JPH029382A - Novel antibiotic ikd-8344 substance and production thereof and antitumor agent with the same substance as active ingredient - Google Patents

Abstract

PURPOSE:To provide the title substance of specific structural formula, low toxic and giving marked antitumor activity and having significant effect in various kinds of tumor therapies. CONSTITUTION:The objective substance of formula. This substance can be produced by culture of the antibiotic IKD-8344 substance-productive bacteria classified as Alkalomyces sp. and by separation and collection from the resulting cultured product. For said bacteria, Alkalomyces sp. NO 8344 (FERM NO. 10063), for example, can be advantageously used.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a new antibiotic, a method for producing the same, and an antitumor agent containing the same as an active ingredient.

[Conventional technology]

Although various antitumor agents have been developed so far, there are currently very few that exhibit low toxicity and significant antitumor activity.

[Problem to be solved by the invention]

Therefore, one of the objects of the present invention is to provide a substance that exhibits significant antitumor activity with low toxicity and can exert remarkable effects on the treatment of various tumors.

Another object of the invention is to provide a method for producing the above substances by fermentation.

Another object of this invention is to provide an antitumor agent containing the above-mentioned substance as an active ingredient.

[Means to solve the problem]

The object of the present invention is achieved by the novel antibiotic I substance D-8344.

Hereinafter, the IKD-8344 substance having antitumor activity of the present invention and its production method will be described.

The microorganism used in the method of the present invention is a new antibiotic, IKD.
This is a bacterial species belonging to a new species of actinomycetes named Alcalomyces that has the ability to produce -8344 substance.

As an example, Alcalomyces sp. Na33
44 (Alcalomyces sp, N1183
．． A microorganism called 44) is a new strain having the above characteristics and advantageously produces the new antibiotic IKD-8344 substance of the present invention, and is mentioned as a microorganism that can be effectively used in the method of the present invention.

In addition to artificial mutant strains of Alcalomyces sp. Nα8344, including natural and genetic engineering methods, microorganisms belonging to the genus Alcalomyces that have the ability to produce the new antibiotic IKD-8344 described below are also included. All can be used in the method of the invention.

The Alcalomyces sp. Nα8344 strain (hereinafter simply referred to as r8344 strain) is a microorganism discovered by the present inventors in a soil sample collected in Numazu City, Shizuoka Prefecture, and was developed by the Ministry of International Trade and Industry, Agency of Industrial Science and Technology. It was deposited with the Technical Research Institute on June 15, 1986, and its microorganism deposit number is "Choken Bacteria Deposit No. 10063."

Morphological characteristics, culture properties, and physiological properties of strain 8344 The study of the morphological characteristics, culture properties, and physiological properties of strain 8344 was carried out using the Schersing and Gottlieb method (Shi
rling, B, B, and D, Gottl
ieb :Methods for character
Rization of Streptomycess
pecies, Int, J, 5yst, Ba
cteriol, 16: 313-340.
(1966), Japanese Actinomycetes Research Complete Edition, “Experimental Methods for Identification of Actinobacteria” (Japan Actinomycetes Research Society Secretariat, 1985) and Kazuo Komagata (ed.), “Experimental Methods for Chemical Classification of Microorganisms” (Gakkai Publishing Center, 1982). I went there.

Morphological observations were made using light and electron microscopy on cultures grown on a modified tyrosine agar medium (pH 7) at 27°C for one week.

Observation of culture properties can be made using sycoclose/nitrate agar, glucose/asparagine agar, glycerin/asparagine agar, starch/inorganic salt agar, tyrosine agar, nutrient agar, yeast/malt agar, oatmeal agar, and improved agar. The experiment was conducted using 11 types of tyrosine agar medium, Bennett agar medium, and starch yeast agar medium. Each medium was incubated at 27°C for 30 minutes under two conditions: pH 7 and Plllo with 1% sodium carbonate.
The cells were cultured for a week and examined. The color marking method used in this research is based on JIS Z 8721 [Standard Color Standard, No. 7
(Japan Standards Association)].

For analysis of cell wall composition, the method of Haseyo and Seino (Experimental Methods for Identification of Actinobacteria, 47-67, 1985) was cited. For bacterial cells, yeast extract/glucose medium is used, pH 7 or p
It was obtained by culturing at 27° C. for 3 days under H10 conditions.

Carbon source availability was determined using the method of Bridham and Gottlieb (
Pridham, T, G, and D, Go
LLlieb: Theut 1zation of
carbon compounds by
some Actinomyces as a
aid for species determination
ation, J. Bacteriol,, 5 6,
107-1i4.1948).

The growth temperature was examined on a Bennett's slant agar medium at pH 7 and pH 10 supplemented with 1% IJum. Growth pH was examined by culturing at 27°C using Bennett and starch yeast slant agar media. Gelatin liquefaction is performed using glucose/peptone/gelatin medium, simple gelatin medium, and eXto malt/gelatin medium.
(pH 7, pHlo) at 20°C and 27°C. Starch hydrolysis was observed on a starch/inorganic salt agar medium. Peptonization of milk in skimmed milk medium (pt17)
Observation was made at 27°C and 35°C. Melanin pigment production is achieved using tyrosine agar, improved tyrosine agar, and tryptone.
Observation was made using yeast extract broth (p! (7, p) 110).

[I] Morphological characteristics Strain 8344 forms aerial hyphae, and mature spore chains are linear or wavy (Rectiflexibiles).
A long chain of 20 or more spores was observed. The spores are oblong and the size is 0.5-0.7μ x 0.7μ x 1
．． The surface resistance was approximately Ωμ, and the surface was smooth.

[L] Culture characteristics Strain 8344 grew under both conditions of pH 7 and pH 10, and the growth conditions were almost the same, but the attachment of aerial mycelium was considerably better at pH 10. However, on the improved tyrosine agar medium, abundant aerobic filaments were observed under pH 7 conditions. Also, some soluble pigment was observed only on pH 7 tyrosine and modified tyrosine media. The observation results for each medium were as shown in Table-1.

Table - Culture properties of strain 18344 972 no 9/1 sucrose/nitrate agar Glucose/asparagine agar Glycerin/asparagine agar Starch/mineral salt agar Tyrosine agar 6: Nutrient agar 7: Yeast/malt agar 8: Oatmeal agar 9: Improved tyrosine agar 10: Bennett agar 11: Starch yeast agar
Color of back side), SP: Soluble pigment The numbers in parentheses in the table are JIS Z 8721? $ basis [a
Represents the color display method based on the Quasi Color Standard, 7th Edition (Japanese Standards Association).

[[[E Composition of cell wall The presence of LL type and meso type 2,6-diaminopimelic acid was observed as a cell wall component.

Meso-type diaminopimelic acid was observed in both pH 7 and pH 1O cultures, but the amount tended to be particularly high when cultured at 11810, and was approximately 1/2 to 2 times that of the LL type.
/3.

Furthermore, the presence of glucose and ribose was observed as cellular sugars.

CIV'l Physiological Properties Strain 8344 grows at 15°C to 35°C under both I]H7 and ρ)110 conditions;
It did not grow at ℃.

When the growth pH is below 5.5, there is almost no growth.
It grew at 6-1O15 and did not grow at 11. Additionally, starch was hydrolyzed and milk was coagulated into peptone.

Different results were obtained for gelatin liquefaction depending on the medium and pH, but in yeast/malt gelatin medium, p
Slight liquefaction was observed for both f(7 and ptHO. Also,
No production of melanin pigment was observed.

Available carbon sources were L-arabinose, D-xylose, D-glucose, D-fructose and D-mannitol.

Physiological properties are summarized in Table 2, and carbon source utilization is summarized in Table 3.

Table 2: Physiological properties of strain 8344 Table: Utilization of carbon sources by strain 38344 ■Raffinose D7nitol l ++7+: Considerably used, +: Used, -: Not used.

In addition to the IKD-8344 substance, the 8344 strain capable of producing antibiotics (LV) has the ability to produce antimycin antibiotics (Harada,
.

and ^, ^sa+: J, ^nti
biot,, 11Δ, 32. 1958°Li
u, W, and F, M, Strong
: J, Am, Chem, Sac.

81.4387.1959).

From the above properties, the 8344 strain is Streptomyc.
Similar to the genus es. However, as a cell wall component, LL
Strepto, which is characterized by having only the LL type, because it also has the meso type diaminopimelic acid in addition to the LL type.
It is not classified into the genus myces.

As a genus of actinomycetes that has LL-type and meso-type diaminopimelic acid in the cell wall, Kijas, which was discovered by Omura et al.
Only atosporia @ is known. This kit
The genus asatOspor+a includes K, 5etalha
(K, 5etae) (K已asatosporia,
a new genus of the
orderAct +nomyceta les, J
, ^ntibiotics, 3 5, 1013~
1019.1982) and K, phosalac+n
ea and ni9 griseola (Two new 5
pecies or the genusKitasa
tosporia, K+tasatosporia
phosalacineasp, nov, and 1
tasatoaporia gr+5eola sp,
novlJ, General Appl, !
Jicrobio1. , 30, 377-387
．． 1984) have been discovered to date. However, all of these bacteria have galactose as a cellular sugar, and the growth pH range is 9 or less.
It clearly differs from strain 8344 in that it does not utilize D-mannitol as a carbon source and in its ability to produce antibiotics.

Therefore, strain 8344 is not classified into either the genus Streptomyces or the genus Kitasatosporia.

On the other hand, regarding alkaliphilic or alkali-tolerant actinomycetes, Mikami et al. reported (Diaminopimel 1c
ac+d proC+les or alkaloph
ilic and alkaline-resista
nt 5trains of Actionnomyce
tes, J. Gen.

Microbiol,, 128.1709-171
2.1982). They are conventionally streptomy
It has been reported that mesodiaminopimelic acid is contained in the bacterial cells of three species of actinomycetes classified in the genus Ces when they are cultured in alkaline conditions, but there is no mention of the case when they are cultured in a neutral medium. However, the genus has not yet been identified.

Therefore, strain 8344 cannot be classified into any known actinomycete genus and is considered to belong to a new genus.

The 8344 strain is characterized by being an alkaliphilic or alkali-tolerant bacterium that can grow even at pH 10.5, having LL-type and meso-type diaminopimelic acid as cell wall components, and not having any characteristic sugars.

Based on these properties, the genus to which strain 8344 belongs was named the genus Alcalomyces.

Production of IKD-8344 substance The IKD-8344 substance of the present invention is produced by IKD-8344 substance-producing bacterial strains belonging to the genus Alcalomyces (for example, ^Ic
alomyces sp, Nα8344) is grown in a nutrient medium containing available carbon and nitrogen sources under aerobic conditions (e.g., shaking culture, deep culture, etc.)
to be produced by For industrially advantageous cultivation, a spore U suspension or culture solution of the above-mentioned producing bacteria may be inoculated into a medium and cultured with aeration and stirring.

The nutrient source of the medium is not particularly limited, and the medium may contain carbon sources, nitrogen sources, and other sources commonly used for culturing microorganisms. Carbon sources include starch,
Dextrin, glucose, glycerin, mannitol, etc., and nitrogen sources include yeast extract, peptone, soybean flour, meat extract, cornstarch liquor, malt, cottonseed meal, dried yeast, rice bran, bran, etc., and ammonium salts, Organic or inorganic nitrogen compounds such as nitrates, urea, amino acids, etc. are used. Other inorganic salts, such as common salt, phosphates, potassium, calcium, zinc, manganese,
Metal salts such as cobalt and iron may be added as appropriate.
Liquid paraffin, animal oil,
Vegetable oil, mineral oil, silicone, etc. may also be added. In addition, when making the pH of the medium alkaline, sodium carbonate, sodium bicarbonate, sodium sesquicarbonate,
Add 3-sodium phosphate, etc. Furthermore, fatty acids,
Alternatively, esters of fatty acids can also be added.

Conditions such as culture pH 1 culture glue temperature and culture time are selected so as to be suitable for the growth of the bacteria used and to maximize the production of IKD-8344 substance. For example, the pH of the culture is 6-1
1, and the culture temperature is preferably 15 to 35°C, preferably 20 to 30°C.

The culture time is preferably 24 to 168 hours, preferably 48 to 96 hours. However, these culture conditions such as culture composition, hydrogen ion concentration of the medium, culture temperature, and stirring should be adjusted as appropriate to obtain favorable results depending on the type of bacterial strain used and external conditions. It goes without saying that there is. From the culture solution obtained in this way, IKD-83
44 substances can be recovered by various methods commonly used for the recovery and purification of antibiotics, such as solvent extraction using suitable solvents or mixtures of such solvents, chromatography, or recrystallization method from a mixture of such solvents) [KD-8
Since 344 substances exist in both the culture filtrate and the bacterial cells,
By extracting from the culture filtrate using a water-immiscible solvent such as ethyl acetate, chloroform, or ether, and from the bacterial cells using an appropriate solvent such as ethyl acetate, methanol, or acetone, or a mixture thereof. It can be recovered. The culture solution can also be directly subjected to the above extraction operation without separating the bacterial cells. Countercurrent distribution methods can also be included in the scope of extraction.

The extract is processed by conventional methods to obtain IKD-8344 material. For example, the extract is concentrated under reduced pressure and purified by conventional purification methods, such as chromatography or recrystallization from a suitable solvent or mixture thereof.

It can be separated by dissolving the extract containing the IKD-8344 substance in a solvent such as chloroform, subjecting it to silica gel column chromatography, and eluting with a suitable solvent such as chloroform, methanol, or a mixture thereof. The IKD-8344 substance obtained in this way can be further purified by a conventional method (for example, gel filtration using Sephadex LH-20, thin layer chromatography, liquid chromatography). It can be obtained as a pure substance by recrystallization using methanol, acetone, etc.

Physicochemical properties of IKD-8344 substance 1) Shape and color: White powder.

Molecular weight (1 = “A B - M S ): m/
z 845 (M+H)" m/z 867 (M+Na)" 3) Molecular formula: % formula % : : ): Easily soluble: Ethyl acetate, methanol, Ether soluble: Hexane, Petroleum ether Insoluble: Water 8) Coloration Reaction: Positive: 10% sulfuric acid/methanol, iodine vapor negative; Ehrlich reaction, orcinol reaction and ninhydrin reaction 9) Ultraviolet absorption spectrum: No characteristic absorption 10) Infrared absorption spectrum: (Main maxima in KBr tablets ) 2990.2950.2900.1740.1710.
1460S1415.1380°1335.1300.
1270.1230.1180.1140.1080.
1000°970.940.900.815 cm-
'11) Thin layer chromatography (Rf value) Chloroform: methanol lO: 1 0.25 Ethyl acetate: methanol: noroammonia 10:1:0.1 0.2
7 Ethyl acetate; methanol:acetic acid 20:1 co0.1 0.51 12>13 (j*Magnetic resonance spectrum (cD[!3)
δ (Ppm): 10.9.13.8.14.4.21
．． 3.29.5.29.7.30.8.30.9.32
．． 2.34. o, 36.3.41.2.45.6.45
．． 7.53.4.72, 3.74.8.75.0.75
．． 3.79.6.80.1.8o, 6.175.2.2
12.0 The molecular formula of IKD-8344 substance is C*aHtso 12
Since only 24 carbons are observed in the 13C nuclear magnetic resonance spectrum, it is presumed that the IKD-8344 substance is a dimer.

Biological Properties of Substance IKD-8344 The substance IKD-8344 of the present invention has significant antitumor activity and antibacterial activity as shown below, and is a useful compound as a medicine.

1> rKD-8344 substance 1 investigated by antibacterial spectrum paper disk method
The diameter of the inhibition circle at a concentration of 000 μg/- is shown below. Note that bacteria and Candida were evaluated after culturing at 37° C. for 24 hours, and Shiromakan was evaluated after culturing at 27° C. for 48 hours.

Table-41 Antibacterial spectrum of KO-8344 substance
1 inhibition circle diameter IKD-8344 substance showed antibacterial activity against Candida albicans.

2) IKD-8344 substance with growth inhibitory activity against mouse leukemia L5178Y cells was added to a culture medium [added with 10% horse serum].
4”/Fischers medium
(Hinaga Pharmaceutical Co., Ltd.)] to a concentration of 1000μg/-, and further diluted sequentially with culture medium to 0.002μg/-
A test solution was prepared. Using this as the highest concentration, make 2-fold and 4-fold dilutions, and add 1 rnl of each to 1 rnl of L5.
Add to culture solution 1- containing 2 x 10' 178Y cells,
The cells were cultured in an upright stationary state at 7°C for 72 hours.

After the completion of the culture, the number of cells in the culture solution was measured using a Coulter counter (ZBI type), and the ratio of the average number of cells in the sample group (N = 6) and the control group (N = 6) was determined to indicate growth. The inhibition rate was determined. I Cs. was determined from the inhibition rate at each concentration using logarithmic establishment paper.

Table-5 Mouse leukemia cell L5178Y of IKO-8344 substance
It had growth inhibitory activity on cells. The sample is 0°5% CMC
(carboxymethylcellulose) suspended in physiological saline. The control group received 0.5% CMC saline. There were 6 animals per group for both the sample administration group and the control group. After inoculation of P388 cells, the animals were observed for 30 days, and the median survival days (+nedian surv
ivaltime: MST), the life extension rate (ILS) was determined according to the following formula.

From this result, [Cs. is 0.00054 μg/ml
In particular, the IKD-8344 substance had an extremely strong growth-inhibiting activity against tumor cells.

3) Life extension effect on P388 tumor-bearing mice BDF, male mice (average weight 20 g) were intraperitoneally inoculated with 1X10' P388 leukemia cells subcultured in DBA/2 mice, and from 1 day later, once a day. 9 consecutive days or 5
．． After 9 days, winter 1 IKD-8344 substance was administered intraperitoneally to mice. As shown in Table 6, IKD-8344 substance was administered daily for 9 days at 0, 25 mg/kg or more, and 3, Omg/kg. Three intermittent administrations of 1 kg showed an excellent survival effect on P388 leukemia.

Table 61: Survival prolonging effect of KD-8344 substance on P388 leukemia Animals used: BDF, mouse, male tumor Inoculation site: Intraperitoneal (IXIO' pieces 10.1m!
/mouse) Sample administration site: intraperitoneal (0,2rn1/mouse)1
Number of animals in the group: 6 animals, 8344 bottles: IKD-8344 substance 4) Acutely toxic BDF, administered intracavitally to 7 mice at 5 B/kg once or 2 mg/kg once a day for 5 consecutive days. However, no fatal cases were observed.

As described above, the IKD-8344 substance of the present invention exhibits strong anti-tumor activity and is useful as an anti-tumor agent. below,
An antitumor agent containing IKD-8344 substance as an active ingredient will be described.

Therapeutic use of IKD-8344 substance The antitumor agent of the present invention can be administered either orally or parenterally, and when administered orally, it can be administered in soft or hard capsules, tablets, granules, fine granules, Used as powder, IiA solvent, and suspending agent. For parenteral administration, injections,
It can be administered in a dosage form that maintains mucosal or transdermal absorption, such as in the form of drops, tablets, emulsions, or suspensions, or as ointments, creams, solutions, bags, or tapes.

The active ingredient of the present invention can be formulated as appropriate using pharmaceutically acceptable surfactants, excipients, lubricants, adjuvants, and others, and can be formulated into solid, semisolid, liquid, etc. can be obtained. Additionally, stabilizers, colorants and fragrances may be used. The active ingredient is contained in the formulation in an amount sufficient to treat or prevent the disease.

When this preparation is used in humans, it is preferable to use it by intravenous, intramuscular, oral or transdermal administration. The dosage is sufficient to effectively treat the target tumor and depends on the symptoms of the tumor, administration route, dosage form, etc., but in general, in the case of oral administration, it is approximately 0.0001 to 0.0001 per day.
2 mg/kg body weight, the upper limit of which is preferably about 0
．． 5 mg/kg body weight? In the case of parenteral administration, the upper limit is about 1 mg/7 kg body weight, preferably about 0.2 mg/kg body weight or less. Examples and formulation examples are merely for illustrative purposes of the present invention. provided without limiting its scope.

Example 1: Glucose 2.0g, soluble few minutes 1. Og, soy flour LJ
, 0.4 g of yeast, 0.2 g of salt, 0.1 g of beef extract, and 5 g of dipotassium phosphate O, OO (90 mI!, pH 8.5) were placed in a 500-mL Yosaka flask. , autoclaved at 120°C for 20 minutes.

After sterilization, 10 d of 10% aqueous sodium carbonate solution, which had been sterilized under high pressure, was added to bring the final pH to about 1O. One platinum loop of the slant culture of the 8344 strain [@Koken Bacteria No. 10063] was inoculated into this medium, and the amplitude was 7 c+n and the number of rotations was 14.
Shaking culture was carried out at 28° C. for 96 hours on an Orpm reciprocating shaker. This culture solution 1- was incubated in 40 Sakaro flasks containing the medium 100-, and cultured at 28°C for 72 hours.

The obtained culture solution was centrifuged at 9500 revolutions per minute for 10 minutes.
The culture supernatant and the bacterial cells were separated, and the culture supernatant was extracted three times with half the amount of ethyl acetate. The bacterial cells were extracted with 95% aqueous acetone, concentrated, and similarly extracted with ethyl acetate. The ethyl acetate extracts were combined and concentrated to dryness.
150g) was applied to column chromatography. Arakashime chloroform + 5ooy, then // Shiroho 711: methanol mixture (30: l) 1500ml
l! The above oily substance was applied to the column washed with and eluted with a chloroform:methanol mixture (10:1). The active fraction was concentrated to obtain crude material (307 mg).

This crude material was dissolved in a small amount of methanol, and a Sephadex LH-20 column (2.8c
mX 80 cm) and eluted with methanol. The eluate was separated into 8 d portions, and active fractions 38 to 45 were concentrated, concentrated to dryness, and extracted with hexane. Dissolve the hexane soluble content in 2rnl of acetate, o, ao
i Add 2 ml of normal hydrochloric acid, leave to stand at 5°C for 30 minutes, collect the resulting precipitate through a glass filter, wash with cold water and dry to obtain 54.9 ml of purified powder of IKD8344 substance.
I got g. Formulation example 1 (injection, infusion) Add 5 g of powdered glucose to a solution containing 50 mg of IKD-8344 substance, dispense aseptically into vials, seal them, fill with inert gas such as nitrogen or helium, and store in a cool, dark place. Save to. 0°85% physiological saline 100% before use
- to make an intravenous injection, 1 to 10 times a day (lt7
! Administer by intravenous injection or drip depending on symptoms.

Formulation example 2 (tablet) 100 large-sized tablets were prepared with the following ash content composition.

AI IKD-8344 1R2.0g Lactose 17
．． 94g Hydroxypropylcellulose 0.06g
Magnesium stearate 0.2g CB) Cellulose acetate phthalate 0.6g Hydroxypropylmethyl 0.6g Take the ingredients of cellulose phthalate [A], mix well, and pressurize this directly or after kneading well, extrude. Neck granules were formed through the screen of a mold granulator, and after sufficient drying, they were pressed to prepare tablets.

Furthermore, if necessary, well-dissolved CB) can be molded into tablets.
Enteric-coated tablets are prepared by coating the base material.

Formulation example 3 (capsule) One hundred adult capsules were prepared with the following ingredients.

[A] IKD-8344 substance 2. Og milk
Sugar 10.46g Hydroxypropyl cellulose 0.04g [B] Cellulose acetate phthalate 1.0g Hydroxypropyl methyl 1.0g Cellulose phthalate Take the above component [A], mix well, and then form into granules according to a conventional method. This was thoroughly dried and sieved to form granules suitable for capsules, which were then filled into capsules to prepare capsules. Further, if necessary, the I granules are coated with a base of dissolved CB) while floating and fluidized to form enteric-coated granules, which are then filled into capsules to obtain enteric-coated capsules.

〔Effect of the invention〕

The novel antibiotic IKD-8344 substance of the present invention has low toxicity and excellent antitumor activity.

[Brief explanation of the drawing]

FIG. 1 is a drawing showing an infrared absorption spectrum of IKD-8344 material, FIG. 2 is a drawing showing a 1H nuclear magnetic resonance spectrum of IKD-8344 material, and FIG.
The figure shows a 3C nuclear magnetic resonance spectrum of rKD-8344 material. 1. Display of $ patent application No. 159948 of 1988...Achieved. " is amended as follows. "The object of the present invention is achieved by the novel antibiotic IKD-8344 substance represented by the following formula. 3. Relationship with the case of the person making the amendment 4, Representative 7 , Contents of the amendment 3゜4, page 26, line 4, “Estimated. ” is corrected to ``It was estimated, and as a result of further investigation, it was revealed that it has the above-mentioned structural formula.'' On page 34, lines 11 to 15, “In advance...
It eluted. ′ is corrected as follows. "Pour the above oily substance onto the column and apply chloroform!500
d, then chloroform:methanol mixture (30:1)
After washing with 1500-, chloroform:methanol mixture (
10:l). ” Claim (1) Antibiotic IKD-834 represented by the following formula
4 substances. (2) Alcalomyces
Antibiotic IKD-8344, which is characterized by culturing the antibiotic IKD-8344 substance-producing bacteria belonging to the genus, and separating and collecting the antibiotic IKD-8344 substance from the culture.
8344 Substance manufacturing method. (3) Alcalomyces sp. Nα8344 (
Alcalomycas sp, Nα8344). (4) An antitumor agent characterized by containing the antibiotic IKD-8344 substance as an active ingredient. −／／

Claims (4)

[Claims] (1) A new antibiotic IKD-8344 substance characterized by having the following physical and chemical properties. (a) Shape and color: White powder. (b) Molecular weight (FAB-MS): m/z845 (M+H)^+ m/z867 (M+Na)^+ (c) Molecular formula: C_4_8H_7_6O_1_2 (d) Elemental analysis: Carbon: 68.52% Hydrogen: 9.08% Nitrogen: 0% (e) Melting point: 133-136°C (f) Specific optical rotation: [α]^1^7_D=+40.2° (c=1.0, methanol) (g) Solubility: Easy to dissolve: Ethyl acetate, methanol, ether Soluble: Hexane, petroleum ether Insoluble: Water (1) Color reaction: Positive: 10% sulfuric acid/methanol, iodine vapor Negative: Ehrlich reaction, orcinol reaction and ninhydrin reaction (1) Infrared absorption spectrum : (Main maximum values in KBr tablets) 2990, 2950, 2900, 1740, 1710,
1460, 1415, 1380, 1335, 1300,
1270, 1230, 1180, 1140, 1080,
1000, 970, 940, 900, 815cm^-^
1(NU)^1^3C nuclear magnetic resonance spectrum (CDCl_
3) δ (ppm): 10.9, 13.8, 14.4, 2
1.3, 29.5, 29.7, 30.8, 30.9, 3
2.2, 34.0, 36.3, 41.2, 45.6, 4
5.7, 53.4, 72.3, 74.8, 75.0, 7
5.3, 79.6, 80.1, 80.6, 175.2,
212.0 (2) Cultivating antibiotic IKD-8344 substance-producing bacteria belonging to the genus Alcalomyces,
A new antibiotic IKD-834 characterized by separating and collecting the new antibiotic IKD-8344 substance from the culture.
Production method of 4 substances. (3) Alcalomyces sp. no. 8344(A
lcalomyces sp. No. 8344). (4) An antitumor agent characterized by containing the antibiotic IKD-8344 substance as an active ingredient.