JP2000001497A - Benzonaphthacenequinone derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a novel compound that is prepared by covalently bonding a benzonaphthacene quinone to a specific organic compound, thus has an antimicrobial activity of the benzonaphthacenequinones, can increase the solubility in water to attain efficient and selective transportation of medicines by introducing a chemical structure for controlling their physical and chemical properties and has high safety with side-effects reduced. SOLUTION: This novel compound is a benzonaphthacene quinone covalently bonded to a polyether or glycosaminoglycan represented by the formula (R1 is H, a lower alkyl or the like; R2 is a hydroxyl group, amino group or the like; R3 is H, a sulfated D-xylosyl or the like; R4 is H, D-xylosyl or the like), typically monodecyloxy-tetraethylene glycol-benanomicin A. The compound is prepared, for example, by protecting the hydroxyl groups in the compound of the formula, then allowing the carboxyl group in the compound of the formula to react with the amino groups or hydroxyl groups to form the acid amide bonds or ester bonds.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a benzonaphthacenequinone derivative or a salt thereof, in which a benzonaphthacenequinone compound represented by the following formula (I) is covalently bonded to a polyether or glycosaminoglycan.

[0002]

2. Description of the Related Art A benzonaphthacenequinone compound such as benanomycin A distinguishes the absolute structure of D-mannose in a cell wall by binding one molecule of calcium ion to a carboxyl group of two molecules of a benzonaphthacenequinone compound. -It is thought to express the ability to bind and to exhibit antibacterial activity. (Y. Fukagawa et al., Actinom
ycetol. (1993) 7: 1-22). However, benzonaphthacenequinone derivatives, which are antibiotics, have not yet been put to practical use as antifungal and antiviral agents due to undesirable side effects.

[0003]

SUMMARY OF THE INVENTION The present invention provides a benzonaphthacenequinone compound having the antibacterial activity and the physical and chemical properties by covalently bonding the benzonaphthacenequinone compound to a specific organic compound. The present invention relates to introducing a control structure to increase water solubility, efficiently and selectively deliver a drug to a target, reduce side effects and increase safety.

[0004]

The present invention relates to (1) a benzonaphthacenequinone derivative or a benzonaphthacenequinone derivative in which a benzonaphthacenequinone compound represented by the following formula (I) is covalently bonded to a polyether or glycosaminoglycan: Its salt;

[0005]

Embedded image

[Wherein, R ₁ represents a hydrogen atom, a lower alkyl group or a hydroxy lower alkyl group, R ₂ represents a hydroxyl group,
R ₃ represents a hydrogen atom, a non-sulfated or sulfated D-xylosyl group or a D-glucosyl group, and R ₄ represents a hydrogen atom, a lower alkyl group or a D-glucosyl group. Indicates a xylosyl group. ]

(2) The covalent bond is an acid amide bond or an ester bond between a carboxyl group of the benzonaphthacenequinone compound and an amino group or a hydroxyl group of the polyether or glycosaminoglycan. (II)
A derivative according to the above (1) or a salt thereof,

[0008]

Embedded image

Wherein R _{1 to} R ₄ are the same as above. X is NH
And -Y or OY, wherein Y represents a residue obtained by removing amino groups or hydroxyl groups involved in the above-mentioned binding from polyether or glycosaminoglycan. ];

(3) The polyether is a polyalkylene glycol having an amino group, and the glycosaminoglycan is obtained by converting an acetylamino group and / or a sulfoamino group of a constituent amino sugar into an amino group. The derivative or salt thereof according to the above (1) or (2), which is chondroitin sulfate, chondroitin sulfate, dermatan sulfate or keratan sulfate;

(4) A pharmaceutical composition comprising the benzonaphthacenequinone derivative or a salt thereof according to (1) to (3).

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.
In the present invention, a lower alkyl group refers to an alkyl group having 1 to 6 carbon atoms. In the above formulas (I) and (II), the lower alkyl group for R ₁ is methyl, ethyl, propyl,
Isopropyl and the like, and the hydroxy lower alkyl group include hydroxymethyl and the like. Examples of the mono- or di-lower alkylamino group for R ₂ include monomethylamino, dimethylamino and the like. Examples of the sulfated D-xylosyl group and sulfated D-glucosyl group of R ₃ include a 3-sulfate-D-xylosyl group and a 3-sulfate-D-glucosyl group. R ₄ is a methyl group, D
-Xylosyl group and the like.

The benzonaphthacenequinone compound represented by the formula (I) is an antifungal antibiotic derived from an actinomycete having a saccharide-binding protein lectin-like activity, and includes benanomicin A; B, pradimicin.
n) A, B, C, D, E, FA-1, FA-2, L, F
L, FS, FB, and S [Actinomycetol.
93) 7: 1-22].

Examples of the polyether include polyalkylene glycols such as polyethylene glycol and polypropylene glycol. Compounds having a terminal amino group, hydroxyl group or carboxyl group, for example, ω-aminohexaethylene glycol methyl ether, ω-amino Polyethylene glycol methyl ether and copolymer derivatives of monomethoxy polyethylene glycol and maleic anhydride are also included. Their average molecular weight is 2
About 00 to 5,000 is preferable.

Glycosaminoglycans include hyaluronic acid (average molecular weight 2,000 to 1.5 million), chondroitin sulfate (average molecular weight 2,000 to 50,000), and dermatan sulfate (average molecular weight 2,000 to 50,000).
0), heparin (average molecular weight 5,000 to 20,000)
0) and heparan sulfate (average molecular weight 5,000 to 2)
Heparin, heparan sulfate, chondroitin sulfate, chondroitin, dermatan sulfate, and keratan sulfate in which the acetylamino group and / or sulfoamino group of the constituent amino sugar is converted to an amino group.

As a method of covalently bonding a carboxyl group of benzonaphthacenequinone to a polyether or glycosaminoglycan, a hydroxyl group of a benzonaphthacenequinone compound may be used as it is, or an aralkyl group such as a benzyl group;
protected with a protecting group such as an alkyl group such as a t-butyl group,
A carboxyl group of the compound is bonded to an amino group or a hydroxyl group of the polyether or glycosaminoglycan by an acid amide bond or an ester bond, and then, if necessary, by catalytic reduction or by using Na / NH ₃ , HF or HBr. A method for deprotection is exemplified.

In particular, when the polyether or glycosaminoglycan has an amino group, a method of binding using a condensing agent without protecting the hydroxyl group of the benzonaphthacenequinone compound is exemplified. When the carboxyl group of the polyether or glycosaminoglycan is used for bonding, a spacer having an amino group such as an alkylenediamine is introduced into the carboxyl group of the benzonaphthacene quinone compound, and the polyether or glycosaminoglycan is used. A method of bonding to a carboxyl group is also exemplified.

The above reaction may be carried out by a method conventionally used for an acid amide bond or an ester bond. Examples of the condensing agent include 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), -Ethyl-3- (3
Carbodiimide reagents such as -dimethylaminopropyl) carbodiimide methioside, 1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide hydrochloride, and dicyclohexylcarbodiimide can be used. The reaction can be carried out in an organic solvent such as dimethylformamide, pyridine or tetrahydrofuran at 0 ° C. to about 40 ° C., preferably at a temperature around room temperature under acidic conditions.

After the reaction, known purification means by a solvent extraction method using an organic solvent and various types of chromatography (adsorption chromatography, molecular weight fractionation chromatography, ion exchange chromatography, etc.) are appropriately combined,
The derivative (I) of the present invention can be purified and isolated.
Examples of the salt of the derivative (I) of the present invention include salts with cations such as calcium, sodium, potassium, and strontium. The salt is produced by a conventional method, but calcium is preferred.

[Formulation] The derivative (I) of the present invention can be used for various diseases of mammals including humans (eg, microbial infections, HI
V and cancer), and can be used as a preparation for oral or parenteral administration of the derivative (I) of the present invention, together with carriers, excipients and other additives. A powder can be obtained by mixing with excipients such as lactose, starch, crystalline cellulose, calcium lactate, calcium hydrogen phosphate, magnesium aluminate metasilicate, and silicic anhydride.

The granules may further contain, if necessary, a binder such as sucrose, hydroxypropylcellulose and polyvinylpyrrolidone, and a disintegrant such as carboxymethylcellulose and calcium carboxymethylcellulose in addition to the above-mentioned excipients. It can be obtained by dry granulation.

Tablets are prepared by using the above powder or granules as they are,
Alternatively, it can be obtained by adding a lubricant such as magnesium stearate, talc or the like and tableting. Further, the tablet or granule is coated with an enteric base such as hydroxypropylmethylcellulose phthalate, methyl methacrylate copolymer, hydroxypropylmethylcellulose acetate, hydroxypropylmethylcellulose succinate,
Or ethyl cellulose, carnauba wax, hardened oil,
They can be coated with white shellac or the like to make them enteric or sustained-release preparations.

A hard capsule can be obtained by filling the above powder or granule into a hard capsule. In addition, the soft capsule can be obtained by dissolving the derivative (I) of the present invention in glycerin, polyethylene glycol, sesame oil, olive oil or the like, and coating this with a gelatin film.

The syrup can be obtained by dissolving a sweetener such as sucrose, sorbitol, glycerin and the derivative (I) of the present invention in water. In addition to sweeteners and water,
An elixir is prepared by adding an essential oil, ethanol, or the like, or an emulsion or suspension is prepared by adding arabic gum, tragacanth, polysorbates (polysorbate 20, polysorbate 60, polysorbate 80 (Tween 80, etc.)), sodium carboxymethylcellulose, and the like. You can also. Also, if necessary for these liquid preparations,
Flavoring agents, coloring agents, preservatives and the like can be added.

Parenteral preparations include injections, rectal administration, pessaries, skin external preparations, inhalants, aerosols,
Eye drops and the like can be mentioned. Injectables may be the derivative (I) of the present invention, if necessary, nonionic surfactants such as polysorbates; hydrochloric acid, sodium hydroxide, lactic acid, sodium lactate, sodium monohydrogen phosphate, sodium dihydrogen phosphate, etc. PH adjuster; tonicity agent such as sodium chloride and glucose; stabilizer such as amino acids; and distilled water for injection or physiological saline, sterilized and filtered, and then filled into an ampoule. . Further, mannitol, dextran, gelatin and the like are added, and the mixture is freeze-dried under vacuum to give a dissolvable injection at the time of use. In addition, powder-filled injections can be used. Further, an emulsifier such as lecithin, polysorbates, polyoxyethylene hydrogenated castor oil, macrogol or the like may be added to the compound of the present invention, followed by emulsification in water to form an emulsion for injection.

The preparation for rectal administration contains the derivative (I) of the present invention:
After adding a suppository base such as mono-, di- or triglyceride of cocoa fatty acid, polyethylene glycol and the like, heat and melt and pour the mixture into a mold and cool, or prepare the derivative (I) of the present invention, It can be obtained by dissolving in polyethylene glycol, soybean oil or the like, and coating with a gelatin film.

[0027] The external preparation for skin includes the derivative (I) of the present invention,
It can be obtained by adding white petrolatum, beeswax, liquid paraffin, polyethylene glycol and the like, heating if necessary, and kneading.

The tape can be obtained by kneading the derivative (I) of the present invention with an adhesive such as rosin or an alkyl acrylate polymer and spreading it on a nonwoven fabric or the like.

The inhalant can be obtained, for example, by dissolving or dispersing the derivative (I) of the present invention in a propellant such as a pharmaceutically acceptable inert gas and filling the same in a pressure-resistant container.

[Administration method] The administration method of a drug containing the derivative (I) of the present invention or a salt thereof as an active ingredient is not particularly limited. For example, when the agent is used for treatment of microbial infection, HIV, cancer and the like. Injection, intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection and the like, rectal administration, pulmonary administration, eye drops and the like. The dose is appropriately determined according to the age, health condition, weight and the like of the patient, but is generally administered once a day or more. Since the derivative (I) of the present invention expresses a strong anti-yeast activity in the presence of calcium ions, it is preferable to select conditions in which calcium ions coexist when used.

[0031]

EXAMPLES Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples unless it exceeds the gist.

Example 1 Preparation of Benzonaphthacenequinone Derivative-1 Benanomycin A (8.7 mg) was dissolved in dimethylformamide (DMF) (10 ml), and ω-aminohexaethylene glycol methyl ether [methoxyhexaethylene glycol to A. Buckmann , et al. (1981) Makromol. C
hem. 182: 1379; S. Zalipsky, et al. (1983) Eur. Pol
ym. J. 19: 1177] 29.0 mg and dicyclohexylcarbodiimide 20 mg were added and reacted at room temperature for 20 hours. Insoluble materials were removed, and ethyl ether was added to the supernatant to obtain a precipitate. This precipitate was dissolved in methanol and reprecipitated by adding ethyl ether. The obtained precipitate was developed with a mixed solution of methanol and aqueous ammonia (95/5 V / V) through a silica gel column (20 cm long × 1 cm diameter) and purified to obtain 8.5 mg of the product (PEG-BenanoA-1). .
FIG. 1 shows the results of measuring the anti-yeast activity of PEG-BenanoA-1 by the following method in the presence and absence of calcium carbonate at 1 mg / ml.

[Rapid evaluation of antibiotic activity using Candida albicans 3143 as a test bacterium] 10 ml of YPD medium comprising 1% yeast extract, 2% polypeptone, and 2% dextrose
/ Candida albi cultured in a test tube at 28 ° C with shaking for 24 hours.
The cans 3143 culture was used as a seed mother. YPD medium 10 ml / test tube (calcium-containing test series) supplemented with 1 mg / ml calcium carbonate and the indicated concentration of test substance PEG-BenanoA-1, YPD supplemented with only the indicated concentration of PEG-BenanoA-1
Medium 10 ml / test tube (calcium-free test series), and
A comparative culture test group of 10 ml of YPD medium containing PEG-BenanoA-1 and a test tube (control normal growth system) was prepared, and each test tube was inoculated with a drop of a seed seeder, and Candida al.
The antibacterial activity of the test substance over time was evaluated by measuring the growth of bicans 3143 at a wavelength of 600 nm using a visible spectrophotometer.

Example 2 Preparation of Benzonaphthacenequinone Derivative-2 Benanomycin 100 mg was dissolved in 50 ml of a mixed solvent of pyridine and 0.2 M hydrochloric acid (pH 4.75), and ω-aminopolyethylene glycol methyl ether (average MW) was dissolved. 500
0, Shearwater Polymer Inc. USA). 500 mg of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) was added thereto, and the mixture was reacted at room temperature for 20 hours. The reaction mixture was concentrated under reduced pressure, and the resulting precipitate was dissolved in hot acetone, cooled and reprecipitated. This operation was repeated several times to obtain 570 mg of the purified product (PEG-BenanoA-2). The content of benanomycin A in this product was 14.0%. FIG. 2 shows the results of measuring the anti-yeast activity in the same manner as in Example 1.
Shown in

Example 3 Preparation of Benzonaphthacenequinone Derivative-3 100 mg of benanomycin A was dissolved in 20 ml of a mixed solvent of pyridine and 0.2 M hydrochloric acid (pH 4.75), and partially deacetylated heparin (aminated heparin: activity) 80 IU / mg, amino group content 1.1 μmol leucine equivalent / 10 mg) 1,00
0 mg was added. To this, 382 mg of EDC was added and reacted at room temperature for 20 hours. Ethanol was added to the reaction mixture to obtain a precipitate. The obtained precipitate was adsorbed on a DEAE Cellulofine (Seikagaku Corporation) ion exchange resin, eluted with a concentration gradient of 0 to 2 M NaCl, purified, and purified using the product (Hep-Benano A) 1, 000 mg were obtained. The content of benanomycin A in this product was 8.0%. FIG. 3 shows the results of measuring the anti-yeast activity in the same manner as in Example 1.

(Example 4) Preparation of benzonaphthacenequinone derivative-4 (1) Preparation of ω-aminotetraethylene glycol dodecyl ether 2,500 mg of tetraethylene glycol dodecyl ether was dissolved in 2 ml of pyridine and 30 ml of tetrahydrofuran. -Toluenesulfonyl chloride solution in tetrahydrofuran (2,631 mg / 20 ml) was added and
Allowed to react for hours. The precipitate formed was filtered and the precipitate was
Was washed four times with tetrahydrofuran, and the tetrahydrofuran solution was concentrated. The obtained residue was washed three times with 100 ml of n-hexane, purified with a silica gel column developed with a solvent dichloroethane / methanol (97: 3), and purified with tosyltetraethylene glycol dodecyl ether 2,49.
5 mg were obtained. This was dissolved in 20 ml of dimethyl sulfoxide, and 314 mg of sodium azide was added thereto.
The reaction was carried out at a temperature of 3 ° C. for 3 hours in a nitrogen stream. After concentration, extract with dichloromethane, wash with water and saturated brine,
200 mg of azidotetraethylene glycol dodecyl ether were obtained. Dissolve this in dichloromethane and add 50mg
And palladium black was added thereto, and the mixture was reduced by passing through hydrogen gas, and the filtrate was concentrated. The residue was purified with a silica gel column developed with methanol / ammonia (9: 1).
3 mg of ω-aminotetraethylene glycol dodecyl ether were obtained.

(2) Preparation of monododecyloxytetraethylene glycol-benanomycin A 100 mg of benanomycin A and 52.9 mg of ω-aminotetraethylene glycol dodecyl ether are dissolved in pyridine / 0.1 M hydrochloric acid buffer (pH 4.75). And 28
0.7 mg of EDC was added and reacted at room temperature for 24 hours.
Purification by a silica gel column gave 122 mg of monododecyloxytetraethylene glycol-benanomycin A. Benanomycin A content was 65.1%.

[0038]

According to the present invention, by covalently bonding a polyether or glycosaminoglycan to a benzonaphthacenequinone compound, the benzonaphthacenequinone compound retains the antibacterial activity of the benzonaphthacenequinone compound while preventing side effects on the liver and other organs. To reduce.

[Brief description of the drawings]

FIG. 1 is a graph showing the anti-yeast activity of the conjugate of benanomycin A-polyethylene glycol obtained in Example 1.

FIG. 2 is a graph showing the anti-yeast activity of the conjugate of benanomycin A-polyethylene glycol obtained in Example 2.

FIG. 3 is a graph showing the anti-yeast activity of the benanomycin A-aminated heparin conjugate obtained in Example 3.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Tomio Takeuchi 5-1A New Fuji Mansion 701A, Higashi Gotanda, Shinagawa-ku, Tokyo (72) Inventor Shinichi Kondo 3-1-1 Minamirokugo, Ota-ku, Tokyo Royal Heights Minamirokugo 608 (72) Inventor Katsumi Sakurai 2-527-6 Kurashiki, Higashiyamato-shi, Tokyo (72) Inventor Yasuo Fukagawa 3-9-1-2 Imaizumidai, Kamakura-shi, Kanagawa (72) Inventor Akiko Miya Fujisawa-shi, Kanagawa 5244-1, Oba, Shonan Castle, 14-103

Claims (4)

[Claims] 1. A benzonaphthacenequinone derivative or a salt thereof, wherein a benzonaphthacenequinone compound represented by the following formula (I) and a polyether or glycosaminoglycan are covalently bonded. Embedded image [Wherein, R ₁ represents a hydrogen atom, a lower alkyl group or a hydroxy lower alkyl group, R ₂ represents a hydroxyl group, an amino group, or a mono- or di-lower alkylamino group, and R ₃ represents a hydrogen atom, a non-sulfated or R ₄ represents a hydrogen atom, a lower alkyl group or a D-xylosyl group; ] 2. The following formula (II), wherein the covalent bond is an acid amide bond or an ester bond between a carboxyl group of the benzonaphthacenequinone compound and an amino group or a hydroxyl group of the polyether or glycosaminoglycan. 2. The derivative according to claim 1 or a salt thereof. Embedded image [R _{1 to} R ₄ are the same as above, and X is NH—Y or O—
And Y represents a residue obtained by removing amino groups or hydroxyl groups involved in the above-mentioned binding from polyether or glycosaminoglycan. ] 3. The polyether is a polyalkylene glycol having an amino group, wherein the glycosaminoglycan is obtained by converting acetylamino group and / or sulfoamino group of a constituent amino sugar into an amino group, heparin, heparan sulfate, The derivative according to claim 1 or 2, which is chondroitin sulfate, chondroitin, dermatan sulfate or keratan sulfate, or a salt thereof. 4. A pharmaceutical composition comprising the benzonaphthacenequinone derivative or a salt thereof according to claim 1.