JP2654445B2 - Pharmaceutical composition using natural albumin as carrier and process for producing the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a pharmaceutical composition containing albumin as a carrier, which contains one or more drugs exhibiting affinity for albumin in a high concentration by a method described below. More specifically, the present invention relates to a pharmaceutical composition which has reduced side effects unfavorable to the human body, improved absorption of a drug, and improved photostability.

[Prior art and its problems]

Conventionally, as a pharmaceutical composition using albumin as a carrier, 5
A complex of a fluorouracil derivative and albumin (JP-A-54-151988) and a sustained-release albumin complex (JP-A-58158)
No. 162595). However, the method described in this patent publication requires changing the binding property of the carrier albumin in advance using a cyanogen halide or perhalic acid at a pH of 6 to 11 and a temperature of 30 ° C. or less,
Further, it is necessary to improve the affinity by using a spacer such as a functional compound.

On the other hand, the pharmaceutical composition of the present invention using natural albumin as a carrier does not require a cyanogen halide, perhalic acid or a spacer as described below, and is completely different from that described in the above-mentioned patent publication. Cyclodextrin-indomethacin inclusion complexes using cyclodextrin as others (JP-A-54-62313, JP-A-54-117018)
), And a biodegradable carrier selected from collagen, gelatin and the like containing indomethacin (Japanese Patent Application Laid-Open No. 60-84213). It is excellent, but its absorption in the body is poor, and it takes a long time to develop its medicinal effect.

An object of the present invention is to provide a pharmaceutical composition capable of reducing the damage to the skin and mucous membranes of a pharmaceutical, particularly the digestive system, and improving the absorbability, the stability to light, and the like of the pharmaceutical.

[Means for solving the problem]

The present invention has been made to achieve the above-mentioned object, and the pharmaceutical composition of the present invention comprises nalidixic acid, indomethacin, nifedipine, tocophenol, aspirin, piroxicam, sulindac, phenylbutazone, cefaclor, cephalexin, hydrochloric acid Tetracycline, ampicillin, dexamethasone, haloperidol, diltiazem hydrochloride, griseofulvin,
Vincristine sulfate, cimetidine, ketotifen, selected from glitiltin or tranilast, 1 to 200 parts of a protein-binding drug, and 100 parts of natural albumin, characterized by containing, reducing undesirable side effects, absorption of the drug It is improved and has excellent light stability.

JP-A-58-216126 describes the use of human serum albumin to make water-insoluble or poorly soluble drugs soluble. However, the method described in this patent application is based on a conventionally known method for measuring the protein binding rate of a drug, and the drug that can be solubilized is extremely 0.01 to 0.8% relative to serum albumin. It has a low content and is an exceptionally useful technique only at very low clinical doses, as in the case of the prostaglandins shown in this patent publication.

On the other hand, the pharmaceutical composition using the natural albumin of the present invention as a carrier is described in the above-mentioned patent publication in that it can be applied to ordinary high-dose pharmaceuticals because the pharmaceutical content can be increased as described in detail below. Completely different from

Examples of the albumin used in the present invention include ovalbumin, serum albumin, plaque albumin, α
-Lactalbumin, leucosine, phaserin, levumerin and the like can be mentioned, with ovalbumin being particularly preferred. In order to apply the composition obtained according to the present invention to the field of pharmaceuticals in which antigenicity is a problem, it is preferable to use human-derived albumin. The drug used in the present invention preferably has a protein binding ratio of 10% or more, preferably 20% or more. Further, the compounding amount is preferably at least 1 part, preferably 5 to 200 parts, per 100 parts of albumin.

Pharmaceuticals used in the present invention, nalidixic acid, indomethacin, nifedipine, tocophenol, aspirin, piroxicam, sulindac, phenylbutazone, cefaclor, cephalexin, tetracycline hydrochloride, ampicillin, dexamethasone, haloperidol, diltiazem hydrochloride, griseofulvin, vincristine sulfate, cimetidine, cimetidine, It is limited to those selected from ketotifen, glycyltin or tranilast.

The method for producing the pharmaceutical-albumin composition of the present invention will be specifically described with reference to an example of nalidixic acid-albumin composition.

First, natural albumin such as ovalbumin is suspended in an aqueous solution, and nalidixic acid dissolved in a suitable organic solvent having a boiling point of 100 ° C. or lower is added to the suspension, followed by high-speed stirring to form a uniform layer. The temperature at this time is not particularly limited as long as the albumin is not denatured.
It is necessary to perform high-speed stirring in a temperature range of about 1 to 30 hours. For example, high-speed stirring is performed using a biomixer BM-4 (manufactured by Nippon Seiki Seisakusho, maximum rotation speed of 30,000 rpm) or a large high-speed homogenizer (Nisson Medical Science Manufactured by the factory, maximum rotation speed 20000r
pm) under conditions of about 5,000 to 40,000 rpm. The concentration of the albumin suspension at the time of producing this composition is not particularly limited, but it can be used at a concentration of at least 1% by weight, preferably 2 to 25% by weight. In general, it is preferable to use albumin in an amount of 5 to 100 parts of nalidixic acid with respect to 100 parts of albumin. Any aqueous solution can be used as long as it dissolves albumin without denaturation, but water is most preferred. However, a part of the water can be replaced by a water-soluble solvent such as methanol, ethanol, isopropanol, and acetone. Examples of the organic solvent used when dissolving nalidixic acid in an organic solvent include acetone, chloroform, methanol, and ethanol. The concentration when dissolving nalidixic acid is not particularly limited, but it is advantageous to dissolve at least 0.1% by weight, preferably 1 to 50% by weight. The mixing ratio of the nalidixic acid solution and the albumin suspension dissolved in the organic solvent is not particularly limited, but preferably the nalidixic acid solution is 1 to 4 with respect to 100 volumes of the albumin suspension.
Mix in a range of 00 volumes. The organic solvent used is distilled off under reduced pressure at a temperature of 40 ° C. or lower from the reaction solution thus obtained. Thereafter, the desired powdery nalidixic acid-albumin composition can be obtained by removing the aqueous solvent using a means such as spray drying or freeze drying. The composition obtained here reacts almost 100% irrespective of the ratio of both nalidixic acid and albumin, and a composition in which nalidixic acid is bound to albumin in an amount of about 5 to 100% is obtained depending on the ratio of both added.

Although the production example of the nalidixic acid-albumin composition has been described above, the pharmaceutical-albumin composition in which other pharmaceuticals are supported on albumin can be produced by the same method. In addition, not only the nalidixic acid-albumin composition, in the production of the pharmaceutical-albumin composition of the present invention, a fatty oil and / or a fatty acid can be used together with the pharmaceutical and the albumin, and the pharmaceutical-albumin composition obtained thereby. The absorptivity of the object is further improved.

The following experiments demonstrate that the nalidixic acid-albumin composition obtained above improves the absorption of nalidixic acid taken after a meal.

Gastrointestinal absorption test of nalidixic acid and nalidixic acid-albumin composition (nalidixic acid content: 8.9%) Experimental animals Rats, Wistar strains, male, (body weight: about 290 g, Sankyo Lab) were used under fed conditions.

Administration and blood collection The dose was 50 mg / kg as nalidixic acid. It was administered in a suspended state in 1 ml of water using an oral gastric probe. Approximately 0. from the fundus with a capillary (Drummond, 0.1 ml).
2 ml of blood was collected and centrifuged to obtain serum.

Sample processing and quantification method Serum was mixed with an equal volume of methanol to remove proteins.

 The quantification was performed by high performance liquid chromatography.

Quantitative conditions Column: TOYOSODA, ODS-80TM, 4.6 mm × 15 cm Mobile phase: acetonitrile: 50 mM citric acid solution: 1 M ammonium acetate = 35: 64: 1 Flow rate: 0.7 ml / min Detection wavelength: 258 nm Injection volume: 10 μl Column temperature: The room temperature calibration curve was obtained by using a blank serum, adding known nalidixic acid, and performing the same treatment as the sample.

Results The results are shown in FIG.

As is clear from FIG. 1, when the nalidixic acid-albumin composition was administered at 50 mg / kg as nalidixic acid, the maximum blood concentration was reached within 1.5 hours after the administration, and the concentration at this time was about 25 μg / ml. It was estimated. On the other hand, the administration of the same volume of nalidixic acid bulk powder reached the maximum blood concentration within 3 hours after administration, and the concentration at this time was estimated to be about 7 μg.
In addition, the area under the serum concentration-time curve (AUC) of both is 0 to
When the 8-hour value was calculated, the composition was 108.9 μg · hr / ml, and the raw powder was 34.6 μg · hr / ml. From the comparison of these pharmacokinetic parameters, it can be determined that the absorbability of the composition of the present invention is superior to that of the bulk powder. In general, a pharmaceutical composition in which a drug is adsorbed or bound to a carrier is sustained-released and absorption is delayed. However, it can be seen from FIG. 1 that the composition of the present invention is accelerated as compared with the bulk powder.

Next, an example of reducing the side effects on the digestive tract according to the present invention will be described. Non-steroidal anti-inflammatory drugs have a strong anti-inflammatory effect at local sites of inflammation, but are known to cause bleeding and ulcers on the digestive system. The indomethacin-albumin composition produced by the above-mentioned method does not show ulceration with indomethacin, which is a nonsteroidal anti-inflammatory drug, and exhibits an anti-inflammatory action equivalent to indomethacin. This is demonstrated by the following experiment.

Gastric ulcer-forming effect of indomethacin and indomethacin-albumin composition (indomethacin content 8.8%) Test method The test was carried out by the method of Okabe et al. [Japanese Journal
Of Pharmacology (J. Jap. Pharmacol.) 29 670
Page (1979)].

That is, a Wistar rat (eight rats per group) weighing 180 to 200 g was fasted for about 18 hours, and the indomethacin-albumin composition was used as a sample in an amount of 20 mg as indomethacin.
/ kg of 1% sodium carboxymethylcellulose solution (abbreviated as 1% CMCNa)
Oral administration was performed at a rate of 0.5 ml per 00 g. As a control, an indomethacin bulk powder suspended in 1% CMCNa at 20 mg / kg was similarly administered. 7 hours after administration, all rats were killed by ether and 8 ml of 1% buffered formalin aqueous solution
Was injected into the stomach, extracted and immersed in a 1% buffered formalin aqueous solution for 15 minutes. Next, the stomach was incised along the greater curvature, and the gastric mucosa was observed under a stereoscopic microscope. When an injured part (ulcer) was observed, its major axis (mm) was measured. The total was defined as an ulcer occurrence coefficient, and 30 mm or more, 10 to 30 mm, 5 to 10 mm +, and less than that, no occurrence was observed.

 Table 1 shows the results.

As is clear from Table 1, at 7 hours after administration, ulcers were observed in all the indomethacin bulk powder administration groups, whereas the indomethacin-albumin composition administration group showed 8 ulcers.
No ulcer formation was observed in up to six of the cases, and the ulcer was small in two cases slightly observed.

Also, the anti-inflammatory effect of the indomethacin-albumin composition is demonstrated by the following experiment.

Anti-inflammatory effect of indomethacin-albumin composition Judged by carrageenan footpad edema method.

That is, after using a Wistar male rat weighing 150 to 180 g (five rats per group) for about 18 hours, the volume of the rat right hind footpad was determined by the volume of the rat footpad edema volume. It was measured using a measuring device (Natsume Seisakusho, Model TK-101) and was set as the pre-dose value. As a sample, an indomethacin-albumin composition was suspended in 1% CMCNa so as to be about 5 mg / kg as indomethacin, and an indomethacin bulk powder was also suspended so as to be 5 mg / kg.
Oral administration was performed at a rate of 0.5 ml per 100 g of rat. As a control, 1% CMCNa was orally administered at a rate of 0.5 ml per 100 g of rat. Thirty minutes after the oral administration, 0.1 ml of a 1% λ-carrageenin physiological saline solution was subcutaneously administered to the right hind footpad of the rat, and immediately thereafter, 5 ml of water was orally administered. 1% λ-carrageenin administration 3
After time, footpad edema was measured. The carrageenan footpad edema suppression rate was determined from the following formulas (1) and (2).

Table 2 shows the results.

As is evident from Table 2, the indomethacin-albumin composition has almost the same effect as the bulk indomethacin powder and exhibits excellent anti-inflammatory action.

Next, an example in which light stability is improved by the present invention will be described.
The following experiment proves that the nifedipine-albumin composition produced by the above method has excellent stability to light.

Light stability test of nifedipine-albumin composition About 0.1 g of nifedipine-albumin composition containing about 0.01 g of nifedipine and about 0.1 g of nifedipine as a control
And about 0.1 g of commercially available nifedipine fine grains containing about 0.001 g of nifedipine were placed on a flat plate (50 × 50 cm) as shown in FIG.
Spread 2 x 2 cm squares at cm intervals. Cover the plate with a wooden box and turn on the attached fluorescent lamp (Toshiba 10w) at the center of the plate.
Left for hours. Room temperature at the start of the experiment is 12 ° C, relative humidity 30%
Upon completion, the room temperature was 19 ° C. and the relative temperature was 25%.

After the light irradiation, each sample was collected for each spot, the weight was precisely weighed accurately, 10 ml of methanol was added, and ultrasonic waves were irradiated for 30 minutes by an ultrasonic cleaner (high-frequency output 150 W, 28 kHz). The insoluble matter was removed by filtration, the remaining amount on the funnel was washed with methanol, and the filtrate and the washing solution were combined. Approximately 100 mg of each of the nifedipine-albumin composition, raw nifedipine powder and commercially available nifedipine fine granules stored in the same manner under light shielding were accurately weighed, and 19 ml of methanol was added. However, with respect to the bulk nifedipine powder, both the control solution and the sample solution were more precisely diluted to 10-fold volume. A fixed amount of each of the control solution and the sample solution was accurately injected into the high performance liquid chromatography to determine the nifedipine concentration. The quantification was calculated from a calibration curve obtained by simultaneously analyzing two different nifedipine standard solutions with known concentrations. The analysis conditions of the high performance liquid chromatography are as follows.

Column: Waters type, Novapack C18 Moving bed: 55% methanol Flow rate: 1 ml / min Detection wavelength: 350 nm The residual ratio was calculated by the following equation.

Table 3 shows the results.

As is evident from Table 3, the nifedipine-albumin composition suppressed the photodecomposition of the contained nifedipine, and the degree of stability was superior to that of commercially available nifedipine fine granules. Furthermore, this stability is improved by general formulation processing.

Further, one of the compositions of the present invention, fat-soluble vitamin dl-
The following experiments demonstrate that the α-tocopherol-ovalbumin composition has excellent absorption properties.

Absorption property test of tocopherol-ovalbumin composition dl-α-tocopherol alone and dl-α-tocopherol-ovalbumin composition (equivalent to 100 mg as dl-α-tocopherol) were filled into two No. 00 capsules, and the dogs were Oral administration. 3, 6, 9, 12, 24, 36 after oral administration
At the time, blood was collected from the limb vein and centrifuged to obtain serum. Serum 0.
1 ml of methanol and 6 ml of hexane were added to 5 ml, and the mixture was shaken for 10 minutes. After evaporating 5 ml of the hexane phase to dryness, 300 μl of hexane was added, and 25 μl thereof was used as a high performance liquid chromatography sample.

High-performance liquid chromatography conditions Column: Lichrosorb SI-60 (7 μm, 4 mm, id × 25c
m) Mobile phase: Hexane: isopropanol (99.5: 0.5) Flow rate: 1.0 ml / min Detection wavelength: Excitation wavelength 295 nm Fluorescence wavelength 330 nm As shown in FIG. 4, the result is dl-α-tocopherol-ovalbumin composition , An increase in serum concentration of dl-α-tocopherol was observed. The area under the serum concentration-time curve (AUC) was 2.8 times that of ovalbumin: dl-α-tocopherol: stearic acid (5: 1: 0.5) when administered alone, and ovalbumin: dl-α-tocopherol (5: In the case of 1), it increased 1.6 times. In particular, when stearic acid was added, the time to reach the maximum blood concentration was reduced from 12 hours when administered alone to 6 hours. These results indicate that the dl-α-tocopherol-ovalbumin composition is extremely excellent in absorption properties.

Thus, when the composition of the present invention is used as a medicament,
It can be manufactured in various forms suitable for oral administration.
For example, the compositions of the present invention can be formulated as a medicament containing a non-toxic pharmaceutically acceptable carrier material for use in such medicament. Such drugs may be in solid forms (eg, tablets, capsules, granules, powders, fine granules, dragees, lozenges, etc.) and semi-solid forms (eg, ointments, haptics, creams, suppositories) depending on their use. ) And liquid forms (eg, emulsions, suspensions, lotions, tinctures, sprays, syrups, etc.). Thus, non-toxic pharmaceutically acceptable carrier substances that can be used include, for example, starch, gelatin, glucose, lactose, fructose, maltose, magnesium carbonate, talc, magnesium stearate, methylcellulose, carboxymethylcellulose or the like. Salt, gum arabic, polyalkylene glycol, alkyl p-hydroxybenzoate, simple syrup, ethanol, propylene glycol, glycerin, petrolatum, carbox and the like. The drug can also include other therapeutically useful drugs, dispersants, antioxidants, preservatives, stabilizers, flavoring agents, binders, lubricants, and the like. The content of the composition according to the invention in the medicament depends on its form, but is generally in a concentration of 5 to 100% by weight for solid and semi-solid forms and 0.1 to 10% for liquid forms. It is desirable to contain the active compound in a concentration by weight.

 Hereinafter, the present invention will be described in detail with reference to examples.

Example 1 18 g of ovalbumin was added to 400 ml of distilled water, and the mixture was stirred at a high speed of about 10,000 rpm using a biolyzer (BM-4, manufactured by Nippon Seiki Seisakusho) while cooling to obtain a homogeneous phase. 2 g of nifedipine was dissolved in 6 ml of acetone and added to the homogeneous phase. High-speed stirring was continued for 1 hour while further cooling, and the organic solvent was distilled off from the obtained homogeneous suspension under reduced pressure, followed by freeze-drying to obtain a powder. This powder was passed through a 100-mesh sieve to obtain a nifedipine-albumin composition having a uniform particle size. The obtained nifedipine-albumin composition,
As shown in the scanning electron micrograph of FIG. 3, it had a plate-like crystal-like three-dimensional structure with a longest particle diameter of about 15 μm.

Examples 2 to 22 Example 1 was repeated except that the drugs and solvents shown in Table 4 were used and that the high-speed stirring conditions and drying conditions were appropriately changed according to the types and properties of the drugs and solvents used. In the same manner as in the above, powders of various drug-albumin compositions were obtained.

* Dl-α-tocopherol-ovalbumin composition (1:
Dl-α-tocopherol-ovalbumin-stearic acid composition using 1.8 g of stearic acid in addition to 5) (1: 5:
0.5) was also manufactured.

〔The invention's effect〕

Pharmaceutical composition using the natural albumin of the present invention as a carrier,
It has remarkable effects such as improving absorption of drugs that reduce side effects that are unfavorable to the human body, and having excellent photostability.

[Brief description of the drawings]

FIG. 1 is a graph showing the change over time in blood concentration after administration of a nalidixic acid-albumin composition. FIG. 2 shows a plate used for placing various samples in a photostability test of a nifedipine-albumin composition and the like. FIG. 3 is a scanning electron micrograph showing the crystal structure of the nifedipine-albumin composition. FIG. 4 is a graph showing the time course of blood concentration after administration of the dl-α-tocopherol-ovalbumin composition. It is a graph.

Claims (2)

(57) [Claims] 1. Nalidixic acid, indomethacin, nifedipine, tocophenol, aspirin, piroxicam, sulindac, phenylbutazone, cefaclor, cephalexin, tetracycline hydrochloride, ampicillin, dexamethasone, haloperidol, diltiazem hydrochloride, griseofulvin, vincristine sulfate, cimetidine, cimetidine, cimetidine Selected from glycyltin or tranilast, 1 to 200 parts of a protein-binding drug and 100 parts of natural albumin, which reduces undesirable side effects, improves absorption of the drug, and photostable. Pharmaceutical composition having excellent properties. 2. Nalidixic acid, indomethacin, nifedipine, tocophenol, aspirin, piroxicam, sulindac, phenylbutazone, cefaclor, cephalexin, tetracycline hydrochloride, ampicillin, dexamethasone, haloperidol, diltiazem hydrochloride, griseofulvin, vincristine sulfate, cimetidine, cimethidine The method is characterized in that 1 to 200 parts of a drug exhibiting a protein binding property selected from glycyltin or tranilast is added to 100 parts of natural albumin, stirred in a solvent at a rotation speed of 5,000 to 40,000 rpm, and then the solvent is removed. A method for producing a pharmaceutical composition which reduces unwanted side effects, improves absorption of pharmaceuticals, and has excellent photostability.