JPS635078B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing a new anti-cancer antibiotic, NCS-C. When Streptomyces carcinostaticus variant F41 Kuroya is cultured, the polymeric anticarcinogenic substance neocarcinostatin (Special Publication No. 42-21752),
It is already known that an N-1 fraction, an N-2 fraction (Japanese Patent Publication No. 47-680) and a Ma fraction (Japanese Patent Publication No. 13516-1988) can be obtained. The present inventors further investigated the culture and found that neocarzinostatin, N-1 fraction, N-2 fraction,
We discovered that there is a new anticancer antibiotic NCS-C, which has physical properties different from those of the Ma fraction and the Ma fraction, and that it can be obtained by decomposing neocarzinostatin under mild conditions. Completed the invention. That is, the present invention provides a method for producing the anticancer antibiotic NCS-C, which is characterized by extracting neocarzinostatin with a non-aqueous polar organic solvent under acidic conditions. Neocarzinostatin, the raw material of the present invention, is
Streptomyces carcinostaticus variant according to the method described in Japanese Patent Publication No. 42-21752.
F41 Kuroya (National Institute of Microbial Technology, Agency of Industrial Science and Technology,
Accession No. 2257; ATCC
No. 15945). The method of the present invention is carried out as follows. That is, when neocarzinostatin powder is suspended in a non-aqueous polar organic solvent such as methanol, ethanol, propanol, butanol, acetone, chloroform, etc. under acidic conditions and stirred, the NCS-C substance dissolved in the organic solvent and It is degraded to insoluble preneocarzinostatin (described in Journal of Antibiotics, Vol. 27, p. 766). Furthermore, the organic layer containing this NCS-C substance is concentrated,
Adsorption chromatography such as silica gel, alumina, Amberlite XAD, etc., reversed phase partition chromatography such as Microbondapak _C18 , Licrosolv RP-8, gel permeation chromatography such as Cephadex LH-20, TSK gel, etc. NCS-C substance can be obtained by combining and purifying. In this process, the NCS-C substance is obtained as an acid salt, and the hydrochloride exhibits the following physical properties. (1) Physicochemical properties (a) Properties Pale yellow to brown powder (b) Decomposition point 125℃ (c) Specific rotation [α] ²⁰ _D = -171° (c = 1.4 x 10 ^-3 , methanol) . (d) Molecular weight 686.5-688.55 (vapor pressure method) 695 (mass spectrometry) Figure 3 (e) Nuclear magnetic resonance spectrum ¹ H-NMR spectrum Figure 4 ¹³ C-NMR spectrum Figure 5 (f) Ultraviolet absorption spectrum ( Figure 1) 240, 265, 274, 290, 305, 330nm shoulder (G) Infrared absorption spectrum (Figure 2) 3400, 1780, 1610, 1400, 1190, 1080, 1010
It has an absorption at cm ^-1 . (H) Color reaction Permanganic acid reaction, diazo coupling reaction are positive, xanthoprotein reaction, Ehrlich reaction, ninhydrin reaction,
Orcinol reaction and Moritsch reaction are negative (li) Solubility Easily soluble in methanol, ethanol, propanol, and butanol, soluble in acetone, slightly soluble in chloroform and ethyl acetate, slightly soluble in water, ether, and benzene. (N) Stability Thermal stability: 60℃

【table】 Ultraviolet light: 20cm distance from Toshiba germicidal lamp GL-15

【table】 PH: 4 hours later

[Table] (2) Acutely toxic mouse intravenous injection LD ₅₀ = 1.8 mg/Kg Mouse peritoneal LD ₅₀ = 10 mg/Kg or more (3) Biological properties (i) Antitumor effect Yoshida sarcoma (ip-ip)

[Table] Hela S3 cells The NCS-C substance does not show antitumor effects at a concentration of 1.0 mg/ml, but compared with preneocarzinostatin.
A mixture of NCS-C substances at a ratio of 10:1
It showed antitumor effects at a concentration of 0.1 mg/ml. (ii) Antimicrobial action

【table】

[Table] As described above, the NCS-C substance of the present invention has anticancer and antimicrobial effects by itself, but its effects are significantly increased when used in combination with preneocarzinostatin. As mentioned above, the physicochemical and biological properties of the NCS-C substance of the present invention are that of neocarzinostatin, N-1 fraction, N-2 produced from Streptomyces carcinostaticus variant F41 Croya.
It is completely different from carcinostatin complex (Special Publication No. 35-5400) produced from the fraction, Ma fraction and the closely related strain Streptomyces carcinostaticus, as well as other anticancer antibiotics. Therefore, the NCS-C substance of the present invention is recognized as a new substance. Next, an example will be given and explained. Example 1 16 g of neocarzinostatin powder was suspended in 1.5 g of a 95% methanol solution, the pH was adjusted to 2.0 with 1N hydrochloric acid, and the suspension was stirred for 2 hours. After stirring, centrifuge
The supernatant was concentrated to dryness to obtain 920 mg of a crude powder of NCS-C substance. Example 2 920 mg of the crude powder obtained in Example 1 was added to 25 ml of methanol.
Dissolved in 1.0N hydrochloric acid:methanol (1:9) in advance
Chromatography was performed by passing it through a column packed with Sephadex LH-20 1 equilibrated with 1.0N hydrochloric acid:methanol (1:9) as an eluent, and the active fraction was concentrated to dryness. 105 mg of powder of material was obtained.

[Brief explanation of the drawing]

Figure 1 shows the ultraviolet absorption spectrum of the anticancer antibiotic NCS-C of the present invention, Figure 2 shows the infrared absorption spectrum of the same substance, Figure 3 shows the FAB mass spectrum of the same substance, and Figure 4 shows the same substance. ^1H -NMR of a substance
Figure 5 shows the ¹³ C-NMR spectrum of the same substance.

Claims (1)

[Claims] 1. A method for producing the anticancer antibiotic NCS-C, which comprises extracting neocarzinostatin with a non-aqueous polar organic solvent under acidic conditions.