JPS63122698A - 7-hydroxyguanine derivative and production thereof - Google Patents
7-hydroxyguanine derivative and production thereofInfo
- Publication number
- JPS63122698A JPS63122698A JP27062486A JP27062486A JPS63122698A JP S63122698 A JPS63122698 A JP S63122698A JP 27062486 A JP27062486 A JP 27062486A JP 27062486 A JP27062486 A JP 27062486A JP S63122698 A JPS63122698 A JP S63122698A
- Authority
- JP
- Japan
- Prior art keywords
- hydroxyguanine
- structural formula
- salt
- compound
- tables
- 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.)
- Granted
Links
- PIIRLHSJVLBMJN-UHFFFAOYSA-N 2-amino-7-hydroxy-3h-purin-6-one Chemical class N1C(N)=NC(=O)C2=C1N=CN2O PIIRLHSJVLBMJN-UHFFFAOYSA-N 0.000 title claims abstract description 20
- 238000004519 manufacturing process Methods 0.000 title claims description 7
- 150000003839 salts Chemical class 0.000 claims abstract description 19
- 239000000758 substrate Substances 0.000 claims abstract description 10
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 3
- 102000018251 Hypoxanthine Phosphoribosyltransferase Human genes 0.000 claims description 9
- 108010091358 Hypoxanthine Phosphoribosyltransferase Proteins 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 9
- PQGCEDQWHSBAJP-TXICZTDVSA-N 5-O-phosphono-alpha-D-ribofuranosyl diphosphate Chemical compound O[C@H]1[C@@H](O)[C@@H](O[P@](O)(=O)OP(O)(O)=O)O[C@@H]1COP(O)(O)=O PQGCEDQWHSBAJP-TXICZTDVSA-N 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 abstract description 25
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 abstract description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 abstract description 2
- 239000007853 buffer solution Substances 0.000 abstract description 2
- 239000007810 chemical reaction solvent Substances 0.000 abstract description 2
- 238000002360 preparation method Methods 0.000 abstract description 2
- 239000001632 sodium acetate Substances 0.000 abstract description 2
- 235000017281 sodium acetate Nutrition 0.000 abstract description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 abstract 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 abstract 2
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 229940005657 pyrophosphoric acid Drugs 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 10
- 238000000034 method Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- -1 Amine salts Chemical class 0.000 description 4
- 238000000862 absorption spectrum Methods 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 229940125904 compound 1 Drugs 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- QUSNBJAOOMFDIB-UHFFFAOYSA-N Ethylamine Chemical compound CCN QUSNBJAOOMFDIB-UHFFFAOYSA-N 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- 241000187180 Streptomyces sp. Species 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 239000003463 adsorbent Substances 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 2
- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 description 1
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- PQGCEDQWHSBAJP-UHFFFAOYSA-N 5-phospho-d-ribose 1-diphosphate Chemical compound OC1C(O)C(OP(O)(=O)OP(O)(O)=O)OC1COP(O)(O)=O PQGCEDQWHSBAJP-UHFFFAOYSA-N 0.000 description 1
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 description 1
- 229910000013 Ammonium bicarbonate Inorganic materials 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000004254 Ammonium phosphate Substances 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000007093 Leukemia L1210 Diseases 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 150000001340 alkali metals Chemical class 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 150000001342 alkaline earth metals Chemical class 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 235000012538 ammonium bicarbonate Nutrition 0.000 description 1
- 239000001099 ammonium carbonate Substances 0.000 description 1
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 1
- 235000019289 ammonium phosphates Nutrition 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 229940121363 anti-inflammatory agent Drugs 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 238000002143 fast-atom bombardment mass spectrum Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 239000008297 liquid dosage form Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229960005181 morphine Drugs 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000007909 solid dosage form Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、構造式（Ｉ）で表される新規な７−ヒドロキ
ングアニン誘導体およびその造塩可能なものの塩、及び
その製造方法、並びにこれを有効成分とする抗腫瘍剤に
関するものである。Detailed Description of the Invention (Industrial Application Field) The present invention relates to a novel 7-hydroquinanine derivative represented by the structural formula (I), a salt of its salt-formable salt, and a method for producing the same. The present invention relates to an antitumor agent containing this as an active ingredient.
（発明が解決しようとする間圧点）
本発明者等は、下記構造式（ＩＩ）
で示される７−ヒドロキシグアニンを化学的あるいは酵
素的に修飾することによって、いくつかの誘導体を得る
ことができ、その薬理作用を広く試験した結果、構造式
（１）で示される化合物及びその塩が優れた抗腫瘍作用
を有することを見出し、本発明に到達した。(Pressure point to be solved by the invention) The present inventors have discovered that several derivatives can be obtained by chemically or enzymatically modifying 7-hydroxyguanine represented by the following structural formula (II). As a result of extensive testing of its pharmacological effects, it was discovered that the compound represented by structural formula (1) and its salts have excellent antitumor effects, leading to the present invention.
以下に本発明を詳細説明する。The present invention will be explained in detail below.
（問題点を解決するための手段及び作用効果）本発明に
よる新規化合物は、下記の構造式（１）で表わされる。(Means for Solving the Problems and Effects) The novel compound according to the present invention is represented by the following structural formula (1).
本発明による化合物は酸性物質であり、塩基と塩を作る
ことが可能であるが、本発明による化合物の塩としては
造塩可能な任意のものが対象になる。例えば、■アルカ
リ金属、アルカリ土類金属との塩、■アンモニウム塩、
■アミン塩、特にエチルアミン、ジメチルアミン、ピペ
リジン、モルフィリンなどとの塩があげられる。これら
の塩を抗ｊ」瘍剤として使用する場合には、生理的に許
容されるものを選ぶべきである。The compound according to the present invention is an acidic substance and can form a salt with a base, and the salt of the compound according to the present invention may be any substance capable of forming a salt. For example, ■Salts with alkali metals and alkaline earth metals, ■Ammonium salts,
■Amine salts, especially salts with ethylamine, dimethylamine, piperidine, morphine, etc. When using these salts as anti-inflammatory agents, physiologically acceptable salts should be selected.
本発明化合物の２ナトリウム塩の性状は次の通りである
。The properties of the disodium salt of the compound of the present invention are as follows.
融点（分解点） １３５〜１４０°Ｃ分子量 ４２
３
元素分析値
実験値 ０２３．５１ ＩＩ４．４０ ＮＬ８．４
８０４３．７５
ＣＩＱＩ■１２Ｎ５０９ＰＮａ２ ・５Ｈ２Ｑ として
の理論値（％）
Ｃ２３，４０ＩＩ４．８２ Ｎ１８．６５０４３．６
４
比旋光度 〔α）、−−４４，６°（ｃ ＝０．５、）
ｆ２０）本発明化合物の袋外吸収スペクトル、赤外部吸
収スペクトル（臭化カリウム錠剤）を夫々図１、図２に
示す。また本発明化合物はＦＡＢマススペクトルにおい
て、水素化分子イオン（（Ｍ＋Ｈ）＋）がｍ／Ｚ＝４２
４に観察された。また試料のナトリウムが水素と置換し
たイオンがｍ／Ｚ＝４０２゜３８０に観測されたｏ ｍ
／Ｚ＝４０２． ：ｄ ８．０のイオンについて高分
解能マススペクトルを行った結果、それぞれのイオンに
対して、組成式０１０Ｈ１４Ｎ５０ｇＰＮａ、Ｃ１□Ｉ
１１５Ｎ５０ｇＦ が、それぞれ１０．９ ｍｍμ、
１２．３ｍｍμの誤差で得られた。Melting point (decomposition point) 135-140°C Molecular weight 42
3 Elemental analysis value experimental value 023.51 II4.40 NL8.4
8043.75 CIQI■12N509PNa2 ・5H2Q Theoretical value (%) C23,40II4.82 N18.65043.6
4 Specific optical rotation [α), -44,6° (c = 0.5,)
f20) The extra-bag absorption spectrum and infrared absorption spectrum (potassium bromide tablet) of the compound of the present invention are shown in FIGS. 1 and 2, respectively. Furthermore, in the FAB mass spectrum of the compound of the present invention, the molecular hydrogen ion ((M+H)+) is m/Z=42
4 was observed. Also, an ion in which sodium in the sample was replaced with hydrogen was observed at m/Z = 402°380.
/Z=402. As a result of performing high-resolution mass spectra on ions with :d 8.0, each ion has a composition formula of 010H14N50gPNa, C1□I
115N50gF is 10.9 mmμ, respectively.
It was obtained with an error of 12.3 mmμ.
次に本発明の構造式（ｌｌに示される７−ヒドロキシグ
アニン誘導体の製法を説明する。Next, a method for producing the 7-hydroxyguanine derivative represented by the structural formula (ll) of the present invention will be explained.
本発明の化合物を製造するにあたっては、基質として構
造式（ＩＩ）で示される７−ヒドロキシグアニンと、構
造式且で示される５−ホスホリボース−１−ピロリン酸
に、ホスホリボシルトランスフェラーゼを作用させるこ
とにより、構造式（１）に示される７−ヒドロキシグア
ニン誘導体を得ることができる。In producing the compound of the present invention, phosphoribosyltransferase is allowed to act on 7-hydroxyguanine represented by the structural formula (II) and 5-phosphoribose-1-pyrophosphate represented by the structural formula (II) as substrates. Accordingly, a 7-hydroxyguanine derivative represented by structural formula (1) can be obtained.
本方法に基質として用いられる７−ヒドロキシグアニン
は、ストレプトミセス・スピーシズ（Ｓｔｒｅｐｔｏｍ
ｙｃｅｓ ｓｐ、）Ａ＝８４７微工研寄第５４１号（
ＦＥＲΔｔ、ＢＰ−５４１）の培養液から、特開昭６１
−１７８９８４号に記載された方法に従って調製できる
。また５−ホスホリボシル−１−ピロリン酸又はそれら
の塩としては市販品を用いることができる。7-Hydroxyguanine used as a substrate in this method is derived from Streptomyces sp.
yces sp,) A=847 Microtechnology Research Institute No. 541 (
From the culture solution of FERΔt, BP-541), JP-A-61
-178984. Moreover, commercially available products can be used as 5-phosphoribosyl-1-pyrophosphoric acid or salts thereof.
本発明に用いるヒポキサンチン・グアニン・ホスホリボ
シルトランスフェラーゼは、動物、微生物など広く生物
界から得られる精製ヒポキサンチン・グアニン・ホスホ
リボシルトランスフェラーゼ、粗ヒポキサンチン・グア
ニン・ホスホリボシルトランスフェラーゼ、ヒポキサン
チン・グアニン・ホスホリボシルトランスフェラーゼ含
有物などのヒポキサンチン・グアニン・ホスホリボシル
トランスフェラーゼ活性を有するものであり、例えばヒ
ポキサンチン−グアニンホスホリボシルトランスフェラ
ーゼ（米国シグマ社製、酵母由来）などが挙げられる。The hypoxanthine guanine phosphoribosyltransferase used in the present invention includes purified hypoxanthine guanine phosphoribosyltransferase, crude hypoxanthine guanine phosphoribosyltransferase, and hypoxanthine guanine phosphoribosyltransferase obtained from a wide range of biological sources such as animals and microorganisms. Those having hypoxanthine-guanine-phosphoribosyltransferase activity such as ribosyltransferase-containing substances, such as hypoxanthine-guanine phosphoribosyltransferase (manufactured by Sigma, USA, derived from yeast).
酵素反応は、基質の化合物Ｉ（７−ヒドロキシグアニン
）を０．０５〜０．２％（Ｗ／Ｖ ）、および化合物１
［（５−ホスホリボース−１−ピロリン酸）を０．０５
〜０．４％（Ｗ／Ｖ）の範囲で反応液に懸濁し、酵素を
適量、例えば酵素と基質との重量比１：２０乃至１：１
０００の割合で加え、温度１０〜６０’Ｃ，好ましくは
２５〜３５°Ｃの範囲で反応を行ない、高速液体クロマ
トグラフィー（ＨＰ Ｌ Ｃ）によって残基質（化合物
■）と生成物（化合物Ｉ）の量を測定し、生成物の増加
が止った時点で反応を止めれば良い。また、酵素反応を
行なう際の溶媒のｐＨ範囲は６〜９、好ましくは７゜５
〜８．５であり、これらの範囲でｐ Ｉ−ｆの調整を行
えばよい。In the enzymatic reaction, the substrate Compound I (7-hydroxyguanine) was added at 0.05 to 0.2% (W/V), and Compound 1
[0.05 (5-phosphoribose-1-pyrophosphate)
Suspend the enzyme in the reaction solution at a concentration of ~0.4% (W/V), and add an appropriate amount of enzyme, for example, a weight ratio of enzyme to substrate of 1:20 to 1:1.
The reaction was carried out at a temperature of 10 to 60°C, preferably 25 to 35°C, and the residue (compound ■) and product (compound I) were separated by high performance liquid chromatography (HPLC). The reaction can be stopped when the amount of the product stops increasing. In addition, the pH range of the solvent when carrying out the enzyme reaction is 6 to 9, preferably 7°5.
~8.5, and p I-f may be adjusted within these ranges.
反応溶媒としては緩衝液の使用が望ましく、トリス−塩
酸の如き無機酸塩の緩衝液、酢酸ナトリウム、クエン酸
ナトリウムの如き有機酸塩の緩衝液を適宜使用すること
ができる。濃度は緩衝液の種類にもよるが１０ｍＭ〜２
００ｍＭ、 好ましくは２０ｍＭ〜１００ ｍＭを使
用すると良い。It is desirable to use a buffer as the reaction solvent, and buffers of inorganic acid salts such as Tris-hydrochloric acid and buffers of organic acid salts such as sodium acetate and sodium citrate can be used as appropriate. The concentration depends on the type of buffer, but is 10mM to 2
00mM, preferably 20mM to 100mM.
反応液中から生成物を分離する方法としては、その水溶
性、酸性の性質を利用して行えば良い。The product can be separated from the reaction solution by utilizing its water-soluble and acidic properties.
反応液中の生成した化合物Ｉは、種々の吸着剤を用いて
採取することができる。吸着剤として、活性炭、弱塩基
性カチオン交換樹脂などを利用できる。そのほか、セフ
ァデックスＧ−１ｏ、 セファテックスＬＨ−２０（フ
ァルマシア製）などのゲル−適用担体によるカラムクロ
マトグラフィーも有用な精製手段である。Compound I produced in the reaction solution can be collected using various adsorbents. Activated carbon, weakly basic cation exchange resin, etc. can be used as the adsorbent. In addition, column chromatography using gel-applicable carriers such as Sephadex G-1o and Sephadex LH-20 (manufactured by Pharmacia) is also a useful purification method.
７−ヒドロキシグアニン誘導体（化合物ｌ）は上記の分
離法または精製法を適宜組合わせ、あるいは繰り返すこ
とによって純粋に採取できる。純粋な化合物１を得る方
法として、例えば水−アルコールなどの混合溶媒系から
結晶を析出させる方法が適している。The 7-hydroxyguanine derivative (compound 1) can be purified in a pure manner by appropriately combining or repeating the above separation or purification methods. A suitable method for obtaining pure Compound 1 is, for example, a method in which crystals are precipitated from a mixed solvent system such as water-alcohol.
次に本発明の抗腫瘍剤について説明する。Next, the antitumor agent of the present invention will be explained.
本発明による抗腫瘍剤は曲記の構造式（Ｉ）で示される
７−ヒドロキシグアニン誘導体またはその造塩可能なも
のの塩を有効成分とするものである。The antitumor agent according to the present invention contains as an active ingredient a 7-hydroxyguanine derivative represented by the following structural formula (I) or a salt thereof that can be salted.
本化合物の抗腫瘍作用は以下の試験例に示される通りで
ある。抗腫瘍作用は以下の如く試験した。The antitumor effect of this compound is as shown in the following test example. The antitumor effect was tested as follows.
ＢＤＦＩ系雌性マウス（体重１８〜２３ｆ）を用い１群
３匹とした。このマウスに、マウス白血病Ｌ−１２１０
１ＸＩ０５個を腹腔内に移植し、被験化合物を５０ｍＭ
燐酸緩衝液（１）Ｈ７，４） に溶解したものを、翌日
より連続５日間腹腔内投与して延命率を測定した。なお
、５０ｍＭのリン酸緩衝液を腹腔内投与したマウスを対
照とした。BDFI female mice (body weight 18-23f) were used, with three mice per group. This mouse was infected with murine leukemia L-1210.
1XI05 pieces were implanted intraperitoneally, and the test compound was added to 50mM.
A solution dissolved in phosphate buffer (1) H7,4) was administered intraperitoneally for 5 consecutive days starting from the next day, and the survival rate was measured. Note that mice to which 50 mM phosphate buffer was intraperitoneally administered were used as controls.
表１に示された結果から、本発明による化合物は優れた
抗腫瘍作用を有することが明らかである。From the results shown in Table 1, it is clear that the compounds according to the invention have excellent antitumor effects.
なお、ＩＣＣ茶系雄性マウス体重１８〜２３ダ）を用い
た急性毒性試験（腹腔内投与）でのＬＤ５０（’９７ｋ
ｑ）値は１５０〜３００ｍダ／に９であった。In addition, the LD50 ('97k
q) value was 9 in 150-300 mda/.
表 １
調剤および投与量
この発明の化合物及びその造塩可能なものの塩は、所望
の投与方法に応じて、例えば錠剤、火剤、カプセル剤、
液剤、けんだく剤などの固体または液体の投与形態にす
ることができる。投与にあたっては、この発明の化合物
及びそれらの造塩可能なものの塩に加えて、慣用される
医薬用担体または賦形剤を含み、さらに他の医薬を含む
こともできるが、その製剤中の主要部分でなくてもよい
ことはいうまでもない。Table 1 Preparations and Dosage The compounds of this invention and their saltizable salts may be prepared, for example, in tablets, gunpowders, capsules, etc., depending on the desired method of administration.
It can be in solid or liquid dosage forms such as solutions and suspensions. For administration, in addition to the compounds of this invention and their salt-formable salts, commonly used pharmaceutical carriers or excipients may be included, and other drugs may also be included; It goes without saying that it does not have to be a part.
本発明の化合物は、一般に所望の作用が副作用を伴なう
ことなく達成される投与量で決定される。Compounds of the invention are generally determined at dosages that will achieve the desired effect without side effects.
その具体的な値は医師の判断で決定されるべきであるが
、一般に成人１日当り１０〜５００〜投与されるのが普
通であろう。Although the specific value should be determined by a doctor's judgment, it is generally 10 to 500 doses per day for an adult.
（実施例）
次に本発明の化合物の製造例をあげて本発明を具体的に
説明するが、これら実施例は本発明を制限するものでは
ない。(Examples) Next, the present invention will be specifically explained with reference to production examples of the compounds of the present invention, but these Examples are not intended to limit the present invention.
基質（７−ヒドロキシグアニン）の製造倒木発明法に基
質として用いられる７−ヒドロキシグアニンは、ストレ
プトミセス・スピーシズ（Ｓｔｒｅｐｔｏｍｙｃｅｓ
ｓｐ、）Ａ−３４７微工研寄第５４１号（ＦＥＲ八ｉ
へＢＰ−５４１）の培養液から特開昭６１−１７８９８
４号に記載された方法に従って調製した。Production of Substrate (7-Hydroxyguanine) 7-Hydroxyguanine used as a substrate in the fallen tree invention method is produced by Streptomyces sp.
sp,) A-347 Microtechnical Laboratory No. 541 (FER8i
JP-A-61-17898 from the culture solution of BP-541)
It was prepared according to the method described in No. 4.
実施例１ 化合物１１（７−ヒドロキシグアニン）ｉｓｏ＊。Example 1 Compound 11 (7-hydroxyguanine) iso*.
５−ホスホ−α−Ｄ−リボースー１−ピロリン酸（米国
、シグマ社製）１．８ｆを２．５ ｍＭＭ化マグネシウ
ム及び１２．５ｍＭジチオスライトールを含む５０ｍＭ
トリス塩酸緩衝液（ｐＨ７，８） ８５０ｍｌに溶解
し、基質溶液とした。５０，０００ユニツトのヒポキサ
ンチン・グアニン・ホスホリボシルトランスフェラーゼ
（シグマ社製）を５０ｔｔｔｔの上記の緩衝液に溶解後
、これを基質溶液に加え、３０°Ｃで１４時間反応させ
た。5-phospho-α-D-ribose-1-pyrophosphate (manufactured by Sigma, USA) 1.8f in 2.5mM 50mM containing magnesium and 12.5mM dithiothreitol
It was dissolved in 850 ml of Tris-HCl buffer (pH 7, 8) to prepare a substrate solution. After dissolving 50,000 units of hypoxanthine guanine phosphoribosyltransferase (manufactured by Sigma) in 50tttt of the above buffer solution, this was added to the substrate solution and reacted at 30°C for 14 hours.
目的生成物を、ＤＥＡＥ−セファデックスＡ−２５（フ
ァルマシア製）炭酸型に吸着させた後、０．２Δ１の重
炭酸アンモニウムで溶出した。活性画分を５０°Ｃで減
圧濃縮し、８ｔｔｌの蒸留水に溶解してセファデックス
ＬＨ２０（スウェーデン、ファルマシア社製）カラムク
ロマトグラフィーに供し、２０ｍＭのアンモニア水で溶
出した。活性画分を４０’Ｃで減圧ａ縮し、２８０ｑの
７−ヒドロキシグアニン誘導体（１）粉末を得た。この
うちｌ ６０１’ｌ’；／を１．２ ｙｔｔの蒸留水に
溶解した後、ＩＭ水水酸死花ナトリウム溶液ｐ Ｉ−Ｉ
７．２とし、エタノール８．２６ ＊１を加えて生じ
た沈澱をＰ取、乾燥し、１２７ｑの７−ヒドロキシグア
ニン誘導体（化合物Ｉ）を？υた。生成物の確認は、以
下に示す条件の高速液体クロマトグラフィーで行なった
。The target product was adsorbed onto DEAE-Sephadex A-25 (manufactured by Pharmacia) carbonate type, and then eluted with 0.2Δ1 ammonium bicarbonate. The active fraction was concentrated under reduced pressure at 50°C, dissolved in 8 ttl of distilled water, and subjected to Sephadex LH20 (manufactured by Pharmacia, Sweden) column chromatography, and eluted with 20 mM aqueous ammonia. The active fraction was condensed under reduced pressure at 40'C to obtain 280q of 7-hydroxyguanine derivative (1) powder. After dissolving l 601'l';/ in 1.2 ytt of distilled water, IM sodium hydroxide dead flower solution p I-I
7.2, add ethanol 8.26*1, remove the resulting precipitate, dry it, and obtain 127q of 7-hydroxyguanine derivative (compound I). It was. The product was confirmed by high performance liquid chromatography under the conditions shown below.
システム：日本分光製トライローター４型カラム： Ｙ
ＭＣ−ｐａｃｋＡ−６１２６Ｘ１５０ｍ＋ｉ（山村化学
製）
溶 媒 ： ｏ、 ｉ ｈｔリン酸アンモニウムｐ Ｈ
４，０：メタノール（１９：１）
流速＝１．０肩ｌ／分
検出：ＵＶ２５４ｎｍ
上記条件で７−ヒドロキシグアニン誘導体（化合物Ｉ）
は保持時間８．９分、７−ヒドロキシグアニンは４．４
分にピークを示した。System: JASCO Tri-Rotor Type 4 Column: Y
MC-pack A-6126X150m+i (manufactured by Yamamura Chemical) Solvent: o, i ht ammonium phosphate pH
4,0: methanol (19:1) Flow rate = 1.0 l/min Detection: UV 254 nm 7-hydroxyguanine derivative (compound I) under the above conditions
has a retention time of 8.9 minutes, and 7-hydroxyguanine has a retention time of 4.4 minutes.
It showed a peak at 10 minutes.
図１は本発明化合物の紫外吸収スペクトルを示す図、図
２は同化合物の赤外吸収スペクトルを示す図である。FIG. 1 is a diagram showing the ultraviolet absorption spectrum of the compound of the present invention, and FIG. 2 is a diagram showing the infrared absorption spectrum of the same compound.
Claims (4)
シグアニン誘導体およびその塩。 ▲数式、化学式、表等があります▼（ I ）(1) A 7-hydroxyguanine derivative represented by the following structural formula (I) and a salt thereof. ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I)
造式（III） ▲数式、化学式、表等があります▼（III） で示される５−ホスホリボース−１−ピロリン酸を基質
とし、ヒポキサンチン・グアニン・ホスホリボシルトラ
ンスフェラーゼを作用させることを特徴とする、構造式
（ I ） ▲数式、化学式、表等があります▼（ I ） で表わされる７−ヒドロキシグアニン誘導体の製造方法
。(2) Structural formula (II) ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (II) 7-hydroxy guanine or its salt represented by and Structural formula (III) ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ ( III) Structural formula (I) characterized by using 5-phosphoribose-1-pyrophosphate as a substrate and allowing hypoxanthine guanine phosphoribosyltransferase to act ▲Mathematical formulas, chemical formulas, tables, etc. are available▼ A method for producing a 7-hydroxyguanine derivative represented by (I).
ンスフェラーゼが酵母由来である特許請求の範囲第２項
記載の製造方法。(3) The production method according to claim 2, wherein the hypoxanthine guanine phosphoribosyltransferase is derived from yeast.
グアニン誘導体またはその生理的に許容される塩を有効
成分とする抗腫瘍剤。 ▲数式、化学式、表等があります▼（ I ）(4) An antitumor agent containing a 7-hydroxyguanine derivative represented by the following structural formula (I) or a physiologically acceptable salt thereof as an active ingredient. ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP27062486A JPS63122698A (en) | 1986-11-13 | 1986-11-13 | 7-hydroxyguanine derivative and production thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP27062486A JPS63122698A (en) | 1986-11-13 | 1986-11-13 | 7-hydroxyguanine derivative and production thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63122698A true JPS63122698A (en) | 1988-05-26 |
| JPH0588720B2 JPH0588720B2 (en) | 1993-12-24 |
Family
ID=17488681
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP27062486A Granted JPS63122698A (en) | 1986-11-13 | 1986-11-13 | 7-hydroxyguanine derivative and production thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63122698A (en) |
-
1986
- 1986-11-13 JP JP27062486A patent/JPS63122698A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0588720B2 (en) | 1993-12-24 |
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