JP2005154342A - Topoisomerase inhibitor - Google Patents

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JP2005154342A
JP2005154342A JP2003395361A JP2003395361A JP2005154342A JP 2005154342 A JP2005154342 A JP 2005154342A JP 2003395361 A JP2003395361 A JP 2003395361A JP 2003395361 A JP2003395361 A JP 2003395361A JP 2005154342 A JP2005154342 A JP 2005154342A
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topoisomerase
benzenediamine
dna
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Keitaro Suzuki
啓太郎 鈴木
Tadashi Okawara
正 大川原
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Mercian Corp
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Abstract

<P>PROBLEM TO BE SOLVED: To provide a new topoisomerase inhibitor inhibiting an enzyme topoisomerase associated with replication of a DNA, and hardly causing hazard such as scission of a DNA chain. <P>SOLUTION: The topoisomerase inhibitor contains a benzenediamine derivative produced by reacting a benzoyl halide derivative with a benzenediamine, and optionally subjecting the product to deacetylation reaction as an active ingredient. The inhibitor is expected to be a medicament having little side effects because the inhibitor strongly inhibits topoisomerase I and II, and has a property of hardly acting on the DNA. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

本発明はトポイソメラーゼ阻害作用を有するベンゼンジアミン誘導体およびそれらを有効成分とするトポイソメラーゼ阻害剤に関する。 The present invention relates to benzenediamine derivatives having a topoisomerase inhibitory action and topoisomerase inhibitors containing these as active ingredients.

高等生物は細胞の形態を形づくるのに必要な細胞増殖の度合いを調節する能力を有しているが、がん化した細胞ではこの能力に異常な変化がおこっており、正常細胞に比べて急速な増殖をするのが特徴と考えられている。がん細胞では、正常細胞に比べて主に２つの異常、すなわち「細胞分裂の制御性の異常」と「がん細胞の転移」とが存在すると考えられている。正常細胞の分裂は細胞増殖抑制遺伝子により制御されており、必要なときにだけ分裂が誘発される。がん細胞は正常細胞の制御機構から逃れ増殖を繰り返す。つまり、１個のがん病巣は繰り返し遺伝的な変化を受けた単一細胞から発生し、分裂を繰り返し無数の同じように変化した異常な細胞を作り出す。正常細胞は、その個体の全体的な形態を維持するのに必要なルールに従って、それぞれの構成組織の中に限定されている。これに対して、がん細胞はそれが生じた組織にとどまることなく、増殖しながら他の組織へ転移していく。このような性質をもつがん細胞に変化してしまう原因としては、化学物質による遺伝子の損傷、放射線による遺伝子の損傷、ウイルスによる遺伝子の変異および先天的な遺伝子の欠損等が挙げられる。 Higher organisms have the ability to regulate the degree of cell growth necessary to shape cell morphology, but cancerous cells undergo abnormal changes in this ability and are more rapid than normal cells. It is thought to be a characteristic of proliferating. In cancer cells, it is considered that there are mainly two abnormalities as compared to normal cells, namely, “abnormalities in control of cell division” and “metastasis of cancer cells”. Normal cell division is controlled by cytostatic genes and is induced only when necessary. Cancer cells escape from the control mechanism of normal cells and repeat growth. In other words, a single cancer lesion arises from a single cell that has undergone repeated genetic changes, and produces numerous abnormal cells that have changed innumerably in a similar manner. Normal cells are confined within each constituent tissue according to the rules necessary to maintain the overall morphology of the individual. In contrast, cancer cells metastasize to other tissues while proliferating without staying in the tissues where they are produced. The causes of cancer cells having such properties include gene damage due to chemical substances, gene damage due to radiation, gene mutation due to viruses, and congenital gene defects.

がんを治療する戦略として、制がん剤を用いる方法は最も一般的であり、多くの薬剤が研究開発されてきた。中でも増殖細胞に対する抑制機能を指標とし開発された制がん剤としては、アルキル化剤、抗腫瘍性抗生物質、植物アルカロイドおよびキレート剤等が知られている。しかし、これらは殆どが遺伝子に作用するものであるため、選択毒性が低く、またその効果も必ずしも満足できるものではない。また、これらの制がん剤は遺伝子の変異を引き起こすこともあり、二次的ながんを誘発する可能性も高い等の問題がある。 As a strategy for treating cancer, a method using an anticancer drug is the most common, and many drugs have been researched and developed. Among them, alkylating agents, antitumor antibiotics, plant alkaloids, chelating agents, and the like are known as anticancer agents developed using the inhibitory function on proliferating cells as an index. However, since most of these act on genes, their selective toxicity is low and their effects are not always satisfactory. In addition, these anticancer agents may cause gene mutations and have a high possibility of inducing secondary cancer.

一方、がん細胞に関する分子生物学的研究の進展は、従来明らかでなかった制がん剤の作用機構に重大な手掛かりを与えている。中でも、エピポドフィロトキシン(epipodophyllotoxin)、ドキソルビシン(doxorubicin)、ストレプトニグリン(streptonigrin)等の制がん剤は、がん細胞内に多く存在する酵素であるトポイソメラーゼ(topoisomerase)の酵素活性を阻害することで、制がん効果を示すことが明らかになった(非特許文献１参照）。 On the other hand, the progress of molecular biological research on cancer cells has given a serious clue to the mechanism of action of anticancer drugs, which has not been clarified so far. Among them, antipodes such as epipodophyllotoxin, doxorubicin, and streptonigrin inhibit the enzyme activity of topoisomerase, an enzyme that is abundant in cancer cells. By doing so, it became clear that the anticancer effect was shown (refer nonpatent literature 1).

このトポイソメラーゼは、超らせん型DNAを一時的な切断と再結合により弛緩型DNAに変換する酵素であり、DNAの複製、転写、組み換え等の過程で生じる高次構造のひずみを解消し、DNA代謝をスムーズに進行させる機能を有している。トポイソメラーゼには、超らせん２本鎖DNAの１本鎖のみを切断し開環型DNAとした後、再結合して弛緩型DNAとするトポイソメラーゼI(topoisomerase I)と、２本鎖を同時に切断し直鎖DNAとした後、再結合して弛緩型DNAとするトポイソメラーゼII(topoisomerase II)の２種類がある。がん細胞のような増殖の激しい細胞には、正常細胞に比べてこのトポイソメラーゼが多く存在することが報告されている(非特許文献２参照)。従って、トポイソメラーゼの酵素活性を阻害する薬剤は、がん細胞の増殖を抑える有効な制がん剤の一つとなり得る。 This topoisomerase is an enzyme that converts supercoiled DNA into relaxed DNA by temporary cleavage and recombination, eliminating higher-order structural distortions that occur during DNA replication, transcription, recombination, etc. Has a function to smoothly progress. Topoisomerase cuts only one strand of superhelical double-stranded DNA into an open-ended DNA, and then recombines topoisomerase I (topoisomerase I) to form relaxed DNA and double-stranded DNA simultaneously. There are two types of topoisomerase II, which are linear DNA and then recombined to form relaxed DNA. It has been reported that cells with high proliferation such as cancer cells contain more topoisomerase than normal cells (see Non-Patent Document 2). Therefore, a drug that inhibits the enzyme activity of topoisomerase can be an effective anticancer drug that suppresses the growth of cancer cells.

そこで、トポイソメラーゼに作用する制がん剤の探索研究が行われ、カンプトテシン(camptothecin)、テルペンテシン(terpentecin)、セイントピン(saintopin)、ブルガレイン(bulgarein)等、制がん剤の新しいリード化合物となりうる酵素阻害剤が近年次々に報告されている(非特許文献３参照)。しかし、これらの阻害剤は遊離のトポイソメラーゼに結合して阻害するのではなく、酵素と切断されたDNAからなるトポイソメラーゼ−DNA切断複合体(cleavable complex)に結合してトポイソメラーゼの酵素活性を阻害するものであり、そのため、DNAに切れ目や切断等の障害を引き起こし強い毒性を示す。従って、制がん剤としては、トポイソメラーゼに直接作用しDNAには障害を残さない阻害剤、つまり副作用の少ない薬剤が要求されている。 Therefore, exploratory research on anticancer agents that act on topoisomerase was conducted, and enzymes such as camptothecin, terpentecin, saintopin, bulgarein, etc. could become new lead compounds of anticancer agents. Inhibitors have been reported one after another in recent years (see Non-Patent Document 3). However, these inhibitors do not bind to free topoisomerase and inhibit it, but bind to a topoisomerase-cleavable complex consisting of the enzyme and cleaved DNA to inhibit the enzyme activity of topoisomerase. Therefore, it causes damages such as cuts and breaks in DNA and exhibits strong toxicity. Therefore, as an anticancer agent, an inhibitor that directly acts on topoisomerase and does not cause damage to DNA, that is, a drug with few side effects is required.

一方、ある種のベンゼンジアミン誘導体は、がん細胞のテロメラーゼを阻害する性質を持つことが報告されている（非特許文献４参照）。しかし、トポイソメラーゼ阻害活性をもつことは知られていない。 On the other hand, certain benzenediamine derivatives have been reported to have the property of inhibiting telomerase in cancer cells (see Non-Patent Document 4). However, it is not known to have topoisomerase inhibitory activity.

Yamashita,Cancer Res.,50,5841〜5844(1990)Yamashita, Cancer Res., 50, 5841-5844 (1990) Spitzner,Nucleic,Acids,Res.,16,5533〜5556(1988)Spitzner, Nucleic, Acids, Res., 16, 5553-5556 (1988) Yamashita,Biochemistry,30,5838〜5845(1991)Yamashita, Biochemistry, 30, 5838-5845 (1991) Hiroyuki Seimiya et al.,Molecular Cancer Therapeutics,vol.1,657-665,July 2002Hiroyuki Seimiya et al., Molecular Cancer Therapeutics, vol. 1,657-665, July 2002

本発明は、直接トポイソメラーゼに作用しDNAに障害を残さないトポイソメラーゼ阻害物質およびそれらの阻害物質を含有するトポイソメラーゼ阻害剤を提供するものである。 The present invention provides topoisomerase inhibitors that directly act on topoisomerase and leave no damage to DNA, and topoisomerase inhibitors containing these inhibitors.

本発明者らは、上記課題を解決するため、指標酵素としてトポイソメラーゼIを用い、直接トポイソメラーゼに作用しDNAには障害を残さないトポイソメラーゼ阻害物質の探索を行ったところ、ある種のベンゼンジアミン誘導体が優れたトポイソメラーゼ阻害作用を有し、DNAとはほとんど相互作用を示さないことを見出し、本発明を完成した。 In order to solve the above problems, the present inventors conducted a search for a topoisomerase inhibitor that directly acts on topoisomerase and does not cause any damage to DNA using topoisomerase I as an indicator enzyme. The present invention was completed by finding that it has an excellent topoisomerase inhibitory action and shows little interaction with DNA.

すなわち本発明は、式(I)

Figure 2005154342
[式(I)中、Z1およびZ2は、式(II)で示される基 That is, the present invention relates to the formula (I)
Figure 2005154342
 [In the formula (I), Z 1 and Z 2 are groups represented by the formula (II)

Figure 2005154342
（式(II)中、R1、R2およびR3は水素原子、水酸基またはアセトキシ基を表す。但し、R1、R2およびR3が２つ以上水素原子である化合物は除く。）を表す。]で示されるベンゼンジアミン誘導体を有効成分とするトポイソメラーゼ阻害剤に関する。
Figure 2005154342
 (In the formula (II), R 1 , R 2 and R 3 represent a hydrogen atom, a hydroxyl group or an acetoxy group, except for compounds in which R 1 , R 2 and R 3 are two or more hydrogen atoms). Represent. ] The topoisomerase inhibitor which uses the benzenediamine derivative shown by this as an active ingredient.

式(I)における２つの置換基（式(II)で示される基）の配置は、オルト位、メタ位およびパラ位のいずれでもよい。また式(II)において、R1、R2およびR3の少なくとも２つの基が水酸基またはアセトキシ基である化合物は、優れたトポイソメラーゼ阻害活性を有する。しかし、R1、R2およびR3の少なくとも２つの基が水素原子である化合物は、トポイソメラーゼ阻害活性が弱いので、トポイソメラーゼ阻害剤としては適当でない。 The arrangement of the two substituents in the formula (I) (the group represented by the formula (II)) may be any of the ortho, meta and para positions. In the formula (II), a compound in which at least two groups of R 1 , R 2 and R 3 are a hydroxyl group or an acetoxy group has an excellent topoisomerase inhibitory activity. However, a compound in which at least two groups of R 1 , R 2 and R 3 are hydrogen atoms is not suitable as a topoisomerase inhibitor because it has a weak topoisomerase inhibitory activity.

また本発明は、式(I-a)

Figure 2005154342
[式(I-a)中、Z1およびZ2は、式(II-a)で示される基 The present invention also provides a compound of formula (Ia)
Figure 2005154342
 [In the formula (Ia), Z 1 and Z 2 are groups represented by the formula (II-a)

Figure 2005154342
（式(II-a)中、R1、R2およびR3は水素原子、水酸基またはアセトキシ基を表す。但し、R1、R2およびR3が２つ以上水素原子である化合物並びにR1およびR2が水酸基であり、R3が水素原子である化合物は除く。）を表す。]で示されるベンゼンジアミン誘導体に関する。これらの物質は、新規化合物であり優れたトポイソメラーゼ阻害活性を有する。
Figure 2005154342
 (In the formula (II-a), R 1 , R 2 and R 3 represent a hydrogen atom, a hydroxyl group or an acetoxy group, provided that R 1 , R 2 and R 3 are two or more hydrogen atoms, and R 1 And R 2 is a hydroxyl group and R 3 is a hydrogen atom. ] Is shown. These substances are novel compounds and have excellent topoisomerase inhibitory activity.

式(I)で示されるベンゼンジアミン誘導体は、以下の工程に従ってベンゾイルハライド誘導体(III)とベンゼンジアミン（IV）とを反応させ、さらに必要により脱アセチル化反応に付すことにより製造することができる。 The benzenediamine derivative represented by the formula (I) can be produced by reacting the benzoyl halide derivative (III) with benzenediamine (IV) according to the following steps and, if necessary, subjecting to a deacetylation reaction.

Figure 2005154342
上記反応式中、R1'、R2'およびR3'は水素原子またはアセトキシ基を、Xはハロゲン原子を表わす。
Figure 2005154342
 In the above reaction formula, R 1 ′, R 2 ′ and R 3 ′ represent a hydrogen atom or an acetoxy group, and X represents a halogen atom.

この反応は、ジクロロメタン等の不活性溶媒中に溶解したベンゼンジアミンに、トリエチルアミン等の有機塩基の存在下、式(III)で示されるベンゾイルハライド誘導体を徐々に添加してアミド化することにより、R1'、R2'およびR3'が水素原子またはアセトキシ基である式（V）で示されるベンゼンジアミン誘導体を得ることができる。さらにこれをジクロロメタン等の不活性溶媒中、ヒドラジンと反応させ、脱アセチル化することにより、R1'、R2'およびR3'が水素原子または水酸基である式（V）で示されるベンゼンジアミン誘導体を容易に得ることができる。なお、原料の一般式(III)で示されるベンゾイルハライド誘導体は、公知の方法（例えば、J.Med.Chem.,1989,32,2344等）に従って製造することができる。 In this reaction, benzenediamine dissolved in an inert solvent such as dichloromethane is amidated by gradually adding a benzoyl halide derivative represented by the formula (III) in the presence of an organic base such as triethylamine. A benzenediamine derivative represented by the formula (V) in which 1 ′, R 2 ′ and R 3 ′ are a hydrogen atom or an acetoxy group can be obtained. Furthermore, this is reacted with hydrazine in an inert solvent such as dichloromethane, and deacetylated, whereby benzenediamine represented by the formula (V) in which R 1 ′, R 2 ′ and R 3 ′ are hydrogen atoms or hydroxyl groups. Derivatives can be easily obtained. The benzoyl halide derivative represented by the general formula (III) as a raw material can be produced according to a known method (for example, J. Med. Chem., 1989, 32, 2344).

このようにして得られる本発明のトポイソメラーゼ阻害剤は、後述する実施例で示すとおり、トポイソメラーゼIに対して優れた阻害作用を示す。またトポイソメラーゼIIに対しても優れた阻害作用を示す。例えば、N,N'-Bis(3,4,5-trihydroxybenzoyl)-1,2-benzenediamineは、トポイソメラーゼIIに対する阻害作用IC50が２．６×１０-7Ｍであり、公知の阻害剤であるドキソルビシン（ＩＣ50：１×１０-6Ｍ）やエトポシド（ＩＣ50：８．７×１０-5Ｍ）よりも強い阻害活性を示す。また、その阻害形式は非拮抗阻害であり、DNA自体には全く作用しない性質を示した。また、本発明のトポイソメラーゼ阻害剤はある種の培養腫瘍細胞の増殖を阻害する（例えば、PDO-h3（後記例２参照）は、30μMの濃度でHeLa細胞の増殖を50％阻害する）が、細胞周期は変化させない性質を有する。 The topoisomerase inhibitor of the present invention thus obtained exhibits an excellent inhibitory action against topoisomerase I as shown in the examples described later. It also exhibits an excellent inhibitory effect on topoisomerase II. For example, N, N′-Bis (3,4,5-trihydroxybenzoyl) -1,2-benzenediamine is a known inhibitor having an inhibitory action IC 50 against topoisomerase II of 2.6 × 10 −7 M. It shows stronger inhibitory activity than doxorubicin (IC 50 : 1 × 10 −6 M) and etoposide (IC 50 : 8.7 × 10 −5 M). Moreover, the inhibition form was non-competitive inhibition, and showed the property that it did not act at all on DNA itself. Further, the topoisomerase inhibitor of the present invention inhibits the growth of certain cultured tumor cells (for example, PDO-h3 (see Example 2 below) inhibits the growth of HeLa cells by 50% at a concentration of 30 μM), The cell cycle has the property of not changing.

以上のとおり本発明のトポイソメラーゼ阻害剤は、直接トポイソメラーゼに作用し、DNAへの作用が少ない阻害剤であり、DNAに障害を与えないことから副作用の少ない薬剤となることが期待される。 As described above, the topoisomerase inhibitor of the present invention is an inhibitor that acts directly on topoisomerase and has little action on DNA, and since it does not damage DNA, it is expected to be a drug with few side effects.

以下、本発明について具体例を挙げてより詳細に説明するが、本発明をこれらの例に限定することを意図するものではない。なお、下記の例中のパーセント(％)は、特に断りのない限り、容量パーセントを示す。 Hereinafter, the present invention will be described in more detail with specific examples, but the present invention is not intended to be limited to these examples. In the following examples, the percentage (%) indicates a volume percentage unless otherwise specified.

例１
N,N'-Bis(3,4,5-triacetoxybenzoyl)-1,2-benzenediamine（以下、PDO-a3と略称するときがある）の製造
2mmolの1,2-ベンゼンジアミン(1,2-benzenediamine)を10mlのジクロロメタンに溶解し、これにトリエチルアミン6mmolを加えた。この溶液に氷冷下、J.Med.Chem.,1989,32,2344の記載に従って調製した3,4,5-triacetoxybenzoyl chloride 4mmolを少量ずつ加え、室温で２時間撹拌した。ジクロロメタン層を水洗して、無水硫酸マグネシウムで乾燥した後、ジクロロメタンを留去し、EtOAc/n-hexaneから再結晶した。融点 228 ℃，収量0.99g（収率75％） Example 1
Production of N, N'-Bis (3,4,5-triacetoxybenzoyl) -1,2-benzenediamine (hereinafter sometimes abbreviated as PDO-a3)
2 mmol of 1,2-benzenediamine was dissolved in 10 ml of dichloromethane, and 6 mmol of triethylamine was added thereto. Under ice-cooling, 4 mmol of 3,4,5-triacetoxybenzoyl chloride prepared according to the description of J. Med. Chem., 1989, 32, 2344 was added little by little, and the mixture was stirred at room temperature for 2 hours. The dichloromethane layer was washed with water and dried over anhydrous magnesium sulfate, and then the dichloromethane was distilled off and recrystallized from EtOAc / n-hexane. Melting point 228 ℃, Yield 0.99g (Yield 75%)

1H核磁気共鳴スペクトル(300MHz,CDCl3),ppm
2.26(12H,s,acetyl at 3',5',3'' and 5''),2.27(6H,s,acetyl at 4',4''),6.99(2H,q,J=3.6Hz,H 4,5),7.36(2H,q,J=3.6Hz,H 3,6),7.70(4H,s,H 2',6',2'',6''),9.64(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, CDCl 3 ), ppm
2.26 (12H, s, acetyl at 3 ', 5', 3 '' and 5``), 2.27 (6H, s, acetyl at 4 ', 4''), 6.99 (2H, q, J = 3.6Hz, H 4,5), 7.36 (2H, q, J = 3.6Hz, H 3,6), 7.70 (4H, s, H 2 ', 6', 2``, 6 ''), 9.64 (2H, s , 2 NH)

13C核磁気共鳴スペクトル(75MHz,CDCl3),ppm
20.07(CH3),20.45(CH3),120.61(CH),125.61(CH),126.67(CH),130.08(C),132.06(C),137.83(C),143.49(C),164.14(C),166.54(C),167.79(C) 13 C nuclear magnetic resonance spectrum (75 MHz, CDCl 3 ), ppm
20.07 (CH3), 20.45 (CH3), 120.61 (CH), 125.61 (CH), 126.67 (CH), 130.08 (C), 132.06 (C), 137.83 (C), 143.49 (C), 164.14 (C), 166.54 (C), 167.79 (C)

マススペクトル
FAB-MAS:m/z 665(M+1) Mass spectrum
FAB-MAS: m / z 665 (M + 1)

例２
N,N'-Bis(3,4,5-trihydroxybenzoyl)-1,2-benzenediamine（以下、PDO-h3と略称するときがある）の製造
例１で調製したPDO-a3 1mmolを10mlのジクロロメタンに溶解し、これにヒドラジン・１水和物7mmolを加え、室温で１時間撹拌した。ジクロロメタン層を水洗して、無水硫酸マグネシウムで乾燥した後、ジクロロメタンを留去し、これに適当量のメタノールを加えて再結晶した。融点 265 〜 266 ℃，収量0.50g（収率61％） Example 2
1 mmol of PDO-a3 prepared in Production Example 1 of N, N'-Bis (3,4,5-trihydroxybenzoyl) -1,2-benzenediamine (hereinafter sometimes abbreviated as PDO-h3) in 10 ml of dichloromethane After dissolution, 7 mmol of hydrazine monohydrate was added thereto and stirred at room temperature for 1 hour. The dichloromethane layer was washed with water and dried over anhydrous magnesium sulfate, the dichloromethane was distilled off, and an appropriate amount of methanol was added thereto for recrystallization. Melting point: 265-266 ° C, yield 0.50g (61% yield)

1H核磁気共鳴スペクトル(300MHz,CD3OD),ppm
6.87(4H,s,H 2',6',2'',6''),7.16-7.19(2H,q,J=3.6Hz,H 4,5),7.45-7.48(2H,q,J=3.6Hz,H 3,6) 1 H nuclear magnetic resonance spectrum (300 MHz, CD 3 OD), ppm
6.87 (4H, s, H 2 ', 6', 2``, 6 ''), 7.16-7.19 (2H, q, J = 3.6Hz, H 4,5), 7.45-7.48 (2H, q, J = 3.6Hz, H 3,6)

13C核磁気共鳴スペクトル(75MHz,CD3OD),ppm
108.34(CH),125.78(C),126.87(CH),127.18(CH),132.85(C),138.85(C),146.84(C),169.04(C) 13 C nuclear magnetic resonance spectrum (75 MHz, CD 3 OD), ppm
108.34 (CH), 125.78 (C), 126.87 (CH), 127.18 (CH), 132.85 (C), 138.85 (C), 146.84 (C), 169.04 (C)

マススペクトル
FAB-MAS:m/z 413(M+1) Mass spectrum
FAB-MAS: m / z 413 (M + 1)

例３
N,N'-Bis(3,4-diacetoxybenzoyl)-1,2-benzenediamine（以下、PDO-a2と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りに3,4-diacetoxybenzoyl chlorideを用いた以外は、例１と同様に行い、PDO-a2を製造した。
融点 206 〜 207 ℃，収量0.81g（収率74％） Example 3
Production of N, N'-Bis (3,4-diacetoxybenzoyl) -1,2-benzenediamine (hereinafter sometimes abbreviated as PDO-a2)
PDO-a2 was produced in the same manner as in Example 1 except that 3,4-diacetoxybenzoyl chloride was used instead of 3,4,5-triacetoxybenzoyl chloride.
Melting point 206-207 ° C, Yield 0.81g (Yield 74%)

1H核磁気共鳴スペクトル(300MHz,CDCl3),ppm
2.31(6H,s,acetyl at 3' and 3''),2.33(6H,s,acetyl at 4',4''),6.87(2H,q,J=3.6Hz,H 4,5),7.25-7.34(4H,m,H 3,6,5',5''),7.83(2H,d,J=2.1Hz,H 6',6''),7.86(2H,s,H 2',2''),9.49(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, CDCl 3 ), ppm
2.31 (6H, s, acetyl at 3 'and 3``), 2.33 (6H, s, acetyl at 4', 4 ''), 6.87 (2H, q, J = 3.6Hz, H 4,5), 7.25 -7.34 (4H, m, H 3,6,5 ', 5``), 7.83 (2H, d, J = 2.1Hz, H 6', 6 ''), 7.86 (2H, s, H 2 ', 2``), 9.49 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,CD3Cl),ppm
18.75(CH3),18.85(CH3),121.81(CH),121.90(CH),124.01(CH),124.09(CH),124.83(CH),130.40(C),132.09(C),142.29(C),145.32(C),164.81(C),167.64(C),167.89(C) 13 C nuclear magnetic resonance spectrum (75 MHz, CD 3 Cl), ppm
18.75 (CH 3 ), 18.85 (CH 3 ), 121.81 (CH), 121.90 (CH), 124.01 (CH), 124.09 (CH), 124.83 (CH), 130.40 (C), 132.09 (C), 142.29 (C) , 145.32 (C), 164.81 (C), 167.64 (C), 167.89 (C)

マススペクトル
FAB-MAS:m/z 549(M+1) Mass spectrum
FAB-MAS: m / z 549 (M + 1)

例４
N,N'-Bis(3,4-dihydroxybenzoyl)-1,2-benzenediamine（以下、PDO-h2と略称するときがある）の製造
PDO-a3の代りにPDO-a2を用いた以外は、全く例２と同様に行い、PDO-h2を製造した。
融点 276 ℃，収量0.24g（収率63％） Example 4
Production of N, N'-Bis (3,4-dihydroxybenzoyl) -1,2-benzenediamine (hereinafter sometimes abbreviated as PDO-h2)
PDO-h2 was produced in exactly the same manner as in Example 2 except that PDO-a2 was used instead of PDO-a3.
Melting point 276 ℃, Yield 0.24g (Yield 63%)

1H核磁気共鳴スペクトル(300MHz,CD3OD),ppm
6.73(2H,d,J=8.4Hz,H 5',5''),7.16-7.24(4H,m,H 4,5,6',6''),7.31(2H,d,J=2.4Hz,H 2',2''),7.48(2H,q,J=3.3Hz,H 3,6) 1 H nuclear magnetic resonance spectrum (300 MHz, CD 3 OD), ppm
6.73 (2H, d, J = 8.4Hz, H 5 ', 5``), 7.16-7.24 (4H, m, H 4,5,6', 6 ''), 7.31 (2H, d, J = 2.4 Hz, H 2 ', 2``), 7.48 (2H, q, J = 3.3Hz, H 3,6)

13C核磁気共鳴スペクトル(75MHz,CD3OD),ppm
116.06(CH),116.12(CH),121.04(CH),126.63(C),126.88(CH),127.21(CH),132.85(C),146.54(C),150.82(C),168.68(C);FAB-MAS:m/z(M+1) 13 C nuclear magnetic resonance spectrum (75 MHz, CD 3 OD), ppm
116.06 (CH), 116.12 (CH), 121.04 (CH), 126.63 (C), 126.88 (CH), 127.21 (CH), 132.85 (C), 146.54 (C), 150.82 (C), 168.68 (C); FAB-MAS: m / z (M + 1)

マススペクトル
FAB-MAS:m/z 381(M+1) Mass spectrum
FAB-MAS: m / z 381 (M + 1)

例５
N,N'-Bis(4-acetoxybenzoyl)-1,2-benzenediamine（以下、PDO-a1と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りに4-acetoxybenzoyl chlorideを用いた以外は、例１と同様に行い、PDO-a1を製造した。
融点 192 〜 194 ℃，収量0.61g（収率71％） Example 5
Production of N, N'-Bis (4-acetoxybenzoyl) -1,2-benzenediamine (hereinafter sometimes abbreviated as PDO-a1)
PDO-a1 was produced in the same manner as in Example 1 except that 4-acetoxybenzoyl chloride was used instead of 3,4,5-triacetoxybenzoyl chloride.
Melting point 192-194 ° C, Yield 0.61g (Yield 71%)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
2.28(6H,s,acetyl at 4',4''),7.26-7.30(6H,m,H 4,5,3',5',3'',5''),7.66(2H,q,J=3.6Hz,H 3,6),7.98(4H,d,J=8.4Hz,H 2',6',2'',6''),10.04(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
2.28 (6H, s, acetyl at 4 ', 4``), 7.26-7.30 (6H, m, H 4,5,3', 5 ', 3``, 5''), 7.66 (2H, q, J = 3.6Hz, H 3,6), 7.98 (4H, d, J = 8.4Hz, H 2 ', 6', 2``, 6 ''), 10.04 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
20.85(CH3),121.99(CH),125.56(CH),125.88(CH),129.08(CH),131.30(C),131.79(C),153.08(C),164.75(C),168.94(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
20.85 (CH3), 121.99 (CH), 125.56 (CH), 125.88 (CH), 129.08 (CH), 131.30 (C), 131.79 (C), 153.08 (C), 164.75 (C), 168.94 (C)

マススペクトル
FAB-MAS:m/z 433(M+1) Mass spectrum
FAB-MAS: m / z 433 (M + 1)

例６
N,N'-Bis(4-hydroxybenzoyl)-1,2-benzenediamine（以下、PDO-h1と略称するときがある）の製造
PDO-a3の代りにPDO-a1を用いた以外は、全く例２と同様に行い、PDO-h1を製造した。
融点 300 ℃，収量0.20g（収率58％） Example 6
Production of N, N'-Bis (4-hydroxybenzoyl) -1,2-benzenediamine (hereinafter sometimes abbreviated as PDO-h1)
PDO-h1 was produced in exactly the same manner as in Example 2 except that PDO-a1 was used instead of PDO-a3.
Melting point 300 ° C, yield 0.20g (58% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
6.84(4H,d,J=8.7Hz,H 3',5',3'',5''),7.24(2H,q,J=3.45Hz,H 4,5),7.60(2H,q,J=3.6Hz,H 3,6),7.81(4H,d,J=8.4Hz,H 2',6',2'',6''),9.87(2H,s,2 OH),10.14(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
6.84 (4H, d, J = 8.7Hz, H 3 ', 5', 3``, 5 ''), 7.24 (2H, q, J = 3.45Hz, H 4,5), 7.60 (2H, q, J = 3.6Hz, H 3,6), 7.81 (4H, d, J = 8.4Hz, H 2 ', 6', 2``, 6 ''), 9.87 (2H, s, 2 OH), 10.14 ( (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
114.45(CH),124.47(CH),124.83(CH),125.18(C),128.74(CH),131.05(C),160.38(C),164.61(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
114.45 (CH), 124.47 (CH), 124.83 (CH), 125.18 (C), 128.74 (CH), 131.05 (C), 160.38 (C), 164.61 (C)

マススペクトル
FAB-MAS:m/z 349(M+1) Mass spectrum
FAB-MAS: m / z 349 (M + 1)

例７
N,N'-dibenzoyl-1,2-benzenediamine（以下、PDO-h0と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りにbenzoyl chlorideを用いた以外は、例１と同様に行い、PDO-h0を製造した。
融点 300 ℃，収量0.48g（収率76.5％） Example 7
Manufacture of N, N'-dibenzoyl-1,2-benzenediamine (hereinafter sometimes abbreviated as PDO-h0)
PDO-h0 was produced in the same manner as in Example 1 except that benzoyl chloride was used instead of 3,4,5-triacetoxybenzoyl chloride.
Melting point 300 ° C, yield 0.48g (yield 76.5%)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
7.30-7.35(4H,m,H 4,5,3,6),7.50-7.63(6H,m,H 3',5',3'',5'',4',4''),7.93(4H,d,J=7.2,H 2',6',2'',6''),10.05(2H,s,2NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
7.30-7.35 (4H, m, H 4,5,3,6), 7.50-7.63 (6H, m, H 3 ', 5', 3``, 5 '', 4 ', 4''), 7.93 (4H, d, J = 7.2, H 2 ', 6', 2``, 6 ''), 10.05 (2H, s, 2NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
125.34(CH),125.57(CH),127.24(CH),128.33(CH),131.15(C),131.58(CH),134.07(C),165.31(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
125.34 (CH), 125.57 (CH), 127.24 (CH), 128.33 (CH), 131.15 (C), 131.58 (CH), 134.07 (C), 165.31 (C)

マススペクトル
FAB-MAS:m/z 317(M+1) Mass spectrum
FAB-MAS: m / z 317 (M + 1)

例８
N,N'-Bis(3,4,5-triacetoxybenzoyl)-1,3-benzenediamine（以下、PDM-a3と略称するときがある）の製造
1,2-ベンゼンジアミンの代りに1,3-ベンゼンジアミンを用いた以外は、例１と同様に行い、PDM-a3を製造した。
融点 250 〜 251 ℃，収量1.00g（収率75％） Example 8
Production of N, N'-Bis (3,4,5-triacetoxybenzoyl) -1,3-benzenediamine (hereinafter sometimes abbreviated as PDM-a3)
PDM-a3 was produced in the same manner as in Example 1 except that 1,3-benzenediamine was used instead of 1,2-benzenediamine.
Melting point 250 ~ 251 ℃, Yield 1.00g (Yield 75%)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
2.32(12H,s,acetyl at 3',5',3'' and 5''),2.33(6H,s,acetyl at 4',4''),7.34(1H,t,J=8.4Hz,H 5),7.48(2H,d,J=8.4Hz,H 4,6),7.83(4H,s,H 2',6',2'',6''),8.27(1H,s,H 2),10.40(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
2.32 (12H, s, acetyl at 3 ', 5', 3 '' and 5``), 2.33 (6H, s, acetyl at 4 ', 4''), 7.34 (1H, t, J = 8.4Hz, H 5), 7.48 (2H, d, J = 8.4Hz, H 4,6), 7.83 (4H, s, H 2 ', 6', 2``, 6 ''), 8.27 (1H, s, H 2), 10.40 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
19.7(CH3),20.15(CH3),112.92(CH),116.33(CH),120.39(CH),128.56(CH),132.75(C),137.17(C),138.87(C),143.00(C),162.97(C),166.75(C),167.80(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
19.7 (CH3), 20.15 (CH3), 112.92 (CH), 116.33 (CH), 120.39 (CH), 128.56 (CH), 132.75 (C), 137.17 (C), 138.87 (C), 143.00 (C), 162.97 (C), 166.75 (C), 167.80 (C)

マススペクトル
FAB-MAS:m/z 665(M+1) Mass spectrum
FAB-MAS: m / z 665 (M + 1)

例９
N,N'-Bis(3,4,5-trihydroxybenzoyl)-1,3-benzenediamine（以下、PDM-h3と略称するときがある）の製造
PDO-a3の代りにPDM-a3を用いた以外は、全く例２と同様に行い、PDM-h3を製造した。
融点 > 300 ℃，収量0.25g（収率61％） Example 9
Production of N, N'-Bis (3,4,5-trihydroxybenzoyl) -1,3-benzenediamine (hereinafter sometimes abbreviated as PDM-h3)
PDM-h3 was produced in exactly the same manner as in Example 2 except that PDM-a3 was used instead of PDO-a3.
Melting point> 300 ° C, yield 0.25 g (61% yield)

1H核磁気共鳴スペクトル(300MHz,CD3OD),ppm
6.87(4H,s,H 2',6',2'',6''),7.20(1H,q,J=7.2Hz,H 5),7.29-7.33(2H,m,H 4,6),7.83(1H,t,J=1.95Hz,H 2) 1 H nuclear magnetic resonance spectrum (300 MHz, CD 3 OD), ppm
6.87 (4H, s, H 2 ', 6', 2``, 6 ''), 7.20 (1H, q, J = 7.2Hz, H 5), 7.29-7.33 (2H, m, H 4,6) , 7.83 (1H, t, J = 1.95Hz, H 2)

13C核磁気共鳴スペクトル(75MHz,CD3OD),ppm
108.20(CH),115.37(CH),118.33(CH),126.62(C),129.89(CH),138.44(C),140.39(C),146.72(C),169.15(C);FAB-MAS:m/z 413(M+1) 13 C nuclear magnetic resonance spectrum (75 MHz, CD 3 OD), ppm
108.20 (CH), 115.37 (CH), 118.33 (CH), 126.62 (C), 129.89 (CH), 138.44 (C), 140.39 (C), 146.72 (C), 169.15 (C); FAB-MAS: m / z 413 (M + 1)

マススペクトル
FAB-MAS:m/z 413(M+1) Mass spectrum
FAB-MAS: m / z 413 (M + 1)

例１０
N,N'-Bis(3,4-diacetoxybenzoyl)-1,3-benzenediamine（以下、PDM-a2と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りに3,4-diacetoxybenzoyl chlorideを用いた以外は、例８と同様に行い、PDM-a2を製造した。
融点 203 〜 204 ℃，収量0.83g（収率76％） Example 10
Production of N, N'-Bis (3,4-diacetoxybenzoyl) -1,3-benzenediamine (hereinafter sometimes abbreviated as PDM-a2)
PDM-a2 was produced in the same manner as in Example 8 except that 3,4-diacetoxybenzoyl chloride was used in place of 3,4,5-triacetoxybenzoyl chloride.
Melting point 203-204 ° C, yield 0.83g (76% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
2.31(6H,s,acetyl at 3'and 3''),2.32(6H,s,acetyl at 4',4''),7.32(1H,t,J=8.0Hz,H 5),7.44(2H,d,J=8.7Hz,H 5',5''),7.49(2H,dd,J=9.6,1.46Hz,H 4,6),7.87(2H,s,H 2',2''),7.92(2H,dd,J=8.4,1.83Hz,H 6',6''),8.30(1H,s,H 2),10.38(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
2.31 (6H, s, acetyl at 3'and 3``), 2.32 (6H, s, acetyl at 4 ', 4''), 7.32 (1H, t, J = 8.0Hz, H 5), 7.44 (2H , d, J = 8.7Hz, H 5 ', 5``), 7.49 (2H, dd, J = 9.6,1.46Hz, H 4,6), 7.87 (2H, s, H 2', 2 '') , 7.92 (2H, dd, J = 8.4,1.83Hz, H 6 ', 6``), 8.30 (1H, s, H 2), 10.38 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
20.30(CH3),20.38(CH3),112.87(CH),116.22(CH),123.21(CH),123.70(CH),126.22(CH),128.68(CH),133.39(C),139.15(C),141.83(C),144.59(C),163.90(C),168.03(C),168.23(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
20.30 (CH3), 20.38 (CH3), 112.87 (CH), 116.22 (CH), 123.21 (CH), 123.70 (CH), 126.22 (CH), 128.68 (CH), 133.39 (C), 139.15 (C), 141.83 (C), 144.59 (C), 163.90 (C), 168.03 (C), 168.23 (C)

マススペクトル
FAB-MAS:m/z 549(M+1) Mass spectrum
FAB-MAS: m / z 549 (M + 1)

例１１
N,N'-Bis(3,4-dihydroxybenzoyl)-1,3-benzenediamine（以下、PDM-h2と略称するときがある）の製造
PDO-a3の代りにPDM-a2を用いた以外は、全く例２と同様に行い、PDM-h2を製造した。
融点 280 〜 281 ℃，収量0.27g（収率71％） Example 11
Production of N, N'-Bis (3,4-dihydroxybenzoyl) -1,3-benzenediamine (hereinafter sometimes abbreviated as PDM-h2)
PDM-h2 was produced in exactly the same manner as in Example 2, except that PDM-a2 was used instead of PDO-a3.
Melting point 280-281 ° C, Yield 0.27g (Yield 71%)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
6.81(2H,d,J=8.1Hz,H 5',5''),7.23(1H,t,J=7.95Hz,H 5),7.33-7.42(6H,m,H 4,6,2',6',2'',6''),8.22(1H,s,H 2),9.41(2H,s,2 OH),9.93(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
6.81 (2H, d, J = 8.1Hz, H 5 ', 5``), 7.23 (1H, t, J = 7.95Hz, H 5), 7.33-7.42 (6H, m, H 4,6,2' , 6 ', 2``, 6''), 8.22 (1H, s, H 2), 9.41 (2H, s, 2 OH), 9.93 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
112.94(CH),114.87(CH),115.49(CH),115.67(CH),119.66(CH),125.98(C),128.29(CH),139.57(C),144.92(C),148.82(C),165.21(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
112.94 (CH), 114.87 (CH), 115.49 (CH), 115.67 (CH), 119.66 (CH), 125.98 (C), 128.29 (CH), 139.57 (C), 144.92 (C), 148.82 (C), 165.21 (C)

マススペクトル
FAB-MAS:m/z 381(M+1) Mass spectrum
FAB-MAS: m / z 381 (M + 1)

例１２
N,N'-Bis(4-acetoxybenzoyl)-1,3-benzenediamine（以下、PDM-a1と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りに4-acetoxybenzoyl chlorideを用いた以外は、例８と同様に行い、PDM-a1を製造した。
融点 275 〜 276 ℃，収量0.70g（収率81％） Example 12
Production of N, N'-Bis (4-acetoxybenzoyl) -1,3-benzenediamine (hereinafter sometimes abbreviated as PDM-a1)
PDM-a1 was produced in the same manner as in Example 8 except that 4-acetoxybenzoyl chloride was used instead of 3,4,5-triacetoxybenzoyl chloride.
Melting point 275-276 ° C, Yield 0.70g (Yield 81%)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
2.30(6H,s,acetyl at 4',4''),7.28(4H,d,J=8.7Hz,H 3',5',3'',5''),7.32-7.35(1H,m,H 5),7.49(2H,d,J=8.1Hz,H 4,6),8.00(4H,d,J=8.7Hz,H 2',6',2'',6''),8.30(1H,s,H 2)10.31(2H,s,2NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
2.30 (6H, s, acetyl at 4 ', 4''), 7.28 (4H, d, J = 8.7Hz, H 3', 5 ', 3``, 5''), 7.32-7.35 (1H, m , H 5), 7.49 (2H, d, J = 8.1Hz, H 4, 6), 8.00 (4H, d, J = 8.7Hz, H 2 ', 6', 2``, 6 ''), 8.30 (1H, s, H 2) 10.31 (2H, s, 2NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
20.86(CH3),112.81(CH),116.04(CH),121.79(CH),128.59(CH),129.21(CH),132.47(C),139.30(C),152.88(C),164.79(C),168.99(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
20.86 (CH3), 112.81 (CH), 116.04 (CH), 121.79 (CH), 128.59 (CH), 129.21 (CH), 132.47 (C), 139.30 (C), 152.88 (C), 164.79 (C), 168.99 (C)

マススペクトル
FAB-MAS:m/z 433(M+1) Mass spectrum
FAB-MAS: m / z 433 (M + 1)

例１３
N,N'-Bis(4-hydroxybenzoyl)-1,3-benzenediamine（以下、PDM-h1と略称するときがある）の製造
PDO-a3の代りにPDM-a1を用いた以外は、全く例２と同様に行い、PDM-h1を製造した。
融点 300 ℃，収量0.23g（収率66％） Example 13
Production of N, N'-Bis (4-hydroxybenzoyl) -1,3-benzenediamine (hereinafter sometimes abbreviated as PDM-h1)
PDM-h1 was produced in exactly the same manner as in Example 2 except that PDM-a1 was used instead of PDO-a3.
Melting point 300 ° C, yield 0.23g (66% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
6.86(4H,d,J=8.4Hz,H 3',5',3'',5''),7.26(1H,t,J=8.1Hz,H 5),7.44(2H,dd,J=9.9Hz,H 4,6),7.87(4H,d,J=8.7Hz,H 2',6',2'',6''),8.25(1H,s,H 2),10.00(2H,s,2 OH),10.10(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
6.86 (4H, d, J = 8.4Hz, H 3 ', 5', 3``, 5 ''), 7.26 (1H, t, J = 8.1Hz, H 5), 7.44 (2H, dd, J = 9.9Hz, H 4,6), 7.87 (4H, d, J = 8.7Hz, H 2 ', 6', 2``, 6 ''), 8.25 (1H, s, H 2), 10.00 (2H, s, 2 OH), 10.10 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
112.95(CH),114.90(CH),115.76(CH)125.46(C),128.39(CH),129.76(CH),139.57(C),160.53(C),165.09(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
112.95 (CH), 114.90 (CH), 115.76 (CH) 125.46 (C), 128.39 (CH), 129.76 (CH), 139.57 (C), 160.53 (C), 165.09 (C)

マススペクトル
FAB-MAS:m/z 349(M+1) Mass spectrum
FAB-MAS: m / z 349 (M + 1)

例１４
N,N'-dibenzoyl-1,3-benzenediamine（以下、PDM-h0と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りにbenzoyl chlorideを用いた以外は、例８と同様に行い、PDM-h0を製造した。
融点 > 300 ℃，収量0.48g（収率76％） Example 14
Production of N, N'-dibenzoyl-1,3-benzenediamine (hereinafter sometimes abbreviated as PDM-h0)
PDM-h0 was produced in the same manner as in Example 8 except that benzoyl chloride was used instead of 3,4,5-triacetoxybenzoyl chloride.
Melting point> 300 ° C, yield 0.48 g (76% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6)
7.31(1H,t,J=8.1Hz,H 5),7.48-7.58(8H,m,H 4,6,3',3',4',5',3'',4'',5''),7.96(4H,d,J=6.9Hz,H 2',6',2'',6''),8.33(1H,s,H 2)10.30(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 )
7.31 (1H, t, J = 8.1Hz, H 5), 7.48-7.58 (8H, m, H 4,6,3 ', 3', 4 ', 5', 3``, 4 '', 5 ''), 7.96 (4H, d, J = 6.9Hz, H 2', 6 ', 2``, 6''), 8.33 (1H, s, H 2) 10.30 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6)112.95(CH),116.08(CH),127.67(CH),128.34(CH),128.55(CH),131.52(CH),134.94(C),139.34(C),165.54(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ) 112.95 (CH), 116.08 (CH), 127.67 (CH), 128.34 (CH), 128.55 (CH), 131.52 (CH), 134.94 (C), 139.34 (C), 165.54 (C)

マススペクトル
FAB-MAS:m/z 317(M+1) Mass spectrum
FAB-MAS: m / z 317 (M + 1)

例１５
N,N'-Bis(3,4,5-triacetoxybenzoyl)-1,4-benzenediamine（以下、PDP-a3と略称するときがある）の製造
1,2-ベンゼンジアミンの代りに1,4-ベンゼンジアミンを用いた以外は、例１と同様に行い、PDP-a3を製造した。
融点 273 ℃，収量0.90g（収率68％） Example 15
Production of N, N'-Bis (3,4,5-triacetoxybenzoyl) -1,4-benzenediamine (hereinafter sometimes abbreviated as PDP-a3)
PDP-a3 was produced in the same manner as in Example 1 except that 1,4-benzenediamine was used instead of 1,2-benzenediamine.
Melting point 273 ℃, yield 0.90g (68% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
2.32(12H,s,acetyl at 3',5',3''and 5''),2.33(6H,s,acetyl at 4',4''),7.73(4H,s,H 2,3,5,6),7.82(4H,s,H 2',6',2'',6'')10.36(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
2.32 (12H, s, acetyl at 3 ', 5', 3``and 5 ''), 2.33 (6H, s, acetyl at 4 ', 4''), 7.73 (4H, s, H 2,3, 5,6), 7.82 (4H, s, H 2 ', 6', 2``, 6 '') 10.36 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
20.27(CH3),20.72(CH3)120.86(CH),121.27(CH),133.33(C),135.27(C),137.69(C),143.58(C),163.32(C),167.32(C),168.37(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
20.27 (CH3), 20.72 (CH3) 120.86 (CH), 121.27 (CH), 133.33 (C), 135.27 (C), 137.69 (C), 143.58 (C), 163.32 (C), 167.32 (C), 168.37 (C)

マススペクトル
FAB-MAS:m/z 665(M+1) Mass spectrum
FAB-MAS: m / z 665 (M + 1)

例１６
N,N'-Bis(3,4,5-trihydroxybenzoyl)-1,4-benzenediamine（以下、PDP-h3と略称するときがある）の製造
PDO-a3の代りにPDP-a3を用いた以外は、全く例２と同様に行い、PDP-h3を製造した。
融点 > 300 ℃，収量0.23g（収率56％） Example 16
Production of N, N'-Bis (3,4,5-trihydroxybenzoyl) -1,4-benzenediamine (hereinafter sometimes abbreviated as PDP-h3)
PDP-h3 was produced in exactly the same manner as in Example 2, except that PDP-a3 was used instead of PDO-a3.
Melting point> 300 ° C, yield 0.23g (56% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
6.94(4H,s,H 2',6',2'',6''),7.65(4H,s,H 2,3,5,6) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
6.94 (4H, s, H 2 ', 6', 2``, 6 ''), 7.65 (4H, s, H 2,3,5,6)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
106.90(CH),119.95(CH),120.05(CH),124.86(C),134.50(C),136.14(C),144.99(C),164.76(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
106.90 (CH), 119.95 (CH), 120.05 (CH), 124.86 (C), 134.50 (C), 136.14 (C), 144.99 (C), 164.76 (C)

マススペクトル
FAB-MAS:m/z 413(M+1) Mass spectrum
FAB-MAS: m / z 413 (M + 1)

例１７
N,N'-Bis(3,4-diacetoxybenzoyl)-1,4-benzenediamine（以下、PDP-a2と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りに3,4-diacetoxybenzoyl chlorideを用いた以外は、例１５と同様に行い、PDP-a2を製造した。
融点 288 〜 290 ℃，収量0.85g（収率78％） Example 17
Production of N, N'-Bis (3,4-diacetoxybenzoyl) -1,4-benzenediamine (hereinafter sometimes abbreviated as PDP-a2)
PDP-a2 was produced in the same manner as in Example 15 except that 3,4-diacetoxybenzoyl chloride was used in place of 3,4,5-triacetoxybenzoyl chloride.
Melting point 288-290 ° C, Yield 0.85g (Yield 78%)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
2.31(6H,s,acetyl at 3'and 3''),2.32(6H,s,acetyl at 4',4''),7.44(2H,d,J=8.4Hz,H 5',5''),7.74(4H,s,H 2,3,5,6),7.86(2H,s,H 2',2''),7.92(2H,dd,J=8.1Hz,H 6',6''),10.32(2H,s,2NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
2.31 (6H, s, acetyl at 3'and 3``), 2.32 (6H, s, acetyl at 4 ', 4''), 7.44 (2H, d, J = 8.4Hz, H 5', 5 '' ), 7.74 (4H, s, H 2, 3, 5, 6), 7.86 (2H, s, H 2 ', 2''), 7.92 (2H, dd, J = 8.1Hz, H 6', 6 ''), 10.32 (2H, s, 2NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
20.29(CH3),20.35(CH3),120.60(CH),120.70(CH),123.13(CH),123.68(CH),126.08(CH),133.40(C),134.86(C),141.83(C),144.51(C),163.64(C),168.00(C),168.19(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
20.29 (CH3), 20.35 (CH3), 120.60 (CH), 120.70 (CH), 123.13 (CH), 123.68 (CH), 126.08 (CH), 133.40 (C), 134.86 (C), 141.83 (C), 144.51 (C), 163.64 (C), 168.00 (C), 168.19 (C)

マススペクトル
FAB-MAS:m/z 549(M+1) Mass spectrum
FAB-MAS: m / z 549 (M + 1)

例１８
N,N'-Bis(3,4-dihydroxybenzoyl)-1,4-benzenediamine（以下、PDP-h2と略称するときがある）の製造
PDP-a3の代りにPDP-a2を用いた以外は、全く例２と同様に行い、PDP-h2を製造した。
融点 > 300 ℃，収量0.26g（収率68％） Example 18
Production of N, N'-Bis (3,4-dihydroxybenzoyl) -1,4-benzenediamine (hereinafter sometimes abbreviated as PDP-h2)
PDP-h2 was produced in exactly the same manner as in Example 2, except that PDP-a2 was used instead of PDP-a3.
Melting point> 300 ° C, yield 0.26 g (68% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
6.81(2H,d,J=8.4Hz,H 4',4''),7.32(2H,dd,J=2.25Hz,H 6',6''),7.37(2H,s,H 2',2''),7.66(4H,s,H 2,3,5,6),9.25(2H,s,2 OH),9.87(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
6.81 (2H, d, J = 8.4Hz, H 4 ', 4``), 7.32 (2H, dd, J = 2.25Hz, H 6', 6 ''), 7.37 (2H, s, H 2 ', 2``), 7.66 (4H, s, H 2,3,5,6), 9.25 (2H, s, 2 OH), 9.87 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
114.87(CH),115.38(CH),119.52(CH),120.43(CH),125.99(C),134.93(C),144.92(C),148.73(C),164.99(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d6), ppm
114.87 (CH), 115.38 (CH), 119.52 (CH), 120.43 (CH), 125.99 (C), 134.93 (C), 144.92 (C), 148.73 (C), 164.99 (C)

マススペクトルFAB-MAS:m/z 381(M+1) Mass spectrum FAB-MAS: m / z 381 (M + 1)

例１９
N,N'-Bis(4-acetoxybenzoyl)-1,4-benzenediamine（以下、PDP-a1と略称するときがある）の製造3,4,5-triacetoxybenzoyl chlorideの代りに4-acetoxybenzoyl chlorideを用いた以外は、例１５と同様に行い、PDP-h0を製造した。
融点 > 300 ℃，収量0.69g（収率80％） Example 19
Production of N, N'-Bis (4-acetoxybenzoyl) -1,4-benzenediamine (hereinafter sometimes abbreviated as PDP-a1) 4-acetoxybenzoyl chloride was used in place of 3,4,5-triacetoxybenzoyl chloride Except that, PDP-h0 was produced in the same manner as in Example 15.
Melting point> 300 ° C, yield 0.69 g (yield 80%)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
2.30(6H,s,acetyl at 4',4''),7.28(4H,d,J=8.4Hz,H 3',5',3'',5''),7.74(4H,s,H 2,3,5,6),8.00(4H,d,J=8.4Hz,H 2',6',2'',6''),10.25(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
2.30 (6H, s, acetyl at 4 ', 4``), 7.28 (4H, d, J = 8.4Hz, H 3', 5 ', 3``, 5''), 7.74 (4H, s, H 2,3,5,6), 8.00 (4H, d, J = 8.4Hz, H 2 ', 6', 2``, 6 ''), 10.25 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
20.85(CH3),120.49(CH),120.59(CH),121.79(CH),129.11(CH),132.48(C),134.92(C),152.83(C),164.53(C),168.96(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d6), ppm
20.85 (CH3), 120.49 (CH), 120.59 (CH), 121.79 (CH), 129.11 (CH), 132.48 (C), 134.92 (C), 152.83 (C), 164.53 (C), 168.96 (C)

マススペクトル
FAB-MAS:m/z 433(M+1) Mass spectrum
FAB-MAS: m / z 433 (M + 1)

例２０
N,N'-Bis(4-hydroxybenzoyl)-1,4-benzenediamine（以下、PDP-h1と略称するときがある）の製造
PDO-a3の代りにPDP-a1を用いた以外は、全く例２と同様に行い、PDP-h1を製造した。
融点 > 300 ℃，収量0.23g（収率66％） Example 20
Production of N, N'-Bis (4-hydroxybenzoyl) -1,4-benzenediamine (hereinafter sometimes abbreviated as PDP-h1)
PDP-h1 was produced in exactly the same manner as in Example 2 except that PDP-a1 was used instead of PDO-a3.
Melting point> 300 ° C, yield 0.23 g (66% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
6.85(4H,d,J=8.7Hz,H 3',5',3'',5''),7.68(4H,s,H 2,3,5,6),7.84(4H,d,J=8.7Hz,H 2',6',2'',6''),9.93(2H,s,2 OH),10.07(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
6.85 (4H, d, J = 8.7Hz, H 3 ', 5', 3``, 5 ''), 7.68 (4H, s, H 2,3,5,6), 7.84 (4H, d, J = 8.7Hz, H 2 ', 6', 2``, 6 ''), 9.93 (2H, s, 2 OH), 10.07 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
114.88(CH),120.51(CH),125.46(C),129.61(CH),134.94(C),160.44(C),164.85(C); 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
114.88 (CH), 120.51 (CH), 125.46 (C), 129.61 (CH), 134.94 (C), 160.44 (C), 164.85 (C);

マススペクトル
FAB-MAS:m/z 349(M+1) Mass spectrum
FAB-MAS: m / z 349 (M + 1)

例２１
N,N'-dibenzoyl-1,4-benzenediamine（以下、PDP-h0と略称するときがある）の製造
3,4,5-triacetoxybenzoyl chlorideの代りにbenzoyl chlorideを用いた以外は、例１５と同様に行い、PDP-h0を製造した。
融点 > 300 ℃，収量0.48g（収率76％) Example 21
Manufacture of N, N'-dibenzoyl-1,4-benzenediamine (hereinafter sometimes referred to as PDP-h0)
PDP-h0 was produced in the same manner as in Example 15 except that benzoyl chloride was used instead of 3,4,5-triacetoxybenzoyl chloride.
Melting point> 300 ° C, yield 0.48 g (76% yield)

1H核磁気共鳴スペクトル(300MHz,DMSO-d6),ppm
7.49-7.58(6H,m,H 3',4',5',3'',4'',5''),7.75(4H,s,H 2,3,5,6),7.94-7.96(4H,m,H 2',6',2'',6''),10.23(2H,s,2 NH) 1 H nuclear magnetic resonance spectrum (300 MHz, DMSO-d 6 ), ppm
7.49-7.58 (6H, m, H 3 ', 4', 5 ', 3``, 4'',5''), 7.75 (4H, s, H 2,3,5,6), 7.94-7.96 (4H, m, H 2 ', 6', 2``, 6 ''), 10.23 (2H, s, 2 NH)

13C核磁気共鳴スペクトル(75MHz,DMSO-d6),ppm
120.64(CH),127.57(CH),128.35(CH),131.46(CH),134.94(C),134.97(C),165.30(C) 13 C nuclear magnetic resonance spectrum (75 MHz, DMSO-d 6 ), ppm
120.64 (CH), 127.57 (CH), 128.35 (CH), 131.46 (CH), 134.94 (C), 134.97 (C), 165.30 (C)

マススペクトル
FAB-MAS:m/z 317(M+1) Mass spectrum
FAB-MAS: m / z 317 (M + 1)

例２２
トポイソメラーゼI阻害活性測定
トポイソメラーゼＩは、二重鎖ＤＮＡに切断を導入することにより超らせんを弛緩する活性を有しているので、トポイソメラーゼＩによるＤＮＡ弛緩（ｒｅｌａｘａｔｉｏｎ）活性の減少を測定して、例１〜２１において製造したベンゼンジアミン誘導体の阻害活性を算出した。９６穴プレートに２倍濃度のトポイソメラーゼＩ活性測定用緩衝液（２×ＴｏｐｏＩ ａｓｓａｙ ｂｕｆｆｅｒ；１００ｍＭ Ｔｒｉｓ−ＨＣｌ緩衝液（ｐＨ７．５）、２４０ｍＭ ＫＣｌ、２０ｍＭ ＭｇＣｌ2、１ｍＭ ＥＤＴＡ、１ｍＭジチオスレイトール、６０μｇ／ｍｌ 牛血清アルブミン（ＢＳＡ））１０μｌ、ベンゼンジアミン誘導体試料溶液８．３μｌ（ＤＭＳＯを１０％含むＭｅＯＨに溶解）、子牛胸腺由来のトポイソメラーゼＩ（２ｕｎｉｔ／μｌ）０．５μｌを添加後、３７℃で５分間プレインキュベーションする。次に、基質ＤＮＡであるｐＢＲ３２２（０．１２５μｇ／μｌ）１．２μｌを添加し、３７℃、４０分間反応させる。反応後、電気泳動試料用緩衝液（２倍濃度のＴＢＥ緩衝液（２００ｍＭ Ｔｒｉｓ、２００ｍＭホウ酸、５ｍＭ ＥＤＴＡ）、１０％ブロムフェノールブルー、５０％グリシン）１０μｌを添加し、その反応液１２μｌをアガロースゲル電気泳動に用いた。 Example 22
Measurement of Topoisomerase I Inhibitory Activity Topoisomerase I has the activity of relaxing a super helix by introducing a break into double-stranded DNA. Therefore, the decrease in DNA relaxation activity by topoisomerase I was measured. The inhibitory activity of the benzenediamine derivatives produced in 1-21 was calculated. Buffer for measuring topoisomerase I activity (2 × Topo assay buffer; 100 mM Tris-HCl buffer (pH 7.5), 240 mM KCl, 20 mM MgCl 2 , 1 mM EDTA, 1 mM dithiothreitol, 60 μg in 96-well plate) / Ml bovine serum albumin (BSA) 10 μl, benzenediamine derivative sample solution 8.3 μl (dissolved in MeOH containing 10% DMSO), calf thymus-derived topoisomerase I (2 units / μl) 0.5 μl Pre-incubate at 5 ° C for 5 minutes. Next, 1.2 μl of substrate DNA pBR322 (0.125 μg / μl) is added and reacted at 37 ° C. for 40 minutes. After the reaction, 10 μl of electrophoresis sample buffer solution (2 times TBE buffer solution (200 mM Tris, 200 mM boric acid, 5 mM EDTA), 10% bromophenol blue, 50% glycine) was added, and 12 μl of the reaction solution was added to agarose. Used for gel electrophoresis.

電気泳動は、次のようにして行った。アガロース０．３ｇにＴＢＥ緩衝液３０ｍｌを加え、加熱溶解する。電気泳動用プレートにアガロース液を流し入れ、固化後、アガロースゲルの溝に試料溶液を添加し、１００Ｖ、１時間、ＴＢＥ緩衝液中で電気泳動を行う。泳動後、ゲルをエチジウムブロマイド溶液（５０μｇ／ｍｌ）２００ｍｌで１分間染色し、次に水５００ｍｌで５分、１０分および１０分間隔で３回脱色する。泳動および染色後、ｐＢＲ３２２の弛緩ＤＮＡ（ｒｅｌａｘｅｄ ＤＮＡ）のバンドと超らせんＤＮＡ（ｓｕｐｅｒｃｏｉｌｅｄ ＤＮＡ）のバンドとは移動度の違いにより別バンドとして現れるので、各バンドをＵＶトランスイルミネーター（ＴＭ３６（商品名、フナコシ（株））とデンシトメーター（ＡＥ−６９００Ｍ（商品名、アトー（株）））で測定し、反応前後における各ＤＮＡ量の変化を求める。すなわち、ベンゼンジアミン誘導体によってトポイソメラーゼＩが阻害されるために現れる未反応のｐＢＲ３２２のバンド（超らせんＤＮＡ量）から、トポイソメラーゼＩのＤＮＡ弛緩（ｒｅｌａｘａｔｉｏｎ）活性の減少を求め、阻害活性を算出した。例１〜２１において製造したベンゼンジアミン誘導体のトポイソメラーゼＩ阻害活性を表１に示す。 Electrophoresis was performed as follows. Add 30 ml of TBE buffer to 0.3 g of agarose and dissolve by heating. The agarose solution is poured into the electrophoresis plate, solidified, the sample solution is added to the groove of the agarose gel, and electrophoresis is performed in TBE buffer solution at 100 V for 1 hour. After electrophoresis, the gel is stained with 200 ml of ethidium bromide solution (50 μg / ml) for 1 minute and then decolorized three times with 500 ml of water at 5 minutes, 10 minutes and 10 minute intervals. After electrophoresis and staining, the band of relaxed DNA (re relaxed DNA) of pBR322 and the band of supercoiled DNA (supercoiled DNA) appear as separate bands due to the difference in mobility. Therefore, each band is expressed as a UV transilluminator (TM36 (trade name). , Funakoshi Co., Ltd.) and a densitometer (AE-6900M (trade name, Ato Co., Ltd.)) to determine the change in the amount of each DNA before and after the reaction. From the band of unreacted pBR322 (the amount of supercoiled DNA) that appears, the decrease in the DNA relaxation activity of topoisomerase I was determined, and the inhibitory activity was calculated.Topoisomerase of the benzenediamine derivative produced in Examples 1-21 I inhibition The activity is shown in Table 1.

Figure 2005154342
Figure 2005154342

例２３
トポイソメラーゼＩＩ阻害活性測定
トポイソメラーゼＩＩによるＤＮＡ弛緩（ｒｅｌａｘａｔｉｏｎ）活性が、ベンゼンジアミン誘導体によって減少する程度を測定し、ベンゼンジアミン誘導体の阻害活性を算出した。阻害活性の測定は以下のようにして行った。ＤＮＡ弛緩活性の阻害を測定するための反応混合液は、９６穴プレートに２倍濃度のトポイソメラーゼＩＩ活性測定用緩衝液（２×Ｔｏｐｏ ＩＩ ａｓｓａｙ ｂｕｆｆｅｒ（商品名、トポゲン（ＴｏｐｏＧｅｎ）社）；１０ｍＭ Ｔｒｉｓ−ＨＣｌ，ｐΗ８．０、２４ｍＭ ＫＣｌ、２ｍＭ ΜｇＣｌ2、０．１ｍＭ ＡＴＰ、０．１ｍΜ ジチオスレイトール、６μｇ／ｍｌ ＢＳＡ）１０μｌ、ベンゼンジアミン誘導体試料溶液８．３μｌ、ヒト胎盤由来のトポイソメラーゼＩＩ（２ｕｎｉｔ／μｌ）０．５μｌおよびｐＢＲ３２２（０．１２５μｇ／μｌ）１．２μｌを添加後、３７℃で４０分間反応させたものである。反応後、前記の電気泳動試料用緩衝液４μｌを添加し、その１２μｌをアガロースゲル電気泳動に用いた。泳動後、ｐＢＲ３２２の超らせんＤＮＡ（ｓｕｐｅｒｃｏｉｌｅｄ ＤＮＡ）量の変化をデンシトメーターで測定し、ＤＮＡ弛緩（ｒｅｌａｘａｔｉｏｎ）活性の減少から、ベンゼンジアミン誘導体の阻害活性を求めた。 Example 23
Measurement of Topoisomerase II Inhibitory Activity The degree to which the DNA relaxation activity by topoisomerase II was reduced by the benzenediamine derivative was measured, and the inhibitory activity of the benzenediamine derivative was calculated. Inhibitory activity was measured as follows. A reaction mixture for measuring the inhibition of DNA relaxation activity was prepared in a 96-well plate with a buffer solution for measuring topoisomerase II activity (2 × Topo II assay buffer (trade name, TopoGen)); 10 mM Tris -HCl, p Η 8.0, 24 mM KCl, 2 mM ΜgCl 2 , 0.1 mM ATP, 0.1 mM チ オ dithiothreitol, 6 µg / ml BSA) 10 µl, benzenediamine derivative sample solution 8.3 µl, human placenta-derived topoisomerase II (2 units / Μl) 0.5 μl and pBR322 (0.125 μg / μl) 1.2 μl were added and reacted at 37 ° C. for 40 minutes. After the reaction, 4 μl of the electrophoresis sample buffer solution was added, and 12 μl thereof was used for agarose gel electrophoresis. After the electrophoresis, the change in the amount of supercoiled DNA of pBR322 was measured with a densitometer, and the inhibitory activity of the benzenediamine derivative was determined from the decrease in the DNA relaxation activity.

ベンゼンジアミン誘導体の、トポイソメラーゼＩＩのＤＮＡ弛緩（ｒｅｌａｘａｔｉｏｎ）活性に対する阻害活性（ＩＣ50）は以下のとおりである。ベンゼンジアミン誘導体はトポイソメラーゼＩＩのＤＮＡ弛緩活性を強く阻害する。ＰＤＯ−ｈ３ ０．０９μＭ、ＰＤＭ−ｈ３ ０．２２μＭ、ＰＤＰ−ｈ３ ０．１５μＭ
The inhibitory activity (IC 50 ) of the benzenediamine derivative on the DNA relaxation activity of topoisomerase II is as follows. Benzenediamine derivatives strongly inhibit the DNA relaxing activity of topoisomerase II. PDO-h3 0.09 μM, PDM-h3 0.22 μM, PDP-h3 0.15 μM

Claims (2)

式(I)
Figure 2005154342
 [式(I)中、Z1およびZ2は、式(II)で示される基
Figure 2005154342
 （式(II)中、R1、R2およびR3は水素原子、水酸基またはアセトキシ基を表す。但し、R1、R2およびR3が２つ以上水素原子である化合物は除く。）を表す。]で示されるベンゼンジアミン誘導体を有効成分とするトポイソメラーゼ阻害剤。 Formula (I)
Figure 2005154342
 [In the formula (I), Z 1 and Z 2 are groups represented by the formula (II)
Figure 2005154342
 (In the formula (II), R 1 , R 2 and R 3 represent a hydrogen atom, a hydroxyl group or an acetoxy group, except for compounds in which R 1 , R 2 and R 3 are two or more hydrogen atoms). Represent. ] The topoisomerase inhibitor which uses the benzenediamine derivative shown by this as an active ingredient. 式(I-a)
Figure 2005154342
 [式(I-a)中、Z1およびZ2は、式(II-a)で示される基
Figure 2005154342
 （式(II-a)中、R1、R2およびR3は水素原子、水酸基またはアセトキシ基を表す。但し、R1、R2およびR3が２つ以上水素原子である化合物並びにR1およびR2が水酸基であり、R3が水素原子である化合物は除く。）を表す。]で示されるベンゼンジアミン誘導体。
Formula (Ia)
Figure 2005154342
 [In the formula (Ia), Z 1 and Z 2 are groups represented by the formula (II-a)
Figure 2005154342
 (In the formula (II-a), R 1 , R 2 and R 3 represent a hydrogen atom, a hydroxyl group or an acetoxy group, provided that R 1 , R 2 and R 3 are two or more hydrogen atoms, and R 1 And R 2 is a hydroxyl group and R 3 is a hydrogen atom. ] The benzenediamine derivative shown by this.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100427507C (en) * 2006-10-17 2008-10-22 中国林业科学研究院林产化学工业研究所 Nutgall acylation modified fibre an dits synthetic method
EP2439194A1 (en) * 2010-10-05 2012-04-11 Rheinische Friedrich-Wilhelms Universität Phenyl diamides and a pharmaceutical preparation comprising phenyl diamides
WO2014175136A1 (en) * 2013-04-23 2014-10-30 コニカミノルタ株式会社 Optical film, circularly polarizing plate and organic electroluminescent display device

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100427507C (en) * 2006-10-17 2008-10-22 中国林业科学研究院林产化学工业研究所 Nutgall acylation modified fibre an dits synthetic method
EP2439194A1 (en) * 2010-10-05 2012-04-11 Rheinische Friedrich-Wilhelms Universität Phenyl diamides and a pharmaceutical preparation comprising phenyl diamides
WO2012045734A1 (en) * 2010-10-05 2012-04-12 Rheinische Friedrich-Wilhelms Universität Phenyl diamides and a pharmaceutical preparation comprising phenyl diamides
WO2014175136A1 (en) * 2013-04-23 2014-10-30 コニカミノルタ株式会社 Optical film, circularly polarizing plate and organic electroluminescent display device
US9720140B2 (en) 2013-04-23 2017-08-01 Konica Minolta, Inc. Optical film, circularly polarizing plate and organic electroluminescent display device
KR101792374B1 (en) 2013-04-23 2017-10-31 코니카 미놀타 가부시키가이샤 Optical film, circularly polarizing plate and organic electroluminescent display device

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