JP2003201235A - Medicine containing neuraminic acid compound - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide the hydrate and crystal of a neuraminic acid compound having good storage stability and useful as a medicine. <P>SOLUTION: This hydrate or crystal of the neuraminic acid compound is shown by the formula. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a hydrate or crystal of neuraminic acid compound (I) having excellent sialidase inhibitory activity, and a medicine containing the hydrate or crystal as an active ingredient, particularly anti-influenza. Regarding medicine.

[0002]

2. Description of the Related Art Japanese Patent Laid-Open No. 10-330373 discloses
A neuraminic acid compound represented by formula (I) (hereinafter referred to as compound (I)) is disclosed. Compound (I) exhibits an excellent sialidase inhibitory action, and can be expected to be useful as a therapeutic drug for influenza and a preventive drug. However, in order to put this compound into practical use as a drug substance, storage stability and easy handling are required.

[0003]

[Patent Document 1] Japanese Unexamined Patent Publication No. 10-330373 (No. 9)
(Page 8, Example 35)

[0004]

DISCLOSURE OF THE INVENTION As a result of intensive studies on the compound (I), the inventors have found that a hydrate of the compound (I) can be obtained as a crystal exhibiting excellent stability. A hydrate crystal of compound (I) is disclosed in JP-A-10-
Compared with the trifluoroacetate salt of compound (I) described as Example 35 in JP-A-330373, storage stability and easiness of handling are remarkably improved, and it is extremely useful for practical use as a medicine. The present invention has been completed and the present invention has been completed.

[0005]

The present invention provides (1) Formula (I)

[0006]

[Chemical 3]

A hydrate of a compound represented by the formula (2): Formula (I)

[0008]

[Chemical 4]

A crystal containing a compound represented by: (3)
In (2), a hydrate crystal of the compound represented by formula (I), and (4) In (2) and (3), K of copper
A compound represented by the formula (I) having main peaks at interplanar spacings of 4.0, 4.4, 4.7, 7.5 and 10.2 angstroms in powder X-ray diffraction obtained by irradiation with α rays. It is a hydrate crystal. The crystal of the present invention is a solid whose internal structure is three-dimensionally made up of regular repeating atoms (or a group thereof), and is distinguished from an amorphous solid which does not have such regular internal structure. .

Even if crystals of the same compound, a plurality of crystals (polymorphism) having different internal structures and physicochemical properties may be produced depending on the crystallization conditions. Any of these crystal polymorphs may be used, and a mixture of two or more crystal polymorphs may be used.

Since the compound (I) has a guanidino group and a carboxyl group in the molecule, it can form a pharmacologically acceptable salt by binding with an acid or base which is not pharmacologically toxic.

Examples of pharmaceutically acceptable salts include hydrohalides such as hydrofluoric acid salts, hydrochloric acid salts, hydrobromic acid salts, and hydroiodic acid salts; nitrates, perchlorates. Inorganic acid salts such as sulphate and phosphate; alkane sulphonates such as methane sulphonate, ethane sulphonate and trifluoromethane sulphonate; benzene sulphonate, p-toluene sulphonate Aryl sulfonates; organic acid salts such as acetate, trifluoroacetate, citrate, tartrate, oxalate, maleate; glycine, lysine, arginine, ornithine, glutamate, Amino acid salt such as aspartate; Alkali metal salt such as lithium salt, sodium salt, potassium salt; Alkaline earth metal such as calcium salt, magnesium salt Salts; metal salts such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts; ammonium salts, t-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, ethylenediamine salts, guanidine salts. , Diethylamine salt, triethylamine salt, dicyclohexylamine salt, procaine salt, ethanolamine salt, diethanolamine salt, piperazine salt, organic amines such as tetramethylammonium salt, or organic ammonium salt, and the like, preferably lithium salt, It is an alkali metal salt such as sodium salt or potassium salt; an organic acid salt such as acetate or trifluoroacetate; or an inorganic acid salt such as hydrochloride or sulfate.

Compound (I) and its pharmaceutically acceptable salts absorb water or solvent by leaving it in the air or mixing with water or an organic solvent to form a hydrate or solvate. May be formed.

A crystal containing the compound (I) of the present invention is
In addition to the crystal consisting of compound (I), a crystal consisting of a pharmacologically acceptable salt, hydrate or solvate of compound (I) is included. Of the crystals containing these compounds (I), hydrate crystals are preferable.

As one form of the crystal containing the compound (I) of the present invention, for example, in the powder X-ray diffraction obtained by irradiating copper with Kα rays (wavelength λ = 1.54 Å), the interplanar spacing d = 4. .0, 4.4, 4.7, 7.5 and 1
A crystal exhibiting a main peak at 0.2 Å can be mentioned. Here, the main peak is a peak having a relative intensity of 80 or more when the intensity of the peak showing the interplanar spacing d = 4.7 Å is 100. In the powder X-ray analysis pattern of FIG. 1, the vertical axis represents the diffraction intensity /
It is shown in units of seconds (cps), and the horizontal axis shows the diffraction angle 2θ (degrees). The surface spacing d (unit: angstrom) is
It can be calculated as n = 1 in the formula 2d sin θ = nλ.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION Compound (I) or a pharmaceutically acceptable salt thereof can be produced according to the method disclosed in JP-A-10-330373 or a method analogous thereto.

Crystals containing the compound (I) are prepared by dissolving the compound (I) or a pharmacologically acceptable salt thereof in a suitable solvent, adjusting the pH, concentrating the solution, mixing a good solvent and a poor solvent, and the like. Then, the compound (I), a pharmacologically acceptable salt or solvate thereof (including a hydrate) is brought to a supersaturated state and precipitated.

The crystal precipitation can be initiated spontaneously in the reaction vessel, but it can also be initiated or promoted by applying mechanical stimuli such as seed crystal inoculation, ultrasonic stimulation, and rubbing the surface of the reactor. it can.

More specifically, the crystal (particularly hydrate crystal) containing the compound (I) of the present invention is an aqueous solution containing the compound (I) or a pharmacologically acceptable salt thereof, if necessary. It can be produced by adjusting the pH to a suitable value, concentrating and precipitating crystals by cooling, and then isolating the precipitated crystals.

Crystals containing the compound (I) (particularly hydrate crystals) are purified by an aqueous solution containing the crystals using a reverse phase silica gel column, and the fraction containing the compound (I) is collected as necessary. It can also be produced by precipitating crystals by concentrating, cooling and then isolating the precipitated crystals.

The crystals obtained can be improved in purity and quality by recrystallization or slurry refining.

The temperature for crystallizing the crystal containing the compound (I) (particularly the hydrate crystal) is 0 to 40.
C. is preferable, and more preferably 20 to 30.degree.

The pH suitable for crystallizing the crystal containing the compound (I) (particularly the hydrate crystal) is preferably pH 4 to 9 in which the solubility of the hydrate of the compound (I) in water is small, More preferably, it is 5 to 7.

The precipitated crystals can be isolated, for example, by filtration, centrifugation, or decantation. The isolated crystals can be washed with a suitable solvent as needed.

When the hydrate crystals are washed, for example, water, ethanol, isopropanol, acetone, ethyl acetate,
Solvents such as toluene, acetonitrile, methyl acetate or ether can be used, preferably water, acetone, ethyl acetate or toluene.

The isolated crystals (particularly hydrate crystals) are usually at a temperature of 10 to 100 ° C., preferably 30 to 50 ° C.
It can be dried at a temperature of 1 to a constant weight. The crystals can be dried, if necessary, in the presence of a desiccant such as silica gel or calcium chloride, or under reduced pressure.

The dried crystals (particularly hydrate crystals) are usually at a temperature of 10 to 30 ° C. and a relative humidity of 20% to 90%, preferably at a temperature of 20 to 30 ° C. and a relative humidity of 50% to 80%. Moisture may be absorbed until the weight becomes almost constant.

The eluent used for the purification of the reversed phase silica gel column of compound (I) or a pharmaceutically acceptable salt thereof is water, acetone, acetonitrile, methanol, ethanol and a mixed solvent thereof, preferably water, methanol and It is the mixed solvent. As the reverse phase silica gel, silica gel whose surface is modified with an organic residue such as an alkyl group can be used, but is preferably modified with an octadecyl group.

Recrystallization can be achieved by a method usually used in the field of synthetic organic chemistry, such as dissolving a crystal containing compound (I) in a good solvent and adding a poor solvent to precipitate the crystal.

Examples of the good solvent used for recrystallizing the hydrate crystal of the compound (I) include alcohols such as methanol and ethanol, and preferably methanol.

Examples of the poor solvent used for recrystallizing the hydrate crystal of compound (I) include water, hexane, diisopropyl ether, t-butyl methyl ether and the like, preferably water, hexane or diisopropyl. Ether, more preferably water or diisopropyl ether.

Slurry refining is the operation of suspending the crystal of the compound in a suitable solvent and stirring, and then isolating the crystal again. As a solvent used for slurry purification of hydrate crystals, for example, acetone, methyl ethyl ketone, methyl acetate, ethyl acetate, acetonitrile, methylene chloride, toluene, ethanol, isopropanol, tetrahydrofuran, N, N-dimethylformamide, water, hexane,
Examples thereof include diisopropyl ether and ether, preferably acetone, methyl ethyl ketone, methyl acetate, ethyl acetate, acetonitrile, ethanol or isopropanol, and more preferably acetone or ethyl acetate.

The crystals obtained by recrystallization and slurry purification can also be isolated in the same manner as above.

When a crystal containing compound (I) (particularly a hydrate crystal) is used as a medicine, particularly as a therapeutic or prophylactic agent for influenza, itself or an appropriate pharmacologically acceptable agent. Mixing with excipients, diluents, etc., tablets, capsules, granules, powders, syrups, injections,
It can be administered by an ointment, a liquid, a suspension, an aerosol, a troche, or the like. The medicament of the present invention can be administered orally or parenterally, but by a method in which the active ingredient, compound (I), can be directly delivered to the lungs or respiratory tract (including intraoral and intranasal). Preferably.

These formulations include excipients such as lactose,
Sugars such as sucrose, glucose, mannitol and sorbitol; corn starch, potato starch, α-starch, dextrin, starch derivatives such as carboxymethyl starch; crystalline cellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose. Cellulose derivatives such as carboxymethyl cellulose calcium, sodium carboxymethyl cellulose internally crosslinked; gum arabic; dextran; pullulan; light anhydrous silicic acid, synthetic aluminum silicates, silicates such as magnesium aluminometasilicate; phosphates such as calcium phosphate; Carbonates such as calcium carbonate; sulfates such as calcium sulfate, etc.), binders (for example, the above-mentioned excipients; gelatin; polyvinyl chloride) Pyrrolidone; Maglogol, etc.), disintegrants (for example, the aforementioned excipients; croscarmellose sodium, sodium carboxymethyl starch, chemically modified starch or cellulose derivatives such as cross-linked polyvinylpyrrolidone, etc.), lubricants ( For example, talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; bee gum; waxes such as beeswax and gallow; boric acid; glycols; carboxylic acids such as fumaric acid and adipic acid. Carboxylic acid sodium salt such as sodium benzoate; sulfuric acid salt such as sodium sulfate; leucine; lauryl sulfate such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acid anhydride such as silicic acid hydrate and silicic acid hydrate; Excipients mentioned above In starch, etc.), stabilizers (eg, parahydroxybenzoic acid esters such as methylparaben, propylparaben; alcohols such as chlorobutanol, benzyl alcohol, phenylethyl alcohol; benzalkonium chloride; phenol, cresol, etc. Phenols; thimerosal; acetic anhydride; sorbic acid, etc.), flavoring agents (eg, commonly used sweeteners, acidulants, flavors, etc.), suspending agents (eg polysorbate 80, sodium carboxymethylcellulose, etc.) , Diluents, formulation solvents (eg water, ethanol,
It is produced by a known method using additives such as glycerin).

The amount used depends on the symptoms, weight, and
The lower limit is 0.1 mg (preferably 1 mg) and the upper limit is 1000 mg (preferably 500), depending on age and the like.
It is desirable to administer mg) once to several times a day depending on the symptoms.

[0037]

EXAMPLES The present invention will be described in more detail by showing Examples, Formulation Examples and Test Examples below. (Production Example 1) 5-acetamido-4-guanidino-9-O-octanoyl-2,3,4,5-tetradeoxy-7-O-methyl-D-glycero-D-galacto-non-2-enopyrazonic acid Hydrate Crystal (1) Example 35 of JP-A-10-330373
Compound of (i), 5-acetamido-4- (N, N'-bis-t-butyroxycarbonyl) guanidino-9-O-
Octanoyl-2,3,4,5-tetradeoxy-7-
O-methyl-D-glycero-D-galacto-non-2-
Diphenylmethyl enopyranosonate (3.46 g, 4.
1 mmol) was dissolved in methylene chloride (27 ml) and trifluoroacetic acid (14 ml), and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated to dryness under reduced pressure, and then 3 times with toluene (5 ml).
Azeotropically dried up. The obtained oily matter was treated with ethyl acetate (10 m
It was dissolved in 1). On the other hand, add this solution to saturated sodium bicarbonate water (50 m
l), add 20% sodium carbonate aqueous solution and add pH
It was set to 8.5. After stirring at room temperature for 3 hours, hydrochloric acid (3 m
The pH was adjusted to 5.0 with l) and the mixture was stirred at room temperature for 1 hour. After stirring for 1 hour under ice-cooling, the crystals are suction-filtered and the external temperature is 50 ° C.
It was vacuum dried for 10 hours. It was left in the air for 1 day to obtain the title object compound as crystals. (0.97g, 51
%) Infrared absorption spectrum (KBr) ν max cm ^-1 : 3412, 2929, 2
856, 1676, 1401, 1320, 1285, 1205, 1137, 722. ¹ H nuclear magnetic resonance spectrum (400MHz, CD ₃ OD) δppm: 5.88
(1H, d, J = 2.5 Hz), 4.45 (3H, m), 4.27 (1H, dd, J = 1
0.0 Hz, 10.0 Hz), 4.15 (1H, m), 3.47 (2H, m), 3.42
(3H, s), 2.37 (2H, t, J = 7.4 Hz), 2.10 (3H, s), 1.
31 (2H, m), 1.20-1.40 (8H, m), 0.85-0.95 (3H, m). ¹³ C nuclear magnetic resonance spectrum (100MHz, CD ₃ OD) δppm: 176.
5, 173.7, 164.7, 158.9, 146.7, 108.7, 80.1, 78.0,
69.3, 66.8, 61.4, 52.4, 35.1, 32.8, 30.2, 30.1, 26.
0, 23.7, 22.8, 14.4. FIG. 1 shows a powder X-ray diffraction pattern obtained by irradiating copper with Kα rays (wavelength λ = 1.54 Å) of this crystal. The vertical axis of the powder X-ray analysis pattern represents the diffraction intensity in units of counts / second (cps), and the horizontal axis represents the diffraction angle 2θ (degrees). (2) The title compound was also obtained by the following method.

The compound of Example 35 (ii), 5-acetamido-4-guanidino-9-O-octanoyl-2,3,4,5-tetradeoxy-7-O-methyl, disclosed in JP-A-10-330373. -D-glycero-D-galacto-non-2-enopyrazonic acid trifluoroacetate (3.
0 g, 5.1 mmol) by reverse phase column chromatography (Nacalai Tesque Cosmosil 75C18PREP, 100)
g), and methanol-water (0: 1, 1: 1, 2 :).
Elution was performed using 1) while increasing the methanol content.
The fraction containing the target compound was concentrated under reduced pressure, and the precipitated crystals were suction filtered and dried under reduced pressure. The mixture was allowed to stand in air for 1 day to give the title object compound as crystals (1.2 g, 49%). The physical property data of the obtained compound were completely in agreement with those obtained in (1) above. (Test Example 1) Stability Test Crystal of compound (I) of Production Example 1 of the present invention and amorphous powder of compound (I) described as Example 35 in JP-A-10-330373 as a control (tri (Fluoroacetate) was stored in a desiccator at 40 ° C. and 75% relative humidity, and the residual rate was measured after 14 days (2 weeks), 28 days (4 weeks) and 56 days (8 weeks). The results are shown in Table 1. Separately, 14 ° C in a silica gel desiccator at 60 ° C.
The survival rate after day (2 weeks), 28 days (4 weeks), 56 days (8 weeks) was measured. The results are shown in Table 2.

The residual rate (%) of the compound was determined from the residual rate of the compound measured by high performance liquid chromatography (HPLC). The HPLC measurement conditions are as follows.

Column: L-colum manufactured by Chemicals Evaluation and Research Institute
n ODS (4.6mm ID x 250mm) Mobile phase: 0.1mol / phosphate buffer (pH3): acetonitrile
(7: 3) Flow rate: 1 mL / min Detection: 230 nm Column temperature: 30 ° C. [Table 1] 40 ° C., relative humidity 75% Stability in desiccator (residual rate) ---------- −−−−−−−−−−−−−−−−−−−−−−−− Compound Days Elapsed 14 days 28 days 56 days ---------- -------------------- Crystal of hydrate of compound (I) 99.8% 100.2% 100.3% Amorphous powder of compound (I) 38.5 % 27.2% 10.2% −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−− As shown in Table 1. The amorphous powder of compound (I) (trifluoroacetate salt) had extremely low stability in a humidified state at 40 ° C., and after 56 days, the content decreased to about 10.2%. On the other hand, the crystal (hydrate crystal) of the compound (I) of the present invention shows a residual rate of 100% under the same conditions and has an extremely high storage stability. [Table 2] Stability (residual rate) in a silica gel desiccator at 60 ° C -------------------------------------. Compound Elapsed days 14 days later 28 days 56 days later ------------------ Crystal of hydrate of compound (I) 99.9% 100.6% 99.6% Amorphous powder of compound (I) 95.7% 92.9% 89.0% ---------------. As shown in Table 2, the amorphous powder of compound (I) (trifluoroacetate salt) has low stability even at 60 ° C. in a dry state, and is stored for 56 days. After that, the content decreased to about 89%. On the other hand, the crystal (hydrate crystal) of the compound (I) of the present invention is
Under the same conditions, it shows a high residual rate of 99% or more and has extremely high storage stability. (Test Example 2) 5-acetamide-4-guanidino-9-O-octanoyl-
Anti-influenza effect of 2,3,4,5-tetradeoxy-7-methoxy-D-glycero-D-galacto-non-2-enopyrazonic acid hydrate crystals 5-6 week old Balb / C mice (female) In an anesthesia bottle filled with chloroform / ether (1: 1),
Anesthetized. Anesthetized mice were administered with 50 μL of a solution of a compound (hydrate crystal) dissolved in physiological saline, and the dose was 0.3 μm.
It was dripped into the nasal cavity so that it became ol / kg. Mice similarly anesthetized 10 days, 7 days, 5 days, 3 days, or 4 hours after the administration of influenza virus A / PR / 8/34 strain (1,500 plaque formati)
on units). Mice were observed to be alive or dead until 20 days after infection.

As a result, all non-administered mice died by 6 days after infection, whereas all the 10-day-pretreated mice and 7-day-pretreated mice died, 8 days and 11 days, respectively.
It took a day. Mice administered 5 days, 3 days, and 4 hours before were still alive at 20 days after infection, and the survival rates at that time were 50%, 83%, and 100%, respectively. These results show that the present compound (hydrate crystal) has a therapeutic effect on influenza and also a preventive effect. (Formulation Example 1) Liquid formulation 1 The compound of Production Example 1 is 10% (W / W), benzalkonium chloride is 0.04% (W / W), phenethyl alcohol is 0.40% (W / W), and is purified. Adjust the solution so that the water content is 89.56% (W / W). (Formulation Example 2) Liquid formulation 2 The compound of Production Example 1 is 10% (W / W), benzalkonium chloride is 0.04% (W / W), polyethylene glycol 4
00 is 10% (W / W), propylene glycol is 30%
(W / W) and purified water are adjusted to 39.96% (W / W). (Formulation Example 3) 40% (W / W) of the compound of Powder Production Example 1 and 60 of lactose
Adjust the powder so that it is% (W / W). (Formulation Example 4) 10% (W / W) of the compound of Aerosol Preparation Example 1 and 0.5 of lecithin
% (W / W), CFC 11 is 34.5% (W / W), CFC 1
Adjust the aerosol agent so that 2 becomes 55% (W / W).

[0042]

EFFECTS OF THE INVENTION The crystal containing the compound (I) of the present invention (particularly a hydrate crystal) is a crystal which is excellent in storage stability and practically easy to handle as compared with an amorphous trifluoroacetate salt. Therefore, it is useful as an active ingredient of a medicine (in particular, a therapeutic or prophylactic agent for influenza). In addition, the hydrate of compound (I) is useful as a molecular form that gives excellent crystals.

[Brief description of drawings]

FIG. 1 shows 5-acetamide-obtained in Production Example 1.
4-guanidino-9-O-octanoyl-2,3,4,
5-tetradeoxy-7-O-methyl-D-glycero-
It is a powder X-ray-diffraction pattern obtained by irradiating a copper K (alpha) ray (wavelength (lambda) = 1.54 angstrom) about a D-galacto-non-2-enopyrazonic acid hydrate crystal.
The vertical axis of the powder X-ray analysis pattern shows the diffraction intensity in units of counts / second (cps), and the horizontal axis shows the diffraction angle 2θ (degrees).
Indicates. The surface spacing d (angstrom) is calculated by the formula
It can be calculated as n = 1 in 2d sin θ = nλ.

[Procedure amendment]

[Submission date] October 25, 2002 (2002.10.
25)

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be corrected] 0005

[Correction method] Change

[Correction content]

[0005]

The present invention provides (1) Formula (I)

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Makoto Yamaoka             1-12-1, Shinomiya, Hiratsuka City, Kanagawa Prefecture             Kyo Co., Ltd. (72) Inventor Yuu Honda             1-2-2 Hiromachi, Shinagawa-ku, Tokyo Sankyo Co., Ltd.             Inside the company F-term (reference) 4C062 BB16 BB19                 4C086 AA01 AA04 BA07 MA01 MA17                       MA52 MA55 MA66 NA03 ZB33

Claims (5)

[Claims] 1. A compound represented by the formula (I): An anti-influenza drug containing a hydrate of a compound represented by: 2. Formula (I): An anti-influenza drug containing, as an active ingredient, a crystal containing a compound represented by: 3. An anti-influenza drug according to claim 2, which comprises a hydrate crystal of the compound represented by formula (I) as an active ingredient. 4. The powder X-ray diffraction obtained by irradiating copper with Kα rays according to claim 2 or 3, wherein the interplanar spacing is 4.0,
An anti-influenza drug containing, as an active ingredient, a hydrate crystal of a compound represented by the formula (I) having major peaks at 4.4, 4.7, 7.5 and 10.2 angstroms. 5. A medicine containing the active ingredient according to any one of claims 1 to 4.