JP3683003B2 - Anti-tumor substance epolactaene - Google Patents

Description

で示されるエポラクタエンを提供するものである。本発明の別の態様によれば、上記エポラクタエンを有効成分とする神経分化誘導剤；神経疾患の治療・予防剤；及び抗腫瘍剤、並びに神経腫瘍の治療及び／又は予防に用いる上記抗腫瘍剤が提供される。
【０００７】
本発明のさらに別の態様によれば、ペニシリウム属に属し上記のエポラクタエンを産生するカビが提供される。その好ましい態様として、ペニシリウム・エスピー・BM1689-P株が提供される。さらに、上記産生菌を培養し培養物からエポラクタエンを採取することを特徴とする、上記エポラクタエンの製造方法も提供される。ペニシリウム・エスピー・BM1689-P株を産生菌として用いる方法は上記発明の好ましい態様である。
【０００８】
本発明のエポラクタエンは３個の不斉炭素を有しており、これら３個の不斉中心に基づく光学異性体またはジアステレオ異性体が存在するが、本発明のエポラクタエンには上記立体異性体の全てが包含される。また、光学的あるいはジアステレオ的に純粋な上記立体異性体のほか、光学異性体の任意の割合の混合物やラセミ体、あるいはジアステレオ異性体の任意の割合の混合物なども全て本発明の範囲に包含される。
【０００９】
本発明のエポラクタエンは、内浦湾の海底102 m から採取された土壌から分離されたカビBM1689-P株を培養した培養物から分離採取された。下記の生物学的性質に基づき、本発明の放線菌BM1689-P株はペニシリウム(Penicillium) 属に属するカビであることが判明した。なお、ペニシリウム・エスピー・BM1689-P株は、平成７年３月24日付けで工業技術院生命工学工業技術研究所に受託番号FERM P-14859 として寄託されている。
【００１０】
本発明のエポラクタエンを生産するカビ BM1689-P 株は、以下の生物学的性質を有する。
(1) 形態学的特徴
(i) 分生子柄
110 〜200 × 1.8〜3.5 μm で滑面、微細な突起を有する。また、複輪生のペニシリを形成する。
(ii) メトレ
10.0〜13.0×2.3 〜3.0 μm で4 〜8 本が束生する。
(iii) フィアライド
11.0〜15.0×1.8 〜2.5 μm で輪生体となる。
(iv) 分生子
3.5 〜4.2 ×2.6 〜3.5 μm で滑面、亜球形から楕円型である。
(2) 培養性状
【００１１】
各種寒天培地を用い、25℃で７日間培養した場合の生育的特徴を示す。
【表１】

【００１２】
いずれの培地でも分生子形成は良好で、分泌液は認められない。また、37℃での培養時、未熟な菌核状組織が形成される場合がある。なお、本菌株の生育温度範囲は10〜38℃、生育至適温度は32℃である。
以上の菌学的諸性質より、BM1689-P株をペニシリウム・エスピー(Penicillium
sp.) と同定した。
【００１３】
本発明のエポラクタエンを製造するには、上記のペニシリウム・エスピー・BM1689-P株の他、ペニシリウム属に属するエポラクタエン産生菌を用いることができる。このような菌のうち、BM1689-P株は好ましい菌株である。このようなペニシリウム属に属するエポラクタエン産生菌を培養して培養物から本発明のを分離採取するには、当業者に周知の方法によればよい。その一例として以下に実施例を示すが、本発明のエポラクタエンの製造方法はこれらの例に限定されることはなく、当業者により種々の改変や修飾が可能であることはいうまでもない。
【００１４】
本発明のエポラクタエンは抗腫瘍作用を有しているので、本発明のエポラクタエンを有効成分として含む医薬は、ヒトを含む哺乳類の各種の腫瘍の治療に有用である。特に、本発明の抗腫瘍剤は脳神経などの神経系の腫瘍の治療に有用である。また、本発明のエポラクタエンは、脳神経などの神経系細胞に対して分化誘導作用を有するので、神経分化誘導剤として有用である。なお、本明細書で神経という場合には、脳神経などの中枢神経の他、末梢神経などの神経系の全てが含まれる。
【００１５】
本発明の医薬としては、有効成分である上記のエポラクタエンをそのまま患者に投与してもよいが、上記の化合物の１種あるいはそれ以上を有効成分として含む医薬組成物を製剤添加物を用いて製造し、医薬組成物の形態で患者に投与することが好適である。本発明の医薬の有効成分としては、上記の一般式に包含される遊離形態の化合物の他、任意の水和物を用いてもよい。本発明の医薬の投与形態は特に制限されず、経口的又は非経口的に投与することができる。上記の医薬組成物の製造に用いる薬理学的、製剤学的に許容しうる添加物としては、例えば、賦形剤、崩壊剤ないし崩壊補助剤、結合剤、滑沢剤、コーティング剤、色素、希釈剤、基剤、溶解剤ないし溶解補助剤、等張化剤、pH調節剤、安定化剤、噴射剤、及び粘着剤等を挙げることができる。
【００１６】
経口投与に適する製剤の例としては、例えば、錠剤、カプセル剤、散剤、細粒剤、顆粒剤、液剤、又はシロップ剤等を挙げることができる。また、非経口投与に適する製剤としては、例えば、注射剤、点滴剤、坐剤、吸入剤、又は貼付剤等を挙げることができる。経口投与に適する製剤には、薬理学的、製剤学的に許容しうる添加物として、例えば、ブドウ糖、乳糖、D-マンニトール、デンプン、又は結晶セルロース等の賦形剤；カルボキシメチルセルロース、デンプン、又はカルボキシメチルセルロースカルシウム等の崩壊剤又は崩壊補助剤；ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロース、ポリビニルピロリドン、又はゼラチン等の結合剤；ステアリン酸マグネシウム又はタルク等の滑沢剤；ヒドロキシプロピルメチルセルロース、白糖、ポリエチレングリコール又は酸化チタン等のコーティング剤；ワセリン、流動パラフィン、ポリエチレングリコール、ゼラチン、カオリン、グリセリン、精製水、又はハードファット等の基剤を用いることができる。
【００１７】
経皮又は経粘膜投与に適する製剤は、例えば、フロン，ジエチルエーテル、又は圧縮ガス等の噴射剤；ポリアクリル酸ナトリウム、ポリビニルアルコール、メチルセルロース、ポリイソブチレン、ポリブテン等の粘着剤；木綿布又はプラスチックシート等の基布等の製剤用添加物を用いて製造することができる。注射あるいは点滴用に適する製剤には、注射用蒸留水、生理食塩水、プロピレングリコール等の水性あるいは用時溶解型注射剤を構成しうる溶解剤又は溶解補助剤；ブドウ糖、塩化ナトリウム、D-マンニトール、グリセリン等の等張化剤；無機酸、有機酸、無機塩基又は有機塩基等のpH調節剤等の製剤用添加物を添加して製造してもよい。
【００１８】
本発明の抗腫瘍剤は、胃癌、肺癌、肝臓癌、腎臓癌、脳腫瘍、大腸癌、食道癌などの固形癌、あるいは悪性リンパ腫、白血病などのあらゆる種類の癌の治療にに用いることができるが、特に、神経腫瘍、例えば脳腫瘍の治療に用いることが好ましい。また、本発明の神経分化誘導剤は、老人痴呆症などの脳疾患の治療及び／又は予防に有用である。もっとも、本発明の医薬の適用対象は上記のものに限定されることはなく、また、一般的に悪性腫瘍や神経疾患の治療に用いられる他の薬剤と併用してもさしつかえない。本発明の抗腫瘍剤及び神経分化誘導剤の投与量は特に制限されず、投与形態、年齢、体重、症状等に応じて適宜選択すればよい。例えば、経鼻投与の場合には、成人１日あたり有効成分量として1 〜100 mg/Kg 、好ましくは 2〜10 mg/Kgを投与すればよい。投与は１日あたり１回もしくは数回に分けてもよく、投与期間も、年齢、症状等に応じて任意に定めることができる。
【００１９】
【実施例】
以下、実施例により本発明をさらに具体的に説明するが、本発明の範囲は下記の実施例に限定されることはない。
(A) 製造例
グルコース(3.5%)、可溶性澱粉(1%)、肉エキス(0.5%)、ポリペプトン(0.5%)、酵母抽出液(0.3%)、食塩(0.2%)、リン酸第二カリウム(0.05%) 、硫酸マグネシウム(0.05%) 、及び大豆粉(2%)の組成の培地（pH 5.8）にペニシリウム・エスピー・BM1689-P株を接種し、 28 ℃で48時間振蘯培養を行った。この培養液140 mlを同組成の培地 18 リットルに接種し、28℃で50時間にわたって毎分７リットルの通気を行いながら、毎分 300回転の振盪培養を行った。
【００２０】
上記培養液を遠心分離機（6,000 回転／分）で菌体と上清に分離し、菌体を80% アセトン 5リットルを用いて抽出した。減圧濃縮後、得られた水溶液をpH 7.0に調整し、5 リットルの酢酸エチルで2 回抽出を繰り返した。抽出後、全ての酢酸エチルを合わせて減圧濃縮し、褐色のシロップ 4.7 gを得た。
このシロップをクロロホルム 10 mlに溶解し、クロロホルムで調製したシリカゲルカラム (直径 4 cm, 長さ 60 cm) に浸潤させた。最初にクロロホルム 1リットルで溶出した後、配合割合を順次変えたクロロホルム−メタノール溶液 (100:1, 100:2, 100:5, 100:10, 100:20, 100:50)各 500 ml ずつで溶出し、最後にメタノール 1リットルで溶出した。活性成分はクロロホルム−メタノール溶液 (100:5 から100:10) の画分に溶出した。この分画を減圧濃縮することにより、1.6 g の褐色シロップを得た。
【００２１】
さらにこの褐色シロップ 0.8 gをクロロホルム−メタノール溶液(1:1) に溶解し、同じ溶液で調製したセファデックス LH-20 (ファルマシア社製）カラム（直径 2 cm, 長さ 65 cm) に吸着させ、クロロホルム−メタノール溶液(1:1) で溶出した。各10 ml ずつ分取し、主な活性画分であるフラクションを集めて減圧濃縮し、260 mgの褐色のシロップを得た。
次に ODSカラム (直径 2 cm, 長さ 25 cm; カプセルパック, 資生堂社製）を用いて高速液体クロマトグラフィーによる分取（80% メタノール、流速 8 ml/分) を行い、無色アモルファス状物質 15 mgを得た。最後に ODSカラム (直径 2 cm, 長さ 25 cm; カプセルパック, 資生堂社製）を用いて高速液体クロマトグラフィーによる分取（60% メタノール、流速 8 ml/分) を行い、無色アモルファス状物質 5 mg を得た。
【００２２】
1.物質の性状：無色アモルファス状
2.比旋光度： [α] _D +32 °(26 ℃, c=0.1, MeOH)
3.分子量、分子式: 高分解能EIマススペクトル分析により MW=389, C₂₁H₂₇NO₆
4.溶解性: メタノール、エタノール、ジメチルスルホキシドに易溶。ヘキサン、水に不溶
5.呈色反応: 硫酸、ニンヒドリン、ヨウ素に陽性
6.紫外線吸収スペクトル：λ_max (MeOH) (ε): 232 nm(21800), 280 nm(sh,15600)
7.赤外線吸収スペクトル：ν(KBr): 3420, 2920, 1720, 1640, 1435, 1270, 1135, 970, 965
8.核磁気共鳴スペクトル(¹H-NMR 及び¹³C-NMR)：CD₃OD 中, ¹H-NMR: 600 MHz, ¹³C-NMR: 100 MHz, δ ppm, 内部標準 TMS
【００２３】
【化３】

【００２４】
【表２】

【００２５】
(B) 試験例
マウス神経芽腫細胞 Neuro 2A 細胞は、公知の神経分化誘導剤であるジブチリルサイクリックAMP により分化が誘導され、神経突起を伸長する(J. Antibiotics, 44, 113, 1991) 。一方、ヒト神経芽腫細胞 SH-SY5Y細胞においても、ジブチリルサイクリック AMPを添加すると神経分化特有の樹状突起を形成した。そこで、本発明のエポラクタエンについて、ヒト神経芽腫細胞 SH-SY5Y細胞に対する樹状突起形成促進作用を上記の方法により検討した。結果を表３に示す。表中、樹状突起形成率は、全細胞数に対する樹状突起形成細胞数の割合を示す。
【００２６】
【表３】

【発明の効果】
本発明のエポラクタエンは、例えば神経腫瘍などの治療に用いる抗腫瘍剤、並びに、例えば神経疾患等の治療及び／又は予防に用いる神経分化誘導剤として有用である。本発明により提供される新規微生物ならびに製造方法は、上記のエポラクタエンを効率的に製造するために有用である。[0001]
[Industrial application fields]
The present invention relates to a novel compound, epolactaene, a method for producing the same, and use thereof. More specifically, the present invention relates to a novel epolactaene, a mold belonging to the genus Penicillium, which produces the epolactaene, a method for producing the epolactaene using the fungus, a neuronal differentiation inducer and an antitumor agent comprising the epolactaene as an active ingredient About.
[0002]
[Prior art]
Conventionally, adriamycin, mitomycin C, bleomycin, and the like are known as antitumor agents. These inhibit the nucleic acid synthesis system by acting directly on the structure and function of DNA by the mechanism of action such as intercalation, cross-linking, or DNA strand breakage. It exhibits cytotoxicity. However, these drugs are not sufficiently specific for tumor cells and are toxic to normal cells such as bone marrow cells with proliferative proliferation. It was.
[0003]
On the other hand, a compound having a differentiation-inducing action specifically for tumor cells is expected to be useful as an antitumor agent, and conventionally, many studies on differentiation-inducing therapy for blood cell tumors have been made. For neural tumors, dibutyryl cyclic AMP is known to have an action of promoting the formation of dendrites unique to neural differentiation on nerve cells. However, at present, there are almost no studies on differentiation induction therapy for tumors of the nervous system other than those described above. In recent years, with the increase of the elderly population, establishment of effective treatment methods for brain diseases such as senile dementia has been demanded, but no effective drugs for these diseases are known.
[0004]
[Problems to be solved by the invention]
An object of the present invention is to provide a novel anti-tumor agent exhibiting high specificity for tumor cells. From another viewpoint, an anti-tumor based on differentiation-inducing action particularly on tumors of the nervous system. An object of the present invention is to provide a novel antitumor agent that exerts an action. Another object of the present invention is to provide a neuronal differentiation inducer that has a differentiation-inducing action on nerve cells such as cranial nerves and is effective against brain diseases such as senile dementia.
Furthermore, this invention aims at providing the manufacturing method of the compound which is an active ingredient of said antitumor agent and nerve differentiation-inducing agent. More specifically, it is also an object of the present invention to provide a novel mold that produces the above compound and a method for producing the compound using the mold.
[0005]
[Means for Solving the Problems]
As a result of diligent efforts to solve the above problems, the present inventors have found that a novel compound produced by a novel fungus belonging to the genus Penicillium has a cell differentiation-inducing action, and is particularly excellent in inducing differentiation against nerve cells. It was found to have an effect. The present invention has also found that the above compounds are extremely useful as an antitumor agent and a neuronal differentiation inducer. The present invention has been completed based on the above findings.
[0006]
That is, the present invention provides the following formula:
[Chemical formula 2]

It provides epolactaene represented by the following. According to another aspect of the present invention, a neuronal differentiation-inducing agent comprising the above epolactaene as an active ingredient; a therapeutic / preventive agent for neurological diseases; and an antitumor agent; and the antitumor agent used for treating and / or preventing neuronal tumors. Is provided.
[0007]
According to still another aspect of the present invention, there is provided a mold that belongs to the genus Penicillium and produces the above-mentioned epolactaene. As a preferred embodiment thereof, Penicillium sp. BM1689-P strain is provided. Furthermore, a method for producing the epolactaene is also provided, wherein the bacterium is cultured and epolactaene is collected from the culture. A method of using Penicillium sp. BM1689-P strain as a producing bacterium is a preferred embodiment of the above invention.
[0008]
The epolactaene of the present invention has three asymmetric carbons, and there exist optical isomers or diastereoisomers based on these three asymmetric centers. All are included. In addition to the above-mentioned stereoisomers that are optically or diastereomerically pure, mixtures of optical isomers in arbitrary proportions, racemates, or mixtures of diastereoisomers in arbitrary proportions are all within the scope of the present invention. Is included.
[0009]
The epolactaene of the present invention was isolated and collected from a culture obtained by culturing mold BM1689-P strain isolated from soil collected from the sea floor 102 m in Uchiura Bay. Based on the following biological properties, the actinomycete BM1689-P strain of the present invention was found to be a mold belonging to the genus Penicillium. Penicillium sp. BM1689-P was deposited at the National Institute of Advanced Industrial Science and Technology as of March 24, 1995 under the accession number FERM P-14859 .
[0010]
The mold BM1689-P strain producing the epolactaene of the present invention has the following biological properties.
(1) Morphological features
(i) Conidial pattern
110-200 × 1.8-3.5 μm with smooth surface and fine protrusions. In addition, it forms a double-wheeled penicillary.
(ii) Metre
10.0 to 13.0 × 2.3 to 3.0 μm with 4 to 8 bundles.
(iii) Fear Ride
11.0-15.0 × 1.8-2.5 μm becomes a living body.
(iv) Conidia
It is 3.5 to 4.2 × 2.6 to 3.5 μm, and is smooth, subspherical to elliptical.
(2) Culture characteristics [0011]
The growth characteristics when cultured for 7 days at 25 ° C. using various agar media are shown.
[Table 1]

[0012]
In any medium, conidia formation is good and no secretion is observed. In addition, immature fungal nuclei may be formed during culture at 37 ° C. In addition, the growth temperature range of this strain is 10 to 38 ° C, and the optimum growth temperature is 32 ° C.
Based on the above bacteriological properties, the strain BM1689-P is penicillium sp.
sp.).
[0013]
In addition to the above-mentioned Penicillium sp. BM1689-P strain, an epollactaene-producing bacterium belonging to the genus Penicillium can be used for producing the epolactaene of the present invention. Of these bacteria, the BM1689-P strain is a preferred strain. In order to cultivate such an epolactaene-producing bacterium belonging to the genus Penicillium and to separate and collect the present invention from the culture, a method well known to those skilled in the art may be used. Examples are shown below as examples, but the method for producing epolactaene of the present invention is not limited to these examples, and it goes without saying that various alterations and modifications can be made by those skilled in the art.
[0014]
Since the epolactaene of the present invention has an antitumor action, the medicament containing the epolactaene of the present invention as an active ingredient is useful for the treatment of various tumors in mammals including humans. In particular, the antitumor agent of the present invention is useful for the treatment of tumors of the nervous system such as cranial nerves. Moreover, since the epolaractaene of this invention has a differentiation-inducing action with respect to nervous system cells, such as a cranial nerve, it is useful as a nerve differentiation-inducing agent. In the present specification, the term “nerve” includes not only the central nerve such as the cranial nerve but also the entire nervous system such as the peripheral nerve.
[0015]
As the medicament of the present invention, the above-mentioned epolactaene, which is an active ingredient, may be administered to a patient as it is, but a pharmaceutical composition containing one or more of the above-mentioned compounds as an active ingredient is prepared using a formulation additive. However, it is preferable to administer to a patient in the form of a pharmaceutical composition. As the active ingredient of the medicament of the present invention, any hydrate may be used in addition to the free form compound included in the above general formula. The administration form of the medicament of the present invention is not particularly limited, and can be administered orally or parenterally. Examples of pharmacologically and pharmaceutically acceptable additives used in the production of the above pharmaceutical composition include excipients, disintegrants or disintegrants, binders, lubricants, coating agents, dyes, Diluents, bases, solubilizers or solubilizers, isotonic agents, pH adjusters, stabilizers, propellants, adhesives and the like can be mentioned.
[0016]
Examples of preparations suitable for oral administration include, for example, tablets, capsules, powders, fine granules, granules, liquids, syrups and the like. Examples of preparations suitable for parenteral administration include injections, instillations, suppositories, inhalants, or patches. For formulations suitable for oral administration, pharmacologically and pharmaceutically acceptable additives include, for example, excipients such as glucose, lactose, D-mannitol, starch, or crystalline cellulose; carboxymethylcellulose, starch, or Disintegrants or disintegration aids such as carboxymethylcellulose calcium; binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, or gelatin; lubricants such as magnesium stearate or talc; hydroxypropylmethylcellulose, sucrose, polyethylene glycol or Coating agents such as titanium oxide; bases such as petrolatum, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water, or hard fat can be used.
[0017]
Preparations suitable for transdermal or transmucosal administration include, for example, propellants such as Freon, diethyl ether, or compressed gas; adhesives such as sodium polyacrylate, polyvinyl alcohol, methyl cellulose, polyisobutylene, polybutene; cotton cloth or plastic sheet It can manufacture using additives for preparations, such as foundation cloth. Preparations suitable for injection or infusion include aqueous solutions such as distilled water for injection, physiological saline, and propylene glycol, or solubilizers or solubilizers that can constitute injectable solutions for use; glucose, sodium chloride, D-mannitol And an isotonic agent such as glycerin; an additive for pharmaceutical preparation such as a pH regulator such as an inorganic acid, an organic acid, an inorganic base or an organic base may be added.
[0018]
The antitumor agent of the present invention can be used for the treatment of solid cancers such as stomach cancer, lung cancer, liver cancer, kidney cancer, brain tumor, colon cancer, and esophageal cancer, or all types of cancer such as malignant lymphoma and leukemia. In particular, it is preferably used for the treatment of nerve tumors such as brain tumors. The neuronal differentiation inducer of the present invention is useful for the treatment and / or prevention of brain diseases such as senile dementia. However, the application target of the medicament of the present invention is not limited to the above, and may be used in combination with other drugs generally used for the treatment of malignant tumors and neurological diseases. The dosage of the antitumor agent and neuronal differentiation inducer of the present invention is not particularly limited, and may be appropriately selected depending on the administration form, age, weight, symptoms and the like. For example, in the case of nasal administration, 1 to 100 mg / Kg, preferably 2 to 10 mg / Kg as an active ingredient amount per day for an adult may be administered. The administration may be divided once or several times per day, and the administration period can be arbitrarily determined according to age, symptoms and the like.
[0019]
【Example】
EXAMPLES Hereinafter, although an Example demonstrates this invention further more concretely, the scope of the present invention is not limited to the following Example.
(A) Production Example Glucose (3.5%), soluble starch (1%), meat extract (0.5%), polypeptone (0.5%), yeast extract (0.3%), salt (0.2%), dipotassium phosphate (0.05%), Magnesium sulfate (0.05%), and soy flour (2%) medium (pH 5.8) was inoculated with Penicillium sp. BM1689-P and incubated at 28 ° C for 48 hours It was. 140 ml of this culture solution was inoculated into 18 liters of the medium having the same composition, and shaking culture was performed at 300 revolutions per minute while aeration of 7 liters per minute was performed at 28 ° C. for 50 hours.
[0020]
The culture broth was separated into cells and supernatant with a centrifuge (6,000 rpm), and the cells were extracted with 5 liters of 80% acetone. After concentration under reduced pressure, the resulting aqueous solution was adjusted to pH 7.0, and extraction was repeated twice with 5 liters of ethyl acetate. After extraction, all the ethyl acetate was combined and concentrated under reduced pressure to obtain 4.7 g of a brown syrup.
This syrup was dissolved in 10 ml of chloroform and infiltrated into a silica gel column (diameter 4 cm, length 60 cm) prepared with chloroform. First elute with 1 liter of chloroform, and then gradually change the mixing ratio of chloroform-methanol solution (100: 1, 100: 2, 100: 5, 100: 10, 100: 20, 100: 50) in 500 ml each. Elute and finally elute with 1 liter of methanol. The active ingredient was eluted in a fraction of chloroform-methanol solution (100: 5 to 100: 10). This fraction was concentrated under reduced pressure to obtain 1.6 g of a brown syrup.
[0021]
Further, 0.8 g of this brown syrup was dissolved in a chloroform-methanol solution (1: 1) and adsorbed on a Sephadex LH-20 (Pharmacia) column (diameter 2 cm, length 65 cm) prepared in the same solution. Elution was performed with chloroform-methanol solution (1: 1). 10 ml each was collected, and the main active fraction was collected and concentrated under reduced pressure to obtain 260 mg of brown syrup.
Next, fractionation by high performance liquid chromatography (80% methanol, flow rate 8 ml / min) was performed using an ODS column (diameter 2 cm, length 25 cm; capsule pack, manufactured by Shiseido Co., Ltd.). mg was obtained. Finally, a high-performance liquid chromatographic fractionation (60% methanol, flow rate 8 ml / min) was performed using an ODS column (diameter 2 cm, length 25 cm; capsule pack, manufactured by Shiseido Co., Ltd.). mg was obtained.
[0022]
1. Material properties: colorless and amorphous
2. Specific rotation: [α] _D +32 ° (26 ° C, c = 0.1, MeOH)
3.Molecular weight, molecular formula: MW = 389, C ₂₁ H ₂₇ NO ₆ by high resolution EI mass spectral analysis
4. Solubility: Easily soluble in methanol, ethanol and dimethyl sulfoxide. Insoluble in hexane and water
5. Color reaction: positive for sulfuric acid, ninhydrin, iodine
6. UV absorption spectrum: λ _max (MeOH) (ε): 232 nm (21800), 280 nm (sh, 15600)
7. Infrared absorption spectrum: ν (KBr): 3420, 2920, 1720, 1640, 1435, 1270, 1135, 970, 965
8. Nuclear magnetic resonance spectrum ( ¹ H-NMR and ¹³ C-NMR): in CD ₃ OD, ¹ H-NMR: 600 MHz, ¹³ C-NMR: 100 MHz, δ ppm, internal standard TMS
[0023]
[Chemical 3]

[0024]
[Table 2]

[0025]
(B) Test Example Mouse neuroblastoma cells Neuro 2A cells are induced to differentiate by dibutyryl cyclic AMP, a known neuronal differentiation inducer, and extend neurites (J. Antibiotics, 44, 113, 1991). . On the other hand, human neuroblastoma cells SH-SY5Y cells also formed dendrites peculiar to neuronal differentiation when dibutyryl cyclic AMP was added. Therefore, the epolaractaene of the present invention was examined for dendrite formation promoting action on human neuroblastoma cells SH-SY5Y cells by the above method. The results are shown in Table 3. In the table, the dendrite formation rate indicates the ratio of the number of dendrite-forming cells to the total number of cells.
[0026]
[Table 3]

【The invention's effect】
The epolactaene of the present invention is useful, for example, as an antitumor agent for use in the treatment of neurological tumors, and as a neuronal differentiation inducer for use in the treatment and / or prevention of neurological diseases. The novel microorganism and the production method provided by the present invention are useful for efficiently producing the above-mentioned epolactaene.

Claims (5)

The following formula:

Epolactaene indicated by Penicillium sp. Producing the epolactaene according to claim 1 FERM P-14859 stock. Penicillium sp. FERM P-14859 shares. The method for producing epolactaene according to claim 1, wherein Penicillium sp. FERM P-14859 strain is cultured, and epolactaene according to claim 1 is collected from the culture.  A neuronal differentiation-inducing agent comprising the epolactaene according to claim 1 as an active ingredient.