JPH08259589A - New saccharide derivative - Google Patents

Abstract

PURPOSE: To obtain a new saccharide derivative by glycosylating a lankacidin produced by a microorganism, showing activity against methicillin-resistant Staphylococcus aureus and Haemophilus influenzae, having high safety and stability in blood, useful as an antimicrobial agent. CONSTITUTION: This new lankacidin derivative is shown by formula I (either one of R1 and R2 is a glycosyl which may be protected or 1,2-ortho esterified and the other is H, an acyl or glycosyl which may be protected or 1,2-ortho esterified), exhibits activity against methicillin-resistant Staphylococcus aureus and Haemophilus influenzae and has high safety and stability in blood, useful as an antimicrobial agent, etc. The saccharide derivative is obtained by glycosylating a compound of formula II (one of R1 ' and R2 ' is H and the other is an acyl group, a protecting group or glycosyl group) produced by acylating a part of a hydroxyl group of a lankacidin produced by a microorganism.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to novel lancacidin sugar derivatives.

[0002]

BACKGROUND OF THE INVENTION Lankacidins are antibiotics having a structure represented by R ₁ , R ₂ ═H of the general formula (I) produced by microorganisms and antibiotics Lankacidin K and It is a generic term for L and constitutes the antibiotic T-2636 group. The structures, physicochemical and biological properties of these antibiotics have already been clarified in several documents [The Journal of Antibiotics, Vol. 26, p. 1 (1971);
Ibid., Vol. 26, p. 647 (1973); Chemical Pharmaceutical Bulletin, vol. 22, p. 99.
(1974); Vol. 23, p. 2201 (1975).
Year)]]. In addition, studies on the use of lancasidin compounds have revealed that they are effective as antibacterial infection agents, anti-porcine dysentery preventive / therapeutic agents, and antitumor agents, and some of them have been put to practical use. . In particular, a compound in which the hydroxyl groups at the 8th and 14th positions of lancasidine are modified with an alkanoyl ester or a carbamoyl group is already known. [JP-A-59-183695; JP-A-58-1
79496]. However, there is a demand for a derivative which is as active as the known lancacidins or more, has excellent pharmacokinetics, and is low in toxicity and is superior as a pharmaceutical.

[0003]

In view of the above circumstances, the present inventors have diligently studied the creation of a lancacidin derivative which is more stable in blood and has low toxicity, which is more desirable as a human drug. As a result, 8 of Lancacidin
Succeeded in obtaining a sugar-modified derivative at position 14 or position 14,
It was found that these derivatives had unexpected physiological activities, and further studies were conducted to complete the present invention. That is, the present invention provides the formula (I):

[0004]

Embedded image

(In the formula, _one of R ₁ and R ₂ is a glycosyl group which may be protected or is 1,2-orthoesterified, and the other is a hydrogen atom,
An acyl group or an optionally protected 1,2-orthoesterified glycosyl group), a lancascidin sugar derivative (2) formula (I ′):

[0006]

[Chemical 4]

(In the formula, _one of R ₁ 'and R ₂ ' represents a hydrogen atom, and the other is a hydrogen atom, an acyl group or may be protected and may be 1,2-orthoesterified. A compound having a good glycosyl group) is subjected to a glycosylation reaction, and (3) a method for producing a lancacidin sugar derivative, and (3) the lancascidin sugar derivative according to (1) above. The present invention relates to an antibacterial agent.

In the formula (I) of the present invention, the optionally protected 1,2-orthoesterified glycosyl group represented by R ₁ and R ₂ is, for example, glucose, galactose or mannose. , Sugars such as fucose, ribose, cructose, glucosamine, N-acetylglucosamine, glucuronic acid, maltose, maltotriose, and cellobiose from monosaccharides to trisaccharides and their hydroxyl groups are protected or 1,2-orthoesterified Included are glycosyl groups. The protected glycosyl group means that all or a part of the hydroxyl groups of the glycosyl group are protected by a usual hydroxyl group-protecting group such as a carboxylate acyl group and an aralkyl group. Examples of the carboxylic acid acyl group include an acyl group derived from an aliphatic carboxylic acid or an aromatic carbocyclic carboxylic acid. Examples of the aliphatic carboxylic acid include saturated aliphatic carboxylic acids having 2 to 7 carbon atoms such as acetic acid and propionic acid, and examples of the aromatic carbocyclic carboxylic acid include benzoic acid. Examples of the aralkyl group include a benzyl group and a phenethyl group.

In addition, the 1,2-orthoesterified glycosyl group has the above-mentioned glycosyl group having, for example, 2 to 6 carbon atoms.
Aliphatic carboxylic acids (eg, acetic acid, propionic acid, butyric acid,
Isobutyric acid, valeric acid, caproic acid, etc.), carbon number 7-10
Aromatic carbocyclic carboxylic acids (eg, benzoic acid, etc.), aromatic carbocyclic substituted aliphatic carboxylic acids having 8 to 11 carbon atoms (eg,
1,2-with phenylacetic acid, phenylpropionic acid, etc.
It means ortho-esterified. Specific examples include glucose 1,2-orthoacetate, glucose 1,2-orthopropionate, galactose 1,2.
-Orthoacetate, mannose 1,2-orthoacetate, glucuronic acid 1,2-orthoacetate, glucuronic acid methyl ester 1,2-orthoacetate, maltose 1,2-orthoacetate and the like.

Examples of the acyl group represented by R ₁ or R ₂ include aliphatic carboxylic acid acyl groups having 2 to 6 carbon atoms (eg, acetyl, propionyl, butyryl, isobutyryl, valeryl, caproyl, etc.).

The lancassidine derivative of the present invention is obtained by subjecting a compound represented by the above formula (I ′) (eg, lancassidine A or C etc.) to a glycosylation reaction in which a sugar derivative in which the 1-position is activated is reacted. Can be manufactured by. This reaction makes it possible to obtain a lancacidin sugar derivative in which either or both of the 8- and 14-positions of lancasidin are substituted with a group obtained from the sugar derivative. In formula (I ′), the acyl group represented by R ₁ ′ or R ₂ ′ and the optionally protected 1,2-orthoesterified glycosyl group are represented by R ₁ or R ₂ . The groups defined as these groups are included. Examples of the sugar derivative in which the 1-position is activated include glycosyl halides (eg, glycosyl chloride, glycosyl bromide, glycosyl fluoride, etc.), glycosylimidate, thioglycoside and the like.

The reaction is usually carried out in a solvent. As the solvent used in such a reaction, any solvent may be used so long as it does not inhibit the reaction, preferably chloroform,
Halogenated hydrocarbons such as dichloromethane, toluene,
Aromatic hydrocarbons such as benzene, solvents such as nitromethane, acetonitrile and ethers can also be used. This reaction is preferably carried out under anhydrous conditions, but it can also be carried out by adding molecular sieves, anhydrous calcium sulfate or the like. Mercury salts and silver salts are generally used as auxiliaries for activating this reaction, but for substrates with low reactivity, use trifluoromethanesulfonic acid silver salt or silver perchlorate salt. You can also The reaction temperature can be a wide range of -70 to 100 ° C depending on the reactivity of the substrate, but it is usually desirable to carry out the reaction at a low temperature unless it interferes with the progress of the reaction.

From the reaction mixture thus obtained,
The target product can be easily separated and purified by a usual method such as chromatographic separation or recrystallization. Among the lancassidin sugar derivatives thus obtained, for those in which the hydroxyl group of the sugar is protected by an acyl group etc., the acyl group can be eliminated by hydrolysis with an alkali metal to obtain the corresponding lancassidine sugar derivative. it can. As the alkali metal used, sodium and potassium are desirable, and anhydrous solvent systems such as sodium methoxide / methanol and sodium ethoxide / ethanol are particularly desirable. The reaction temperature is −30 to 60 ° C., and it is preferable to carry out the reaction at a low temperature unless the reaction is usually hindered.
On the other hand, an enzymatic method is preferable as a method for removing the protective group attached to the hydroxyl group of the sugar among the lancacidin sugar derivatives.
As an enzyme, Streptomyces rochei
Particularly preferred is a carboxylesterase from yces rochei). The reaction using this enzyme can be carried out in an organic solvent, such as methanol, ethanol, 1-propanol,
Acetonitrile, acetone, etc. can be used,
Of these, methanol is particularly preferred. The compound represented by the formula (I ′) used as a starting material for this reaction is microbiologically according to a known method (for example, the method described in JP-A-59-183695), or a combination thereof with a chemical synthesis method known per se. Can be manufactured.

It has been found that the desired lancacidin sugar derivative obtained as described above exhibits excellent antibacterial activity against methicillin-resistant Staphylococcus aureus and is useful as an anti-MRSA agent. Furthermore, it has an antibacterial action against Haemophilus influenzae. Furthermore, since the lancacidin sugar derivative of the present invention has low toxicity, mammals including human (eg,
It is useful as an antibacterial agent for pigs, cows, sheep, etc.).

The antibacterial agent containing the lancacidin sugar derivative of the present invention can be obtained by a conventional method. In addition to the lancacidin sugar derivative of the present invention, other active ingredients may be contained. For example, in the case of an anti-MRSA agent, it may contain other compounds having anti-MRSA activity.

The form of the antibacterial agent of the present invention is not particularly limited,
For example, solid preparations such as tablets, granules, fine granules, pills, powders, capsules, troches, chewable preparations, elixirs, syrups, suspensions, emulsions, injections, infusions, and other liquids. May be.

For the preparation of the preparation, a conventional carrier component can be used depending on the kind of the preparation. For example, for the preparation of solid formulations,
Conventional ingredients such as starch, lactose, sucrose, mannitol, sugars such as corn starch, crystalline cellulose, carboxymethyl cellulose, excipients such as light anhydrous silicic acid; polyvinyl alcohol, polyvinylpyrrolidone, polyvinyl ether, ethyl cellulose, gum arabic,
Binders such as tragacanth, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, calcium citrate, dextrin, pectin; lubricants such as magnesium stearate, calcium stearate, talc, polyethylene glycol, colloidal silica; starch, carboxymethylcellulose, carboxy A disintegrant such as methylcellulose calcium and croscarmellose sodium, a disintegration aid, a moisturizer, and a surfactant can be used.

For the preparation of liquid preparations, conventional ingredients such as water for injection, water, solvents such as ethyl alcohol and ethylene glycol, ethanol, polyethylene glycol, propylene glycol, D-mannitol, cholesterol,
Solubilizing agents such as triethanolamine, sodium carbonate, sodium citrate, etc., surfactants such as stearyltriethanolamine, sodium lauryl sulfate, lecithin, glyceryl monostearate; polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxy. Suspending agents such as hydrophilic polymers such as methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, isotonic agents such as sodium chloride, glycerin, D-mannitol, phosphates, acetates, carbonates, citrates, etc. Buffers, soothing agents such as benzyl alcohol, glucose, amino acids and the like can be used. In the above-mentioned solid preparation and liquid preparation, if necessary, a preservative, a solubilizer, an emulsifier, a dispersant, a thickener, a plasticizer, an adsorbent, a flavor, a coloring agent, a flavoring agent, a sweetener, a preservative, Antioxidants and the like can be used. The antibacterial agent of the present invention can be produced by a conventional method such as kneading, kneading, granulation, tableting, coating, sterilization, emulsification, etc. depending on the form of the preparation. Regarding the production of the preparation, each section of the Japanese Pharmacopoeia General Rules for Preparations can be referred to.

In the antibacterial agent of the present invention, the content of the lancacidin sugar derivative may be any effective amount that exhibits antibacterial activity against MRSA or Haemophilus influenzae. The administration method may be oral or parenteral. The dose of the lancasidin sugar derivative varies depending on the dosage form, administration route, age, degree and type of gastrointestinal disease, but when orally administered to humans, it is usually about 0.001 to 10 g, preferably 0.01 per day for an adult. ~ 5g, usually 1-3g, can be administered in 1 to several divided doses. In the case of parenteral administration, the daily dose for an adult is about 0.0001 to 10 g, usually about 0.001 to 5 g, preferably 0.1 to 3 g, which is administered in 2 to 4 divided doses. .

[0020]

The present invention will be described in more detail with reference to the following examples, but these examples do not limit the scope of the present invention. Example 1 10 g of Lancacidin A was added to 160 ml of dry dichloromethane.
Then, 12 g of α-acetobrom-D-glucose and 8 g of silver carbonate were added, and further 30 g of Molecular Sieves-4A was added, and the mixture was reacted for 18 hours under stirring at room temperature under shading. The reaction solution was filtered through a filter paper, and the mother liquor was evaporated under reduced pressure to remove the solvent. The obtained residue was chromatographed on silica gel (φ5c
m × 46 cm), chloroform-methanol (100
0:12), elution, elution, collection of the target fractions and evaporation of the solvent, a pale yellow powder of Lancacidin A-8- (3,4,6-tri-O-acetyl-β-D-glucose 1, 12.0 g (yield 70%) of 2-orthoacetate) was obtained. N of this product
MR (200 MHz, CDCl ₃ ) δ; 1.32 (3H, d, J =
6.5), 1.39 (3H, s), 1.51 (3H, s), 1.67
(3H, s), 1.90 (3H, s), 2.03, 2.07, 2.0
8, 2.10 (4 × 3H, s), 2.47 (3H, s), 6.25
(1H, d, J = 15.5), UVλmax MeOH: 226.5
nm.

Example 2 Lancacidin A-8- (3,4,6-tri-O-acetyl-β-D-glucose-1,2-orthoacetate) 12
g was dissolved in 1 liter of dry methanol, and then under ice cooling (0 to 2 ° C.), 28 g of sodium methoxide 1.4 g /
A solution consisting of 100 ml of methanol was added dropwise with stirring. Two
After reacting for 0 minutes, 32 g of silica gel 60 was added to stop the reaction, and then the solvent was distilled off under reduced pressure. The target fractions were collected and the solvent was distilled off to obtain a pale yellow powder of Lancacidin A-8.
-(Β-D-glucose 1,2-orthoacetate)
4.88 g (yield 42%) was obtained. NMR of this product (200M
Hz, CDCl ₃ ) δ: 1.30 (3H, d, J = 6.5), 1.2
9 (3H, s), 1.39 (3H, s), 1.64 (3H, s), 2.
03 (3H, s), 2.47 (3H, s), 6.26 (1H, d, J =
15.5), UV [lambda] max MeOH: 226.5 nm.

Example 3 Lancacidin A-8- (β-D-Glucose 1,2-
0.3 g of orthoacetate) was dissolved in 30 ml of methanol and cooled to 10 ° C., then 35.5 mg (87 units / mg) of carboxyesterase (manufactured by Wako Pure Chemical Industries, Ltd.) and 3 ml of female buffer solution were added and combined with the above reaction solution. , 8-12 ℃
At 20 minutes with stirring, then 60 mg phenol /
After adding a solution of 1.2 ml of methanol and stirring for 2 minutes, 3.6 g of silica gel 60 was added to form a slurry, and the solvent was distilled off under reduced pressure. The obtained powder was subjected to silica gel column chromatography (silica gel 100 ml), developed and eluted with ethyl acetate / acetone (2: 1), the target fractions were collected, and the solvent was evaporated under reduced pressure to give a pale yellow powder. 0.19 g of crude product was obtained. This product is a medium pressure preparative column chromatograph (YAMAZ
The fraction was eluted with chloroform / methanol (15: 1) and then chloroform / methanol (10: 1) with EN, silica gel, φ1.5 cm × 30 cm, and the target Lancacidin C-8- (β- D-glucose 1,2-
37 mg of orthoacetate) was obtained. NMR of this product (200
MHz, CDCl ₃ ) δ: 1.27 (3H, d, J = 6.5), 1.
29 (3H, s), 1.38 (3H, s), 1.53 (3H, s),
1.69 (3H, s), 2.47 (3H, s), 6.14 (1H, d,
J = 15.5), UV λ max MeOH: 226.4 nm.

Example 4 In the same manner as in Examples 1 to 3, the following compounds were synthesized. Lancacidin A-8- (3,4,6-tri-O-acetyl-β-D-galactose 1,2-orthoacetate) NMR (200 MHz, CDCl ₃ ) δ: 1.31 (3H, d, J
= 6.5), 1.38 (3H, s), 1.52 (3H, s), 1.9
3 (3H, s), 1.99, 2.03, 2.04, 2.05 (4
× 3H, s), 2.47 (3H, s), 6.25 (1H, d, J = 1)
5.5). UV λ max MeOH; 226.5 nm.

Lancacidin A-8- (3,6,8,9,1
0,12-hexa-O-acetyl-β-D-maltose 1,2-orthoacetate) NMR (200 MHz, CDCl ₃ ) δ: 1.33 (3H, d, J
= 6.5), 1.40 (3H, s), 1.52 (3H, s), 1.7
0 (3H, s), 1.90 (3H, s), 2.02, 2.03, 2.
04, 2.05, 2.08, 2.09, 2.11 (7 × 3H,
s), 2.48 (3H, s), 6.26 (1H, d, J = 15.5).
UV λ max MeOH: 226.5 nm.

Lancacidin C-8- (β-D-galactose 1,2-orthoacetate) NMR (200 MHz, CDCl ₃ ) δ: 1.28 (3H, d, J
= 6.5), 1.39 (3H, s), 1.53 (3H, s), 1.6
7 (3H, s), 1.89 (3H, s), 2.46 (3H, s), 6.
09 (1H, d, J = 15.5). UV λ max MeOH: 22
6.5 nm.

Lancacidin A-8- (3,4-di-O-
Acetyl-β-D-glucuronide 1,2-orthoacetate) methyl ester NMR (200 MHz, CDCl ₃ ) δ: 1.32 (3H, d, J
= 6.5), 1.39 (3H, s), 1.52 (3H, s), 1.6
4 (3H, s), 1.70 (3H, s), 1.93 (3H, s), 2.
04, 2.09, 2.10 (3 × 3H, s), 2.48 (3H,
s), 3.77 (3H, s), 6.26 (1H, d, J = 15.5).

Lancacidin A-8- (β-D-glucuronide 1,2-orthoacetate) methyl ester NMR (200 MHz, CDCl ₃ ) δ: 1.32 (3H, d, J
= 6.5), 1.39 (3H, s), 1.52 (3H, s), 1.6
9 (3H, s), 1.89 (3H, s), 2.04 (3H, s), 2.
48 (3H, s), 3.81 (3H, s), 6.16 (1H, d, J =
15.5). UVλmax MeOH; 226.5nm

Lancacidin C-8- (β-D-glucuronide 1,2-orthoacetate) methyl ester NMR (200 MHz, CDCl ₃ ) δ: 1.27 (3H, d, J
= 6.5), 1.39 (3H, s), 1.53 (3H, s), 1.6
9 (3H, s), 1.90 (3H, s), 2.48 (3H, s), 3.
82 (3H, s), 6.14 (1H, d, J = 15.5).

Lancacidin A-8- (β-D-mannose 1,2-orthoacetate) NMR (200 MHz, CDCl ₃ ) δ: 1.24 (3H, d, J
= 6.5), 1.28 (3H, s), 1.53 (3H, s), 1.6
6 (3H, s), 1.89 (3H, s), 2.04 (3H, s), 2.
47 (3H, s), 6.26 (1H, d, J = 15.5).

Lancacidin C-8- (β-D-mannose 1,2-orthoacetate) NMR (200 MHz, CDCl ₃ ) δ: 2.61 (3H, d, J
= 6.5), 1.39 (3H, s), 1.53 (3H, s), 1.6
7 (3H, s), 1.89 (3H, s), 2.48 (3H, s), 6.
16 (1H, d, J = 15.5).

Lancacidin A-8- (3,4-di-O-
Acetyl-β-D-fucose 1,2-orthoacetate) NMR (200 MHz, CDCl ₃ ) δ: 1.31 (3H, d, J
= 6.5), 1.39 (3H, s), 1.52 (3H, s), 1.6
8 (3H, s), 2.03 (3H, s), 2.07, 2.09, 2.
10 (3 × 3H, s)), 2.48 (3H, s), 6.23 (1H,
d, J = 15.5).

Example 5 Lancacidin C-8- (β-D-Glucose 1,2-
Orthoacetate) (Compound A), Lancacidin C-8
-(Β-D-galactose 1,2-orthoacetate)
(Compound B) and Lancacidin C-8- (β-D-
For the anti-MRSA effect of glucuronide 1,2-orthoacetate) methyl ester (Compound C), the minimum inhibitory concentration (MIC) was determined by the agar dilution method. Table 1 shows the results.

[0033]

[Table 1]

Example 6 Lancacidin C 4.6 g (10 mmol), acetobromo-
α-D-glucose 12.3 g (30 mmol), silver carbonate 1
0.7 g (39 mmol) and 30 g of MS4A were shielded from light in 160 ml of dichloromethane, stirred under an argon stream, and reacted at room temperature for 16 hours. The resulting reaction solution was filtered and the filtrate was concentrated to dryness. This was subjected to silica gel chromatography (chloroform / methanol = 100 /
12) The desired product, Lancacidin C-8- (2,3,4,6-tetra-O-acetyl-β-D-glucoside), was obtained (3.8
g, 48% Y). δ _H (CDCl ₃ : 90 MHz) 1.26 (3
H, d, J = 7Hz), 1.40 (3H, s), 1.55 (3H,
s), 1.93 (3H, s), 2.01 (3H, s), 2.02 (3
H, s), 2.03 (3H, s), 2.07 (3H, s), 2.10
(3H, s), 2.45 (3H, s). C-8- obtained above
3.8 g (4.8 mmol) of (2,3,4,6-tetra-O-acetyl-β-D-glucoside) was dissolved in 750 ml of methanol, and 0.23 g (4.3 mmo of sodium methylate was added thereto.
150 ml of a methanol solution of l) was added dropwise at 0 ° C. for 5 minutes under a nitrogen stream. After continuously stirring at 0 ° C. for 20 minutes, 60 g of silica gel was added and the mixture was further stirred for 5 minutes. The resulting reaction solution was filtered and the filtrate was concentrated to dryness. This was subjected to silica gel chromatography (chloroform / methanol = 5/1 and acetonitrile) lancasidine C-8.
-Β-D-Glucoside was obtained (0.5 g, 17%). ¹³ C
_{-NMR (125MHz, CD 3 CN)} δ: 212.8
(s), 197.8 (s), 171.6 (s), 160.9 (s), 1
39.3 (s), 138.1 (d), 137.4 (s), 137.3
(d), 131.1 (d), 130.2 (d), 128.6 (d), 1
26.2 (d), 101.3 (d), 80.4 (d), 77.7 (d),
77.1 (d), 76.7 (d), 74.9 (d), 71.7 (d), 7
0.0 (d), 63.0 (t), 57.5 (s), 52.9 (d), 4
7.5 (d), 38.1 (t), 35.6 (t), 24.7 (q), 2
1.5 (q), 13.2 (q), 12.9 (q), 9.7 (q); High resolution-FABMS (positive) m / z: 622.2858
[MH] ⁺ (calculated value: C ₃₁ H ₄₄ NO ₁₂ as 622.286
4).

[0035]

INDUSTRIAL APPLICABILITY The lancacidin sugar derivative of the present invention has antibacterial activity against methicillin-resistant Staphylococcus aureus and has anti-M activity.
It is useful as an RSA agent. Furthermore, it also has an antibacterial action against Haemophilus influenzae. Further, the compound of the present invention is
Highly safe and stable in blood. Therefore, it has few side effects and is useful as a highly active antibacterial agent.

Claims (3)

[Claims] 1. Formula (I): (In the formula, _one of R ₁ and R ₂ represents a glycosyl group which may be protected and may be 1,2-orthoesterified, and the other is a hydrogen atom, an acyl group or a protected group. Or a 1,2-orthoesterified glycosyl group which is optionally present). 2. Formula (I ′): (In the formula, _one of R ₁ ′ and R ₂ ′ represents a hydrogen atom, and the other represents a hydrogen atom, an acyl group or an optionally protected 1,2-orthoesterified glycosyl group. The method for producing a lancacidin sugar derivative according to claim 1, wherein the compound represented by (1) is subjected to a glycosylation reaction. 3. An antibacterial agent, which comprises the lancacidin sugar derivative according to claim 1.