JP2024039787A - Malignant tumor treatment agent - Google Patents
Malignant tumor treatment agent Download PDFInfo
- Publication number
- JP2024039787A JP2024039787A JP2022144405A JP2022144405A JP2024039787A JP 2024039787 A JP2024039787 A JP 2024039787A JP 2022144405 A JP2022144405 A JP 2022144405A JP 2022144405 A JP2022144405 A JP 2022144405A JP 2024039787 A JP2024039787 A JP 2024039787A
- Authority
- JP
- Japan
- Prior art keywords
- compound
- represent
- hydrogen atom
- cancer
- same
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 201000011510 cancer Diseases 0.000 title claims abstract description 31
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims abstract description 34
- 102000007537 Type II DNA Topoisomerases Human genes 0.000 claims abstract description 26
- 108010046308 Type II DNA Topoisomerases Proteins 0.000 claims abstract description 26
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 20
- 125000004432 carbon atom Chemical group C* 0.000 claims abstract description 20
- 125000000561 purinyl group Chemical class N1=C(N=C2N=CNC2=C1)* 0.000 claims abstract description 19
- 150000003839 salts Chemical class 0.000 claims abstract description 19
- 239000003814 drug Substances 0.000 claims abstract description 18
- 125000005843 halogen group Chemical group 0.000 claims abstract description 17
- 229940124597 therapeutic agent Drugs 0.000 claims abstract description 12
- 239000003112 inhibitor Substances 0.000 claims description 9
- 210000004027 cell Anatomy 0.000 description 48
- 229940126062 Compound A Drugs 0.000 description 40
- NLDMNSXOCDLTTB-UHFFFAOYSA-N Heterophylliin A Natural products O1C2COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC2C(OC(=O)C=2C=C(O)C(O)=C(O)C=2)C(O)C1OC(=O)C1=CC(O)=C(O)C(O)=C1 NLDMNSXOCDLTTB-UHFFFAOYSA-N 0.000 description 40
- 206010028980 Neoplasm Diseases 0.000 description 26
- 150000001875 compounds Chemical class 0.000 description 19
- 230000005782 double-strand break Effects 0.000 description 19
- 230000000694 effects Effects 0.000 description 18
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 description 18
- 229960005420 etoposide Drugs 0.000 description 18
- 238000000034 method Methods 0.000 description 12
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 10
- 238000010586 diagram Methods 0.000 description 9
- 229910052801 chlorine Inorganic materials 0.000 description 8
- 125000001309 chloro group Chemical group Cl* 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 230000035945 sensitivity Effects 0.000 description 7
- 229940124087 DNA topoisomerase II inhibitor Drugs 0.000 description 6
- 239000000317 Topoisomerase II Inhibitor Substances 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 238000003197 gene knockdown Methods 0.000 description 6
- 230000006957 competitive inhibition Effects 0.000 description 5
- 238000012217 deletion Methods 0.000 description 5
- 230000037430 deletion Effects 0.000 description 5
- 229960004679 doxorubicin Drugs 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 229910052731 fluorine Inorganic materials 0.000 description 5
- 125000001153 fluoro group Chemical group F* 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- OBYGAPWKTPDTAS-OCAPTIKFSA-N ICRF-193 Chemical compound N1([C@H](C)[C@H](C)N2CC(=O)NC(=O)C2)CC(=O)NC(=O)C1 OBYGAPWKTPDTAS-OCAPTIKFSA-N 0.000 description 4
- 108020004459 Small interfering RNA Proteins 0.000 description 4
- 230000022131 cell cycle Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000002552 dosage form Substances 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 230000001093 anti-cancer Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 239000000460 chlorine Substances 0.000 description 3
- 239000003086 colorant Substances 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 102100033350 ATP-dependent translocase ABCB1 Human genes 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical group [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 2
- 206010048610 Cardiotoxicity Diseases 0.000 description 2
- 206010009944 Colon cancer Diseases 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 208000008839 Kidney Neoplasms Diseases 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 108010047230 Member 1 Subfamily B ATP Binding Cassette Transporter Proteins 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 description 2
- 206010060862 Prostate cancer Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- 206010038389 Renal cancer Diseases 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 231100000259 cardiotoxicity Toxicity 0.000 description 2
- 230000025084 cell cycle arrest Effects 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000012054 celltiter-glo Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 208000029742 colonic neoplasm Diseases 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000007884 disintegrant Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003534 dna topoisomerase inhibitor Substances 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- 230000009422 growth inhibiting effect Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 201000010982 kidney cancer Diseases 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 230000010534 mechanism of action Effects 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 208000016691 refractory malignant neoplasm Diseases 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000829 suppository Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 229940044693 topoisomerase inhibitor Drugs 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical group [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 238000011729 BALB/c nude mouse Methods 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 102000003915 DNA Topoisomerases Human genes 0.000 description 1
- 108090000323 DNA Topoisomerases Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- 101710195517 Histone H2AX Proteins 0.000 description 1
- 102100034533 Histone H2AX Human genes 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 206010027406 Mesothelioma Diseases 0.000 description 1
- 208000034578 Multiple myelomas Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- 208000021712 Soft tissue sarcoma Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 101150019239 TOP2B gene Proteins 0.000 description 1
- 208000024313 Testicular Neoplasms Diseases 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- 101150107801 Top2a gene Proteins 0.000 description 1
- 101710183280 Topoisomerase Proteins 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 201000009036 biliary tract cancer Diseases 0.000 description 1
- 208000020790 biliary tract neoplasm Diseases 0.000 description 1
- 206010006007 bone sarcoma Diseases 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 230000009702 cancer cell proliferation Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 210000004413 cardiac myocyte Anatomy 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 201000007455 central nervous system cancer Diseases 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 208000006990 cholangiocarcinoma Diseases 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 238000010835 comparative analysis Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 201000004101 esophageal cancer Diseases 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 238000010230 functional analysis Methods 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 201000010175 gallbladder cancer Diseases 0.000 description 1
- 201000007487 gallbladder carcinoma Diseases 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 201000010536 head and neck cancer Diseases 0.000 description 1
- 208000014829 head and neck neoplasm Diseases 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000003589 local anesthetic agent Substances 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- SYSQUGFVNFXIIT-UHFFFAOYSA-N n-[4-(1,3-benzoxazol-2-yl)phenyl]-4-nitrobenzenesulfonamide Chemical class C1=CC([N+](=O)[O-])=CC=C1S(=O)(=O)NC1=CC=C(C=2OC3=CC=CC=C3N=2)C=C1 SYSQUGFVNFXIIT-UHFFFAOYSA-N 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001280 n-hexyl group Chemical group C(CCCCC)* 0.000 description 1
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- -1 organic acid salts Chemical class 0.000 description 1
- 230000002611 ovarian Effects 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- LVTJOONKWUXEFR-FZRMHRINSA-N protoneodioscin Natural products O(C[C@@H](CC[C@]1(O)[C@H](C)[C@@H]2[C@]3(C)[C@H]([C@H]4[C@@H]([C@]5(C)C(=CC4)C[C@@H](O[C@@H]4[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@@H](O)[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@H](CO)O4)CC5)CC3)C[C@@H]2O1)C)[C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1 LVTJOONKWUXEFR-FZRMHRINSA-N 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 206010038038 rectal cancer Diseases 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000028617 response to DNA damage stimulus Effects 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 201000003120 testicular cancer Diseases 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000012929 tonicity agent Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
【課題】トポイソメラーゼＩＩαを特異的に阻害する悪性腫瘍治療剤を提供する。【解決手段】次の一般式（１）JPEG2024039787000013.jpg91170（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）で表されるプリン誘導体又はその塩を含有する悪性腫瘍治療剤。【選択図】図１[Problem] To provide a malignant tumor therapeutic agent that specifically inhibits topoisomerase IIα. [Solution] A malignant tumor therapeutic agent containing a purine derivative represented by the following general formula (1): JPEG2024039787000013.jpg91170 (wherein X1 and X2 are the same or different and represent a hydrogen atom or a halogen atom, and R1 and R2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms) or a salt thereof. [Selected Figure] Figure 1
Description
本発明は、悪性腫瘍治療剤に関する。 The present invention relates to a therapeutic agent for malignant tumor.
がん細胞は無秩序な増殖を繰り返し、正常な細胞を障害し組織を壊したり、転移を行うことで本来がんのかたまりがない組織でも増殖を行う。細胞の増殖には遺伝情報が刻まれたＤＮＡの複製が必要となり、複製時にはＤＮＡが持つらせん構造のねじれやひずみを一度解消させる必要がある。このらせん構造を変化させる酵素にトポイソメラーゼがあり、Ｉ型とＩＩ型のタイプが存在する。トポイソメラーゼＩはＤＮＡの２重らせん構造のうち１本を切断し再結合に関与する。トポイソメラーゼＩＩは２重らせん構造の両方を切断し再結合に関与する。このトポイソメラーゼＩＩを阻害することによる悪性腫瘍治療剤としては、エトポシドが知られている（非特許文献１）。また、ドキソルビシンなどのアントラサイクリン系悪性腫瘍治療剤も、トポイソメラーゼＩＩ阻害作用を有することが知られている。 Cancer cells repeatedly proliferate in a disorderly manner, damaging normal cells and destroying tissues, and by metastasizing, they can proliferate even in tissues that are not originally cancerous. Cell proliferation requires the replication of DNA, which contains genetic information, and during replication, the twists and distortions in the helical structure of DNA must be resolved once. Topoisomerases are enzymes that change this helical structure, and there are two types: type I and type II. Topoisomerase I cleaves one of the double helix structures of DNA and is involved in recombination. Topoisomerase II cleaves both sides of the double helix and participates in recombination. Etoposide is known as a therapeutic agent for malignant tumors by inhibiting topoisomerase II (Non-Patent Document 1). Furthermore, anthracycline therapeutic agents for malignant tumors such as doxorubicin are also known to have topoisomerase II inhibitory effects.
トポイソメラーゼＩＩには、α型とβ型の二つのタイプがあり、エトポシドやドキソルビシンが阻害するのは、トポイソメラーゼＩＩα及びＩＩβの両方であることがわかってきた。そして、ドキソルビシンによる心毒性は、心筋細胞におけるトポイソメラーゼＩＩβの機能障害を介して起こることもわかってきた（非特許文献２）。
従って、本発明の課題は、トポイソメラーゼＩＩαを特異的に阻害する悪性腫瘍治療剤を提供することにある。
There are two types of topoisomerase II, α type and β type, and it has been found that etoposide and doxorubicin inhibit both topoisomerase IIα and IIβ. It has also been found that cardiotoxicity caused by doxorubicin occurs through dysfunction of topoisomerase IIβ in cardiomyocytes (Non-Patent Document 2).
Therefore, an object of the present invention is to provide a therapeutic agent for malignant tumors that specifically inhibits topoisomerase IIα.
そこで本発明者は、ＤＮＡ２本鎖切断（ＤＮＡ ｄｏｕｂｌｅ－ｓｔｒａｎｄ ｂｒｅａｋ：ＤＳＢ）定量システム（非特許文献３）を用いて、種々の癌細胞におけるＤＳＢ形成を生じさせる化合物をスクリーニングした。その結果、特定のプリン誘導体が多様な癌細胞株でＤＳＢ形成作用を示し、そのプリン誘導体は、細胞増殖抑制及びＤＮＡ損傷応答として特異的に起こるＧ２／Ｍ細胞周期アレストを引き起こすことも見出した。次に、多種の癌細胞株に対する細胞毒性作用を検討し、得られたフィンガープリントを用いて、既知の２５０化合物のデータベースをもとに類似性についてドライ解析した結果、そのプリン誘導体はトポイソメラーゼ阻害作用により抗がん効果を示していると予測された。そこで、そのプリン誘導体のトポイソメラーゼ阻害剤との競合作用を検討したところ、そのプリン誘導体はトポイソメラーゼ阻害剤との競合阻害を示し、さらに機能欠失実験により、その作用はトポイソメラーゼＩＩα特異的阻害剤であることが判明した。さらに、そのプリン誘導体は、ｉｎ ｖｉｖｏでも優れた抗癌効果を有すること確認し、本発明を完成した。 Therefore, the present inventor used a DNA double-strand break (DSB) quantification system (Non-Patent Document 3) to screen for compounds that cause DSB formation in various cancer cells. As a result, it was found that a specific purine derivative exhibits a DSB-forming effect in various cancer cell lines, and that the purine derivative induces G2/M cell cycle arrest that specifically occurs as a cell growth inhibition and DNA damage response. Next, we examined the cytotoxic effects on various cancer cell lines, and using the obtained fingerprints, we conducted a dry analysis of similarities based on a database of 250 known compounds. It was predicted that it would exhibit anticancer effects. Therefore, when we investigated the competitive effect of this purine derivative with topoisomerase inhibitors, we found that this purine derivative exhibited competitive inhibition with topoisomerase inhibitors, and further, through functional deletion experiments, we found that the effect was that of a topoisomerase IIα-specific inhibitor. It has been found. Furthermore, it was confirmed that the purine derivative has excellent anticancer effects even in vivo, and the present invention was completed.
すなわち、本発明は、次の発明［１］～［８］を提供するものである。
［１］次の一般式（１）
That is, the present invention provides the following inventions [1] to [8].
[1] The following general formula (1)
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩を含有する悪性腫瘍治療剤。
［２］次の一般式（１）
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
A therapeutic agent for malignant tumors containing a purine derivative represented by or a salt thereof.
[2] The following general formula (1)
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩を含有するトポイソメラーゼＩＩα阻害剤。
［３］悪性腫瘍治療剤製造のための、次の一般式（１）
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
A topoisomerase IIα inhibitor containing a purine derivative represented by or a salt thereof.
[3] The following general formula (1) for producing a therapeutic agent for malignant tumor
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩の使用。
［４］トポイソメラーゼＩＩα阻害剤製造のための、次の一般式（１）
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
Use of a purine derivative represented by or a salt thereof.
[4] The following general formula (1) for producing topoisomerase IIα inhibitor
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩の使用。
［５］悪性腫瘍を治療するための次の一般式（１）
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
Use of a purine derivative represented by or a salt thereof.
[5] The following general formula (1) for treating malignant tumors
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩。
［６］トポイソメラーゼＩＩαを阻害するための次の一般式（１）
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
A purine derivative represented by or a salt thereof.
[6] The following general formula (1) for inhibiting topoisomerase IIα
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩。
［７］次の一般式（１）
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
A purine derivative represented by or a salt thereof.
[7] The following general formula (1)
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩を投与することを特徴とする悪性腫瘍の治療方法。
［８］次の一般式（１）
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
A method for treating malignant tumors, which comprises administering a purine derivative represented by the formula or a salt thereof.
[8] The following general formula (1)
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩を投与することを特徴とするトポイソメラーゼＩＩαの阻害方法。
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
A method for inhibiting topoisomerase IIα, which comprises administering a purine derivative or a salt thereof.
本発明のプリン誘導体又はその塩を含有する医薬組成物を用いれば、癌患者のトポイソメラーゼＩＩαが特異的に阻害され、安全かつ確実に悪性腫瘍を治療することができる。 By using a pharmaceutical composition containing the purine derivative or a salt thereof of the present invention, topoisomerase IIα in cancer patients can be specifically inhibited, and malignant tumors can be safely and reliably treated.
本発明は、プリン誘導体又はその塩を含有する悪性腫瘍治療剤又はトポイソメラーゼＩＩα阻害剤であり、本発明の一態様は、次の一般式（１） The present invention is a malignant tumor therapeutic agent or topoisomerase IIα inhibitor containing a purine derivative or a salt thereof, and one aspect of the present invention is a compound represented by the following general formula (1).
（式中、Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示し、Ｒ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す）
で表されるプリン誘導体又はその塩を含有する悪性腫瘍治療剤又はトポイソメラーゼＩＩα阻害剤である。
(In the formula, X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, and R 1 and R 2 are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.)
A malignant tumor therapeutic agent or topoisomerase IIα inhibitor containing a purine derivative or a salt thereof represented by
式（１）中のＸ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示す。ここで、ハロゲン原子としては、塩素原子、臭素原子、フッ素原子、ヨウ素原子が挙げられるが、塩素原子、臭素原子又はフッ素原子が好ましく、塩素原子又はフッ素原子がより好ましく、塩素原子がさらに好ましい。
Ｘ１及びＸ２は、同一又は異なって、水素原子又はハロゲン原子を示すが、Ｘ１がハロゲン原子、Ｘ２が水素原子又はハロゲン原子であるのが好ましい。より好ましくは、Ｘ１が塩素原子又はフッ素原子であり、Ｘ２水素原子、塩素原子又はフッ素原子である場合であり、さらに好ましくは、Ｘ１が塩素原子であり、Ｘ２水素原子又は塩素原子である場合である。
X 1 and X 2 in formula (1) are the same or different and represent a hydrogen atom or a halogen atom. Here, examples of the halogen atom include a chlorine atom, a bromine atom, a fluorine atom, and an iodine atom, and a chlorine atom, a bromine atom, or a fluorine atom is preferable, a chlorine atom or a fluorine atom is more preferable, and a chlorine atom is even more preferable.
X 1 and X 2 are the same or different and represent a hydrogen atom or a halogen atom, but it is preferable that X 1 is a halogen atom and X 2 is a hydrogen atom or a halogen atom. More preferably, X 1 is a chlorine atom or a fluorine atom, and X 2 is a hydrogen atom, a chlorine atom, or a fluorine atom, and even more preferably, X 1 is a chlorine atom, and X 2 is a hydrogen atom or a chlorine atom. This is the case.
式（１）中のＲ１及びＲ２は、同一又は異なって、水素原子又は炭素数１～６のアルキル基を示す。ここで、炭素数１～６のアルキル基としては、メチル基、エチル基、ｎ－プロピル基、イソプロピル基、ｎ－ブチル基、ｔ－ブチル基、イソブチル基、ｎ－ペンチル基、ｎ－ヘキシル基などが挙げられる。このうち、炭素数１～４のアルキル基が好ましく、炭素数１～３のアルキル基がより好ましく、メチル基、エチル基がさらに好ましい。
Ｒ１とＲ２の組み合わせとしては、Ｒ１及びＲ２が水素原子の場合、Ｒ１及びＲ２が炭素数１～６のアルキル基の場合がより好ましい。Ｒ１及びＲ２が水素原子の場合、Ｒ１及びＲ２が炭素数１～４のアルキル基の場合がさらに好ましい。Ｒ１及びＲ２が水素原子の場合、Ｒ１及びＲ２が炭素数１～３のアルキル基の場合がよりさらに好ましい。Ｒ１及びＲ２が水素原子の場合、Ｒ１及びＲ２が炭素数１～２のアルキル基の場合がよりさらに好ましい。
R 1 and R 2 in formula (1) are the same or different and represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms. Here, as the alkyl group having 1 to 6 carbon atoms, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, t-butyl group, isobutyl group, n-pentyl group, n-hexyl group Examples include. Among these, an alkyl group having 1 to 4 carbon atoms is preferred, an alkyl group having 1 to 3 carbon atoms is more preferred, and a methyl group and an ethyl group are even more preferred.
As for the combination of R 1 and R 2 , when R 1 and R 2 are hydrogen atoms, it is more preferable when R 1 and R 2 are alkyl groups having 1 to 6 carbon atoms. When R 1 and R 2 are hydrogen atoms, it is more preferable that R 1 and R 2 are alkyl groups having 1 to 4 carbon atoms. When R 1 and R 2 are hydrogen atoms, it is even more preferable that R 1 and R 2 are alkyl groups having 1 to 3 carbon atoms. When R 1 and R 2 are hydrogen atoms, it is even more preferable that R 1 and R 2 are alkyl groups having 1 to 2 carbon atoms.
式（１）で表される化合物の塩としては、塩酸塩、硫酸塩、硝酸塩などの鉱酸塩、酢酸塩、シュウ酸塩、コハク酸塩などの有機酸塩が挙げられる。また、式（１）で表される化合物には、種々の異性体が存在する場合があり、それらの異性体、異性体の混合物が含まれる。さらに、式（１）で表される化合物又はその塩の水和物も含まれる。 Examples of the salt of the compound represented by formula (1) include mineral acid salts such as hydrochloride, sulfate, and nitrate, and organic acid salts such as acetate, oxalate, and succinate. Further, the compound represented by formula (1) may exist in various isomers, and includes such isomers and mixtures of isomers. Furthermore, hydrates of the compound represented by formula (1) or a salt thereof are also included.
式（１）の化合物又はその塩は、既に知られている化合物であり、それ自体公知の方法により製造することもできるし、市販品を購入することもできる。 The compound of formula (1) or a salt thereof is a known compound, and can be produced by a method known per se, or can be purchased commercially.
式（１）の化合物は、後記実施例に示すように、トポイソメラーゼＩＩα特異的阻害剤である。従って、多様な癌細胞に対してＤＮＡ二本鎖切断（ＤＳＢ）を形成し、広範囲の癌に対して優れた抗癌作用を示す。また、トポイソメラーゼＩＩα特異的阻害剤であるから、ドキソルビシンのような心毒性を示さず、安全性も高いものと考えられる。さらに、多くの抗癌剤に対して耐性を誘発する、薬剤排出Ｐ糖タンパク陽性癌細胞株に対して増殖抑制効果を示すことから、薬剤耐性の癌に対しても有効である。 The compound of formula (1) is a topoisomerase IIα-specific inhibitor, as shown in the Examples below. Therefore, it forms DNA double-strand breaks (DSB) in various cancer cells and exhibits excellent anticancer effects against a wide range of cancers. Furthermore, since it is a topoisomerase IIα-specific inhibitor, it does not exhibit cardiotoxicity like doxorubicin and is considered to be highly safe. Furthermore, it is effective against drug-resistant cancers as it exhibits a growth-suppressing effect on drug-extracting P-glycoprotein-positive cancer cell lines that induce resistance to many anticancer drugs.
本発明の悪性腫瘍治療剤の対象癌としては、例えば頭頚部癌、食道癌、胃癌、結腸癌、直腸癌、肝臓癌、胆嚢・胆管癌、胆道癌、膵臓癌、肺癌、乳癌、卵巣癌、子宮頚癌、子宮体癌、腎癌、膀胱癌、前立腺癌、精巣腫瘍、骨・軟部肉腫、白血病、悪性リンパ腫、多発性骨髄腫、皮膚癌、脳腫瘍、中皮腫等が挙げられる。 Target cancers for the malignant tumor therapeutic agent of the present invention include, for example, head and neck cancer, esophageal cancer, gastric cancer, colon cancer, rectal cancer, liver cancer, gallbladder/cholangiocarcinoma, biliary tract cancer, pancreatic cancer, lung cancer, breast cancer, ovarian cancer, Examples include cervical cancer, uterine body cancer, renal cancer, bladder cancer, prostate cancer, testicular cancer, bone/soft tissue sarcoma, leukemia, malignant lymphoma, multiple myeloma, skin cancer, brain tumor, mesothelioma, and the like.
式（１）の化合物又はその塩を医薬として用いるにあたっては、必要に応じて薬学的担体と配合し、治療目的に応じて各種の投与形態（医薬組成物）を採用可能であり、該形態としては、例えば、経口剤、注射剤、坐剤、軟膏剤、貼付剤等のいずれでもよい。これらの投与形態は、各々当業者に公知慣用の製剤方法により製造できる。 When using the compound of formula (1) or a salt thereof as a medicine, it is possible to mix it with a pharmaceutical carrier as necessary and adopt various dosage forms (pharmaceutical compositions) depending on the therapeutic purpose. may be, for example, an oral preparation, an injection, a suppository, an ointment, a patch, etc. Each of these dosage forms can be manufactured by conventional formulation methods known to those skilled in the art.
薬学的担体としては、製剤素材として慣用の各種有機或いは無機担体物質が用いられ、固形製剤における賦形剤、結合剤、崩壊剤、滑沢剤、着色剤、液状製剤における溶剤、溶解補助剤、懸濁化剤、等張化剤、緩衝剤、無痛化剤等として配合される。また、必要に応じて防腐剤、抗酸化剤、着色剤、甘味剤、安定化剤等の製剤添加物を用いることもできる。
経口用固形製剤を調製する場合は、式（１）の化合物に賦形剤、必要に応じて賦形剤、結合剤、崩壊剤、滑沢剤、着色剤、矯味・矯臭剤等を加えた後、常法により錠剤、被覆錠剤、顆粒剤、散剤、カプセル剤等を製造することができる。
注射剤を調製する場合は、式（１）の化合物にｐＨ調節剤、緩衝剤、安定化剤、等張化剤、局所麻酔剤等を添加し、常法により皮下、筋肉内及び静脈内用注射剤を製造することができる。
上記の各投与単位形態中に配合されるべき式（１）化合物の量は、これを適用すべき患者の症状により、或いはその剤形等により一定ではないが、一般に投与単位形態あたり、経口剤では約０．０５～１０００ｍｇ、注射剤では約０．０１～５００ｍｇ、坐剤では約１～１０００ｍｇとするのが望ましい。
また、上記投与形態を有する薬剤の１日あたりの投与量は、患者の症状、体重、年齢、性別等によって異なり一概には決定できないが、通常成人（体重５０ｋｇ）１日あたり約０．０５～５０００ｍｇ、好ましくは０．１～１０００ｍｇとすればよく、これを１日１回又は２～３回程度に分けて投与するのが好ましい。
As pharmaceutical carriers, various organic or inorganic carrier substances commonly used as formulation materials are used, including excipients, binders, disintegrants, lubricants, and colorants in solid formulations, solvents and solubilizing agents in liquid formulations, It is blended as a suspending agent, tonicity agent, buffering agent, soothing agent, etc. Further, formulation additives such as preservatives, antioxidants, coloring agents, sweeteners, and stabilizers can also be used as necessary.
When preparing a solid preparation for oral use, an excipient is added to the compound of formula (1), and if necessary, an excipient, a binder, a disintegrant, a lubricant, a coloring agent, a flavoring agent, etc. Thereafter, tablets, coated tablets, granules, powders, capsules, etc. can be manufactured by conventional methods.
When preparing an injection, add a pH adjuster, a buffer, a stabilizer, an isotonic agent, a local anesthetic, etc. to the compound of formula (1), and prepare it for subcutaneous, intramuscular, or intravenous administration using a conventional method. Injections can be manufactured.
The amount of the compound of formula (1) to be incorporated into each of the above dosage unit forms is not constant depending on the symptoms of the patient to whom it is applied or the dosage form, but generally per dosage unit form, the amount of the compound of formula (1) For example, the amount is preferably about 0.05 to 1000 mg, for injections it is about 0.01 to 500 mg, and for suppositories it is about 1 to 1000 mg.
In addition, the daily dosage of the drug having the above dosage form varies depending on the patient's symptoms, weight, age, gender, etc., and cannot be determined unconditionally, but it is usually about 0.05 to 0.05 per day for an adult (weight 50 kg). The amount may be 5000 mg, preferably 0.1 to 1000 mg, and it is preferable to administer this once a day or in divided doses about 2 to 3 times a day.
次に実施例を挙げて、本発明をさらに詳細に説明するが、本発明はこれら実施例に限定されるものではない。 EXAMPLES Next, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited to these Examples.
実施例１
（方法）
ＤＳＢの検出には、ＤＳＢが形成された際に、損傷周辺箇所で起こるヒストンＨ２ＡＸのＳｅｒ１３９のリン酸化であるγＨ２ＡＸをフローサイトメトリー法により検出した。すなわち、接着細胞に化合物Ａを２４時間処理後、細胞をトリプシンにより剥離、洗浄後、γＨ２ＡＸ抗体混合の染色バッファーにより４℃条件でオーバーナイト染色した。フローサイトメーターにより細胞内のγＨ２ＡＸ量を測定、解析した。
Example 1
(Method)
To detect DSBs, γH2AX, which is phosphorylation of Ser139 of histone H2AX, which occurs in the vicinity of damage when DSBs are formed, was detected by flow cytometry. That is, after treating adherent cells with Compound A for 24 hours, the cells were detached with trypsin, washed, and then stained overnight at 4° C. with a staining buffer containing γH2AX antibody. The amount of γH2AX in the cells was measured and analyzed using a flow cytometer.
（結果）
化合物Ａ（式（１）中、Ｘ１，Ｘ２＝Ｃｌ、Ｒ１，Ｒ２＝Ｈ）のＤＮＡ二本鎖切断（ＤＳＢ）形成作用を図１に示す。
図１より、化合物Ａは、ＨｅＬａ細胞、Ａ５４９細胞及びＤＬＤ－１細胞のいずれの癌細胞に対して濃度依存的にＤＳＢ形成作用を有することがわかる。
(result)
FIG. 1 shows the DNA double-strand break (DSB) forming effect of compound A (in formula (1), X 1 , X 2 =Cl, R 1 , R 2 =H).
From FIG. 1, it can be seen that Compound A has a DSB-forming effect on all cancer cells, HeLa cells, A549 cells, and DLD-1 cells, in a concentration-dependent manner.
実施例２
（方法）
細胞増殖解析には、代謝活性の指標であるＡＴＰを測定するＣｅｌｌＴｉｔｅｒ－Ｇｌｏアッセイを用いた。９６ウェルプレートに播種した細胞に化合物Ａを５日間処理した。ＣｅｌｌＴｉｔｅｒ－ＧｌｏＴＭ ２．０溶液を細胞に混合、プレートリーダーにより発光を検出、用量反応曲線を解析した。
細胞周期解析は、以下の方法により実施した。細胞に化合物Ａを２４時間処理、トリプシンにより細胞を剥離、洗浄後、７０％エタノールで細胞固定した。さらに洗浄後、ＲＮＡｓｅを処理、Ｐｒｏｐｉｄｉｕｍ Ｉｏｄｉｄｅ（ＰＩ；ヨウ化プロピジウム）により染色した。フローサイトメーターにより細胞内のＰＩ量の測定、Ｇ１、Ｓ、Ｇ２／Ｍ細胞周期分布を解析した。
Example 2
(Method)
Cell Titer-Glo assay, which measures ATP, which is an index of metabolic activity, was used for cell proliferation analysis. Cells seeded in 96-well plates were treated with compound A for 5 days. CellTiter-Glo ™ 2.0 solution was mixed with cells, luminescence was detected using a plate reader, and a dose-response curve was analyzed.
Cell cycle analysis was performed by the following method. Cells were treated with Compound A for 24 hours, detached with trypsin, washed, and then fixed with 70% ethanol. After further washing, the cells were treated with RNAse and stained with propidium iodide (PI). The intracellular PI amount was measured using a flow cytometer, and the G1, S, and G2/M cell cycle distributions were analyzed.
（結果）
化合物Ａの癌細胞増殖抑制作用を図２に示す。図２より、化合物Ａは、ＨｅＬａ細胞、Ａ５４９細胞及びＤＬＤ－１細胞のいずれの癌細胞に対しても強い増殖抑制作用を有することがわかる。
化合物Ａの細胞周期に対する分布状態を示す。図３より、化合物Ａは、ＨｅＬａ細胞、Ａ５４９細胞及びＤＬＤ－１細胞においてＤＳＢの応答として起こるＧ２／Ｍ細胞周期アレストを引き起こすことがわかる。
(result)
The cancer cell proliferation inhibitory effect of Compound A is shown in FIG. 2. From FIG. 2, it can be seen that Compound A has a strong growth-inhibiting effect on all cancer cells, HeLa cells, A549 cells, and DLD-1 cells.
The distribution state of compound A with respect to the cell cycle is shown. FIG. 3 shows that Compound A causes G2/M cell cycle arrest that occurs in response to DSB in HeLa cells, A549 cells, and DLD-1 cells.
実施例３
ＪＦＣＲ３９細胞パネル試験
（方法）
３９種類のがん細胞株から構成される細胞パネルＪＦＣＲ３９に対する化合物Ａの５０％細胞増殖阻害濃度（ＧＩ５０）を解析、細胞増殖阻害のスペクトル（フィンガープリント）を解析した。その後、化合物Ａのフィンガープリントを、２５０種類の標準物質のフィンガープリントデータベースと比較解析（ＣＯＭＰＡＲＥ解析）することで、作用機序の予測評価を実施した。
Example 3
JFCR39 cell panel test (method)
The 50% cell growth inhibition concentration (GI50) of Compound A against JFCR39, a cell panel composed of 39 types of cancer cell lines, was analyzed, and the spectrum (fingerprint) of cell growth inhibition was analyzed. Thereafter, a predictive evaluation of the mechanism of action was performed by performing a comparative analysis (COMPARE analysis) of the fingerprint of Compound A with a fingerprint database of 250 types of standard substances.
ＪＦＣＲ３９細胞パネル試験は、以下の文献に従った。
Ｙａｍｏｒｉ Ｔ． Ｐａｎｅｌ ｏｆ ｈｕｍａｎ ｃａｎｃｅｒ ｃｅｌｌ ｌｉｎｅｓ ｐｒｏｖｉｄｅｓ ｖａｌｕａｂｌｅ ｄａｔａｂａｓｅ ｆｏｒ ｄｒｕｇ ｄｉｓｃｏｖｅｒｙ ａｎｄ ｂｉｏｉｎｆｏｒｍａｔｉｃｓ． Ｃａｎｃｅｒ Ｃｈｅｍｏｔｈｅｒ Ｐｈａｒｍａｃｏｌ ２００３；５２ Ｓｕｐｐｌ １：Ｓ７４－９．
ｈｔｔｐｓ：／／ｈｔｔｐｓ：／／ｄｏｉ．ｏｒｇ／１０．１００７／ｓ００２８０－００３－０６４９－１
ＣＯＭＰＡＲＥ解析は、以下の文献に従った。
Ｐａｕｌｌ ＫＤ， Ｓｈｏｅｍａｋｅｒ ＲＨ， Ｈｏｄｅｓ Ｌ， ｅｔ ａｌ． Ｄｉｓｐｌａｙ ａｎｄ ａｎａｌｙｓｉｓ ｏｆ ｐａｔｔｅｒｎｓ ｏｆ ｄｉｆｆｅｒｅｎｔｉａｌ ａｃｔｉｖｉｔｙ ｏｆ ｄｒｕｇｓ ａｇａｉｎｓｔ ｈｕｍａｎ ｔｕｍｏｒ ｃｅｌｌ ｌｉｎｅｓ： ｄｅｖｅｌｏｐｍｅｎｔ ｏｆ ｍｅａｎ ｇｒａｐｈ ａｎｄ ＣＯＭＰＡＲＥ ａｌｇｏｒｉｔｈｍ． Ｊ Ｎａｔｌ Ｃａｎｃｅｒ Ｉｎｓｔ １９８９；８１：１０８８－９２．
ｈｔｔｐｓ：／／ｄｏｉ．ｏｒｇ／１０．１０９３／ｊｎｃｉ／８１．１４．１０８８
The JFCR39 cell panel test was conducted according to the following literature.
Yamori T. Panel of human cancer cell lines provides valuable database for drug discovery and bioinformatics. Cancer Chemother Pharmacol 2003;52 Suppl 1:S74-9.
https://https://doi. org/10.1007/s00280-003-0649-1
COMPARE analysis followed the following literature.
Paul KD, Shoemaker RH, Hodes L, et al. Display and analysis of patterns of differential activity of drugs against human tumor cell lines: development of mean graft h and COMPARE algorithm. J Natl Cancer Inst 1989;81:1088-92.
https://doi. org/10.1093/jnci/81.14.1088
（結果）
各癌細胞株に対する化合物Ａのフィンガープリント（ＧＩ５０）を図４に示す。
図４中、Ｂｒｅｓｔは、乳癌細胞株に対する感受性を示す。ＣＮＳは、中枢神経系癌細胞株に対する感受性を示す。Ｃｏｌｏｎは、大腸癌細胞株に対する感受性を示す。Ｌｕｎｇは、肺癌細胞株に対する感受性を示す。Ｍｅｌａｎｏｍａは、メラノーマ細胞株に対する感受性を示す。Ｏｖａｒｉａｎは、卵巣癌細胞株に対する感受性を示す。Ｒｅｎａｌは、腎臓癌細胞株に対する感受性を示す。Ｓｔｏｍａｃｈは、胃癌細胞株に対する感受性を示す。Ｐｒｏｓｔａｔｅは、前立腺癌細胞株に対する感受性を示す。ＭＧ－ＭＩＤは、ＬｏｇＧＩ５０の平均値を示す。ＤｅｌｔａはＭＧ－ＭＩＤと最も感受性の高い細胞株のＬｏｇＧＩ５０の差を示す。Ｒａｎｇｅは、最も感受性の低い細胞株と感受性の高い細胞株のＬｏｇＧＩ５０の差を示す。
図４から、化合物Ａは、ほとんどすべての癌細胞に対して類似した濃度領域で細胞毒性を引き起こすことがわかる。また、化合物Ａは、薬剤排出Ｐ糖タンパク陽性株ＨＣＴ－１５（基質薬物：エトポシド、ドキソルビシン等）に対しても他の細胞株と同等の毒性作用を示し、薬剤耐性癌に対しても有効であることが示唆された。
化合物Ａに対する標準化合物とのＣＯＭＰＡＲＥ解析の結果を図５に示す。
図５から、化合物Ａは、トポイソメラーゼＩＩ阻害剤である可能性が示唆された。
(result)
The fingerprint (GI50) of Compound A for each cancer cell line is shown in FIG. 4.
In FIG. 4, Brest indicates sensitivity to breast cancer cell lines. The CNS exhibits susceptibility to central nervous system cancer cell lines. Colon shows sensitivity to colon cancer cell lines. Lung shows sensitivity to lung cancer cell lines. Melanoma exhibits susceptibility to melanoma cell lines. Ovarian shows sensitivity to ovarian cancer cell lines. Renal shows sensitivity to kidney cancer cell lines. Stomach shows sensitivity to gastric cancer cell lines. Prostate exhibits sensitivity to prostate cancer cell lines. MG-MID indicates the average value of LogGI50. Delta indicates the difference in LogGI50 between MG-MID and the most sensitive cell line. Range indicates the difference in LogGI50 between the least sensitive and most sensitive cell lines.
From FIG. 4, it can be seen that Compound A causes cytotoxicity to almost all cancer cells in a similar concentration range. Compound A also shows the same toxic effect on drug-extracting P-glycoprotein-positive cell line HCT-15 (substrate drugs: etoposide, doxorubicin, etc.) as other cell lines, and is also effective against drug-resistant cancers. Something was suggested.
The results of COMPARE analysis of Compound A with standard compounds are shown in FIG.
FIG. 5 suggested that Compound A may be a topoisomerase II inhibitor.
実施例４
（方法）
化合物ＡがトポイソメラーゼＩＩ阻害剤である可能性を検証するため、トポイソメラーゼＩＩ阻害剤との競合阻害実験とトポイソメラーゼＩＩの機能欠失実験による機能解析を実施した。
競合阻害実験は、以下の方法により実施した。エトポシドと作用機序の異なるトポイソメラーゼＩＩ阻害剤のＩＣＲＦ－１９３処理１時間後、１０μＭのエトポシドもしくは化合物Ａを添加し、さらに２４時間処理した。その後、ＤＳＢを実施例１と同様の方法により検出した。
機能欠失実験は、はじめにＴｏｐ２α（ＴＯＰ２Ａの遺伝子産物）及びＴｏｐ２β（ＴＯＰ２Ｂの遺伝子産物）を標的としたｓｉＲＮＡによる遺伝子ノックダウン効率をウエスタンブロッティングにより解析した。その後、ノックダウン下において、１０μＭの化合物ＡによるＤＳＢ形成を解析した。
Example 4
(Method)
In order to verify the possibility that Compound A is a topoisomerase II inhibitor, functional analysis was performed using a competitive inhibition experiment with a topoisomerase II inhibitor and a topoisomerase II functional deletion experiment.
Competitive inhibition experiments were conducted using the following method. One hour after treatment with ICRF-193, a topoisomerase II inhibitor with a different mechanism of action than etoposide, 10 μM etoposide or compound A was added and the treatment was continued for an additional 24 hours. Thereafter, DSB was detected by the same method as in Example 1.
In the functional deletion experiment, first, the efficiency of gene knockdown by siRNA targeting Top2α (TOP2A gene product) and Top2β (TOP2B gene product) was analyzed by Western blotting. Thereafter, DSB formation by 10 μM Compound A was analyzed under knockdown.
（結果）
化合物ＡとトポイソメラーゼＩＩ阻害剤（ＩＣＲＦ－１９３）の競合阻害実験の結果を図６に示す。単剤処理と比べ、ＩＣＲＦ－１９３との併用時では、化合物Ａにより形成されるＤＳＢ量は低下することを示した。この効果はエトポシドと同様であった。つまり、化合物ＡはトポイソメラーゼＩＩを標的としていることが示唆された。
図７に、ｓｉＲＮＡによるノックダウン効率の結果を示す。ｓｉＲＮＡにより、Ｔｏｐ２α及びＴｏｐ２βの発現量は有意に低下していることが示された。
図８に、トポイソメラーゼＩＩに対する機能欠失実験の結果を示す。Ｔｏｐ２α及びＴｏｐ２βのノックダウン下における化合物ＡのＤＳＢ形成作用を調べたところ、Ｔｏｐ２αのノックダウン下で、顕著に当該作用が抑制されることを示した。一方で、Ｔｏｐ２βに対してはそのような作用は見られなかった。したがって、化合物Ａは、Ｔｏｐ２αの特異的な阻害剤であることが示唆された。
(result)
The results of a competitive inhibition experiment between Compound A and topoisomerase II inhibitor (ICRF-193) are shown in FIG. It was shown that the amount of DSB formed by Compound A was reduced when used in combination with ICRF-193 compared to single agent treatment. This effect was similar to etoposide. In other words, it was suggested that Compound A targets topoisomerase II.
FIG. 7 shows the results of knockdown efficiency by siRNA. It was shown that the expression levels of Top2α and Top2β were significantly reduced by siRNA.
FIG. 8 shows the results of a functional deletion experiment for topoisomerase II. When the DSB-forming effect of Compound A was investigated under knockdown of Top2α and Top2β, it was shown that the effect was significantly suppressed under knockdown of Top2α. On the other hand, no such effect was observed on Top2β. Therefore, Compound A was suggested to be a specific inhibitor of Top2α.
実施例５
実施例１と同様にして、化合物Ａと類似の化合物について、ＤＳＢ形成作用を検討した。
その結果を、図９に示す。図９より、化合物Ａと類似の化合物Ｂ（式（１）中、Ｘ１，Ｘ２＝Ｃｌ、Ｒ１，Ｒ２＝Ｃ２Ｈ５）も、強力なＤＳＢ形成作用を示した。なお、化合物Ｃ（式（１）中、Ｘ１＝Ｃｌ、Ｘ２＝Ｈ、Ｒ１，Ｒ２＝Ｃ２Ｈ５）も、ＤＳＢ形成作用を示した。
Example 5
In the same manner as in Example 1, compounds similar to Compound A were examined for their DSB-forming effects.
The results are shown in FIG. From FIG. 9, Compound B (in formula (1), X 1 , X 2 =Cl, R 1 , R 2 =C 2 H 5 ), which is similar to Compound A, also showed a strong DSB-forming effect. In addition, compound C (in formula (1), X 1 =Cl, X 2 =H, R 1 , R 2 =C 2 H 5 ) also showed a DSB-forming effect.
実施例６
Ｉｎ ｖｉｖｏ
（方法）
腫瘍移植モデルとして、ＢＡＬＢ／ｃヌードマウスの下肢部皮下に、ＤＬＤ－１細胞を移植した。腫瘍体積が約３００ｍｍ３に達した個体に対し、平日の週５日間連続で、化合物Ａ（５０ｍｇ／ｋｇ）もしくはエトポシド（２０ｍｇ／ｋｇ）を腹腔内投与した。マウスの体重及び腫瘍体積は、週３回（１日もしくは２日おき）、測定した。観察終了後、処死したマウスから腫瘍を摘出し、腫瘍重量を測定した。
Example 6
In vivo
(Method)
As a tumor transplant model, DLD-1 cells were subcutaneously transplanted into the lower limbs of BALB/c nude mice. Compound A (50 mg/kg) or etoposide (20 mg/kg) was administered intraperitoneally to individuals whose tumor volume reached approximately 300 mm 3 for 5 consecutive weekdays. The body weight and tumor volume of mice were measured three times a week (every 1 or 2 days). After the observation was completed, the tumor was removed from the sacrificed mouse and the weight of the tumor was measured.
（結果）
化合物Ａは、エトポシドと同等の体重推移を示した（図１０）。化合物Ａは、エトポシドと同等の腫瘍体積抑制効果を示した（図１１）。また、化合物Ａは、エトポシドよりも強い腫瘍重量抑制効果を示した（図１２）。
(result)
Compound A showed a weight change similar to that of etoposide (FIG. 10). Compound A showed a tumor volume suppressing effect equivalent to that of etoposide (FIG. 11). Compound A also showed a stronger tumor weight suppressing effect than etoposide (FIG. 12).
Claims (2)
で表されるプリン誘導体又はその塩を含有する悪性腫瘍治療剤。 The following general formula (1)
A therapeutic agent for malignant tumors containing a purine derivative represented by or a salt thereof.
で表されるプリン誘導体又はその塩を含有するトポイソメラーゼＩＩα阻害剤。
The following general formula (1)
A topoisomerase IIα inhibitor containing a purine derivative represented by or a salt thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2022144405A JP2024039787A (en) | 2022-09-12 | 2022-09-12 | Malignant tumor treatment agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2022144405A JP2024039787A (en) | 2022-09-12 | 2022-09-12 | Malignant tumor treatment agent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2024039787A true JP2024039787A (en) | 2024-03-25 |
Family
ID=90367197
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2022144405A Pending JP2024039787A (en) | 2022-09-12 | 2022-09-12 | Malignant tumor treatment agent |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2024039787A (en) |
-
2022
- 2022-09-12 JP JP2022144405A patent/JP2024039787A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10183035B2 (en) | Etoposide and prodrugs thereof for use in targeting cancer stem cells | |
| Lai et al. | A STAT inhibitor patent review: progress since 2011 | |
| TW201615196A (en) | The new cancer therapy indication of the cellcept | |
| CN104363913A (en) | CDK8/CDK19 selective inhibitors and their use in anti-metastatic and chemopreventative methods for cancer | |
| JP2008514726A (en) | Combination therapy with hedgehog inhibitor, radiation and chemotherapy | |
| KR20240014585A (en) | Preparation and composition for treatment of malignant tumors | |
| EP3213752B1 (en) | Composition for treating cancer stem cells | |
| Lu et al. | Up-regulation of hypoxia-inducible factor antisense as a novel approach to treat ovarian cancer | |
| US20220168333A1 (en) | Combination Treatment for Hematological Cancers | |
| KR20230016158A (en) | KRAS mutation specific inhibitor and composition for preventing or treating cancer comprising the same | |
| Lee et al. | A novel imidazopyridine analogue as a phosphatidylinositol 3-kinase inhibitor against human breast cancer | |
| JP2024023269A (en) | Antitumor agent and compounding agent | |
| KR20140044362A (en) | Composition for preventing and treating non-small cell lung cancer, containing pyrazino-triazine derivatives | |
| JP2024039787A (en) | Malignant tumor treatment agent | |
| Zhang et al. | Kaempferol 3‐O‐gentiobioside, an ALK5 inhibitor, affects the proliferation, migration, and invasion of tumor cells via blockade of the TGF‐β/ALK5/Smad signaling pathway | |
| WO2022188491A1 (en) | Tumor chemotherapy pharmaceutical composition | |
| KR102591642B1 (en) | Targets and their applications for drug treatment of tumor metastases | |
| CN109602734A (en) | For treating the Compounds and methods for of leukaemia | |
| US10758501B2 (en) | Use of histone acetyltransferase inhibitor amidoximes as anti-proliferative agents | |
| CN115996716A (en) | EIF4A inhibitor combinations | |
| CN113230249A (en) | Application of pseudolaric acid B in serving as or preparing Hedgehog signal path inhibitor | |
| US11648217B2 (en) | Use of histone acetyltransferase inhibitor amidoximes as anti-proliferative agents | |
| US10314831B2 (en) | RAC1 inhibitors of neurofibroma formation | |
| Wahi et al. | Targeting the epigenetic reader “BET” as a therapeutic strategy for cancer | |
| US11696904B2 (en) | Prodrug for therapeutic applications |