JPH07506332A - Phosphate esters of heterocyclic compounds as medicines - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention]

Phosphate ester technology of heterocyclic compounds as medicine This invention provides a new biologically active compound (hereinafter referred to as WF11231A substance). related. More specifically, this invention describes the novel compound WF11231 which has an immune reaction suppressing effect. Substance A or its salt, its manufacturing method, and its usefulness as an immune reaction suppressing medium containing them Concerning their effective medicinal uses. Therefore, one purpose of this invention is to prevent rejection caused by transplantation and to prevent graft damage caused by bone marrow transplantation. WF, a new compound used to suppress host diseases, autoimmune diseases, etc.] 1231 The goal is to provide substance A and its salts. Another object of the present invention is that Clatopotrium (C1adoI) belongs to the genus Otrum. WF11231A substance-producing bacteria are cultured, and WF11231A is produced from the culture. An object of the present invention is to provide a method for producing a substance or a salt thereof. A further object of this invention is to use the WF11231A substance or its salt as an active ingredient. An object of the present invention is to provide a pharmaceutical composition comprising: Another object of this invention is to remove the WF11231A substance or its salt by transplantation. For prevention and treatment of absolute reactions, graft-versus-host disease caused by bone marrow transplantation, autoimmune diseases, etc. It is to provide access.

[Disclosure of the invention]

The WF11231A substance of this invention can be represented by the following chemical formula. The WF11231A substance (1) of this invention may have one or more stereoisomers such as optical isomers based on asymmetric carbon atoms, but these isomers also have this emission. Included in the scope of light. Suitable salts of WF11231A substance (I) are conventional non-toxic pharmaceutically acceptable salts. salts such as alkali metal salts (e.g., sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g., calcium salts, magnesium salts, etc.), salts with inorganic bases such as ammonium salts, organic amine salts (e.g., trimethylamine salts, trimethylamine salts, etc.); ethylamine salts, pyridine salts, picoline salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, etc.), organic acid salts (e.g. acetate salts, triflic acid salts, etc.), fluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate inorganic acid salts (e.g. hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, etc.), Salts with amino acids (eg, arginine, aspartic acid, glutamic acid, etc.), and the like. The WF11231A substance can be produced by fermenting a WF11231A substance-producing bacterium belonging to Clatopotrium sp. No. 11231 in a medium containing a nutrient source. The method of manufacturing WF11231A is based on the specific product shown here for illustrative purposes only. It should not be understood that it is limited to the use of objects (organisms). This invention includes the use of any variant, natural or artificial, capable of producing WF1 1231A material. Such variants can be produced from the aforementioned organisms by conventional means such as irradiation, ultraviolet radiation, N-methyl-N-nitrosoguanidine, 2-aminopurine, and the like. The details of Clatopotrium sp. No. 11231 are as follows. Mycological properties of strain No. 11231 Strain No. 11231 was originally isolated from litter collected in Iwaki City, Fukushima. The organism reproduced rapidly on a variety of media, forming yellow to red colonies. Strain No. 11231 formed deformed metamorphoses on various agar plates, including conidiophores with short filaments branching into almost whorls with regressively formed hyaline and conidia separated by septa. . In this case of conidia, the eggs were completely cracked. This strain did not produce long range mutant constructs. Due to its morphological characteristics, this strain is a ligated form of the fungus, Clatopotrium Neesex 5teud. 1924”’. be done. The characteristics of the surname are as described below. Culture characteristics on various agar media are summarized in Table 1. When the malt extract agar medium was widely spread and two weeks had passed at 25°C, the seeds had grown to a diameter of 8.5 cm or more. The surface of this colony was flat, felt-like, and reddish-yellow. The back is brown It was a deep yellow color. Conidial structures were observed. Colo on cornmeal agar Under the same conditions, Knee spread widely and spread to a diameter of 8.5 cm or more. The surface was flat, thin, and reddish white. The back was reddish white. Conidia Child structures were abundantly formed. Morphological characteristics were determined by mushroom agar (1). The conidial structure is filamentous, contains a single group, is transparent, smooth, and separated by septa. Conidiophores are fragile, transparent and smooth, often 1 mm long and up to 15 μm wide. The conidiophores are branched almost in whorls, with 2.3 branches wrapped around the main stem, and the primary branches bear fruit in whorls with 2 to 7 fertile cells. Conidial cells are straight, acupuncture-shaped cylinders 20-45 μm long, 3.0-6.0 μm wide, with a 1.0-3.0 μm wide tip on the base. It is slightly tapered towards the end and forms conidia in a retrograde manner. The conidia are separated by 1 to 3 septa, straight and smooth, with a rounded tip and a pointed tip measuring 18.0-28.5 x 7.0-11.0 μm on the i-rod towards the cylinder. There is a strong foundation. The viable hyphae are smooth, separated by septa, and transparent and branched. The hyphal cells are cylindrical and have a diameter of 2.0 to 8.0 μm. No. 11231 strain was incubated at a temperature range of 4°C to 32°C, with the optimum condition being 23°C to 26°C. I was able to reproduce. Data in this temperature range was confirmed on potato dextrose agar medium (by Ninosui). Clatopotrium (1) Genus No. 11231 According to the taxonomic standards of fungi, it is a family of fungi. It appears to be similar to the Clatopotorium stage of Dactylarioides G, Arnold 1971. death However, since this strain did not generate long-range structures, we It was named Latopotorium sp. No. 11231. And this stock is an industrial technology Deposited as FERM BP-3665 at the Institute of Microbial Technology (Higashi 1-1-3, Tsukuba City, Ibaraki Prefecture) (Deposit date: December 4, 1991) Table 1. No. 11231 strain culture characteristics Abbreviation 2 G: Growth rate, colony size measured by diameter S: Colony surface R: Back * MY20 agar medium: 5 g of peptone 1 3 g of yeast extract per water II. 3g of malt extract 1 200g of glucose and 20g of agar medium These characteristics last 14 days It was observed during cultivation at 25°C. Color descriptions are based on the Methuen color handbook (2). Reference: (1) De Hoog, G, S: Hyphomycetes, a fungus that reproduces Notes on Hyphomycete S and its related fungi, Palsoonia, 10 (1), pp. 33-81, 1978° (2) Cornerup, A and J, Il, Wanshi + - (W Anscher): Messue Color Handbook (3rd Edition), 525 pages, Messue London, 1978. Production of WF11231A substance This WF11231A substance is a WF11231A substance-producing bacterial strain belonging to the genus Radobotryum that contains nutrients containing carbon and nitrogen. It is produced when grown under aerobic conditions in a culture medium containing the same substance (e.g., shaking culture, deep culture, etc.). The medium composition includes carbohydrates such as glucose, sucrose, and starch as preferred carbon sources. Plum, fructose, glycerin, etc. are used. Nitrogen sources include yeast extract, peptone, gluten meal, cottonseed flour, soybean flour, and coke. bleach, dried yeast, wheat germ, etc., and ammonium salts as inorganic and organic nitrogen compounds (e.g. ammonium nitrate, ammonium sulfate, ammonium phosphate). urea, amino acids, etc.) are used. Carbon and nitrogen sources can be beneficially used in combination, but it is not necessary to use highly pure sources. There's no need. Less pure products have more evidence of growth factors and are richer in mineral nutrients. It is suitable for use because there are many. If necessary, add mineral salts to the mineral culture such as sodium, calcium carbonate, sodium phosphate, potassium phosphate, sodium chloride, or potassium chloride, sodium iodide, or potassium iodide, magnesium salts, copper lead, zinc salts, cobalt salts. etc. are added to the medium. If necessary, if there is significant foaming during culturing, add liquid parafine, fatty oil, vegetable oil, etc. An antifoaming agent such as oil or silicone may be added. Agitation and aeration of the culture mixture can be accomplished in a variety of ways, such as propeller or similar mechanical agitation, rotation, or shaking. . Cultivation is carried out at a temperature of about 10°C to 40°C, preferably 20°C to 30°C, for 50 to 150 hours, but this may vary depending on culture conditions and culture capacity. When the culture is completed, the WF11231A substance is separated from the liquid medium. Separation and purification are performed by means used in the production of general biologically active substances. In other words, a suitable solution It is separated and purified by solvent extraction, chromatography, or recrystallization from an appropriate solvent. WF11231A substance is separated or purified during the separation or purification of WF11231A substance. After production, the salt of the WF11231A substance can be obtained by a conventional method. Physical and chemical properties of the hydrochloride of the obtained WF11231A substance: Appearance: Colorless needle-like: Molecular formula: % Formula %: : : Ultraviolet absorption spectrum λ□, (methanol) + 225.275.285 (shoulder) nm Solubility: Easy to dissolve : Methanol poorly soluble Water insoluble: Acetone, ethyl acetate Color reaction: Positive: cerium sulfate, iodine vapor, ninhydrin Negative: Molish, Ehrlich Thin layer chromatography (TLC): High performance liquid chromatography (HPLC): Conditions Mobile phase Niacetonitrile: Water Nitrifluoroacetic acid (8:9220, 1, v/v) Column: YMCODS AM-303** (S-5, 120 people, 4.6 mm 1 D x 150 mm) Flow rate 1.0ml/min Detection: UV230nm Retention time near, 0 min **Trademark, manufactured by Yamamura Kagaku Kenkyusho Co., Ltd. Infrared absorption spectrum (KBr): 3350.3200.2960.2760.24 80.1660.1610.1510.1460.1430.1360.132 0.1290.1240.1180.1140.1080.1040.980.950 .940cm-' 1H nuclear magnetic resonance absorption spectrum (400MHz, CD, OD) δ□ 7.31 (2Hd, J = 8Hz), 6.88 (2Hd, J = 8Hz), 4.45 (IH, m) , 4.24-4.13 (2HCm). 3.84-3.76 (2H, m), 3.78 (3H, s), 3.59 (I H, m), 3.28 (IH, m), 3.03 (IH, mj, 2 .70 (3H, s), 2.60 (LH, m), 2.43 (LH, m), 2.28- 2.06 (6H, m), 1.89 (IH, m) Figure 1 As seen -13C nuclear magnetic resonance absorption spectrum (100MHz, CD, OD) δ0 160.4 (s), 131.7 (d) x2,128.7 (s), 115.3 (d) x2,70 .9 (d), 68.2 (s) A64.1 (d), 61° 6 (d), 55.8 (q), 54.9 (d), 51.8 (t), 42.9 (d), 41.9 (t), 34.0 (t), 32.R (q), 28.3 (t). 28.0 (t), 22.4 (t) seen in Figure 2 Is it the physical and chemical properties mentioned above? Therefore, the planar structural formula of the WF11231A substance is estimated as shown below. WF11231A substance (I) has pharmacological activities such as immune reaction suppressing activity. So Therefore, the heart, kidneys, liver, bone marrow, skin, cornea, lungs, pancreas, intestines, small intestines, limbs, and muscles. Rejection reactions due to transplants of organs and tissues such as meat and nerves; graft-versus-host diseases due to bone marrow transplants: rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, It is effective in treating and preventing immune-mediated diseases such as autoimmune diseases such as diabetes. Furthermore, WF11231A substance (I) has been shown to be effective in psoriasis, atopic dermatitis, contact dermatitis, as well as eczematous dermatitis, seborrheic dermatitis, young age, epidermolysis, water eruption, epidermolysis, and epidermolysis. Measles, angioedema, vasculitis, erythema, eosinophilia affecting the skin, erythema lupus, inflammatory and hyperproliferative skin diseases such as acne and alopecia areata, and immunologically mediated skin diseases; and various eye diseases such as autoimmune diseases (e.g. keratoconjunctivitis, Seasonal conjunctivitis, Behcet's disease-related enteritis, keratitis, keratoconus, dystrophic epithelial cornea, corneal vitiligo, optic ejaculation, Moren's ulcer, enterocolitis, and keratoconus. Regis disease, Vogt-Koyanagi-Harada syndrome, sarcoidosis, etc.); also reversible Conditions included in obstructive airway diseases include asthma (e.g., bronchial asthma, allergic asthma, reversible obstructive airway diseases such as bronchitis, particularly chronic or addictive asthma (e.g., late-onset asthma, airway hyperresponsiveness); also gastric ulcers, Vascular damage due to ischemic diseases and thrombosis, ischemic intestinal diseases, diseases caused by intestinal inflammation, enterocolitis causing necrosis, intestinal trauma related to burns, leukocyte triene B, vector-mediated diseases Inflammation of the mucous membranes and blood vessels, such as childhood steatorrhea, proctitis, gastroenteritis with eosinophilia, hypertrophy, Crohn's disease, ulcerative colitis, and other intestinal inflammations and allergies. : Food-related allergic diseases that manifest symptoms in areas far from the gastrointestinal tract, such as migraines, rhinitis, and eczema; and intestinal nephritis and gutpasture disease. Kidney diseases such as syndrome, hemolytic uremic syndrome, diabetic nephropathy; also polymyositis, Lanbarre syndrome, Meniere's disease, polyneuritis, mononeuritis multiplex, mononeuritis, neuropathy Nervous diseases such as transradicular inflammation; also endocrine diseases such as hyperthyroidism and Graves' disease; blood-related diseases such as cavities; and bone diseases such as osteoporosis. qi; also respiratory diseases such as sarcoidosis, lacrimal fibroma lung, idiopathic interstitial pneumonia, as well as dermatomyositis, vitiligo, ichthyosis vulgaris, sun allergy sensitivity, and T-cell lymph nodes that affect the skin. Skin diseases such as: also arterial lamina, atherosclerosis, aorta such as phlebitis syndrome, polyarteritis, and cardiomyopathy (e.g., autoimmune myocarditis, viral myocarditis). circulatory diseases such as scleroderma, Wegener's granuloma, and collagen diseases such as Siegren's syndrome; fatty degeneration; eosinophilic fasciitis; periodontal disease; nephrotic syndrome such as glomerulonephritis; androgenic alopecia, senile alopecia: muscular dystrophy; pyoderma, Sézary syndrome: Adi Also, organ damage (e.g., heart, kidneys, liver, gastrointestinal tract) due to ischemia-reperfusion that occurs, for example, during storage, transplantation, or ischemic disease (e.g., thrombosis or myocardial infarction). ROS-mediated diseases such as endotoxin-shock, pseudomembranous hyperplasia, Intestinal diseases such as colitis, colitis caused by drugs or radiation, acute kidney disease, chronic Kidney diseases such as kidney disease, as well as lung oxygen and medications (e.g. paraquat, pleomycin) poisoning, lung cancer, pulmonary diseases such as emphysema, visual organ diseases such as cataracts, cibrosis, retinitis, hyperpigmentation, deterioration of plaques due to aging, Vitrile scarring, corneal alkali burns; Dermatitis such as erythema plastica, IgA dermatitis, cement dermatitis; also gingivitis, periodontitis, septicemia, pancreatitis due to environmental pollution (e.g. air pollution), aging, carcinogenesis, cancer metastasis, altitude sickness. Other diseases such as; also histamine and leukemia Diseases caused by the release of bulbar triene C4: treatment and prevention of Behcet's disease of the intestines, blood vessels, nerves, and Behcet's disease of the oral cavity, skin, eyes, genitals, bite, epidermis, lungs, and kidneys. It can also be used for processing. Furthermore, WF11231A substance (I) has a liver regeneration effect and a stimulating effect on hyperplasia and hypertrophy of liver incision cells. Therefore, this substance may be used to treat immunogenic diseases (e.g., chronic autoimmune liver diseases such as autoimmune hepatitis, early biliary cirrhosis and schizocholial hepatitis), partial liver resection, acute liver necrosis (e.g., necrosis caused by toxins, and viruses). sexual hepatitis, shock Hepatitis B, non-A/non-B hepatitis, severe hepatitis as cirrhosis or liver weakness, late-onset liver disease, and liver function disease that has transitioned from acute to chronic (chronic liver function disease) It is useful in the treatment and prevention of liver diseases such as acute liver function diseases). Furthermore, WF11231A substance (1) has the effect of enhancing the effect of chemotherapy, For the prevention and treatment of Tomegao virus infection and pharmacological effects such as anti-inflammatory effects, etc. It is more useful for various diseases. Biological data is recorded below as an example showing the biological effects of the WF11231A substance. Describe. Test 1 Concanavalin A-WF1 in induced lymphocyte proliferation], 231A substance Quality Effects Spleen from Bulb/C rats were removed under aseptic conditions and gently dissociated into RPMI 1640 mice supplemented with penicillin (100 units/ml) and streptomycin (100 μg/ml). Cells 1. It was made into a medium sphere by centrifugation at OOOrpm for 5 minutes. The contained red blood cells are dissociated and washed with a lysing buffer containing ammonium chloride for 2 minutes at room temperature. The washed splenic cells were finally mixed with 10% fetoscopic lymph, 50 μM 2-mercabotethanol, and penicillin (100 units). nits/ml) and streptomycin (100 μg/ml). Add concanavalin A at 10 μg/ml. The cell suspension is immediately distributed into 96 round bottom microculture plates at 50 μm/tube. The hydrochloride of the WF11231A substance (hereinafter referred to as WF11231A-HCI) is dissolved in water. Furthermore, a thin layer of 50 μm/tube was added to the RPMi1640 medium. Add to the heavy tube to obtain the final concentration shown below. The media plates are incubated at 37° C. in air under a 5% carbon dioxide, 95% humid atmosphere for 72 hours. After incubation, lymphocyte proliferation was assessed using the MTT [3(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium potassium bromide] dye reduction assay. MTT was dissolved in 5 mg/ml phosphate buffer. 10 μl of MTT solution is added to all tubes in one assay and the plates are incubated at 37° C. for 4 hours. At the final stage of culture, remove 75 μm of culture medium, add isopropaturic acid (100 μm of isopropanol and 0.04N hydrochloric acid) to all tubes, and mix thoroughly to dissolve the dark blue crystals. match. Mix at room temperature for 10 minutes to ensure all crystals have melted, then plate. Read the sample using a microplate photometer using a test wavelength of 550 nm (630 nm wavelength for reference). The absorbance average of the triplicate tubes is calculated and the results are expressed as a percent (inhibition) of chemical reaction termination of the amount of dye reduction as follows: : Inhibition rate %=100 Δ/B X 100A Absorbance of test sample B, absorbance of control medium The results are shown in Table 2. WF11231A-HCI inhibits concanavalin A-induced lymphocyte proliferation Table 2 Effect of WF11231A substance on concanavalin A-induced lymphocyte proliferation WF11231A-HCI Inhibition ICS at 550 nm. 1.25 0.185±0.00370.70・625 0.177±0.00 1 72.00・3130.186 Sat 0.003 70.50.156 0.1 97±0.001 68.80・078 0.202±0.007 68.0 0.0270・039 0.259±0.020 58.90.020 0.3 90±0.049 38.20.010 0.609±0.045 3. 40 0.630±0.007 0 Ventral allografts from donor (Lewis) rats were implanted in the lateral thoracic position of host (F344) rats. Wound dressings are removed 5 days after skin grafting. 90% of transplanted parts The graft is examined daily until rejection, defined as epithelial necrosis. WF11231A-HCI was dissolved in physiological saline and administered intraperitoneally at doses of 0.0.1 and 0.3 mg/kg daily for 14 consecutive days, or at a dose of 1 mg/kg daily for 9 consecutive days. or at a dose of 3 mg/kg daily for 5 consecutive days. Treat. Start from the day you transplant it. As seen in Table 3, all skin allografts that received physiological saline intraperitoneally in mice exhibited rejection within 11 days. On the other hand, treatment with WF11231A-HCI at doses of 1 and 3 mg/kg prolonged skin allograft survival. Numbers in parentheses indicate the number of days it took for the host mouse to die from the active allograft. Table 3 Skin allograft survival effect of WF11231A-HCI Treatment Dose Number of animals Skin allograft survival WF11231A 0.1 3 91010 The pharmacological compositions of the invention can be used in the form of pharmacological formulations. For example, a solid or semi-solid containing the WF11231A substance or its pharmacologically acceptable salt as an active ingredient and accompanied by an organic or inorganic base or binder suitable for external, internal, or parenteral administration; Can be used in liquid form. The active ingredient may be used, for example, in tablets, granules, capsules, herbal medicines, solutions, emulsions, suspensions, injections, ointments, liniments, eye drops, etc. in the usual non-toxic pharmacologically acceptable bases. It can be used as a lotion, gel, cream, or any other suitable form. Available bases include water, glucose, lactose, acacia, gelatin, and manila. Tall, starch paste, magnesium trisilicate, talc, cornstarch and keratin Chin, cornstarch, colloidal silica, urea and other bases suitable for production in solid, semi-solid and liquid form, as well as additives, stabilizers, thickeners, solubilizers, colorants and the like. You can also use fragrances. To administer this composition to humans, intravenous, intramuscular, local, or oral administration is recommended. It is preferable to use Therapeutically, the effective dose of the substance WF11231A varies depending on the age and condition of the individual patient being treated, so in the case of treatment of individual patients, when administered intravenously, 0 per 1 kg of human body weight. 0.01 to 10 mg of WF11 231A substance is administered daily; for intramuscular injection, 0.1 to 10 mg of WF11231A substance is administered daily per kg of human body weight, and for oral administration, 0.01 to 10 mg of WF11231A substance per kg of human body weight is administered daily. .5 to 50 mg of WF] 1231A material is typically given. The invention will be explained in more detail with the following examples. Roasted I, [LL (1) Fermentation: 4% Sanoiroku loin, 296% cottonseed flour, 1% dry yeast, 0.2% potassium dihydrogen phosphate, 0.2% calcium carbonate and 800% Tween. Contains 196 Seed medium (120 ml) of transparent tissue was added to 20 500 ml Erlenmeyer flasks. Pour into each lask and sterilize at 121°C for 30 minutes. Platinum-eared Kratopotli Sp. No. 11231 is inoculated from a slant and poured into each flask. child Shake the flask on a rotary stirrer at 25°C for 3 days. Inoculate 150 liters of sterile production medium in a 200 liter jar fan meter with the Elareta seed culture. Ru. This medium contains 4% of various starches, 1% glucose, 2% cottonseed flour, 2% soybean flour, 1% ammonium phosphate, 0.7% sodium hydrogen phosphate decahydrate, and sulfur. Contains 0.001% of zinc acid heptahydrate, 0.025% of Adekanol LG109 (antifoaming agent manufactured by Asahi Denka Co., Ltd.) and 0.025% of silicone KM70 (manufactured by Shin-Etsu Chemical Co., Ltd.). nothing. Fermentation is carried out at 25° C. for 4 days with aeration of 100 liters per minute and stirring at 200 rpm. The amount of WF11231A in the fermentation fluid medium was determined in vitro for its immunosuppressive effect. Determined by concanavarin A-induced lymphocyte proliferation and HPLC analysis. (2) Isolation The cultured fluid medium (1601J Sotolu) was filtered with the aid of diatomaceous earth (1 kg). Ru. Discard the filtrate. Mix 80 liters of methanol with the hard precipitate of mycelium. Add it completely. The mixture is allowed to sit for one hour and filtered. 20 liters of filtrate under reduced pressure! - Concentrate the solution to a volume of 100 liters, and apply the liquid to a column (6 liters) using an adsorbent of 5EPABEADS SP-207 (trade name, manufactured by Mitsubishi Chemical Corporation). Wash the column with 18 liters of water, 18 liters of 50% methanol, and 18 liters of 0.14% aqueous solution. Elute with 50% methanol water containing ammonium hydroxide. The eluate is concentrated to 10 liters under reduced pressure. Adjust the pH to 3.0 with IN hydrochloric acid and then click the aqueous solution. Apply to a column (0.35 liters) using on-exchange resin Diaion 5K-IB (NH4" type, product name manufactured by Mitsubishi Kasei Corporation). Wash the column with 3.5 liters of water, and add 0.IN ammonium water. eluted with oxides Ru. Ammonium hydroxide was removed under reduced pressure, and the solution was poured into a column (0.12 liters) using an anion exchange resin DOWEX 1-X2 (OH type, trade name, manufactured by Mitsubishi Chemical Corporation). tor). Wash the column with 3.5 liters of water and elute with 0.6 liters of 0.025N acetic acid and 1 liter of 0.25N acetic acid. Evaporate the solvent to obtain a coarse powder. The obtained coarse powder was further mixed with YMC gel (ODS-AM 120-350.1.5 cm 1D 50 cm manufactured by Yamamura Kagaku Kenkyujo Co., Ltd.), mobile phase, acetonitrile-water-trifluoroacetic acid. Subject to pre-prepared HPLC using a mixture of acids (6:94:0.1, v/v). The eluate is monitored with a UV detector at 230% m, and the appropriate fractions are combined and lyophilized. Freeze-dried powder to obtain 5 ml of 97.5% acetonitrate (130 mg). Brief description of the drawings Figure 1. 'H-NMR spectrum of WF11231A hydrochloride 2. "C-NMR spectrum diagram of WF11 231A hydrochloride" Figure 2 Continuation of front page (72) Inventor Okuhara Kingdom 2-14-10 Umezono, Tsukuba City, Ibaraki System

Claims (1)

[Claims] 1. formula ▲ Contains mathematical formulas, chemical formulas, tables, etc. ▲ A WF11231A substance represented by or a salt thereof. 2. 2. The compound according to claim 1, which is a hydrochloride of substance WF11231A. 3. WF11231A belonging to the genus Cladobtryum The special method involves cultivating substance-producing bacteria and collecting the WF11231A substance from the culture. A method for producing a WF11231A substance or a salt thereof. 4. Pharmaceutical composition containing WF11231A substance or its salt5. WF11231A Use of the substance or its salts as a medicine. 6. Do not administer the WF11231A substance or its salt to humans or animals. transplant rejection, graft-versus-host disease caused by bone marrow transplantation, and autoimmune disease. Prevention and treatment methods. ▲ Contains mathematical formulas, chemical formulas, tables, etc. ▲