WO1992011275A1

WO1992011275A1 - 13-dimethyl fr-900506 derivative and the use as an immunosuppressive agent

Info

Publication number: WO1992011275A1
Application number: PCT/JP1991/001726
Authority: WO
Inventors: Kanji Hayami; Kazuto Yonetani; Takeshi Kawagita
Original assignee: Fujisawa Pharmaceutical Co., Ltd.
Priority date: 1990-12-19
Filing date: 1991-12-18
Publication date: 1992-07-09
Also published as: JPH06503327A; GB9027471D0

Abstract

A compound of formula (I) is disclosed. And a process for its production, composition containing it, and its use as immunosuppressive agent are also described.

Description

DESCRIPTION 13-Demethyl FR-900506 derivative and the use as an immunosuppresslve agent

This invention relates to a novel compound having pharmacological activities, hereinafter entitled FR-901429 substance, to a process for its production, to a pharmaceutical composition containing the same and to a use thereof.

More particularly, it relates to a novel compound, FR-901429 substance which has pharmacological activities such as immunosuppressive activity, antimicrobial activity, and the like, to a process for its production, to a

pharmaceutical composition containing the same and to a use thereof as a medicament.

Accordingly, one object of this invention is to provid the novel compound, FR-901429 substance which is of use for treating and preventing rejection by transplantation, graft-versus-host diseases by medulla ossium

transplantation, autoimmune diseases, infectious diseases, and the like.

Another object of this invention is to provide a process for production of the FR-901429 substance by fermen tation of a FR-901429 substance-producing strain belonging to the genus Streptomyces in a nutrient medium.

A further object of this invention is to provide a pharmaceutical composition containing, as an active

ingredient, the FR-901429 substance.

Still further object of this invention is to provide a use of the FR-901429 substance for treating and preventing rejection by transplantation, graft-versus-host diseases by medulla ossium transplantation, autoimmune diseases, infectious diseases, and the like. This invention is originated from the newly discovered

FR-900506 substance [chemical name: 17-allyl-1,14-dihydroxy- 12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25- dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo- [22.3.1.0^4,9]octacos-18-ene-2,3,10,16-tetraone] which was newly isolated in pure form from culture broth obtained by fermentation of new species belonging to genus Streptomyces.

Based on the discovery of the FR-900506 substance, the inventors of the present invention studied to produce the FR-900506 substance by fermentation in high yield, and during its study they have found that the FR-901429

substance has also been produced in the culture broth.

The physical and chemical properties of the FR-901429 substance of the present invention, its production and its pharmacological activities are explained hereinbelow.

(continued to the next page)

PHYSICAL AND CHEMICAL PROPERTIES OF

FR-901429 SUBSTANCE

The FR-901429 substance of the present invention sses the following physical and chemical properties,

(1) Form and Color: white powder

(2) Mass Spectrum:

FAB-MS : m/z 812 (M+Na)⁺ HRFAB-MS: 812.4584

(3) Molecular Formula:

C₄₃H₆₇NO₁₂

(4) Specific Rotation:

[α]_D : -79.6° (c=1.1, CHCl₃) (5) Infrared Absorption Spectrum: CHCl₃·3570' 3500, 2940, 2860, 2820, 1730, 1640, vmax 1450, 1380, 1280, 1260, 1190, 1170, 1090,

910, 860 cm^-1 (6) ¹³C Nuclear Magnetic Resonance Spectrum: δ(ppm, CDCl₃, 100MHz):

212.2 (s) 194.9 (s) 169.3 (s)

212.1 (s), 194.2 (s), 168.9 (s). 165.3 (s) 139.9 (s) 135.5 (d)

164.3 (s), 139.7 (s), 135.3 (d),

132.9 (s) 132.1 (d) 122.7 (d)

132.3 (s), 131.4 (d), 122.3 (d),

116.7 (t) 98.8 (s) 84.2 (d)

116.5 (t), 97.9 (s), 84.1 (d),

76.8 (d) 76.2 (d) 73.7 (d)

76.4 (d), 75.3 (d), 73.6 (d),

73.5 (d) 72.7 (d) 65.7 (d)

73.4 (d), 72.4 (d), 65.2 (d),

57.5 (q) 56.39 (q) 56.2 (q)

57.1 (g), 56.36 (q), 56. 0 (q),

52.9 (d) 49.3 (t) 47.0 (t)

52.7 (d), 48.9 (t), 46.7 (t),

44.2 (t) 39.3 (t)

39.6 (t), 38.9 (t)

34.85 (d) , 34.5 (t) 34.4 (d)

34.83 (d), 34. 4 (t) 34.3 (d).

33.8 (t) 32.3 (t)

32.6 (t), (31.5 (t)

31.2 (t) 30.4 (t) 27.2 (t)

31.2 (t), 30.3 (t), 26.3 (t),

26.1 (d) 24.6 (t) 20.9 (t),

25.9 (d), 24.4 (t), 20.7 (t),

19.9 (q) 16.2 (q) 15.7 (q)

19.5 (q), (15.8 (q). 15.6 (q),

12.38 (q)

12.36 (q), the chart of which being shown in Figure 1,

(7) H Nuclear Magnetic Resonance Spectrum: the chart of which being shown in Figure 2, (8) Thin Layer Chromatography: Developing

Stationary Phase Solvent Rf Values silica gel plate n-hexane and ethyl

acetate (1:2) 0.23 silica gel plate n-hexane and acetone

(2:1) 0.21

(9) Property of the Substance: neutral substance

With regard to the FR-901429 substance, it is to be noted that in case of measurement of 13C nuclear magnetic resonance spectrum, this substance showed pairs of the signals on the various chemical shifts, but in case of the measurements of the thin layer chromatography, the FR-9014 substance showed a single spot.

From the above physical and chemical properties, the FR-901429 substance is inferred to have the following plan structural formula.

PRODUCTION OF FR-901429 SUBSTANCE

The FR-901429 substance of this invention can be produced by fermentation of a FR-901429 substance-producing strain belonging to the genus Streptomyces such as

Streptomyces tsukubaensis No. 9993 in a nutrient medium.

A lyophilized sample of the Streptomyces tsukubaensis No. 9993 was deposited with the Fermentation Research

Institute, Agency of Industrial Science and Technology (No. 1-3, Higashi 1-chome, Tsukuba-shi, Ibaraki Prefecture, Japan) under the number of FERM P-7886 (deposited date:

October 5th, 1984), and was converted to the Budapest Treaty route of the same depository on October 19, 1985 under the new deposit number of FERM BP-927.

It is to be understood that the production of the novel FR-901429 substance is not limited to the use of the particular organism described above, which is given for the illustrative purpose only. This invention also includes the use of any mutants which are capable of producing the

FR-901429 substance including natural mutants as well as artificial mutants which can be produced from the.described organism by conventional means such as irradiation of

X-rays, ultra-violet radiation, treatment with

N-methyl-N'-nitro-N-nitrosoguanidine, 2-aminopurine, and the like.

In general, the FR-901429 substance can be produced by culturing the FR-901429 substance-producing strain in an aqueous nutrient medium containing sources of assimilable carbon and nitrogen, preferably under aerobic conditions (e.g. shaking culture, submerged culture, etc.). The preferred sources of carbon in the nutrient mediu are carbohydrates such as glucose, xylose, galactose, glycerin, starch, dextrin, and the like. Other sources whi may be included are maltose, rhamnose, raffinose, arabinos mannose, salicin, sodium succinate, and the like.

The preferred sources of nitrogen are yeast extract, peptone, gluten meal, cottonseed meal, soybean meal, corn steep liquor, dried yeast, wheat germ, feather meal, peanut powder etc., as well as inorganic and organic nitrogen compounds such as ammonium salts (e.g. ammonium nitrate, ammonium sulfate, ammonium phosphate, etc.), urea, amino acid, and the like. The carbon and nitrogen sources, though advantageously employed in combination, need not be used in their pure form, because less pure materials which contain traces of growth factors and considerable quantities of mineral nutrients, are also suitable for use. When desired, there may be added to the medium mineral salts such as sodium or calcium carbonate, sodium or potassium phosphate, sodium or potassium chloride, sodium or potassium iodide, magnesium salts, copper salts, cobalt salt and the like. If .necessary especially when the culture medium foams seriously, a defoaming agent, such as liquid paraffin, fatty oil, plant oil, mineral oil or silicone may be added.

As the conditions for the production of the FR-901429 substance in massive amounts, submerged aerobic cultural conditions are preferred therefor. For the production in small amounts, a shaking or surface culture in a flask or bottle is employed. Furthermore, when the growth is carried out in large tanks, it is preferable to use the vegetative form of the organism for inoculation in the production tanks in order to avoid growth lag in the process of production of the FR-901429 substance. Accordingly, it is desirable first to produce a vegetative inoculum of the organism by

inoculating a relatively small quantity of culture medium with spores or mycelia of the organism and culturing said inoculated medium, and then to transfer the cultured vegetative inoculum aseptically to large tanks. The medium, in which the vegetative inoculum is produced, is

substantially the same as or different from the medium utilized for the production of the FR-901429 substance.

Agitation and aeration of the culture mixture may be accomplished in a variety of ways. Agitation may be provided by a propeller or similar mechanical agitation equipment, by revolving or shaking the fermentor, by various pumping equipment or by the passage of sterile air through the medium. Aeration may be effected by passing sterile air through the fermentation mixture.

The fermentation is usually conducted at a temperature between about 20°C and 40°C, preferably 25-35°C, for a period of about 50 hours to 250 hours, preferably 100 hours to 200 hours which may be varied according to fermentation conditions and scales. Thus produced FR-901429 substance can be recovered from the culture medium by conventional means which are commonly used for the recovery of other known biologically active substances. The FR-901429 substance produced is found in the cultured mycelium and filtrate, and accordingly the

FR-901429 substance can be isolated and purified from the mycelium and the filtrate, which are obtained by filtering or centrifuging the cultured broth, by a conventional method such as concentration under reduced pressure,

lyophilization, extraction with a conventional solvent, pH adjustment, treatment with a conventional resin (e.g. anion or cation exchange resin, non-ionic adsorption resin, etc. treatment with a conventional adsorbent (e.g. activated charcoal, silicic acid, silica gel, cellulose, alumina, etc.), crystallization, recrystallization, and the like.

PHARMACOLOGICAL ACTIVITIES OF THE FR-901429 SUBSTANCE

The FR-901429 substance possesses pharmacological activities such as immunosuppressive activity, antimicrobial activity, and the like, and therefore are useful for the treatment and prevention of immune-mediated diseases such as the resistance by transplantation of organs or tissue such as heart, kidney, liver, medulla ossium, skin, cornea, lung, pancreas, intestinum tenue, limb, muscle, nervus, etc.;

graft-versus-host diseases by medulla ossium

transplantation; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's

thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, and the like; and further infectious diseases caused by pathogenic microorganisms.

Further, the FR-901429 substance is also useful for the treatment and the prophylaxis of inflammatory and

hyperproliferative skin diseases and cutaneous

manifestations of immunologically-mediated illnesses, such as, psoriasis, atopical dermatitis, contact dermatitis and further eczematous dermatitises, seborrhoeis dermatitis, Lichen planus. Pemphigus, bullous Pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Lupus erythematosus, acne and

Alopecia areata;

various eye diseases such as autoimmune diseases and so on (e.g. keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, conical cornea, dystrophia epithelialis corneae, corneal leukoma, ocular pemphigus, Mooren's ulcer,

Scleritis, Graves' ophthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, etc.);

reversible obstructive airways disease, which includes conditions such as asthma ( e.g. bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma and dust asthma ), particularly chronic or inveterate asthma (e.g. late asthma and airway hyper-responsiveness), bronchitis and the like;

inflammation of mucosa and blood vessels such as gastric ulcers, vascular damage caused by ischemic diseases and thrombosis, ischemic bowel disease, inflammatory bowel disease, necrotizing enterocolitis, intestinal lesions associated with thermal burns, leukotriene B₄-mediated diseases;

intestinal inflammations/allergies such as Coeliac disease, proctitis, eosnophilic gastroenteritis, mastocytosis,

Crohn's disease and ulcerative colitis;

food related allergic diseases which have symptomatic manifestation remote from the gastro-intestinal tract, for example migraine, rhinitis and eczema;

renal diseases such as interstitial nephritis,

Goodpasture's syndrome, hemolytic-uremic syndrome and diabetic nephropathy;

nervous diseases such as multiple myositis,

Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple neuritis, mononeuritis and radiculopathy;

endocrine diseases such as hyperthyroidism and

Basedow's disease;

hematic diseases such as pure red cell aplasia,

aplastic anemia, hypoplastic anemia, idiopathic

thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia and anerythroplasia; bone diseases such as osteoporosis;

respiratory diseases such as sarcoidosis, fibroid lung and idiopathic interstitial pneumonia;

skin diseases such as dermatomyositis, leukoderma

vulgaris, ichthyosis vulgaris, photoallergic sensitivity and cutaneous T cell lymphoma;

circulatory diseases such as arteriosclerosis,

atherosclerosis, aortitis syndrome, polyarteritis nodosa and myocardosis;

collagen diseases such as scleroderma, Wegener's

granuloma and Sjogren's syndrome;

adiposis;

eosinophilic fasciitis;

periodontal disease such as lesion of gingiva, periodontium, alveolar bone, substantia ossea dentis;

nephrotic syndrome such as glomerulonephritis;

male pattern alopecia or alopecia senilis;

muscular dystrophy;

Pyoderma and Sezary's syndrome;

Addison's disease;

active oxgen-mediated diseases, for example, organ injury such as ischemia-reperfusion injury of organs

(e.g. heart, liver, kidney, digestive tract) which occurs on preservation, transplantation or ischemic diseases (e.g.

thrombosis, cardiac infarction): intestinal diseases such as endotoxin-shock, pseudomembranous colitis, colitis caused by drug or radiation: renal diseases such as ischemic acute renal insufficiency, chronic renal insufficiency: pulmonary diseases such as toxinosis caused by lung-oxygen or drug (e.g. paracort,bleomycins), lung cancer, pulmonary

emphysema: ocular diseases such as cataracta, siderosis, retinitis, pigmentosa, senile macular degeneration, vitreal scarring, corneal alkali burn: dermatitis such as erythema multiforme, linear IgA ballous dermatitis, cement

dermatitis: and others such as gingvatis, periodontitis. sepsis, pancreatitis, diseases caused by environmental pollution(e.g. air pollution), aging, carcinogenis,

metastasis of carcinoma, hypobaropathy;

diseases caused by histamine or leukotriene C₄ release;

Behcet's disease such as intestinal-, vasculo- or

neuro-Behcet ' s disease , and also the one which affects oral cavity, skin, eye, vulva, articulation, epididymis, lung, kidney; and so on. And further, the FR-901429 substance has liver

regenerating activity and/or activities of stimulating hypertrophy and hyperplasia of hepatocytes. Therefore, it is useful for the treatment and prevention of hepatic diseases such as immunogenic diseases (e.g. chronic autoimmune liver diseases such as the group consisting of autoimmune

hepatitis, primary biliary cirrhosis and sclerosing

σholangitis), partial liver resection, acute liver necrosis (e.g. necrosis caused by toxins, viral hepatitis, shock or anoxia), B-virus hepatitis, non-A/non-B hepatitis, cirrhosis (e.g. alcoholic cirrhosis) and hepatic failure such as fulminant hepatic failure, late-onset hepatic failure and "acute-on-chronic" liver failure (acute liver failure on chronic liver diseases). And further, the FR-901429 substance is useful for various diseases because of its useful pharmaceutical activity such as augmenting activity of chemotherapeutic effect, preventing or treating activity of cytomegalovirus infection, anti-inflammatory activity, and so on.

The pharmaceutical composition of this invention can be used in the form of a pharmaceutical preparation, for example, in solid, semisolid or liquid form, which contains the FR-901429 substance, as an active ingredient, in admixture with an organic or inorganic carrier or excipient suitable for external, enteral or parenteral applications. The active ingredient may be compounded, for example, with the usual non-toxic, pharmaceutically acceptable carriers for tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions, injections, ointments, liniments, eye drops, lotion, gel, creme and any other form suitable for use. The carriers which can be used are water, glucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloida silica, potato starch, urea and other carriers suitable for use in manufacturing preparations, in solid, semisolid, or liquid form, and in addition auxiliary, stabilizing,

solubilizing, thickening and coloring agents and perfumes may be used. The active object compound is included in the pharmaceutical composition in an amount sufficient to produce the desired effect upon the process or condition of diseases. For applying this composition to human, it is

preferable to apply it by intravenous, intramuscular, topical or oral administration. While the dosage of

therapeutically effective amount of the FR-901429 substance varies from and also depends upon the age and condition of each individual patient to be treated, a daily dose of about 0.01-1000 mg, preferably 0.1-500 mg and more preferably 0.5-100 mg, of the active ingredient is generally given for treating diseases, and an average single dose of about 0.5 mg, 1 mg, 5 mg, 10 mg, 50 mg, 100 mg, 250 mg and 500 mg is generally administered.

The following examples are given for the purpose of illustrating the present invention. Example 1

Fermentation A culture medium (100 ml) containing corn starch (1%), glycerin (1%), glucose (0.5%), cottonseed meal (1%), dried yeast (0.5%), corn steep liquor (0.5%) and calcium carbonate (0.2%) (adjusted to pH 6.5) was poured into each of eight 500 ml-Erlenmeyer flasks and sterilized at 120°C for 30 minutes. A loopful of slant culture of Streptomyces

tsukubaensis No.9993, FERM P-7886 was inoculated to each of the media and cultured at 30°C for 72 hours on a rotary shaker. The seed culture was transferred to the same seed medium (160 liters) added Adekanol (defoaming agent. Trade Mark, maker; Asahi Denka Co.) (0.05%) and Silicone

(Shinetsu Chemical Co.) (0.05%), in a 200 liter

jar-fermentor which had been sterilized at 120°C for 30 minutes in advance. After the culture was performed at 30°C for 48 hours under aeration of 160 liters/minute and

agitation at 200 rpm, 30 liters of the preculture was inoculated to a production medium (3000 liters) containing soluble starch (3%), wheat germ (0.8%), dried yeast (0.4%), corn steep liquor (0.6%), calcium carbonate (0.1%), Adekanol (0.05%) and Silicone (0.05%) at pH 6.8 in a 4 ton tank, which had been sterilized at 120°C for 30 minutes in

advance. The fermentation was carried out at 25°C for 168 hours under aeration of 1500 liters/minute and agitation at 140 rpm.

Isolation and Purification About 7 m³ of the fermentation broth thus obtained was mixed with acetone (7 m³) and perlite (150 kg). The mixture was filtered using diatomaceous earth as a filter aid.

3

About 15 m of the filtrate was passed through a column containing non-ionic adsorption resin "Diaion HP-20"

(trademark, maker: Mitsubishi Chemical Industries) (2 m³). After washing with 50 % aqueous acetone (4 m³), the elution was carried out with 75 % aqueous acetone (4 m³). The eluate was diluted with water to make a concentrate of acetone to 30 v/v % and passed through a column containing activated carbon (500 liters). After washing with 30 % aqueous acetone (1 m³) and water (1 m³), the elution was carried out with ethyl acetate (1 m³). The eluate was concentrated in vacuo to give an oily residue (15 liters) and dehydrated with anhydrous sodium sulfate. After the concentrate was mixed with two times volume of n-hexane, the mixture was subjected to column chromatography of silica gel (70-230 mesh) (200 liters). The column was developed witha mixture of n-hexane and ethyl acetate (1:1 v/v, 200 liters and 1:2 v/v, 1200 liters). The fractions containing the desired compound were collected and concentrated in vacuo. To this concentrate was added ethyl alcohol (70 liters) and then FR-900506 substance was precipitated by slow addition of water (100 liters). The obtained mother liquor (170 liters) with separation of FR-900506 substance crystals contained about 25 g of FR-901429 substance. 30 Liters of mother liquor was concentrated to dryness in vacuo and then the residue was dissolved in ethyl acetate (1 liter). FR-900506 substance was precipitated by addition of n-hexane (3 liters). After separation of FR-900506 substance crystals, the mother liquor was concentrated to dryness in vacuo and the residue was dissolved in 100 ml of ethyl acetate. After addition of 250 ml of n-nexane, the mixture was applied onto silica gel (70-230 mesh) (2.2 liters). Development was carried out with the mixture of n-hexane and ethyl acetate (1:1 v/v, 6.6 liters and 1:2 v/v, 22 liters). The effluent was fractionated each 500 ml.

Fractions containing the desired compound (elution volume from 17.5 to 18.5 liters) were collected and concentrated to dryness in vacuo. The obtained residue (737 mg) was

dissolved in the mixture of n-hexane and ethyl acetate (2:1 v/v, 35 ml). The solution was applied onto Lobar column Si60 (Trademark, maker: Merck & Co.) (350 ml) packed with the same solvent. Development was carried out with the mixture of n-hexane and ethyl acetate (2:1 v/v, 4.5 liters and 1:1 v/v, 1.8 liters). The effluent was fractionated each 15 ml. Fractions containing the desired compound

(elution volume from 3.9 to 4.2 liters) were collected and concentrated to dryness in vacuo. The obtained residue was dissolved in chloroform (35 ml) and applied onto Lobar column Si60 (350 ml) packed with the same solvent.

Development was carried out with chloroform (350 ml) and the mixture of chroloform and isopropanol (30:1 v/v, 1050 ml). The effluent was fractionated each 15 ml. Fractions

containing the desired compound (elution volume from 540 to 570 ml) were collected and concentrated to dryness in vacuo. The obtained residue (73 mg) was dissolved in 60 % aqueous acetonitrile (10 ml) and applied onto Lobar column RP-18 (Trademark, maker: Merck & Co.) (80 ml) conditioned with the same solvent. Development was carried out with 60 % aqueous acetonitrile (800 ml). The effluent was fractionated each 8 ml. Fractions containing the desired compound (elution volume from 560 to 660 ml) were collected and extracted with 100 ml of ethyl acetate three times. The organic layer was collected and dehydrated with anhydrous magnesium.sulfate. After filtration, the extract was concentrated to dryness in vacuo to give 33 mg of the purified FR-901429 substance in a form of white powder.

Brief Description of Drawings Figure 1 shows ¹³C Nuclear Magnetic Resonance Spectrum of the FR-901429 substance.

Figure 2 shows ¹H Nuclear Magnetic Resonance Spectrum of the FR-901429 substance.

Claims

1. FR-901429 substance possessing the following physical an chemical properties:

(1) Form and Color: white powder

(2) Mass Spectrum: FAB-MS : m/z 812 (M+Na)⁺

HRFAB-MS: 812.4584

(3) Molecular Formula:

C₄₃H₆₇NO₁₂

(4) Specific Rotation: [α]_D : -79.6° (c=1.1, CHCl₃)

(5) Infrared Absorption Spectrum: CHCl₃·3570, 3500, 2940, 2860, 2820, 1730, 1640, vmax

1450, 1380, 1280, 1260, 1190, 1170, 1090,

910, 860 cm^-1

(6) ¹³C Nuclear Magnetic Resonance Spectrum: δ(ppm, CDCl₃, 100MHz):

212.2 (s) 194.9 (s) 69.3 (s)

212.1 (s), 194.2 (s), 68.9 (s),

165.3 (s) 139.9 (s) 135.5 (d) 164.3 (s), 139.7 (s), 135.3 (d), 132.9 (s) 132.1 (d) 122.7 (d) 132.3 (s), 131.4 (d), 122.3 (d), 116.7 (t) 98.8 (s) 84.2 (d) 116.5 (t), 97.9 (s), 84.1 (d),

76.8 (d) 76.2 (d) 73.7 (d)

76.4 (d), 75.3 (d), (73.6 (d), 73.5 (d) 72.7 (d) 65.7 (d) 73.4 (d), 72.4 (d), 65.2 (d),

57.5 (q) 56.39 (q) 56.2 (q) 57.1 (q), 56.36 (q), 56.0 (q),

52.9 (d) 49.3 (t) 47.0 (t) 52.7 (d), 48.9 (t), 46.7 (t), 44.2 (t) 39.3 (t)

39.6 (t), (38.9 (t),

34.85 (d) 34.5 (t) 34.4 (d) 34.83 (d), 34.4 (t), 34.3 (d), 33.8 (t) 32.3 (t) 32.6 (t), 131.5 (t),

31.2 (t) 30.4 (t) 27.2 (t) 31.2 (t), 30.3 (t), 26.3 (t), 26.1 (d) 24.6 (t) 20.9 (t) 25.9 (d), 24.4 (t), 20.7 (t), 19.9 (q) 16.2 (q) 15.7 (q) 19.5 (q), 15.8 (q), 15.6 (q), 12.38 (q)

12.36 (q), the chart of which being shown in Figure 1,

(7) ¹H Nuclear Magnetic Resonance Spectrum: the chart of which being shown in Figure 2, (8) Thin Layer Chromatography:

Developing

Stationary Phase Solvent Rf Values silica gel plate n-hexane and ethyl

acetate (1:2) 0.23 silica gel plate n-hexane and acetone

(2:1) 0.21

(9) Property of the Substance: neutral substance

2. A process for production of FR-901429 substance which comprises culturing a FR-901429 substance-producing strain belonging to the genus Streptomyces in a nutrient medium and recovering the same.

3. A pharmaceutical composition containing FR-901429

substance.

4. A use of FR-901429 substance as a medicament.

5. A method for treating or preventing rejection by

transplantation, graft-versus-host diseases by medulla ossium, autoimmune diseases and infectious diseases which comprises administering FR-901429 substance to human or animal.