsa-miR-346, and hsa-miR-361-3p in Table 6 above are as follows: The effect on cell proliferation after introduction into various cancer cells was evaluated under the following conditions.
- GFP-SAS and GFP-ACCM which were established by introducing the green fluorescent protein (GFP) gene into the human oral squamous cell carcinoma cell line SAS and the human salivary gland cancer cell line ACCM, human oral squamous epithelium Cancer cell line B88, human pancreatic cancer cell line MIAPaCa-2, human biliary tract cancer cell line HuCCT1, human lung squamous cell carcinoma cell line EBC-1, human lung adenocarcinoma cell line A549, human prostate cancer cell Strains LNCaP (androgen dependent), PC-3 (androgen independent) were used.
- GFP green fluorescent protein
- Dulbecco's modified Eagle medium containing 10% fetal bovine serum (FBS; manufactured by Biosource International), 100 ⁇ g / ml streptomycin, 100 U / ml penicillin, 0.25 mg / ml amphotericin B (produced by Invitrogen) was used for culturing these cell lines.
- FBS fetal bovine serum
- DMEM fetal bovine serum
- RPMI1640 medium manufactured by Sigma-Aldrich
- Opti-MEM containing 80 ⁇ l of complete medium containing 2 ⁇ 10 3 various human cancer cells in each well of a 96-well plastic plate, knockdown probe 2.5 pmol of the sequence shown in Table 3 above for miRNA, and 0.2 ⁇ l of RNAiMAX (Invitrogen) 20 ⁇ l (Invitrogen) was mixed and added. After culturing for 80 hours, the number of cells in each well was quantified using Cell Counting Kit-8 (manufactured by Dojindo). As a negative control, a trade name: miRCURY Knockdown control probe (SEQ ID NO: 32, manufactured by EXIQON) was used. The result is shown in FIG.
- antisense oligonucleotides for hsa-miR-133a / b, hsa-miR-346, and hsa-miR-361-3p in Table 6 above are used for cancer cells other than head and neck cancer. However, it showed a growth inhibitory effect.
- FIGS. 3 is a graph showing the tumor size calculated under the following conditions after 3, 6, 9, and 12 days after administration of antisense oligonucleotide and control
- FIG. 4 is a photograph of the tumor after 12 days. .
- Tumor model mice were prepared by transplanting 1 ⁇ 10 6 GFP-SAS cells subcutaneously on the back of 6-week-old male Balb / C nude mice. On day 12 after transplantation, tumor formation was confirmed, and an LNA / DNA antisense oligonucleotide and a control LNA / DNA oligonucleotide against hsa-miR-361-3p (both phosphorothioate-type oligonucleotides having the base sequences described in Table 6 above) About 20 bases of DNA (containing about 8 bases of LNA) was systemically administered via the tail vein, and systemically administered 7 days later.
- the antitumor activity of nucleotides was evaluated.
- the antisense oligonucleotide for hsa-miR-361-3p showed a cancer cell growth inhibitory effect even in vivo.
- the present invention is useful in drug development related to cancer, cancer medical fields, cancer research fields, and the like.
- SEQ ID NOs: 1-4 MicroRNA sequences registered in miRBase (hsa-miR-133a / b MIMAT0000427 / MIMAT0000770, hsa-miR-346 MIMAT0000773, hsa-miR-361-3p MIMAT0004682)
- SEQ ID NO: 5-8 Complementary sequence of SEQ ID NO: 1-4
- SEQ ID NO: 9-18 MicroRNA sequence registered in miRBase (hsa-miR-92a MIMAT0000092, hsa-miR-139-5p MIMAT0000250, hsa-miR -197 MIMAT0000227, hsa-miR-328 MIMAT0000752, hsa-miR-605 MIMAT0003273, hsa-miR-766 MIMAT0003888, hsa-miR-1228 MIMAT0005583, hsa-miR-1252 MIMAT0005944, hsa-m
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Abstract
Description
[1]マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトがん細胞の生育を抑制するための組成物;
[2]マイクロRNAのアンチセンスオリゴヌクレオチドを含む医薬組成物であって、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトのがんの治療又は予防のための医薬組成物;
[3]マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、miR-133、miR-346、及びmiR-361からなる群から選択される、非ヒトのがん細胞の生育を抑制するための組成物;
[4]前記がんが、頭頸部がん、肺がん、胆道がん、膵臓がん、及び前立腺がんからなる群から選択される、[1]から[3]のいずれかに記載の組成物;
[5]マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒト頭頸部がん細胞の生育を抑制するための組成物;
[6]マイクロRNAのアンチセンスオリゴヌクレオチドを含む医薬組成物であって、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒトの頭頸部がんの治療又は予防のための医薬組成物;
[7]マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、miR-92、miR-133、miR-139、miR-197、miR-328、miR-346、miR-361、miR-605、miR-766、miR-1228、miR-1252、miR-1260、及びmiR-1271からなる群から選択される、非ヒトの頭頸部がん細胞の生育を抑制するための組成物;
[8]ヒトがん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトがん細胞の生育を抑制する方法;
[9]ヒトのがんの治療又は予防方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を対象に投与することを含み、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトのがんの治療又は予防方法;
[10]非ヒトのがん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、miR-133、miR-346、及びmiR-361からなる群から選択される、非ヒトのがん細胞の生育を抑制する方法;
[11]前記がんが、頭頸部がん、肺がん、胆道がん、膵臓がん、及び前立腺がんからなる群から選択される、[8]から[10]のいずれかに記載の方法;
[12]ヒト頭頸部がん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒト頭頸部がん細胞の生育を抑制する方法;
[13]ヒトの頭頸部がんの治療又は予防方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒトの頭頸部がんの治療又は予防方法;
[14]非ヒトの頭頸部がん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を対象に投与することを含み、前記マイクロRNAが、miR-92、miR-133、miR-139、miR-197、miR-328、miR-346、miR-361、miR-605、miR-766、miR-1228、miR-1252、miR-1260、及びmiR-1271からなる群から選択される、非ヒトの頭頸部がん細胞の生育を抑制する方法;
に関する。
本発明は、一態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトがん細胞の生育を抑制するための組成物(以下、「本発明の第1の組成物」ともいう。)に関する。前記マイクロRNA(成熟型)の配列は、それぞれ、下記表1の配列番号1~4に記載される。
上述した本発明の第1の組成物は、上述したがんの治療又は予防のための医薬組成物として使用できる。したがって、本発明は、その他の態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含む医薬組成物であって、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトのがんの治療又は予防のための医薬組成物(以下、「本発明の第1の医薬組成物」ともいう。)に関する。本明細書において、がんの予防は、がんの再発の予防を含む。
本発明は、さらなる態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、miR-133、miR-346、及びmiR-361からなる群から選択される非ヒトのがん細胞の生育を抑制するための組成物(以下、「本発明の第2の組成物」ともいう。)に関する。例えば、イヌのがん細胞の生育を抑制する場合には、cfa-miR-133a/b/c(MIMIAT0009834, 0009835, 0009833)、cfa-miR-346(MIMIAT0004949)、及びcfa-miR-361(MIMIAT0006751)からなる群から選択されるマイクロRNAのアンチセンスヌクレオチドを使用することができ、他の生物についても同様に対応するマイクロRNA配列についてのアンチセンスヌクレオチドを使用できる。なお、本発明の第2の組成物は、1種類又は複数種類のアンチセンスオリゴヌクレオチドを含みうる。
上述した本発明の第2の組成物は、上述したがんの治療又は予防のための医薬組成物として使用できる。したがって、本発明は、その他の態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含む非ヒトの医薬組成物であって、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、非ヒトのがんの治療又は予防のための医薬組成物(以下、「本発明の第2の医薬組成物」ともいう。)に関する。本発明の第2の医薬組成物に含まれうるマイクロRNAのアンチセンスオリゴヌクレオチドの長さ及び核酸アナログについては、上述のとおりである。なお、本発明の第2の医薬組成物は、1種類又は複数種類のアンチセンスオリゴヌクレオチドを含みうる。
本発明は、その他の態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒト頭頸部がん細胞の生育を抑制するための組成物(以下、「本発明の第3の組成物」ともいう。)に関する。なお、本発明の第3の組成物は、1種類又は複数種類のアンチセンスオリゴヌクレオチドを含みうる。
上述した本発明の第3の組成物は、ヒトの頭頸部がんの治療又は予防のための医薬組成物として使用できる。したがって、本発明は、その他の態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含む医薬組成物であって、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒトの頭頸部がんの治療又は予防のための医薬組成物(以下、「本発明の第3の医薬組成物」ともいう。)に関する。なお、本発明の第3の医薬組成物は、1種類又は複数種類のアンチセンスオリゴヌクレオチドを含みうる。
本発明は、さらなる態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、miR-92、miR-133、miR-139、miR-197、miR-328、miR-346、miR-361、miR-605、miR-766、miR-1228、miR-1252、miR-1260、及びmiR-1271からなる群から選択される非ヒトのがん細胞の生育を抑制するための組成物(以下、「本発明の第4の組成物」ともいう。)に関する。なお、本発明の第4の組成物は、1種類又は複数種類のアンチセンスオリゴヌクレオチドを含みうる。含まれうるオリゴヌクレオチドの長さ及び核酸アナログについては、上述のとおりである。なお、本発明の第4の組成物は、凍結乾燥された形態であってもよい。
上述した本発明の第4の組成物は、非ヒトの頭頸部がんの治療又は予防のための医薬組成物として使用できる。したがって、本発明は、その他の態様として、マイクロRNAのアンチセンスオリゴヌクレオチドを含む非ヒトの医薬組成物であって、前記マイクロRNAが、miR-92、miR-133、miR-139、miR-197、miR-328、miR-346、miR-361、miR-605、miR-766、miR-1228、miR-1252、miR-1260、及びmiR-1271からなる群から選択されるヒトの頭頸部がんの治療又は予防のための医薬組成物(以下、「本発明の第4の医薬組成物」ともいう。)に関する。本発明の第4の医薬組成物に含まれうるマイクロRNAのアンチセンスオリゴヌクレオチドの長さ及び核酸アナログについては、上述のとおりである。なお、本発明の第4の医薬組成物は、1種類又は複数種類のアンチセンスオリゴヌクレオチドを含みうる。
本発明における第1及び第3の組成物によれば、対象とするヒトのがん細胞の生育を抑制できる。したがって、本発明は、その他の態様として、これらの組成物を対象のヒトがん細胞と接触させることを含むがん細胞の生育を抑制する方法に関する。接触方法は特に制限されず、前記組成物中のマイクロRNAのアンチセンスオリゴヌクレオチドを対象とするがん細胞に導入できる方法が採用できる。マイクロRNAのアンチセンスオリゴヌクレオチドをがん細胞に導入する実施形態の一つとしてリポフェクション法が挙げられる。また、リポフェクション法及びその他の導入方法時における前記アンチセンスオリゴヌクレオチドの細胞への投与量としては、例えば、10~25nMが挙げられる。
本発明における第1及び第3の医薬組成物によれば、対象とするヒトのがんの治療や予防が可能となる。したがって、本発明は、その他の態様として、これらの医薬組成物を対象に投与することを含むがん細胞の生育を抑制する方法に関する投与する組成物の剤形は、特に制限されず、例えば、注射剤、クリーム剤、軟膏、錠剤、懸濁剤等が挙げられる。また、投与方法も特に制限されず、例えば、注射、経口、局所、鼻内、直腸、静脈内、動脈内投与等が挙げられる。投与は、治療する病状の重症度と反応性、及び治療経過に依存する。最適な投薬スケジュールは、対象の体内への薬剤の蓄積量の測定値から計算することができる。最適用量は、個々のオリゴヌクレオチドの相対的有効性によって異なりうる。一般的には、in vitro及びin vivo動物モデルで有効であることが分かったEC50に基づいて推定することができる。一般的には、用量は0.01μg~1g/kg体重、好ましくは0.01~100mg/kg体重であり、より好ましくは1~10mg/kg体重である。投与回数としては、1日、1週間、1月間又は1年間に1回又はそれ以上、又は2~10年間に1回投与するか、あるいは数時間~数カ月間連続注入により投与することができる。投与の反復回数は、測定した体液又は組織中の薬剤濃度と滞留時間に基づいて推定することができる。治療の成功後、病状の再発を防ぐために対象は維持療法を受けることが望ましい場合もある。
本発明における第2及び第4の組成物によれば、対象とする非ヒトのがん細胞の生育を抑制できる。したがって、本発明は、その他の態様として、これらの組成物を対象の非ヒトがん細胞と接触させることを含むがん細胞の生育を抑制する方法に関する。接触方法は特に制限されず、前記組成物中のマイクロRNAのアンチセンスオリゴヌクレオチドを対象とするがん細胞に導入できる方法が採用できる。
本発明における第2及び第4の医薬組成物によれば、対象とする非ヒトのがんの治療や予防が可能となる。したがって、本発明は、その他の態様として、これらの医薬組成物を非ヒト対象に投与することを含むがん細胞の生育を抑制する方法に関する投与する組成物の剤形は、特に制限されず、例えば、注射剤、クリーム剤、軟膏、錠剤、懸濁剤等が挙げられる。また、投与方法も特に制限されず、例えば、注射、経口、局所、鼻内、直腸、静脈内、動脈内投与等が挙げられる。投与は、治療する病状の重症度と反応性、及び治療経過に依存する。最適な投薬スケジュールは、対象の体内への薬剤の蓄積量の測定値から計算することができる。最適用量は、個々のオリゴヌクレオチドの相対的有効性によって異なりうる。一般的には、in vitro及びin vivo動物モデルで有効であることが分かったEC50に基づいて推定することができる。一般的には、用量は0.01μg~1g/kg体重、好ましくは0.01~100mg/kg体重であり、より好ましくは1~10mg/kg体重である。投与回数としては、1日、1週間、1月間又は1年間に1回又はそれ以上、又は2~10年間に1回投与するか、あるいは数時間~数カ月間連続注入により投与することができる。投与の反復回数は、測定した体液又は組織中の薬剤濃度と滞留時間に基づいて推定することができる。治療の成功後、病状の再発を防ぐために対象は維持療法を受けることが望ましい場合もある。
miRNA knockdown libraryを用い、下記2種類のヒト頭頸部がん細胞株におけるマイクロRNAを阻害した場合の生育に対する影響を網羅的に解析した。具体的には下記の条件で行った。
〔使用した細胞株〕
ヒト口腔扁平上皮がん細胞株SAS及びヒト唾液腺がん細胞株ACCMにgreen fluorescent protein(GFP)遺伝子を導入して分離樹立したGFP安定発現株GFP-SAS及びGFP-ACCMを使用した。
〔miRNA knockdown library を用いた網羅的機能解析〕
96ウェルプラスチックプレート(商品名:BD Falcon、BD社製)の各ウェルに、ヒト口腔がん細胞株GFP-SASあるいはGFP-ACCMを2x103個含む10%FBS含有DMEM培地160μlと918種類のヒトマイクロRNAに対するLNA(Locked Nucleic Acid)/DNA knockdown probe(商品名:miRCURY LNATM microRNA Knockdown Library-Human v12.0、EXIQON社製)各5pmol、RNAiMAX(Invitrogen社製)0.4μlを含むOpti-MEM(Invitrogen社製)40μlを混合し、加えた。80時間培養した後、各ウェルのGFP蛍光強度をWallac ARVO MX 1420 Multilabel Counter(PerkinElmer社製)にて測定し、つづいてCell Counting Kit-8(Dojindo社製)を用いて細胞数を定量した。
下記のhsa-miR-133a/b、hsa-miR-346、及びhsa-miR-361-3pに対するアンチセンスオリゴヌクレオチドを下記条件でヒト口腔扁平上皮がん細胞GFP-SASに導入し、細胞増殖に与える影響を評価した。
〔使用したアンチセンスオリゴヌクレオチド〕
hsa-miR-133a/b、hsa-miR-346、及びhsa-miR-361-3pに対するアンチセンスオリゴヌクレオチドは、下記表6の配列のknockdown probe(商品名:miRCURY LNA microRNA Inhibitor、EXIQON社製)を用いた。なお、ネガティブコントロールとしては、商品名:miRCURY Knockdown control probe(配列番号32、EXIQON社製)を使用した。これらのオリゴヌクレオチドは、LNA(商品名;Locked Nucleic Acid)を含む(約20塩基のDNAに対して約8塩基程度のLNAを含む)。
〔miRNA knockdown probeの導入と細胞増殖評価法〕
96ウェルプラスチックプレートの各ウェルに、種々のヒトがん細胞を2x103個含むcomplete medium80μlとmiRNAに対する下記表3の配列のknockdown probe2.5pmol、RNAiMAX(Invitrogen社製)0.2μlを含む Opti-MEM(Invitrogen社製)20μlを混合し、加えた。80時間培養した後、各ウェルの生存細胞が発するGFPの蛍光を蛍光顕微鏡観察した。その結果を図1に示す。
上記表6のhsa-miR-133a/b、hsa-miR-346、及びhsa-miR-361-3pに対するアンチセンスオリゴヌクレオチドを、下記のさまざまながん細胞に導入して細胞増殖に与える影響を下記条件で評価した。
〔使用した細胞株〕
ヒト口腔扁平上皮がん細胞株SAS及びヒト唾液腺がん細胞株ACCMにgreen fluorescent protein(GFP)遺伝子を導入して分離樹立したGFP安定発現株GFP-SAS及びGFP-ACCMに加え、ヒト口腔扁平上皮がん細胞株B88、ヒト膵臓がん細胞株MIAPaCa-2、ヒト胆道がん細胞株HuCCT1、ヒト肺扁平上皮がん細胞株EBC-1、ヒト肺腺がん細胞株A549、ヒト前立腺がん細胞株 LNCaP(アンドロゲン依存性)、PC-3(アンドロゲン非依存性)を用いた。これら細胞株の培養には、10%牛胎児血清(FBS;Biosource International社製)、100μg/mlストレプトマイシン、100U/mlペニシリン、0.25mg/mlアンホテリシンB(Invitrogen社製)を含むダルベッコ改変イーグル培地(DMEM)あるいはRPMI1640培地(Sigma-Aldrich社製)を増殖培養液として用い、空気中に5%の割合で炭酸ガスを含む培養器内で、37℃にて行った。
〔miRNA knockdown probe の導入と細胞増殖評価法〕
96ウェルプラスチックプレートの各ウェルに、種々のヒトがん細胞を2x103個含むcomplete medium80μlとmiRNAに対する上記表3の配列のknockdown probe2.5pmol、RNAiMAX(Invitrogen社製)0.2μlを含むOpti-MEM(Invitrogen社製)20μlを混合し、加えた。80時間培養した後、各ウェルの細胞数をCell Counting Kit-8(Dojindo社製)を用いて細胞数を定量した。なお、ネガティブコントロールとしては、商品名:miRCURY Knockdown control probe(配列番号32、EXIQON社製)を使用した。その結果を図2に示す。
hsa-miR-361-3pに対するアンチセンスオリゴヌクレオチド及びネガティブコントロールをそれぞれ、下記条件で腫瘍モデルマウスに投与して腫瘍の増殖(大きさ)の経時変化を観察した。その結果を図3及び図4に示す。図3は、アンチセンスオリゴヌクレオチド及びコントロールを投与して3、6、9、及び12日後の腫瘍の大きさを下記条件で算出したグラフであり、図4は、12日後における腫瘍の写真である。
〔腫瘍モデルマウス、アンチセンスオリゴヌクレオチドの投与方法及び観察方法〕
腫瘍モデルマウスは、GFP-SAS細胞1x106個を6週齢雄のBalb/Cヌードマウス背部皮下に移植して作製した。移植後12日目に腫瘍形成を確認し、hsa-miR-361-3p に対するLNA/DNAアンチセンスオリゴヌクレオチド及びコントロールLNA/DNAオリゴヌクレオチド(いずれも上記表6に記載の塩基配列のホスホロチオエート型オリゴヌクレオチド;約20塩基のDNAに対して約8塩基程度のLNAを含む)を尾静脈から8nmol全身投与し、さらに7日後にも同様に全身投与した。初回投与から3、6、9、及び12日後に腫瘍体積を[腫瘍体積=長径×短径×高さ×0.5236 ]の計算式を用いて算出し、抗hsa-miR-361-3pアンチセンスオリゴヌクレオチドの抗腫瘍活性を評価した。
配列番号5~8:配列番号1~4の相補配列
配列番号9~18:miRBaseに登録されているマイクロRNAの配列(hsa-miR-92a MIMAT0000092、hsa-miR-139-5p MIMAT0000250、hsa-miR-197 MIMAT0000227、hsa-miR-328 MIMAT0000752、hsa-miR-605 MIMAT0003273、hsa-miR-766 MIMAT0003888、hsa-miR-1228 MIMAT0005583、hsa-miR-1252 MIMAT0005944、hsa-miR-1260 MIMAT0005911、hsa-miR-1271 MIMAT0005796)
配列番号19~28:配列番号9~18の相補配列
配列番号29:抗hsa-miR-133a/bアンチセンスオリゴヌクレオチド
配列番号30:抗hsa-miR-346アンチセンスオリゴヌクレオチド
配列番号31:抗hsa-miR-361-3pアンチセンスオリゴヌクレオチド
配列番号32:ネガティブコントロール
Claims (14)
- マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトがん細胞の生育を抑制するための組成物。
- マイクロRNAのアンチセンスオリゴヌクレオチドを含む医薬組成物であって、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトのがんの治療又は予防のための医薬組成物。
- マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、miR-133、miR-346、及びmiR-361からなる群から選択される、非ヒトのがん細胞の生育を抑制するための組成物。
- 前記がんが、頭頸部がん、肺がん、胆道がん、膵臓がん、及び前立腺がんからなる群から選択される、請求項1から3のいずれかに記載の組成物。
- マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒト頭頸部がん細胞の生育を抑制するための組成物。
- マイクロRNAのアンチセンスオリゴヌクレオチドを含む医薬組成物であって、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒトの頭頸部がんの治療又は予防のための医薬組成物。
- マイクロRNAのアンチセンスオリゴヌクレオチドを含み、前記マイクロRNAが、miR-92、miR-133、miR-139、miR-197、miR-328、miR-346、miR-361、miR-605、miR-766、miR-1228、miR-1252、miR-1260、及びmiR-1271からなる群から選択される、非ヒトの頭頸部がん細胞の生育を抑制するための組成物。
- ヒトがん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトがん細胞の生育を抑制する方法。
- ヒトのがんの治療又は予防方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を対象に投与することを含み、前記マイクロRNAが、hsa-miR-133a、hsa-miR-133b、hsa-miR-346、及びhsa-miR-361-3pからなる群から選択される、ヒトのがんの治療又は予防方法。
- 非ヒトのがん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、miR-133、miR-346、及びmiR-361からなる群から選択される、非ヒトのがん細胞の生育を抑制する方法。
- 前記がんが、頭頸部がん、肺がん、胆道がん、膵臓がん、及び前立腺がんからなる群から選択される、請求項8から10のいずれかに記載の方法。
- ヒト頭頸部がん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒト頭頸部がん細胞の生育を抑制する方法。
- ヒトの頭頸部がんの治療又は予防方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を対象に投与することを含み、前記マイクロRNAが、hsa-miR-92a、hsa-miR-133a、hsa-miR-133b、hsa-miR-139-5p、hsa-miR-197、hsa-miR-328、hsa-miR-346、hsa-miR-361-3p、hsa-miR-605、hsa-miR-766、hsa-miR-1228、hsa-miR-1252、hsa-miR-1260、及びhsa-miR-1271からなる群から選択される、ヒトの頭頸部がんの治療又は予防方法。
- 非ヒトの頭頸部がん細胞の生育を抑制する方法であって、マイクロRNAのアンチセンスオリゴヌクレオチドを含む組成物を前記がん細胞に接触させることを含み、前記マイクロRNAが、miR-92、miR-133、miR-139、miR-197、miR-328、miR-346、miR-361、miR-605、miR-766、miR-1228、miR-1252、miR-1260、及びmiR-1271からなる群から選択される、非ヒトの頭頸部がん細胞の生育を抑制する方法。
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JP7340184B2 (ja) * | 2016-03-31 | 2023-09-07 | 東レ株式会社 | 早期膵がん又は膵がん前駆病変の検出キット又はデバイス及び検出方法 |
US20210139895A1 (en) * | 2017-06-16 | 2021-05-13 | Prostemics Co., Ltd. | Pharmaceutical composition for preventing or treating cancer |
CN110354137B (zh) * | 2019-08-20 | 2023-08-04 | 中山大学附属第六医院 | miRNA-197-3p在制备抗前列腺癌药物中的应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007000169A2 (en) | 2005-06-29 | 2007-01-04 | Genmab A/S | Non-human mammalian arthritis model featuring human antibodies against citrullinated proteins |
JP2008500837A (ja) | 2004-05-28 | 2008-01-17 | アンビオン インコーポレーティッド | マイクロrnaに関与する方法および組成物 |
JP2008519606A (ja) * | 2004-11-12 | 2008-06-12 | アンビオン インコーポレーティッド | miRNAおよびmiRNA阻害分子に関する方法および組成物 |
JP2009532392A (ja) | 2006-04-03 | 2009-09-10 | サンタリス ファーマ アー/エス | antimiRNAアンチセンスオリゴヌクレオチドを含む医薬組成物 |
WO2010056737A2 (en) * | 2008-11-11 | 2010-05-20 | Mirna Therapeutics, Inc. | Methods and compositions involving mirnas in cancer stem cells |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003070918A2 (en) | 2002-02-20 | 2003-08-28 | Ribozyme Pharmaceuticals, Incorporated | Rna interference by modified short interfering nucleic acid |
AU2003207708A1 (en) | 2002-02-20 | 2003-09-09 | Sirna Therapeutics, Inc. | Rna interference mediated inhibition of map kinase genes |
WO2009036236A1 (en) * | 2007-09-14 | 2009-03-19 | The Ohio State University Research Foundation | Mirna expression in human peripheral blood microvesicles and uses thereof |
-
2011
- 2011-11-10 WO PCT/JP2011/075924 patent/WO2012063894A1/ja active Application Filing
- 2011-11-10 JP JP2012542971A patent/JPWO2012063894A1/ja active Pending
- 2011-11-10 US US13/884,436 patent/US8889649B2/en active Active
- 2011-11-10 EP EP11840297.3A patent/EP2638912A4/en not_active Withdrawn
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008500837A (ja) | 2004-05-28 | 2008-01-17 | アンビオン インコーポレーティッド | マイクロrnaに関与する方法および組成物 |
JP2008519606A (ja) * | 2004-11-12 | 2008-06-12 | アンビオン インコーポレーティッド | miRNAおよびmiRNA阻害分子に関する方法および組成物 |
WO2007000169A2 (en) | 2005-06-29 | 2007-01-04 | Genmab A/S | Non-human mammalian arthritis model featuring human antibodies against citrullinated proteins |
JP2009532392A (ja) | 2006-04-03 | 2009-09-10 | サンタリス ファーマ アー/エス | antimiRNAアンチセンスオリゴヌクレオチドを含む医薬組成物 |
WO2010056737A2 (en) * | 2008-11-11 | 2010-05-20 | Mirna Therapeutics, Inc. | Methods and compositions involving mirnas in cancer stem cells |
Non-Patent Citations (3)
Title |
---|
SCHICKEL R ET AL., ONCOGENE, vol. 27, 2008, pages 5959 - 5974 |
See also references of EP2638912A4 * |
WEBER F. ET AL.: "A Limited Set of Human MicroRNA Is Deregulated in Follicular Thyroid Carcinoma", J.CLIN.ENDOCRINOL.METAB., vol. 91, no. 9, 2006, pages 3584 - 3591, XP002469100 * |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013088338A1 (en) * | 2011-12-15 | 2013-06-20 | Oncostamen S.R.L. | Micrornas and uses therof |
JP2014128249A (ja) * | 2012-12-28 | 2014-07-10 | Hokkaido Univ | 神経変性疾患の検査と治療に対するmiRNA又はその標的遺伝子の利用 |
JP2015089874A (ja) * | 2013-11-05 | 2015-05-11 | 国立大学法人大阪大学 | miR−340を用いた大腸癌の治療剤 |
WO2015190542A1 (ja) * | 2014-06-11 | 2015-12-17 | 東レ株式会社 | 胆道がんの検出キット又はデバイス及び検出方法 |
US10633708B2 (en) | 2014-06-11 | 2020-04-28 | Toray Industries, Inc. | Biliary tract cancer detection kit or device, and detection method |
US11499198B2 (en) | 2014-06-11 | 2022-11-15 | Toray Industries, Inc. | Biliary tract cancer detection kit or device, and detection method |
US11761046B2 (en) | 2014-06-11 | 2023-09-19 | Toray Industries, Inc. | Biliary tract cancer detection kit or device, and detection method |
JPWO2019240223A1 (ja) * | 2018-06-13 | 2021-07-01 | 公益財団法人川崎市産業振興財団 | 数平均分子量が3kDa〜10kDaであるPEGブロックとカチオン性ポリマーとのブロックコポリマーと、アンチセンスオリゴヌクレオチドとを含む、ポリイオンコンプレックスミセル |
WO2020032228A1 (ja) * | 2018-08-10 | 2020-02-13 | 東レ株式会社 | 前立腺がんの検出のためのキット、デバイス及び方法 |
JP2021080186A (ja) * | 2019-11-15 | 2021-05-27 | 公立大学法人大阪 | 胆道癌又は膵癌治療剤 |
JP7522424B2 (ja) | 2019-11-15 | 2024-07-25 | 公立大学法人大阪 | 胆道癌又は膵癌治療剤 |
Also Published As
Publication number | Publication date |
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EP2638912A1 (en) | 2013-09-18 |
US8889649B2 (en) | 2014-11-18 |
JPWO2012063894A1 (ja) | 2014-05-12 |
US20140045917A1 (en) | 2014-02-13 |
EP2638912A4 (en) | 2015-01-21 |
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