JPH08143570A - Intracellular transfer inhibitor - Google Patents

Abstract

PURPOSE: To obtain an intracellular transfer inhibitor useful as an antiviral and an anti-cancer medicine, having inhibitory action on intracellular transfer, containing a compound prepared from a culture product of a specific bacterium belonging to the genus Streptomyces. CONSTITUTION: This inhibitor contains a compound of the formula (R1 to R4 are each methyl or ethyl) or its salt as an active ingredient. For example, the compound is preferably obtained from a culture product prepared by subjecting a bacterium (FERM P-14,584) isolated from soil in Nishiomotejima Island in Okinawa prefecture to shaking culture or aerated spinner culture in a liquid medium at about 25-30 deg.C at pH5.5 to pH7.5 for 3-6 days.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a medicament containing a macrotetrolide compound or a complex salt thereof as an active ingredient, a novel microorganism capable of producing the macrotetrolide compound or a complex salt thereof, and fermentation using the novel microorganism. The present invention relates to a method for producing the macrotetrolide compound or a complex salt thereof by a method.

[0002]

2. Description of the Related Art In recent years, glycoproteins have been infected with viruses, become cancerous,
It has been clarified that it plays an important role in various cell physiology and disease states such as immunity and allergy. For example, it has been pointed out that a viral coat glycoprotein is involved in a viral infection process and a syncytium formation process of a viral disease. In addition, the relationship between the sugar chain structure of a glycoprotein, canceration, and an immune reaction is also expected, for example, it is revealed that glycoproteins with abnormal sugar chains are synthesized by canceration. Meanwhile, Brefeldin A [Merck, known as an antifungal agent,
Index, 11th edition, 2g8 pages, 1363 Bre
feldin A; 1,6,7,8,9,11
a, 12,13,14,14a-Decahydro-1,13
-Dihydroxy-6-methyl-4-cyclopent [f]
Oxacyclotridecane-4-one] has recently had antiviral, antiallergic and anticancer activities [Aids Res. Hum. Retrovirus
es, 7 (8), 707-712 (1991); I
mmunol. , 147 (10), 3289-3295.
(1991), Proc. Am. Assoc. Canc
er Res. Annu. Meet. , 32 (0), 4
00 (1991)], and was subsequently reported to have the effect of inhibiting the intracellular transfer of glycoproteins through the Golgi organ [J. Gen. Virol. , 73 (10), 2679
-2692 (1992)]. Intracellular transport means that glycoproteins newly synthesized in the endoplasmic reticulum are transferred to the cell surface, extracellular or intracellular organelles via the Golgi organ, and some are accumulated in secretory granules, causing external stimulation. Is a system that is transported to the cell surface or outside the cell in response to Some, such as receptors, are endocytosed after being transferred to the cell surface. Although the flow of these series of transfers has been largely clarified, the mechanism is currently under active research.

On the other hand, the macrotetrolide antibiotic group comprises
A general term for a group of antibiotics having a 32-membered macrocyclic structure and containing four lactones in the ring, specifically nonactin, monactin, dynactin, trinactin, tetranactin and the like. It is already known that these macrotetrolide compounds have antibacterial activity, antifungal activity and insecticidal activity against Gram-positive bacteria [J. Antibi
ot. , 18 , 128 (1965); Antibi
ot. , 17 , 11 (1964); Antibio
t. , 23 , 105 (1965)]. However, it is not known at all that these substances have a glycoprotein intracellular transfer inhibitory action and are useful as antiviral agents, antiallergic agents and anticancer agents.

[0004]

Research on the development of antivirals, antiallergic agents, and anticancer agents is still active, and providing antiviral agents, antiallergic agents, and anticancer agents having excellent clinical effects is still a medical problem. Has become an important issue.

[0005]

Means for Solving the Problems The inventors of the present invention have concluded that, under such a state of the art, a substance that inhibits the intracellular transfer of glycoprotein may exhibit antiviral activity, antiallergic activity, anticancer activity and the like. As a result of vigorous search for active substances by screening the intracellular transfer system of glycoproteins using viruses, a new species of actinomycete produced from the soil has a glycoprotein intracellular transfer inhibitory effect, And so on. The present inventors further identified the substance, and found that the glycoprotein intracellular inhibitor was nonactin, monactin, dynactin, trinactin, and tetranactin, which are conventionally known as macrotetrolide antibiotics. As a result, the present invention has been completed. That is, the present invention relates to a glycoprotein intracellular transfer inhibitor comprising, as an active ingredient, a macrotetrolide compound represented by the following general formula (I) or a complex salt thereof.

[0006]

Embedded image

(Wherein R ₁ , R ₂ , R ₃ , and R ₄ are the same or different and each represents a methyl group or an ethyl group) The present invention also provides a macrotetrolide having a glycoprotein intracellular transfer inhibitory action. The present invention relates to Streptomyces sp. Q30101 having the ability to produce compound (I) or a complex salt thereof. The present invention further comprises culturing a microorganism having the ability to produce a macrotetrolide compound (I) or a complex salt thereof, which belongs to the genus Streptomyces and has a glycoprotein intracellular transfer inhibitory activity, and from the culture, the compound (I) or a compound thereof. The present invention relates to a method for producing the compound, which comprises collecting a complex salt.

Hereinafter, the novel microorganism, production method, and medicine of the present invention will be described in detail. (Microorganism) The microorganism of the present invention belongs to the genus Streptomyces, and is characterized by its ability to produce the macrotetrolide compound (I) or a complex salt thereof. Specific examples of the microorganism include Streptomyces sp .
omyces sp. ) Q30101 strain.
This Q30101 strain was isolated from the soil of Iriomote Island in Okinawa Prefecture. The bacteriological properties of this strain are as follows.

Mycological properties of strain Q30101 Morphology This strain grows well in various organic and inorganic media, and the color of the underlying mycelium changes from yellow-gray to yellow-brown. The aerial mycelium
It is often formed on starch / inorganic salt agar and nutrient agar. The aerial mycelium hardly branches, and its tip shows a wave shape (Flexibilis). A spore chain of 10 to 50 or more spores is formed on a spore stalk branched from an underlying hypha.
No fragmentation of the basal hyphae was observed in liquid culture. Observation with an electron microscope showed that the spores had a cylindrical shape and a size of 0.
It is 4 to 0.6 μm × 0.5 to 0.8 μm, and its surface is smooth. Special organs such as sporangia and motile spores are not observed.

[0010] 2. Properties on various agar media Properties on various agar media are as shown in Tables 1, 2 and 3 below. Unless otherwise specified, the cells were cultured at 28 ° C. for 21 days and observed according to a conventional method. The description of the color tone was based on the color standard (Japan Color Research Institute).

[0011]

[Table 1]

[0012]

[Table 2]

[0013]

[Table 3]

3. Physiological properties

[0015]

[Table 4]

4. Utilization of carbon sources

[0017]

[Table 5]

5. Chemical analysis of bacterial cell components The method of LECHVALIER et al. (LECHVALIE)
R, MP. et al; PP277-238 in D
IETZ, A et al ed. , Actinom
ycete Taxcnomy, SIM Specia
l Publication No. As a result of analysis of the acid hydrolyzate of this strain according to
-Diaminopimelic acid was detected (cell wall type I). Major bacterial menaquinone was MK-9 _{(H 6).} In summary, the properties of the Q30101 strain are as follows: the basic mycelia are yellow to yellow-brown, and the aerial mycelia are white to yellow-gray. On the spore stalk extending from the base hypha, 10 to 50 or more spore chains are formed, and the base hypha does not fragment. Cell wall type is I, major menaquinone is MK-9
(H ₆ ). Based on the above properties, this strain is determined to be a strain belonging to the genus Streptomyces . Various references (Bergey's Manu)
al of Systematic Bacterio
logic vol. 4, 1989), as a result of comparison with a known strain belonging to the genus Streptomyces , no strain similar to the present strain was found. Therefore, this strain was used as Streptomyces sp.
0101. This strain was obtained from the National Institute of Advanced Industrial Science and Technology under the accession number FERM P-14584.
No. has been deposited.

The microorganism of the present invention is liable to be mutated naturally or artificially by ultraviolet rays, X-rays, chemical agents or the like. Accordingly, the microorganism of the present invention includes the above-mentioned strain Q30101 isolated from nature, a microorganism belonging to the same species as strain Q30101, capable of producing glycoprotein intracellular transfer inhibitor (I) or a complex salt thereof, and natural mutants thereof. And artificial variants. Examples of the microorganism used in the production method of the present invention include microorganisms belonging to the genus Streptomyces and capable of producing glycoprotein intracellular transfer inhibitor (I) or a complex salt thereof, natural mutants thereof, and artificial mutants thereof. Included.
The microorganism of the present invention is obtained by separating it from natural soil, and can be easily obtained by restoring the strain deposited at the Biotechnology Industrial Research Institute.

(Production method) The production method of the present invention comprises the steps of culturing a microorganism belonging to the genus Streptomyces and capable of producing a glycoprotein intracellular transfer inhibitor (I) or a complex thereof, and subjecting the culture to a culture of said compound (I) or It is carried out by collecting the complex salt. The cultivation is advantageously performed using a medium containing the nutrient sources utilized by the microorganism. The medium may be any of synthetic, semi-synthetic or natural, solid or liquid medium, but a liquid medium containing a natural nutrient is generally preferred.
Among the nutrient sources added to the medium, the carbon sources include L-arabinose, D-xylose, D-glucose, D-fructose, sucrose, inosit, rhamnose, raffinose, D-mannitol, D-galactose, maltose, Trehalose, lactose, D-sorbit, salicin, glycerin, xanthine, chitin,
Examples include starch, starch, glucose, dextrin, coconut oil, soybean oil and the like, and these are used alone or in combination. Further, alcohols, organic acids and the like can also be used. As the inorganic nitrogen source, ammonium chloride, ammonium sulfate, ammonium nitrate, urea, etc., as the organic nitrogen source, peptone, yeast extract, dried yeast, meat extract, gluten meal, corn steep liquor, soybean powder, fish meal, peanut powder, Cottonseed meal, casamino acid, various amino acids (eg, glutamic acid, alanine, lysine, etc.) are used alone or in combination. In addition, sodium, potassium, magnesium,
Inorganic salts such as sulfates, nitrates, chlorides, carbonates and phosphates of metals such as calcium, zinc, iron and cobalt, and organic acid salts such as sodium citrate can be added. The cultivation is advantageously performed under aerobic conditions, and any of stationary, shaking, and aeration and stirring cultures is possible. Shaking or aeration and stirring culture is particularly preferable. Culture temperature is about 15-5
It is preferably in the range of 0 ° C, especially about 25-30 ° C. The pH of the medium is preferably maintained at about 4.5 to 8.5, especially about 5.5 to 7.5. The culture time varies depending on the culture conditions such as the composition of the medium and the temperature.
It is usually about 2 to 14 days, preferably about 3 to 6 days, and ends at an appropriate time in view of the time when the production substance reaches the maximum titer.

In order to isolate and purify the glycoprotein intracellular transfer inhibitor accumulated in the culture thus cultured, a commonly used isolation and purification means may be applied. Q
When the 30101 strain was cultured under the culture conditions described in Production Examples,
At least five transfer inhibitors accumulate in the culture. Glycoprotein intracellular transfer inhibitor is isolated and purified by removing the cells by centrifugation or filtration of the culture, and then differences in solubility and solubility in an appropriate solvent, differences in the rate of precipitation from the solution, and various other factors. This method is carried out by applying a method utilizing a difference in adsorption affinity to an adsorbent and a difference in distribution between two liquid phases. These methods can be used alone or combined in any order and applied repeatedly as needed. The substance produced by culturing the Q30101 strain is
Utilizing the difference in solubility in a solvent and the affinity for adsorption to various carriers, separation is performed using transfer inhibition activity as an index. In addition, several types of transfer inhibitors can be finally isolated by high-performance liquid chromatography (HPLC).

(Glycoprotein intracellular transfer inhibitor) Q301
By culturing the 01 strain, a glycoprotein intracellular transfer inhibitor was produced, and five kinds of active substances were isolated. As a result of confirming and identifying the physicochemical properties of these substances, Q3
The five kinds of glycoprotein intracellular transfer inhibitors produced by culturing the 0101 strain are nonactin, monactin, dynactin, trinactin, and tetranactin, which have the following structures, which are conventionally known as being included in the macrotetrolide antibiotics group. It was confirmed that.

[0023]

Embedded image

Nonactin, monactin, dynactin,
Among the physicochemical properties of trinactin and tetranactin, their respective mass spectra [FAB (Pos.)]
And ultraviolet absorption spectra (methanol solution) and hydrogen nuclear magnetic resonance spectra (CDCl ₃ ) of nonactin, monactin, and dynactin, respectively, as shown in FIGS.
Shown in In the measurement of the mass spectrum, a complex salt of sodium and potassium of the macrotetrolide compound is formed and measured. In the mass spectrum diagram, Na indicates the peak of the sodium complex salt of the macrotetrolide compound, and K indicates the peak of the macrotetrolide compound. The peak of the potassium complex salt of the ride compound is shown.

It is well known that a macrotetrolide compound contains a metal in its 32-membered ring to form a metal complex, and the metal complex has the same activity as a free macrotetrolide compound. Accordingly, the active ingredient of the medicament of the present invention includes not only free macrotetrolide compound (I), that is, nonactin, monactin, dynactin, trinactin and tetranactin, but also complex salts thereof.
For example, compounds (I) such as sodium, potassium and lithium
And a metal salt capable of forming a metal chelate. Compound (I) or a complex salt thereof may be isolated as a hydrate, various solvates, or a polymorphic substance. Therefore, the active ingredient (I) of the medicament of the present invention or a complex salt thereof also includes these substances.
Further, compound (I) or a complex salt thereof has an asymmetric carbon atom. The active ingredient of the present invention is preferably a natural isomer, but the active ingredient of the medicament of the present invention includes a mixture of optical isomers or all of the isolated isomers. The macrotetrolide compound (I) or a complex salt thereof as a component of the medicament of the present invention should not be limited to those produced by culturing the strain Q30101, but may be a substance conventionally included in the macrotetrolide antibiotic group. Of course, it may be produced by culturing a known strain known to produce E. coli.

Pharmaceutical compositions containing the macrotetrolide compound represented by the general formula (I) or a complex salt thereof as an active ingredient are prepared using carriers, excipients and other additives usually used for formulation. You. Pharmaceutical carriers and excipients include solid or liquid non-toxic pharmaceutical substances. Examples of these include, for example, starch, lactose, sucrose, mannitol, calcium carbonate, calcium phosphate, gum arabic, carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, magnesium stearate, talc, calcium cellulose glycolate, calcium carboxymethylcellulose, gelatin , Agar, pectin, distilled water, ethanol, propylene glycol, polyethylene glycol, olive oil, sesame oil, cocoa butter, polysorbate 80 (trade name: surfactant), and other commonly used ones. Administration can be in any form of oral administration such as tablets, pills, capsules, granules, powders, and liquids, injections such as intravenous injection and intramuscular injection, and parenteral administrations such as suppositories and transdermals. Is also good. The dose is appropriately determined in consideration of the age, sex, weight, disease, symptoms, and the like of the administration subject.

[0027]

The present invention will be described in detail with reference to the following examples. Production Example Glucose 1.0%, potato starch 2.0%, yeast extract (manufactured by Oriental Yeast) 0.5%,
A medium A (pH 7.0) containing 0.5% of polypeptone (manufactured by Nippon Pharmaceutical) and 0.4% of calcium carbonate was prepared. This was dispensed 100 ml at a time into a 500 ml Erlenmeyer flask.
The mycelia of the strain Q30101 grown on a Bennett agar medium were scraped and inoculated to the one sterilized at 20 ° C. for 20 minutes, and shake-cultured at 27 ° C. for 3 days to obtain a seed culture solution. Next, 21 (liter) of medium A was prepared, and 50
Dispense 100 ml at a time into a 0 ml Erlenmeyer flask,
A seed culture solution was inoculated at a rate of 3.0% into a solution sterilized at 0 ° C. for 20 minutes, and cultured at 27 ° C. for 4 days. After adjusting the pH of the thus obtained culture solution to 3 with hydrochloric acid, Radiolite # 600 (manufactured by Showa Chemical Industry Co., Ltd.) was added, followed by filtration.
The cells were separated from the supernatant. After the pH of the supernatant was adjusted to 7, 2 l of ethyl acetate was added for extraction. After separating the ethyl acetate layer, 2 l of ethyl acetate was added again to perform extraction.
The ethyl acetate layers were combined to obtain solution A. Acetone 1 for the cells
1 and stirred well and left for 2 hours. After filtration,
The supernatant was concentrated under reduced pressure to remove acetone. After adding 1 liter of water to dissolve the concentrated solution, 1 liter of ethyl acetate was added to perform extraction. After separating the ethyl acetate layer, 1 l of ethyl acetate was added and extraction was performed again. A solution obtained by combining the ethyl acetate layers was designated as solution B. Anhydrous sodium sulfate was added to the combined solution A and solution B for dehydration. After filtration, the supernatant was concentrated under reduced pressure, and then column chromatography was performed on silica gel. Hexane: ethyl acetate = 1: 0, 9: 1,
Elution was performed in the order of 4: 1, 2: 1 and 1: 2, and the active fractions were collected and concentrated. Column chromatography was performed again on silica gel. Elution was performed with chloroform: methanol = 1: 0, 100: 1, 25: 1 and 3: 1. The active fraction was collected and concentrated under reduced pressure. Finally, fractionation was performed by HPLC.
Column: Sensupak ODS-4251-N, 10
φ × 250 mm, developing solvent: 90% methanol (0.
The sample was collected at a flow rate of 1 ml / min. When the peaks at 39.6 minutes, 44.8 minutes, 52.4 minutes, 61.2 minutes and 72 minutes were collected, nonactin, monactin, dynactin, trinactin, and tetranactin were respectively 5.8 mg, 11.3 mg, and 16.3 min. 8 mg,
26 mg and 2.4 mg were obtained.

The macrotetrolide compound (I) or a complex salt thereof, which is an active ingredient of the medicament of the present invention, has an inhibitory effect on intracellular glycoprotein transfer and exhibits antiviral activity.
It was confirmed by the following test method. That is, the viral glycoprotein synthesized in the endoplasmic reticulum is transferred inside the cell and appears on the cell surface to induce cell fusion. When the intracellular transfer is inhibited, the viral glycoprotein is accumulated in the cell even if synthesized, and its appearance on the cell surface is suppressed to inhibit cell fusion. Therefore, the intracellular transfer inhibitor can be selected by selecting a substance that does not cause cell fusion by the addition of the test sample and does not inhibit the biosynthesis of glycoprotein.

Test Example Assay for Inhibition of Glycoprotein Intracellular Transfer Preparation of Cells The established baby hamster kidney cells (BHK-21) are subcultured using a 700 ml culture colben manufactured by Nunc. The monolayer cultured BHK-21 cells are detached using a 0.25% trypsin solution, and the cells are collected by low-speed centrifugation.
A suspension of 5 × 10 ⁻⁴ cells / ml is prepared using Eagle's minimum essential medium (manufactured by Nissui Pharmaceutical) supplemented with 10% calf serum. 100 μl of the cell suspension was placed in a 96-well Nunc plate.
The mixture was dispensed every 1 and cultured in a carbon dioxide incubator for 3-4 days to prepare dense monolayer cultured cells.

2. Cell fusion assay The medium of the cultured cells prepared by the method of 1 is discarded, and a Newcastle disease virus (NDV) solution diluted with a cell culture medium so as to have a multiplicity of infection of 2 is poured into each well at 100 μl / portion. After incubating for 1 hour at 37 ° C. to adsorb the virus, an equal amount of a medium containing the five test samples obtained in Production Examples or a dilution thereof is added. Thereafter, the virus is propagated by further incubating at 37 ° C. for 20 hours. The presence or absence of syncytia formed as a result of cell fusion is examined using an inverted microscope. Table 5 shows the results. The result of fusion is indicated by ++, and the result of no fusion is indicated by-.

[0031]

[Table 5]

3. Glycoprotein synthesis assay A sample that has been assayed for cell fusion by the method 2 is subjected to sonication to disrupt the cells, and then the hemagglutinin, which is the viral coat glycoprotein, is quantified by a conventional pattern method. Glycoprotein synthesis was assayed. As a result, the specificity is lost and the glycoprotein synthesis is suppressed in the case of high concentration doses of No. 2 or higher in any of the production example compounds, but in the low concentration region at or near the minimum effective concentration, the glycoprotein It was confirmed that the synthesis of was hardly suppressed.

The inhibitory effect of the active ingredient (I) or its complex salt on the appearance of the virus glycoprotein cell surface was also confirmed by another method of staining the fixed infected cells by the secondary fluorescent antibody method.
That is, BHK cells grown on a cover glass were infected with bovine vesicular stomatitis virus (VSV).
After incubation for 1 hour at, the test sample obtained in the production example was added, incubated at 37 ° C. for 8 hours, fixed with formaldehyde, treated with methanol, and treated with anti-VSV-G antibody. The cells were treated with a fluorescent-labeled anti-rabbit Ig antibody and examined under a fluorescence microscope. As a result, as shown in FIG.
As is clear from the results of the g / ml secondary fluorescent antibody method confirmation test, no fluorescence staining based on the presence of glycoprotein was observed in the cells of the control group, but the cells were stained on the cell surface. In contrast, in the dynactin-treated group, fluorescence staining was observed in the cells, but was not stained on the cell surface. Therefore, in the control group, the viral envelope glycoprotein biosynthesized intracellularly appears on the cell surface, whereas in the dynactin-treated group, the viral envelope glycoprotein biosynthesized intracellularly is intracellularly expressed. Since it was accumulated and did not appear on the cell surface, the inhibitory effect of dynactin on intracellular glycoprotein transfer was confirmed.

[0034]

The macrotetrolide compound (I) or its complex salt, which is an active ingredient of the medicament of the present invention, has an inhibitory effect on intracellular transfer of glycoprotein and exhibits antiviral activity.
Therefore, a medicine containing the compound (I) or a complex salt thereof as an active ingredient is a disease in which inhibition of intracellular transfer of a glycoprotein acts as a mechanism to exert a therapeutic activity, such as various viral diseases, various cancers, and the like. It is useful as an agent which can provide a preventive / therapeutic agent for various allergic diseases. Further, the novel microorganism of the present invention is useful as a microorganism which can be used in a production method for producing the active ingredient compound (I) of the medicament of the present invention or a complex salt thereof. Further, the production method of the present invention comprises:
It is useful as a new method for producing the active ingredient compound (I) or a complex salt thereof of the medicament of the present invention.

[Brief description of the drawings]

FIG. 1 shows the mass spectrum of a complex salt of nonactin.

FIG. 2 shows the ultraviolet absorption spectrum of nonactin,

FIG. 3 shows the nuclear magnetic resonance spectrum of nonactin,

FIG. 4 is a mass spectrum of a complex salt of monactin.

FIG. 5 shows the ultraviolet absorption spectrum of monactin.

FIG. 6 shows a nuclear magnetic resonance spectrum of monactin.

FIG. 7 shows a mass spectrum of a complex salt of dynactin,

FIG. 8 shows the ultraviolet absorption spectrum of dynactin,

FIG. 9 shows a nuclear magnetic resonance spectrum of dynactin.

FIG. 10 shows the mass spectrum of the complex salt of trinactin,

FIG. 11 shows the ultraviolet absorption spectrum of trinactin.

FIG. 12 shows a mass spectrum of a complex salt of tetranactin,

FIG. 13 shows an ultraviolet absorption spectrum of tetranactin,

FIG. 14 shows a substitute photograph for a drawing of the confirmation test result of dynactin by the secondary fluorescent antibody method, respectively.

────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] April 10, 1995

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Brief description of drawings

[Correction method] Change

[Correction contents]

[Brief description of the drawings]

FIG. 1 shows a mass spectrum of a complex salt of nonactin.

FIG. 2 shows an ultraviolet absorption spectrum of nonactin.

FIG. 3 shows a nuclear magnetic resonance spectrum of nonactin.

FIG. 4 shows a mass spectrum of a complex salt of monactin.

FIG. 5 shows an ultraviolet absorption spectrum of monactin.

FIG. 6 shows a nuclear magnetic resonance spectrum of monactin.

FIG. 7 shows a mass spectrum of a complex salt of dynactin.

FIG. 8 shows an ultraviolet absorption spectrum of dynactin.

FIG. 9 shows a nuclear magnetic resonance spectrum of dynactin.

FIG. 10 shows a mass spectrum of a complex salt of trinactin.

FIG. 11 shows an ultraviolet absorption spectrum of Trinactin.

FIG. 12 shows a mass spectrum of a complex salt of tetranactin.

FIG. 13 shows an ultraviolet absorption spectrum of tetranactin.

FIG. 14 shows a micrograph instead of a drawing of the result of a confirmation test of dynactin by the secondary fluorescent antibody method.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code Agency reference number FI Technical display location C12N 1/20 A 8828-4B C12P 17/18 D 7432-4B // (C12N 1/20 C12R 1 : 465) (C12P 17/18 C12R 1: 465) (72) Inventor ▲ Takaaki Tsukiaki 2-1 Hirosawa, Wako-shi, Saitama Pref. RIKEN

Claims (6)

[Claims] 1. A glycoprotein intracellular transfer inhibitor comprising a macrotetrolide compound represented by the general formula (I) or a complex salt thereof as an active ingredient. Embedded image (Wherein R ₁ , R ₂ , R ₃ and R ₄ are the same or different and represent a methyl group or an ethyl group) 2. The medicament according to claim 1, which is an antiviral agent. 3. The medicament according to claim 1, which is an anticancer agent. 4. The medicament according to claim 1, which is an antiallergic agent. 5. Streptomyces sp. Q30101, which has an ability to produce the macrotetrolide compound (I) or the complex salt thereof, which is the glycoprotein intracellular transfer inhibitor according to claim 1. 6. A bacterium belonging to the genus Streptomyces, which is capable of producing the macrotetrolide compound (I) or the complex salt thereof according to claim 1,
The glycotetrolide compound (I) or the complex salt thereof according to claim 1, wherein the macrotetrolide compound (I) or the complex salt thereof is collected from the culture. I) or a method for producing a complex salt thereof.