HK1026904A - Kodaistatins a, b, c and d, a process for their production and their use - Google Patents

Description

Costatin A, B, C and D, and preparation method and application thereof

The present invention relates to novel compounds designated Kodaistatin (Kodaistatin) A, B, C and D, their preparation and use.

Increased hepatic glucose output rate is a common feature of diabetes. Especially in non-insulin dependent diabetes mellitus (NIDDM), fasting blood glucose levels have a clear correlation with hepatic glucose output. Glucose is produced in the liver by both gluconeogenesis and glycogenolysis. The last step of both pathways is catalyzed by microsomal glucose-6-phosphatase, a key enzyme in the homeostatic regulation of blood glucose levels. The level of this enzyme is known to be elevated in both experimental and pathological conditions of diabetes. Interference with this enzyme system will result in a decrease in hepatic glucose production.

Hepatic glucose-6-phosphatase is a multi-component system comprising at least three functional active sites: glucose-6-phosphate translocase (T1), glucose-6-phosphate phosphohydrolase and phosphate/pyrophosphate translocase (T2). Glucose-6-phosphate translocase promotes the transfer of glucose-6-phosphate into the lumen of the Endoplasmic Reticulum (ER). Phosphohydrolases, the active site of which is located on the internal surface of the ER, hydrolyze glucose-6-phosphate, releasing glucose and phosphate into the lumen. The phosphate is excreted mediated by phosphate/pyrophosphate translocase, and the exact mechanism of glucose excretion is not known.

The high substrate specificity of glucose-6-phosphate translocase makes it a potential site of action for drugs in the treatment of diabetes. Of the physiologically produced phosphoglycoses, only glucose-6-phosphate is transferred by translocases. In contrast, phosphatases are non-specific and are known to hydrolyze a variety of organophosphates. A) A series of non-specific glucose-6-phosphatase inhibitors were reported in the literature, such as phloridzin [ journal of biochemistry (J.biol. chem.), 242, 1955-1960(1967) ], 5 ' -dithio-bis-2-nitrobenzoic acid [ Biochemist-physical research Command, Biochem.Biophys.Res.Commun ], 48, 694-699(1972) ], 2 ' -diisothiocyanatostilbene and 2-isothiocyanato-2 ' -acetoxystilbene [ journal of biochemistry, 255, 1113-1119(1980) ]. The first inhibitors of the glucose-6-phosphatase system useful as therapeutics are disclosed in European patent publications 587087 (application No. 93114260.8) and 587088 (application No. 93114261.6).

It has now been found that copeptin A, B, C and D have enzyme inhibitor activity, in particular against glucose-6-phosphatase translocase.

The subject of the invention is therefore the following:

1) a codestatin A and a codestatin B of the formula C₃₅H₃₄O₁₁And pharmaceutically acceptable salts, esters, ethers, and obvious chemical equivalents thereof.

Copeptin B has a hitherto unreported new structure, formed from one O-hydroquinone, phenol, unsaturated γ -lactone, dihydroxy-cyclopentenone and an α, β, γ, δ -unsaturated carbonyl moiety, and is one diastereomer of copeptin a.

Codestatins a and B are compounds having the structural formula I:

the invention also relates to:

2) codestin C and codestin D, which are of the formula C₃₅H₃₄O₁₂And pharmaceutically acceptable salts, esters, ethers, and obvious chemical equivalents thereof.

The copeptin C has a hitherto unreported new structure formed from O-hydroquinone, unsaturated gamma-lactone, dihydroxy-cyclopentenone and alpha, beta, gamma, delta-unsaturated carbonyl moieties. Codestatin D is one diastereomer of codestatin C.

The structural formulas of kodaistatin C and kodaistatin D are different from the structural formula I given above, with the addition of (especially most commonly at the 6-position of the terminal phenyl ring A) a hydroxyl group.

The present invention therefore relates to all stereoisomers of the copestine and mixtures thereof. The individual stereoisomers may be separated by known methods such as normal phase chromatography, anion exchange chromatography, HPLC and selective crystallization.

The preparation of physiologically tolerated salts (e.g. sodium, potassium, ammonium salts), esters (e.g. organic acid esters) and chemical equivalents (oxidation products, addition products such as hydrates) is well known to the person skilled in the art.

It is another object of the present invention to provide a method for preparing novel compounds, copestine A, B, C and D, from the culture of HIL-051652 and mutants and variants thereof. The method comprises culturing HIL-051652 culture and its mutants and variants in a medium containing a carbon source, a nitrogen source and nutritive inorganic salts under aerobic conditions, and separating and purifying the compound from the culture filtrate.

The medium comprises a carbon source, a nitrogen source, inorganic salts and optionally trace elements. The carbon source may be, for example, starch, glucose, sucrose, dextrin, fructose, molasses, glycerol, lactose or galactose, preferably starch. The nitrogen source is selected from soybean powder, peanut powder, yeast extract, beef extract, peptone, tryptone, malt extract, corn steep liquor, gelatin or casamino acid, preferably tryptone and yeast extract. The nutritive inorganic salt is, for example, disodium hydrogenphosphate, dipotassium hydrogenphosphate, sodium chloride, calcium carbonate, potassium nitrate, ammonium sulfate or magnesium sulfate, preferably sodium chloride and calcium carbonate.

The HIL-051652 culture was cultured at 25-30 deg.C and pH 6.0-8.0. Preferably, the HIL-051652 culture is cultured at a temperature of 25 ℃ (± 1 ℃) and at a pH of about 7.0.

Preferably, the fermentation is carried out for 40-90 hours, until the best yield of the compounds of the invention is obtained. Submerged fermentations of 45-70 hours, such as shake flask fermentations and fermentations in laboratory fermentors, are particularly preferred. If desired, antifoams such as Desmophen may be used in the fermentation process^(polypropylene oxide, Bayer AG, Leverkusen, Germany). Fermentation and the progression of copestine A, B, C and D formation can be determined by measuring the amount of untreated and TritonX-100 in microtiter plates^The activity of glucose-6-phosphate in disrupted rat liver microsomes was monitored by HPLC using a modification of the colorimetric assay described in enzymology 174, 58-67(1989) at room temperature. In the resulting medium, cotestant B, C and D are minor compounds and cotestant a is the major compound. The crude active product is recovered by extracting the mycelium with a water-miscible solvent such as methanol, ethanol and acetone, extracting the filtrate from the fermentation broth with a water-immiscible solvent such as ethyl acetate, dichloromethane, chloroform or butanol at pH5-8, or with a polymeric resin such as "Diaion HP-20^"(Mitsubishi chemical, Japan)," Amberlite XAD^"(Rohm and Haas Industries, USA) hydrophobic interaction chromatography or ion exchange chromatography on activated carbon at pH 5-8. The preferred method is to use "Diaion HP-20^Adsorbing, and desorbing with an eluent containing water, methanol, acetone or acetonitrile or their mixtureA mixture of (a). Concentration and lyophilization of the active eluate yielded the crude compound.

The crude product can be further purified by any of the following techniques: normal phase chromatography (using alumina or silica gel as stationary phase and ethyl acetate, chloroform, methanol or their mixture as eluent); reverse phase chromatography (using as stationary phase a reverse phase silica gel such as dimethyloctadecylsilyl silica gel, RP-18 or dimethyloctaalkylsilyl silica gel, RP-8, and as eluent a buffer solution such as water, a buffer solution such as phosphate, acetate, citrate (pH2-8), and an organic solvent such as methanol, acetonitrile or acetone, or a mixture of these solvents); gel filtration chromatography (on a resin such as SephadexLH-20)^(Pharmacia chemical industry, Sweden), TSKgel' Toyoperal HW-40F^' (Tosohaas, Tosoh Corp., Japan) "Sephadex  G-10", "Sephadex  G-25" in a solvent such as methanol, chloroform or ethyl acetate and a mixture thereof, or in water; or by ion exchange chromatography, preferably anion exchange chromatography; or countercurrent chromatography, using a biphasic solvent system of two or more solvents such as water and chloroform. These techniques may be reused or a combination of different techniques may be used. A preferred method is first chromatographic separation on Toyopearl followed by reverse phase modified silica gel (RP-18).

The microorganisms used for the preparation of copestine A, B, C and D were isolated from soil samples collected in Kodaikanal, TamiNadu, India under culture number Y-93, 02839(HIL-051652), hereinafter HIL-051652. The microorganism HIL-051652 was identified as Aspergillus terreus. Deposited at 21 months 10 in 1996 at Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Germany, Braunschweig, with the deposit number DSM 11247.

Costatin A, B, C and D were effective in inhibiting the activity of microsomal glucose-6-phosphate translocase enzyme in rat liver. Their approximate IC₅₀The values are as follows:

copeptin a: 0.2. mu.g/ml (about 300nM)

Copeptin B: 0.3. mu.g/ml

Copeptin C: 0.09. mu.g/ml

Copeptin D: 0.5. mu.g/ml

In contrast, it is expected that statin A inhibits the phosphatase activity, IC, in detergent-disrupted microsomes₅₀About 200. mu.g/ml (about 300. mu.M), indicating a high degree of specificity for translocase. Furthermore, cotestastin A does not affect the activity of the phosphate/pyrophosphate translocase. Coxidasatin A is a reversible competitive inhibitor of glucose-6-phosphate translocase.

The effect of copeptin a on glucose output was further investigated in isolated rat hepatocytes. It can inhibit fructose-induced gluconeogenesis and glucagon-induced hepatic glycogenolysis, IC₅₀About 25. mu.g/ml and 50. mu.g/ml, respectively.

Therefore, another aspect of the present application is: the use of copeptin A, B, C and D as medicaments, and the use of copeptin A, B, C and D in the manufacture of medicaments having an anti-diabetic effect. It is also an object of the present application to provide medicaments comprising active amounts of copestine A, B, C and D, respectively.

The pharmaceutical formulation, method of administration and dosage range of the copestine will depend on the type of disease being treated and the specifics of each disease/disorder, and can be optimized by methods well known in the art. In this respect, reference is made to the document cited in paragraph A) above. The copestine A, B, C and D can be administered orally, intramuscularly or intravenously. They can be prepared by mixing the compounds with one or more pharmaceutically acceptable adjuvants and/or excipients, and formulating into suitable administration forms, such as tablets, coated tablets, capsules or solutions or suspensions suitable for parenteral administration. Examples of excipients are fillers, emulsifiers, lubricants, taste-masking agents, colorants or buffer substances.

Examples of adjuvants and/or excipients which may be mentioned are tragacanth, lactose, talc, agar, polyethylene glycol, ethanol and water. Suitable and preferred formulations for parenteral administration are aqueous solutions or suspensions. The active substance may also be administered in a suitable form, such as in a capsule, without the use of a vehicle or diluent.

These compounds can be prepared into their pharmaceutically acceptable derivatives such as esters and ethers. Esters can be prepared by reacting the compound with a carboxylic acid in the presence of a catalyst, or by reacting the compound with an acylating agent, such as an acid chloride. Other methods for preparing esters can be found in the literature, e.g., higher organic synthesis, fourth edition, j.

Ethers can be prepared by reacting the compound with an alkylating agent under basic conditions. Other methods for preparing ethers can be found in the literature, e.g., higher organic synthesis, fourth edition, j.

The following examples are illustrative of the invention, but do not limit its scope: example I isolation of culture from soil HIL-051652(a) composition of isolation medium (saproberts agar) peptone: 10.0g of glucose: 40.0g agar: 13.0g deionized water: 1.0 liter pH: 7.0(b) spreading and separating of soil sample

10 g of a soil sample collected from Kodaikanal, Tami Nadu, India was added to a 250ml conical flask containing 90ml of sterile deionized water and shaken on a rotary shaker for 2 hours (220 rpm). Diluting the soil suspension to 10 degrees by ten times^-5. From the final dilution, 1ml of the suspension was placed in the center of a sterile glass petri dish (15 cm diameter) and about 50ml of the above separation medium supplemented with 50. mu.g/ml chloramphenicol and 0.5% sodium propionate was poured in. The medium was cooled to 45 ℃ and the dish was rotated thoroughly. The soil suspension and medium mixture was allowed to set and incubated at 25 ℃ (± 1 ℃) for 7 days. Observing the culture dish periodically, and separating culture from the grown microorganismSubstance HIL-051652. Example II maintenance of culture HIL-051652 culture HIL-051652 was maintained on Saxabeu's agar as described in example I

Heating to dissolve the above components completely, and subpackaging in test tubes and sterilizing at 121 deg.C for 20 min. The test tube was cooled and placed on an incline until solidification. Cultures grown on agar slants crossed with a metal ring, HIL-051652, incubated at 25 ℃ (+ -1 ℃) until good growth was observed. The grown culture was stored in a refrigerator at 8 ℃. Example III composition of fermentation seed Medium for culture HIL-051652 in Shake flasks: starch: 15.0g of glucose: 5.0g soybean flour: 15.0g yeast extract: 2.0g of corn steep liquor: 1.0g NaCl: 5.0g of CaCO₃: 2.0g deionized water: 1.0 liter pH: 6.8

The seed culture medium is divided into 80ml portions, and the portions are filled into 500ml conical flasks and autoclaved at 121 ℃ for 20 minutes, and cooled to room temperature. Each flask was inoculated with a full circle of the well-grown culture of example II above, and shaken on a rotary shaker at 240 rpm at 25 ℃ (± 1 ℃) for 72 hours to obtain a seed culture. Composition of fermentation medium starch: 24.0g of glucose: 15.0g tryptone: 5.0g of yeast extract: 5.0g beef extract: 3.0g of CaCO₃: 4.0g deionized water: 1.0 liter pH: 6.5

The fermentation medium was divided into 60ml portions, and autoclaved at 121 ℃ for 20 minutes in 500ml Erlenmeyer flasks, cooled to room temperature, and inoculated with the above seed culture (1% v/v). Fermenting at 25 deg.C (+ -1 deg.C) at 240 rpm for 40-48 hr on rotary shaker.

Production of active compound was monitored by measuring inhibition of glucose-6-phosphate translocase. After the fermentation has ended, the medium is centrifuged and the copestine A, B, C and/or D is isolated and purified from the medium filtrate as described in example V. Example IV fermentation of culture HIL-051652 stage 1 in a fermenter: preparation of seed cultures in Shake flasks

The seed medium of example III was divided into 160ml portions and autoclaved in 1L Erlenmeyer flasks for 20 minutes. Seed cultures were grown in these shake flasks as described in example III. And (2) stage: preparation of seed cultures in fermentors

75 l of seed culture were sterilized in situ in a 100 l Marubishi fermenter at 121 ℃ for 45 minutes, cooled to 25. + -. 1 ℃ and inoculated with 3 l of the seed culture described above as described in example III. The fermentation parameters were as follows:

temperature: 25 deg.C (+ -0.5 deg.C)

Stirring: 80 rpm

Ventilating: 50 l/min

Harvesting time: 50 hours stage 3: large-scale fermentation

750 l of the fermentation medium described in example III were placed in a 1000 l Marubishi fermenter, while 175ml Desmophen  (polypropylene oxide) was added as antifoam, sterilized in situ at 121 ℃ for 45 minutes, cooled to 25. + -. 1 ℃ and inoculated with 75 l of the seed culture from stage 2. The fermentation parameters were as follows:

temperature: 25 deg.C (+ -0.5 deg.C)

Stirring: 50 rpm

Ventilating: 350 l/min

Harvesting time: 40-44 hours

Production of active compound was monitored by measuring inhibition of glucose-6-phosphate translocase. The pH of the medium after fermentation is 6.0-7.0. After harvest, the medium was centrifuged and the glucose-6-phosphate translocase inhibitor copestine A, B, C and/or D was isolated from the medium filtrate as described in example V. Example V isolation and purification of copestine A, B, C and/or D:

about 1000 liters of the resulting broth was centrifuged and separated from the mycelium (110 kg). Both mycelium and filtrate contained copestine A, B, C and D. The filtrate (830 l) was combined with a 30% aqueous methanol extract of the mycelium (330 l) and passed through Diaion HP-20^Column (35L, 3% v/v). The column was rinsed thoroughly with deionized water (50 liters) and then with a step gradient of CH₃And (4) eluting with a CN aqueous solution. Elution with 10% CH₃CN (90 liters) and 30% CH₃CN (90L) was completed and fractions were collected in a 15L scale. Combined with 30% CH₃CN eluted the active fraction obtained (3 × 15 l). Concentrating under reduced pressure at 35 deg.C under 10-100 mm Hg, and lyophilizing to obtain crude active product (225g) IC of corbestatin A₅₀25. mu.g/ml.

The crude product obtained was then separated by two successive gel permeation chromatographs on a stationary phase TSKgelToyopearl HW-40F , varying the gel ratio according to the substrate. The crude product was divided into 15 portions, and each 15 g portion was passed through a Latek-S-die M6-48 glass column packed with Toyopearl HW-40F  (1.5L). The mobile phase is 10% CH₃CN aqueous solution, flow rate maintained at 10 ml/min, pressure 3-5 bar. Fractions were collected in a volume of 250 ml. Mixing active eluates, concentrating under reduced pressure at 35 deg.C under 10-100 mm Hg, and lyophilizing to obtain concentrated active substance (3.0g) to be used as IC of statin A₅₀Is 1-1.5 mug/ml.

The concentrated material was further divided into 10 portions of 300 mg each, and passed through a Latek-S-Graulen M4-48 glass column packed with Toyopearl HW-40F  (500 ml). The mobile phase is 10% CH₃CN aqueous solution, the flow rate is maintained between 1.5 and 2.0 ml/min. Fractions were collected in a 20ml measuring vessel. Combining all active fractions, concentrating at 35 deg.C under 10-100 mm Hg, and lyophilizing to obtain a semi-purified substance containing active substance codestine ADaltin A is the main compound, IC₅₀At 0.375. mu.g/ml, copeptin B, C and/or D was the minor compound (0.85 g).

Cofortatins B, C and D were finally separated from Cofortatin A at a mobile phase flow rate of 8 ml/min and detected at 294nm to give pure Cofortatin B (0.004g), Cofortatin C (0.011g) and Cofortatin D (0.004 g).

Purity of copestine B, C and D was checked by HPLC (high pressure liquid chromatography) using LiChrocart-250-4 RP Select B (5. mu. in 20 min at 40 ℃ from 0.1% aqueous phosphoric acid (pH2.5) to CH₃CN was eluted with gradient, flow rate 1 ml/min, UV detection at 294 nm.

The resulting semi-purified statins A were finally purified by preparative HPLC on a column of 16X 250mm Eurospere C-18(10 μ) using 20% CH₃CN water solution is used as a mobile phase, the flow rate is 8 ml/min, the ultraviolet detection wavelength is 294nm, and pure codestatin A (0.14g), IC are obtained₅₀It was 0.2. mu.g/ml.

The physicochemical and spectral properties of copeptin a are summarized in tables 1 and 1A, for copeptin C in table 2, for copeptin B in table 3, and for sitptin D in table 4. Table 1 copeptin a trait: yellow solid solubility: MeOH, CH₃CN, and DMSO melting point: greater than 200 deg.C (decomposition) [ alpha ]]_D: -85.7 ° (c 0.042, methanol) high pressure liquid chromatography: retention time: 7.3 min (HPLC) column: [4mm X (30+250) mm]ODS-Hypersil(5μ)

Eluent: CH (CH)₃CN-H₂O(20∶80)

Flow rate: 1 ml/min; and (3) detection: 294nm

FIG. 1 shows the molecular weights of the appended chromatograms: 630(ESI-MS) formula: c₃₅H₃₄O₁₁[ detection value: m/z 631.2174(M + H)⁺(HR FAB-MS，

Substrate: TFA/NBA, internal standard: PEG 500); c₃₅H₃₄O₁₁The calculated value of (a): 631.2179]UV: FIG. 2 is the accompanying spectrum IR (KBr): FIG. 3 is a chart of the attached spectrum¹HNMR(300MHz，DMSO-d₆): FIG. 4 is a chart of the attached spectrum¹³CNMR(150MHz，DMSO-d₆): FIG. 5 is a chart of the attached spectra, 1A, of Coxidastine A in deuterated methanol¹H and¹³c data (ppm rel. TMS 278K)

	1H	13C
			A1	-	160.14
A2	6.84d	116.89
			A3	7.62d	133.92
A4	-	125.45
			B1	6.20s	110.71
B2	-	142.01
			B3	-	166.51
B4	-	102.86
			B5	-	172.74
C1	-	121.65
			C2	6.87s	118.77
C3	-	147.93
			C4	-	146.95
C5	6.40s	114.76
			C6	-	124.24
D1	-	163.69
			D2	-	139.12
D3	-	202.65
			D4	4.54s	86.32
D5	-	91.69
			D6	-	210.05br
D7	2.46s	28.52br
			E1	3.48d/3.23d	38.69
E2	-	196.65
			E3	6.02d	122.94
E4	6.96d	149.93
			E5	-	133.37
E5-Me	1.75s	12.76
			E6	5.48d	151.51
E7	2.46	36.31
			E7-Me	1.02d	20.85
E8	1.34m/1.22m	31.13
			E9	0.80t	12.51

TABLE 2

Copeptin C trait: yellow solid solubility: MeOH and DMSO melting points: greater than 200 deg.C (decomposition) [ alpha ]]_D: -20.0 ° (c 0.04, methanol) HPLC retention time: 12.81 minutes

FIG. 6 shows the molecular weights of the appended chromatograms: 646(ESI-MS) elemental analysis: detection value: c, 64.52; h, 5.41C₃₅H₃₄O₁₂The calculated value of (a): c, 65.01; h, 5.26 formula: c₃₅H₃₄O₁₂UV: FIG. 7 is the accompanying spectrum IR (KBr): FIG. 8 is a chart of the attached spectrum¹HNMR (300 MHz; FIG. 9 is attached spectrum DMSO-d₆)¹³CNMR(δ，75MHz，： 208.78，201.38，194.58，176.28，172.24，166.94，148.70，DMSO-d₆) 146.83，146.61，145.49，145.10，144.99，142.76，134.56，

131.77，126.03，124.86，122.69，122.46，121.60，116.99，

116.32，115.32，112.48，100.12，90.36，89.75，84.70，

37.29, 34.34, 29.37, 28.17, 19.95, 12.17, and 11.75 table 3

Copeptin B trait: yellow solid solubility: MeOH and DMSO melting points: > 200 ℃ (decomposition) HPLC retention time: 13.45 min molecular weight: 630(ESI-MS) formula: c₃₅H₃₄O₁₁UV (65: 35: 240, 300 and 375 nmCH)₃CN-0.1% phosphoric acid)¹HNMR (300 MHz; FIG. 10 is attached spectrum DMSO-d₆) TABLE 4

Copeptin D trait: yellow solid solubility: MeOH and DMSO melting points: > 200 ℃ (decomposition) HPLC retention time: 12.67 min molecular weight: 646(ESI-MS) formula: c₃₅H₃₄O₁₂UV (65: 35: 285 and 380 nmCH)₃CN-0.1% phosphoric acid)¹HNMR (300 MHz; FIG. 11 is attached spectrum DMSO-d₆)

Claims (14)

1. A compound of formula I, codestine A/B, and stereoisomers, pharmaceutically acceptable salts, esters, ethers and obvious chemical equivalents thereof.

2. A molecule is C₃₅H₃₄O₁₁The compounds of (a) can be rivastigmine A/B and pharmaceutically acceptable salts, esters and ethers thereof.

3. AThe molecular formula is C₃₅H₃₄O₁₁Can be obtained by culturing the microorganism Aspergillus terreus HIL-051652(DSM 11247) in a medium containing a carbon source, a nitrogen source, inorganic salts and trace elements, and isolating and purifying the compound from the culture by a conventional method.

4. A molecule is C₃₅H₃₄O₁₂The compounds of (a) can be rivastigmine C/D and pharmaceutically acceptable salts, esters and ethers thereof.

5. A molecule is C₃₅H₃₄O₁₂Can be obtained by culturing the microorganism Aspergillus terreus HIL-051652(DSM 11247) in a medium containing a carbon source, a nitrogen source, inorganic salts and trace elements, and isolating and purifying the compound from the culture by a conventional method.

6. Copeptin A, B, C or D having any one or more of the physical characteristics set forth in tables 1, 3, 2, or 4, respectively.

7. A process for the preparation of cotropin A, B, C or D, comprising culturing the microorganism aspergillus terreus HIL-051652(DSM 11247) in a medium containing a carbon source, a nitrogen source, inorganic salts and trace elements under aerobic conditions, and isolating and purifying the compound from the culture using conventional methods.

8. The method of claim 7, wherein the culturing is carried out at a temperature of between about 25 and 30 ℃ and a pH of between about 6 and 8.

9. The method of claim 7 or 8, wherein the culturing is carried out at a temperature of 25 ℃ (± 1 ℃) at a pH of about 7.0.

10. The process according to any one of claims 7 to 9, wherein the cultivation is submerged fermentation.

11. A medicament comprising a compound according to any one of claims 1 to 6 together with conventional adjuvants and/or excipients for pharmaceutical use.

12. Use of a compound according to any one of claims 1 to 6 for the preparation of a medicament having glucose-6-phosphate translocase inhibitory activity.

13. The use of a compound according to any one of claims 1 to 6 for the preparation of a medicament having an antidiabetic effect.

14. Aspergillus terreus HIL-051652(DSM 11247).