JPH06329671A - New alkaloid and its use - Google Patents

Abstract

PURPOSE:To obtain a new alkaloid, having remarkable inhibiting activity against saccharide hydrolases and useful as a therapeutic agent for diseases in abnormal ity (lowering) of glucose tolerance. CONSTITUTION:The alkaloid of the formula (R1 is H or OH), e.g. 1, 2alpha, 3beta, 4alpha, 6beta-pentahydroxynortropan. This compound of the formula is obtained by extracting Mori Cortex, dry powder of Mori Folium or Mori Fructus with hot water, then adding ethanol and methanol thereto, separating an alcohol- soluble part with a strong acidic cation exchange resin and a strong basic anion exchange resin and carrying out the fractionation with 0.01-0.0N aqueous ammonia using CM-Sephadex C-25(R) (a ammonium salt type) or with a 0.1M pyridine-acetic acid buffer solution (pH6.0) using Dowex 50WX8(R) (a pyridine salt type).

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel alkaloid. Specifically, it relates to a novel alkaloid useful as a sugar hydrolase inhibitor that can be used as a medicine such as a hypoglycemic agent, and its use.

[0002]

2. Description of the Related Art Ingestion of glucose alone or a carbohydrate constituting the glucose as food and beverage causes hyperglycemia. In the case of healthy humans, this hyperglycemic condition will eventually improve and maintain normal blood glucose levels. This is because glucose in blood is metabolized.

However, hyperglycemic symptoms last for a long time in diseases in which glucose tolerance is abnormal (decreased) typified by diabetic symptoms, and as a result, promotion of progression from prediabetes to diabetes and It gets worse. In addition, there are many known diseases in which this hyperglycemic condition is one of the causes, or diseases in which they are aggravated, and examples thereof include obesity, hypertriglyceridemia, and hyperinsulinemia.

As therapeutic agents for diseases in which abnormal glucose tolerance (decrease) is observed, insulin or an oral hypoglycemic agent is used alone or in combination as necessary. Oral hypoglycemic drugs are widely used to treat non-insulin dependent diabetes. However, conventional oral hypoglycemic agents have strong side effects, and caution is required when using them.
Taking a sulfonylurea drug, which represents an oral hypoglycemic drug, as an example, its side effects include severe gastrointestinal symptoms (anorexia, nausea, diarrhea, etc.), hematopoietic disorders (anemia, leukopenia, etc.), and liver dysfunction. I know what is serious.

Therefore, there has been a demand for the development of a drug useful for treating a disease in which glucose tolerance is abnormal (decreased).

[0006]

Means for Solving the Problems The inventors of the present invention have conducted extensive studies in order to solve the above problems, and as a result, found that a novel alkaloid has a remarkable sugar hydrolase inhibitory activity, and completed the present invention. Came to do.

That is, the present invention provides (1) the following formula (I) (In the formula, R ₁ represents a hydrogen atom or a hydroxyl group.) (Hereinafter, an alkaloid in which R ₁ represents a hydrogen atom is referred to as a compound 01, and an alkaloid in which R ₁ represents a hydroxyl group is referred to as a compound 02). (2) The following formula (II) An alkaloid represented by (hereinafter referred to as compound 03). (3) The following formula (III) (In the formula, R _{2 to} R ₅ are shown in Table 2 below.) Table 2 Is an alkaloid. (4) The following formula (IV) (In the formula, R6 represents a β-glucopyranosyl group.) An alkaloid (hereinafter, referred to as compound 12). (5) A hypoglycemic agent comprising compounds 1 to 12 (hereinafter collectively referred to as the compound of the present invention) as an active ingredient.

The compound of the present invention can be obtained by adding ethanol or methanol to an extract obtained by heat-extracting a mulberry plant, and treating the soluble fraction thereof by column chromatography using various ion exchange resins. .

Specifically, after extracting mulberry bark or dried mulberry leaf powder or mulberry with hot water, ethanol or methal is added, the alcohol-insoluble portion is removed, and the soluble portion is a strongly acidic cation exchange resin or weakly acidic exchange resin. The compound of the present invention can be separated by treatment with (ammonium salt type), a strongly basic anion exchange resin or the like. For the compound of the present invention that is insufficiently separated, fractionation is performed with CM-Sephadex C-25 (ammonium salt type) with 0.01 to 0.05 normal ammonia water, or with Dowex 50W × 8 (pyridine). 0.1 M pyridine-acetic acid buffer (pH 6.
Fractionation in 0) allows complete separation.

Next, experimental examples showing that the compound of the present invention has a remarkable excellent sugar hydrolase inhibitory activity,
The details will be described.

Experimental Example 1 0.05 m of a 0.2 M maleic acid buffer solution (pH 5.8)
1, 3 parts of a reaction solution containing an aqueous solution of the compound of the present invention (0.08 ml) and 0.02 ml of an enzyme solution (rat small intestinal brush border membrane).
Pre-incubated for 10 minutes at 7 ° C. Next, 0.
1M substrate (maltose, sucrose, palatinose, trehalose, lactose, cellobiose) solution 0.05
The reaction was started by adding ml, and the reaction was performed at 37 ° C for 10
Incubated for 30 minutes. After the reaction, 5 at 100 ° C
The reaction was stopped by heating for a minute. After cooling, 0.05 ml of the reaction solution was added to 3 ml of a commercially available glucose B test Wako (Wako Pure Chemical Industries) color developing solution, and the mixture was incubated at 37 ° C. for 20 minutes to release 505 n of D-glucose.
Colorimetric determination was performed by m.

In the case of rice α-glucosidase and porcine kidney trehalase, the same procedure as above was performed using 0.1 M maltose or trehalose aqueous solution as a substrate.

Experimental Example 2 Reaction containing 0.2 ml of 0.2 M acetate buffer (pH 5.0), 0.1 ml of the aqueous solution of the compound of the present invention, 0.05 ml of enzyme (almond β-glucosidase) solution, 0.45 ml of distilled water The solution was preincubated for 30 minutes at 0 ° C. Next, 0.01 M p-nitrophenyl-β-D-
The reaction was started by adding 0.2 ml of an aqueous glucoside solution and incubated at 37 ° C for 30 minutes. After the reaction, 2 ml of 0.4 M sodium carbonate was added to stop the reaction, and the released p-nitrophenol was colorimetrically determined at 400 nm.

Regarding other enzyme (baking yeast α-glucosidase, α-mannosidase, α- or β-galactosidase) inhibitory activity, the appropriate p-nitrophenyl-D-glycoside is used as a substrate according to the above method. went.

The results are shown in Tables 3-5.

Table 3 50% inhibition amount (IC50: M)

Table 4 50% inhibition amount (IC50: M)

Table 5 50% inhibition amount (IC50: M)

As is clear from Tables 3 to 5, the compounds of the present invention have remarkable sugar hydrolase inhibitory activity and are useful as sugar hydrolase inhibitors which can be used as pharmaceuticals such as blood glucose lowering agents. Was confirmed.

Further, the acute toxicity of the compound of the present invention was investigated by oral administration in ICR mice.
No deaths were observed at g / kg, and it was confirmed that the toxicity was low and the safety was high.

Next, the dose and formulation of the compound of the present invention will be explained.

The compound of the present invention can be administered to animals and humans as it is or together with a conventional pharmaceutical carrier. The dosage form is not particularly limited and may be appropriately selected and used as needed, and examples thereof include oral preparations such as tablets, capsules, granules, fine granules and powders, parenteral preparations such as injections and suppositories. To be

In order to exert the intended effect as an oral preparation, it varies depending on the age, body weight and degree of disease of the patient, but it is usually 30 to 1000 m as the weight of the compound of the present invention in adults.
It is considered appropriate to take g in several divided doses a day.

Oral preparations include, for example, starch, lactose, sucrose,
Mannitol, carboxymethyl cellulose, corn starch, inorganic salts and the like are used in a conventional manner.

In addition to the above-mentioned excipients, a binder, a disintegrating agent, a surfactant, a lubricant, a fluidity promoter, a flavoring agent, a coloring agent, a flavoring agent, etc., may be appropriately used in this type of formulation. You can
Specific examples of each are as follows.

[Binder] Starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch,
Methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose, ethyl cellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, carboxymethyl cellulose, low-substituted hydroxypropyl cellulose.

[Surfactant] sodium lauryl sulfate,
Soy lecithin, sucrose fatty acid ester, polysorbate 80.

[Lubricant] Talc, wax, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol.

[Fluidity promoter] Light anhydrous silicic acid, dried aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The compounds of the present invention can also be administered as suspensions, emulsions, syrups and elixirs, and these various dosage forms may also contain flavoring agents and coloring agents. Good.

In order to exert the desired effect as a parenteral agent, it depends on the age, weight and degree of disease of the patient.
1 to 300 per day as the weight of the compound of the present invention in an adult.
Intravenous injections up to mg, intravenous drip, subcutaneous injection, and intramuscular injection seem appropriate.

This parenteral preparation is manufactured according to a conventional method, and is generally used as a diluent in distilled water for injection, physiological saline, glucose aqueous solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol. Etc. can be used. Further, if necessary, a bactericide, a preservative, and a stabilizer may be added. Further, from the viewpoint of stability, this parenteral preparation may be filled in a vial or the like and then frozen, the water content may be removed by a usual freeze-drying technique, and a liquid preparation may be re-prepared from the freeze-dried product immediately before use. Further, an isotonicity agent, a stabilizer, a preservative, a soothing agent and the like may be added as needed.

Examples of other parenteral agents include suppositories for rectal administration and the like, which are manufactured by a conventional method.

Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

Example 1 10 kg of dried mulberry powder was extracted with 20 liters of hot water three times for 2 hours. After the extracts were combined and cooled, the same amount of metall was added and the mixture was allowed to stand overnight at 4 ° C. The insoluble matter was filtered through cheese cloth, the filtrate was subjected to 1.5 l of Amberlyst 15 (H type) column chromatography, the column was washed with 50% methanol, and then eluted with 5 l of 0.5 N ammonia water.
Water was distilled off from the eluate under reduced pressure to obtain 82 g of a brown solid. A 1/4 volume of the column was dissolved in a small amount of distilled water, and a column packed with Amberlite CG-50 (NH ₄ type) (Φ 4 x
(85 cm) and the mixture was developed and eluted with distilled water. The eluate was eluted with 20 g of fractions up to 150 fractions, and then with 0.1 N aqueous ammonia to elute up to 150 fractions.
Fraction 31-48 eluted with distilled water is fraction A, 46-60
The fraction is the B fraction, the 61 to 11 fractions are the C fractions, the 97 to 140 fractions eluted with 0.1N aqueous ammonia are the D fractions,
The same operation was repeated 3 times, and the A, B, C, and D fractions were pooled and concentrated to dryness under reduced pressure to obtain A fraction as 9.6 g, B fraction.
Fractions of 4.3 g, C fractions of 9.6 g and D fractions of 4.0 g were obtained.

The fraction A was passed through 200 ml of strongly basic anion exchange resin Dowex 1 × 2 (OH type) to remove anionic substances. The solution was concentrated under reduced pressure, and the concentrated solution was packed in a column (Φ) filled with Dowex 1 × 2 (OH type).
(2.5 × 90 cm) and poured into distilled water to elute. The eluate was collected in 20 g fractions, and 22 to 30 fractions were A-
1 fraction, 31-60 fraction is A-2 fraction, 61-84 fraction is A-3 fraction, 100-124 fraction is A-4 fraction,
When the respective fractions were concentrated to dryness under reduced pressure, 0.85 g as the A-1 fraction, 0.8 g as the A-2 fraction, 0.3 g as the A-3 fraction, and 0. 22 g of solid was obtained.

The A-1 fraction was dissolved in a small amount of distilled water, poured into the upper end of a column (Φ1.2 × 65 cm) packed with a strongly acidic cation exchange resin Dowex 50W × 8 (pyridine salt type), and 0 Elution was performed with 1 M pyridine acetate buffer (pH 6.0), and the eluate was collected in 10 g fractions. 41-47
Fractions are collected and concentrated under reduced pressure to give a small amount of Dowex 1x2
(OH type) and freeze-drying this solution to give 2-O-α-D- having the following physicochemical properties.
Glucopyranosyl-deoxynojirimycin (compound 0
4) Obtained 50 mg. In addition, after concentrating the 49 to 58 fractions,
2-O-α-D-galactopyranosyl-deoxynojirimycin (Compound 10) having the following physicochemical properties was obtained by freeze-drying a solution treated with a small amount of Dowex 1 × 2 (OH type). 50 mg was obtained. Furthermore, 6
By similarly treating the 1-66 fraction, 7 mg of 6-O-α-D-galactopyranosyl-deoxynojirimycin (Compound 11) having the following physicochemical properties was obtained.

Compound 04 Properties: White powder Specific rotation: [α] _D = + 101.1 ° (c = 0.1,
H ₂ O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.27 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 4 shows a nuclear magnetic resonance spectrum chart.

Compound 10 Properties: White powder Specific optical rotation: [α] _D = + 118.8 ° (c = 0.1,
H ₂ O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.26 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 10 shows a nuclear magnetic resonance spectrum chart.

Compound 11 Properties: White powder Specific rotation: [α] _D = + 107.0 ° (c = 0.1,
H ₂ O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.26 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 11 shows a nuclear magnetic resonance spectrum chart.

The column A-2 fraction was dissolved in a small amount of distilled water, and a column (Φ1.5) filled with Dowex 1 × 2 (OH type) was packed.
(× 95 cm), and the mixture was developed and eluted with distilled water. The eluate was collected in 10 g fractions, and the 29 to 33 fractions were concentrated and freeze-dried to obtain the following physicochemical properties.
10 mg of -O-β-D-glucopyranosyl-deoxynojirimycin (Compound 09) was obtained. By similarly freeze-drying the 37-42 fraction, 37 mg of 4-O-β-D-glucopyranosyl-deoxynojirimycin (Compound 08) having the following physicochemical properties was obtained. 44-4
The 9 fractions were concentrated and freeze-dried to give 2-O-β-D-glucopyranosyl- having the following physicochemical properties.
Deoxynojirimycin (Compound 06) was obtained. 52 ~
The 60 fraction was concentrated and then lyophilized to obtain 15 mg of 3-O-α-D-glucopyranosyl-deoxynojirimycin (Compound 5) having the following physicochemical properties.

Compound 09 Properties: White powder Specific optical rotation: [α] _D = -20.4 ° (c = 0.1, H ₂
O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.30 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 9 shows a nuclear magnetic resonance spectrum chart.

Compound 08 Properties: White powder Specific rotation: [α] _D = -23.4 ° (c = 0.1, H ₂
O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.30 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 8 shows a nuclear magnetic resonance spectrum chart.

Compound 06 Properties: White powder Specific optical rotation: [α] _D = -23.9 ° (c = 0.1, H ₂
O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.32 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 6 shows a nuclear magnetic resonance spectrum chart.

Compound 05 Properties: White powder Specific rotation: [α] _D = + 96.4 ° (c = 0.1, H ₂
O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.30 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 5 shows a nuclear magnetic resonance spectrum chart.

The A-3 fraction was dissolved in a small amount of distilled water, poured into the upper end of a column (Φ1 × 46 cm) packed with Dowex 50 W × 8 (pyridine salt type), and added with 0.1 M pyridine acetate buffer (pH 6). 0.0), and the eluate was collected in 5 g fractions. The 43-58 fraction was concentrated, and a solution treated with a small amount of Dowex 1 × 2 (OH type) was lyophilized to give 1,2α, 3 having the following physicochemical properties.
44 mg of β, 4α, 6β-penta-hydroxynortropane (Compound 2) was obtained.

Compound 02 Properties: White powder Specific rotation: [α] _D = + 23.1 ° (c = 0.8, H ₂
O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.44 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 2 shows a nuclear magnetic resonance spectrum chart.

The A-4 fraction was dissolved in a small amount of distilled water, poured into the upper end of a column (Φ1 × 46 cm) packed with Dowex 50 W × 8 (pyridine salt type), and then added with 0.1 M pyridine buffer (pH 6. It was developed and eluted in 0), and the eluate was collected in 5 g fractions. The 58-76 fraction was concentrated, and a solution treated with a small amount of Dowex 1 × 2 (OH type) was lyophilized to give a 3-O-β-D-glucopyranosyl-deoxynodili having the following physicochemical properties. Mycin (Compound 7)
95 mg was obtained.

Compound 07 Properties: White powder Specific rotation: [α] _D = -10.7 ° (c = 0.1, H ₂
O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.32 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 7 shows a nuclear magnetic resonance spectrum chart.

The B fraction was dissolved in a small amount of distilled water, and a column (Φ2.5 × 9) packed with Dowex 1 × 2 (OH type) was used.
(5 cm), and the mixture was developed and eluted with distilled water, and the eluate was collected in 20 g fractions. By concentrating the 37-46 fraction and lyophilizing it, 1,2 having the following physicochemical properties is obtained.
176 mg of α, 3β, 4α-tetrahydroxynortropane (Compound 1) was obtained.

Compound 01 Properties: White powder Specific optical rotation: [α] _D = + 29.1 ° (c = 1, H ₂ O) HPTLC silica gel 60F254 (Merck) Thin layer chromatography: Rf = 0.44 (n- (Development with propanol: acetic acid: water = 4: 1: 1) Fig. 1 shows a nuclear magnetic resonance spectrum chart.

The D fraction was dissolved in a small amount of water, and a column (Φ2.5 × 95c) filled with Dowex 1 × 2 (OH type) was packed.
m), and then elute with distilled water to elute the eluate with 20
Collected in g fraction. 21-25 fractions to D-1 fractions, 31-
34 fractions are D-2 fractions, 35-42 fractions are D-3 fractions,
Fractions 53 to 68 were designated as D-4 fractions, and each fraction was concentrated to dryness under reduced pressure to give 0.66 g as D-1 fraction, D
-0.22 g as the -2 fraction and 0.58 as the D-3 fraction
g, 0.16 solid is obtained as D-4 fraction.

The D-1 fraction was dissolved in a small amount of distilled water to prepare a CM
It was poured onto the upper end of a column (Φ2.2 × 65 cm) packed with Sephadex C-25 (NH ₄ type), developed and eluted with 0.01 N ammonia water, and the eluate was collected in 10 g fractions. After concentrating the 64-78 fraction, it was freeze-dried to give 40 mg of 3-epifagomine (1,2,5-trideoxy-1,5-imino-D-allo-hexitol, compound 3) having the following physicochemical properties. Got

Compound 03 Property: White crystal Specific optical rotation: [α] _D = + 26.6 ° (c = 0.8, H ₂
O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.37 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 3 shows a nuclear magnetic resonance spectrum chart.

The D-4 fraction was dissolved in a small amount of distilled water, and a column (Φ1.5) packed with Dowex 1 × 2 (OH type) was used.
(95 cm x 95 cm) and developed and eluted with distilled water, and the eluate was collected in 10 g fractions. The 33-44 fraction was concentrated and lyophilized to give 1,4-dideoxy-1,4-imino-2-O-β having the following physicochemical properties.
135 mg of -D-glucopyranosyl-D-arabinitol (Compound 12) was obtained.

Compound 12 Properties: White powder Specific optical rotation: [α] _D = -29.8 ° (c = 0.73,
H ₂ O) HPTLC silica gel 60F254 (Merck) thin layer chromatography: Rf = 0.36 (developed with n-propanol: acetic acid: water = 4: 1: 1) FIG. 12 shows a nuclear magnetic resonance spectrum chart.

Example 2 6 kg of distilled water was added to 1 kg of dried mulberry leaf powder, and the mixture was heated at 100 ° C for 2 hours. After cooling, add the same amount of methanol to 4
After standing overnight at ° C, it was filtered through Celite. The filtrate was subjected to 250 ml of Amberlyst 15 (H type) column chromatography, the column was washed with 50% methanol, and then eluted with 1 liter of 0.5N ammonia water.
g brown solid was obtained. Dissolve 1/2 volume in a small amount of distilled water, pour it on the upper end of a column (Φ2.5 × 90 cm) packed with Amberlite CG-50 (NH ₄ type), develop and elute with distilled water, and elute fraction Were collected in 20 g fractions. 26 ~
31 fractions are A fractions, 32-37 fractions are B fractions, 38-5
Fraction 7 was designated as C fraction, and fraction eluted with 0.5N aqueous ammonia was designated as D fraction. Do the same operation for the remaining 1/2 volume,
Fractions A, B, C and D were pooled and concentrated to dryness to give 0.95 g of A fraction, 0.12 g of B fraction, 1.3 g of C fraction and 0.6 g of solid of fraction. Obtained.

Fraction A was dissolved in a small amount of distilled water, and a column (Φ1.5 × 9) packed with Dowex 1 × 2 (OH type) was used.
5 cm) and the mixture was developed and distilled with distilled water, and the eluate was collected in 10 g fractions. The 18 to 25 fractions were concentrated, poured into the upper end of a column (Φ1 × 46 cm) packed with Dowex 50W × 8 (pyridine salt type), developed and eluted with 0.1M pyridine acetate buffer (pH 6.0), The eluate was collected in 5 g fractions. The 24-36 fractions were concentrated, freeze-dried with a small amount of a solution treated with Dowex 1 × 2 (OH type) to obtain 600 mg of compound 10.

The B fraction was dissolved in a small amount of distilled water, and a column (Φ1.5 × 9) packed with Dowex 1 × 2 (OH type) was used.
5 cm) and the mixture was developed and distilled with distilled water, and the eluate was collected in 10 g fractions. The 26-34 fractions were concentrated and lyophilized to give compound 140 mg.

The D fraction was dissolved in a small amount of water, and a column (Φ1.5 × 95c) packed with Dowex 1 × 2 (OH type) was packed.
m), poured into the upper end of the m), developed and eluted with distilled water, and
Collected in g fraction. Compounds 12 were obtained by concentrating 33-43 fractions and freeze-drying.

Example 3 500 g of dried mulberry (mulberry fruit) was heated in 1 liter of hot water for 2 hours 2
Extracted twice. After the extracts were combined and cooled, the same amount of methanol was added and the mixture was allowed to stand at 4 ° C overnight. Insoluble matter is filtered with cheese cloth, and the filtrate is amberlyst 15 (H type) 100 m.
It was applied to a column chromatograph of 1 and the column was washed with 50% methanol and then eluted with 500 ml of 0.5N ammonia water. The eluate was concentrated to dryness to obtain 2.8 g of a brown solid. This is dissolved in a small amount of distilled water, and a column (Φ 2.5 x) packed with Amberlite CG-50 (NH ₄ type)
90 cm) and the mixture was developed and distilled with distilled water, and the eluate was collected in 20 g fractions up to 50 fractions. The fraction eluted with distilled water was concentrated to dryness to obtain 1 g of a brown solid. This was designated as A fraction. In addition, the same column was used with 0.5N ammonia water 50
The fraction eluted with 0 ml was concentrated to dryness to obtain 150 mg of a brown solid. This was designated as B fraction.

The fraction A was dissolved in a small amount of distilled water, and a column (Φ1.5 × 9) packed with Dowex 1 × 2 (OH type) was used.
5 cm) and the mixture was developed and distilled with distilled water, and the eluate was collected in 10 g fractions. After concentrating the 26 to 30 fractions, a column packed with Amberlite CG-50 (NH ₄ type) (Φ
(1 × 65 cm) and poured into distilled water to elute, and the eluate was collected in 6 g fractions. The 9-12 fraction was concentrated and then lyophilized to obtain Compound 1 (35 mg).

Example 4 Cornstarch 44 g Crystalline cellulose 40 g Carboxymethyl cellulose calcium 5 g Light anhydrous silicic acid 0.5 g Magnesium stearate 0.5 g Compound 1 10 g Total 100 g According to the above formulation, It was compression molded to obtain tablets of 200 mg each. 20 mg of Compound 1 is contained in one tablet, and 3 to 10 tablets for adults are to be taken in several divided doses per day.

Example 5 Crystalline cellulose 84.5 g Magnesium stearate 0.5 g Carboxymethyl cellulose calcium 5 g Compound 4 10 g Total 100 g According to the above formulation, a part of and was uniformly mixed, compression molded, and then crushed, The remaining amounts of and were added and mixed, and the mixture was compression-molded with a tableting machine to obtain 200 mg tablets. This tablet contains 20 mg of compound 4, and 3 to 10 tablets for adults are to be taken in several divided doses.

Example 6 Crystalline Cellulose 79.5 g 10% Hydroxypropyl Cellulose Ethanol Solution 50 g Carboxymethyl Cellulose Calcium 5 g Magnesium Stearate 0.5 g Compound 5 10 g Total 145 g According to the above formulation, and were uniformly mixed, and according to a conventional method. After blending, granulating with an extrusion granulator, drying and crushing, and were mixed and compression-molded with a tableting machine, one tablet of 200 mg was obtained. Each tablet contains 20 mg of Compound 5, and the daily dose for adults is 3 to 3.
Take 10 tablets in several doses.

Example 7 Corn starch 84 g Magnesium stearate 0.5 g Carboxymethyl cellulose calcium 5 g Light anhydrous silicic acid 0.5 g Compound 7 10 g Total 100 g According to the above formulation, the The granules were obtained by crushing with a crusher and sieving. 1 g of this granule contains 100 mg of the compound 7, and 0.6 to 2 g of an adult is taken in several divided doses per day.

Example 8 Crystalline cellulose 86.5 g 10% Hydroxypropyl cellulose ethanol solution 35 g Compound 8 10 g Total 131.5 g According to the above-mentioned prescription, was uniformly mixed and kneaded. After granulating with an extrusion granulator, it was dried and sieved to obtain granules. 1 g of this granule has 100 m of compound 8
g is contained, and 0.6 to 2 g for adults is divided into several doses and taken.

Example 9 Corn starch 89.5 g Light anhydrous silicic acid 0.5 g Compound 10 10 g Total 100 g According to the above-mentioned recipe, 1 to were uniformly mixed to obtain 200 mg.
Was filled in a No. 2 capsule. 20 mg of the compound 10 is contained in 1 capsule of this capsule, and 3 to 10 capsules for an adult are to be taken in several divided doses.

Example 10 Distilled water for injection 89.5 g Soybean oil 5 g Soybean phospholipid 2.5 g Glycerin 2 g Compound 11 1 g Total amount 100 g According to the above formulation, was dissolved in and and the solution of and was added to emulsify, An injection was obtained.

[0071]

The sugar hydrolase inhibitor of the present invention has a remarkable sugar hydrolase inhibitory action and is useful as a blood glucose lowering agent.

In recent years, it has been revealed that glycoprotein chains (receptors, cell adhesion factors, etc.) on the cell surface are involved in the reaction of the immune system, cancer metastasis, and fusion of HIV (AIDS virus) into cells. I was killed. INDUSTRIAL APPLICABILITY The present invention remarkably inhibits the glycohydrolase that acts when this glycoprotein chain is formed, and therefore it is not limited to a hypoglycemic agent, and can be used as a therapeutic agent for diseases associated with this glycoprotein chain. .

[Brief description of drawings]

FIG. 1 is a nuclear magnetic resonance spectrum chart of compound 01.

FIG. 2 is a nuclear magnetic resonance spectrum chart of compound 02.

FIG. 3 is a nuclear magnetic resonance spectrum chart of Compound 03.

FIG. 4 is a nuclear magnetic resonance spectrum chart of compound 04.

FIG. 5 is a nuclear magnetic resonance spectrum chart of Compound 05.

FIG. 6 is a nuclear magnetic resonance spectrum chart of compound 06.

FIG. 7 is a nuclear magnetic resonance spectrum chart of compound 07.

FIG. 8 is a nuclear magnetic resonance spectrum chart of Compound 08.

FIG. 9 is a nuclear magnetic resonance spectrum chart of compound 09.

FIG. 10 is a nuclear magnetic resonance spectrum chart of Compound 10.

11 is a nuclear magnetic resonance spectrum chart of Compound 11. FIG.

FIG. 12 is a nuclear magnetic resonance spectrum chart of Compound 12.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI technical display area C07H 17/02

Claims (5)

[Claims] 1. The following formula (I) (In the formula, R ₁ represents a hydrogen atom or a hydroxyl group.) An alkaloid. 2. The following formula (II) Is an alkaloid. 3. The following formula (III): (In the formula, R _{2 to} R ₅ are shown in Table 1 below.) Table 1 Is an alkaloid. 4. The following formula (IV): (In the formula, R ₆ represents a β-glucopyranosyl group.) An alkaloid. 5. The following formula (I), (In the formula, R ₁ represents a hydrogen atom or a hydroxyl group.) The following formula (II) Formula (III) below, (In the formula, R _{2 to} R ₅ are shown in Table 1 below.) Table 1 Or the following formula (IV) (In the formula, R ₆ represents a β-glucopyranosyl group.) A sugar hydrolase inhibitor containing an alkaloid represented by the formula as an active ingredient.