JP3649989B2 - Test method - Google Patents

Description

[0001]
[Industrial application fields]
The present invention relates to a test method for confirming elution of an active ingredient from a solid preparation such as a tablet in a fraction or extract preparation derived from animals and plants such as a biological preparation or a herbal preparation, and a solid characterized by the test method. The present invention relates to a method for producing a preparation and a pharmaceutical preparation whose quality is ensured by the test method.
[0002]
[Prior art]
The dissolution test method is based on the main component from internal solid preparations (tablets, powders, capsules, dry syrups, etc.) as explained in the General Test Methods section of the 13th revision of the Japanese Pharmacopoeia. This is a method for testing dissolution, and is intended to ensure the quality of internal solid preparations at a certain level and to prevent significant biological inequality.
[0003]
For drugs that are synthesized organically and have a clear active ingredient, the amount of active ingredient in the sampled dissolution test solution was analyzed using high-performance liquid chromatography (HPLC) or a specific reaction reagent. By quantitative analysis, the elution state of the active ingredient from the solid preparation into the aqueous solution can be quantitatively detected and confirmed.
[0004]
[Problems to be solved by the invention]
However, in the case of an extract preparation from animal or plant tissues such as biological preparations and herbal preparations, various active ingredients are included as active ingredients, and these various ingredients work comprehensively. Therefore, the dissolution test of such a drug is very difficult. Therefore, it has been desired to establish a quantitative and simple dissolution test method for an oral solid preparation containing an active ingredient unknown as an active ingredient such as vaccinia virus inoculated rabbit inflammation skin extract.
[0005]
[Means for Solving the Problems]
An object of the present invention is to provide a dissolution test method from a solid preparation such as a biological preparation or a herbal preparation that accurately reflects the dissolution of an active ingredient and can be easily measured, and a method for producing a solid preparation characterized by the test method And providing a pharmaceutical preparation whose quality is guaranteed by the test method.
[0006]
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a test method characterized by confirming elution of an active ingredient from a solid preparation of a fraction derived from animal or plant origin or an extracted preparation into an aqueous solution by measuring the ultraviolet absorbance or the ultraviolet absorbing substance, and the test method And a pharmaceutical preparation whose quality is ensured by the test method.
[0007]
More specifically, the elution of the active ingredient into the aqueous solution from the solid preparation containing vaccinia virus inoculated rabbit inflammatory skin extract as the active ingredient is confirmed by measuring the ultraviolet absorbance or the ultraviolet absorbing substance. A dissolution test method for solid preparations such as tablets, a method for producing a solid preparation characterized by the dissolution test method, and a pharmaceutical preparation whose quality is guaranteed by the dissolution test method.
[0008]
The living body maintains its homeostasis with the dual action of suppressing the excessive response and enhancing the function against the invasion from the outside world such as viruses and the progression of internal pathological conditions, and normalizes the body by adjusting the biological function Therefore, it is known to produce various biological function regulating substances. For example, various biological function regulating substances produced in inflamed tissues inoculated with vaccinia virus, production methods for extracting the substances from pathological tissues, and their pharmacological activities have been reported (Japanese Patent Publication No. 63-39572, Japanese Patent Publication No. Sho). No. 63-25600, Japanese Patent Publication No. 3-43279, Japanese Patent No. 2594422, etc.).
[0009]
As an actual medicine, there is a inflammatory skin extract preparation (trade name: neurotropin) inoculated with vaccinia virus. This preparation is, for example, a house inoculated with vaccinia virus as described on pages 1925 and 1926 of the collection of medical drugs Japan Pharmaceuticals (1998-99, 22nd edition, edited by Japan Pharmaceutical Information Center, published by Yakuho Jihosha). Drug containing non-protein active substance extracted and isolated from inflammatory skin tissue of vagina, low back pain, neck-shoulder arm syndrome, periarthritis, osteoarthritis, symptomatic neuralgia, skin disease (eczema, dermatitis It has been approved for pruritus associated with urticaria, allergic rhinitis, cold feeling of SMON aftereffects, pain, abnormal perception, etc. Injections and tablets for subcutaneous, intramuscular and intravenous injections and tablets It is marketed with the approval of manufacture. One of the objects of the present invention is specifically a tablet elution test for vaccinia virus inoculated rabbit inflammatory skin extract preparation, and a vaccinia virus inoculated rabbit inflammatory skin extract preparation characterized by the dissolution test method. It is a tablet of a vaccinia virus-inoculated rabbit inflammatory skin extract formulation whose quality is ensured by the tablet production method and the dissolution test method.
[0010]
Since the active ingredient of the inflamed tissue extract inoculated with the vaccinia virus is unknown, the amount of the active ingredient may be defined not by weight but by analgesic efficacy (titer). For example, a commercial preparation (neurotropin tablet) contains 4.0 neutropin units (NU) of rabbit skin inoculated vaccinia virus inoculated in one tablet. The analgesic efficacy test is carried out using a disease state model animal (detailed later) in a hyperalgesic state. Therefore, it is considered that the dissolution test from tablets is basically performed by quantitatively confirming the dissolution amount of the inflammatory skin extract inoculated with vaccinia virus using this analgesic efficacy test. However, unlike the quantitative analysis using ordinary HPLC or specific reaction reagents, the method for conducting the dissolution test using the experimental animals is as follows:
(1) It takes days and cost to create the disease model animal.
(2) It is complicated as a dissolution test method to be performed for each production lot.
(3) Efficacy tests using animals are generally not sensitive and require a large amount of test samples (tablets).
It is not necessarily a preferable method in terms of economy and simplicity as a dissolution test method that is repeatedly carried out on a daily basis.
[0011]
Therefore, as a result of various examinations of alternative test methods having a good correlation with the measurement value of the quantitative test based on the above titer, by measuring the ultraviolet absorbance or the ultraviolet absorber of the eluate, the operation is simple and It was clarified that elution of the active ingredient can be accurately reflected, and the present invention has been completed.
[0012]
Preferred embodiments of the present invention are shown below.
(1) A test method characterized by confirming elution of an active ingredient from a solid preparation of a fraction derived from animal or plant origin or an extracted preparation into an aqueous solution by measuring an ultraviolet absorbance or an ultraviolet absorbent.
(2) A test method characterized by confirming the elution of an active ingredient into an aqueous solution from a solid preparation containing a rabbit inflammatory skin inoculated with vaccinia virus as an active ingredient by measuring the ultraviolet absorbance or the ultraviolet absorbing substance. .
(3) The dissolution test method according to (1) or (2), wherein elution of an active ingredient from a solid preparation into an aqueous solution is confirmed by measuring ultraviolet absorbance.
(4) The dissolution test method according to the above (1) or (2), wherein elution of an active ingredient from a solid preparation into an aqueous solution is confirmed by measuring an ultraviolet absorption material.
(5) The elution test method according to the above (4), wherein the ultraviolet absorbing material is urocanic acid, uracil, hypoxanthine, xanthine and / or thymine.
(6) The dissolution test method according to any one of (1) to (5), wherein the solid preparation is a tablet.
[0013]
(7) In the method for producing a solid preparation of animal or plant origin fraction or extract preparation, the elution of the active ingredient from the solid preparation into an aqueous solution is confirmed by measuring the ultraviolet absorbance or the ultraviolet absorbing substance. A method for producing a solid preparation.
(8) In the method for producing a solid preparation containing vaccinia virus-inoculated rabbit inflammation skin extract as an active ingredient, elution of the active ingredient from the solid preparation into an aqueous solution is measured by measuring the ultraviolet absorbance or the ultraviolet-absorbing substance. A method for producing a solid preparation, characterized by confirming.
(9) In a method of producing a solid preparation containing inflammatory tissue of an animal inoculated with vaccinia virus as a raw material and producing a solid preparation containing the active ingredient, a necessary amount of the active ingredient is released from the solid preparation in the digestive tract In order to confirm that this is the case, a method for producing a solid preparation, comprising measuring ultraviolet absorbance or ultraviolet absorbing substance to confirm elution of an active ingredient from the solid preparation into an aqueous solution.
(10) The production method according to (9) above, wherein the inflammatory tissue of the animal is rabbit skin tissue.
(11) The method for producing a solid preparation according to any one of (7) to (10), wherein elution of the active ingredient from the solid preparation into the aqueous solution is confirmed by measuring ultraviolet absorbance.
(12) The method for producing a solid preparation according to any one of the above (7) to (10), wherein elution of an active ingredient from a solid preparation into an aqueous solution is confirmed by measuring an ultraviolet-absorbing substance.
(13) The elution test method according to the above (12), wherein the ultraviolet absorbing material is urocanic acid, uracil, hypoxanthine, xanthine and / or thymine.
(14) The production method according to any one of (7) to (13), wherein the solid preparation is a tablet.
[0014]
(15) A pharmaceutical preparation whose quality is ensured by confirming elution of an active ingredient from a solid preparation of a fraction derived from animal or plant origin or an extracted preparation into an aqueous solution by measuring an ultraviolet absorbance or an ultraviolet absorbing substance.
(16) The quality was ensured by confirming the elution of the active ingredient into the aqueous solution from the solid preparation containing the rabbit skin inoculated with vaccinia virus as an active ingredient by measuring the ultraviolet absorbance or the ultraviolet absorbing substance. Pharmaceutical formulation.
(17) In a method for extracting an active ingredient from inflammatory tissue of an animal inoculated with vaccinia virus as a raw material and producing a solid preparation containing the active ingredient, a necessary amount of the active ingredient is released from the solid preparation in the digestive tract In order to confirm this, a pharmaceutical preparation whose quality is ensured by measuring the ultraviolet absorbance or ultraviolet absorbing substance and confirming the elution of the active ingredient from the solid preparation into the aqueous solution.
(18) The pharmaceutical preparation according to any one of (15) to (17), wherein quality is ensured by measuring elution of the active ingredient from the solid preparation into the aqueous solution by measuring ultraviolet absorbance. .
(19) The pharmaceutical product according to any one of (15) to (17), wherein the quality is ensured by measuring elution of the active ingredient from the solid preparation into the aqueous solution by measuring the ultraviolet absorption material. Formulation.
(20) The pharmaceutical preparation according to the above (19), wherein the ultraviolet-absorbing substance is urocanic acid, uracil, hypoxanthine, xanthine and / or thymine.
(21) The pharmaceutical preparation according to any one of (15) to (20), wherein the solid preparation is a tablet.
[0015]
The tablet dissolution test can be carried out according to a conventional method, for example, a dissolution test method described in a general test method of the Japanese Pharmacopoeia. Vaccinia virus-inoculated rabbit inflammation skin extract commercially available is a film-coated tablet, so perform dissolution test using the second method (paddle method) of the Japanese Pharmacopoeia general test method and dissolution test method. However, an appropriate method may be selected according to the following guidelines depending on the type of solid preparation.
[0016]
The type of eluent is a solution that does not affect the measurement of UV absorbance or the determination of UV absorption materials, that is, a solution that does not absorb near the measurement wavelength (the solvent itself and added reagents such as buffering agents are near the measurement wavelength. Those having no absorption) are preferred. For example, test solutions described in the general test method and disintegration test method of the Japanese Pharmacopoeia can be used. The dissolution test method instructed by the Ministry of Health and Welfare is detailed in, for example, the guideline of Pharmaceutical Examination No. 487 dated December 22, 1997. The test conditions are 900 mL of test solution and the temperature of the test solution. Is to be carried out by the paddle method at 37 ° C., and for formulations containing neutral or basic drugs and coating preparations, the first solution (pH 1.2) described in the general test method and disintegration test method of the Japanese Pharmacopoeia ), {Circle around (2)} second solution (pH 6.8), {circle around (3)} McIlvaine buffer (pH 3.0 to 5.0) and {circle around (4)} water.
[0017]
In the present invention, the wavelength of the ultraviolet absorbance to be measured in order to confirm elution of a vaccinia virus-inoculated rabbit inflammation skin extract from a solid preparation such as a tablet into an aqueous solution can be appropriately set. Since the ultraviolet spectrum of the inflamed skin extract has a maximum absorption within the range of 255 to 275 nm, it is preferable to set the wavelength within the range of 245 to 285 nm. Then, since it can be measured, this is not particularly limited.
[0018]
Further, in the present invention, the ultraviolet-absorbing substance that can serve as an index for the elution test of the inflammatory skin extract inoculated with vaccinia virus in the same manner as the ultraviolet absorbance is exemplified by urocanic acid, uracil, hypoxanthine, and xanthine contained in the extract. , Thymine and the like can be mentioned as preferable substances. As these quantitative test methods, the commonly used HPLC (High Performance Liquid Chromatography) is easy to use. The measurement conditions in HPLC may be appropriately set according to a conventional method depending on the type of the substance to be measured and the eluate. As is common knowledge in this field, use standard reagents for the measurement target substance in advance to set conditions for appropriate separation of each target substance, and confirm the retention time of the target substance under those conditions. The concentration of each substance in the test solution can be quantified using this as an index. Depending on the measurement conditions, urocanic acid and uracil may not be completely separated depending on the pH of the test solution to be charged (eluate), and peaks may overlap. In that case, the sum of both substances may be obtained as the measured value. . In the measurement of the UV-absorbing substance, HPLC is generally used, but naturally, other measurement methods such as a quantitative test method using a color reaction specific to the measurement target substance are used. You may implement this invention. Moreover, it cannot be overemphasized that the method of this invention is applicable also to the dissolution test of internal solid preparations other than a tablet.
The following examples illustrate the invention in more detail.
[0019]
【Example】
(1) Dissolution Test The dissolution test was performed in accordance with the Japanese Pharmacopoeia Manual, 13th Amendment General Test Method and Dissolution Test Method Method 2 (Paddle Method). That is, a tablet sample was placed in a test solution (water 900 mL, solution temperature 37 ± 0.5 ° C.), and a dissolution test was performed with a paddle rotation speed of 50 rpm. The collected test solution was filtered under pressure with a disk filter having a pore diameter of 0.45 μm to obtain a sample solution for ultraviolet absorption measurement. A sample solution for analgesic efficacy test was similarly subjected to a dissolution test and concentrated. That is, the collected dissolution test solution was pressure filtered through a disk filter having a pore size of 0.45 μm in the same manner, except for the first filtrate (100 mL), and the next filtrate (approximately 700 mL) was taken and concentrated to dryness under reduced pressure at 45 ° C. or lower. However, it was subjected to an analgesic efficacy test as exactly 5 mL.
[0020]
(2) UV absorption measurement As a tablet sample, the elution test of (1) was performed using 4 tablets containing the above vaccinia virus-inoculated rabbit inflammation skin extract 4.0 neurotropin unit, and the absorbance of the sample solution at a wavelength of 270 nm was measured. Measured over time.
[0021]
(3) Analgesic efficacy test In the analgesic efficacy test, a pathological model animal exhibiting a chronic hyperalgesic state such as a SART stress mouse was used. SART (Specific Alternation of Rhythm in Temperature) stress, that is, repeated cold stress load, was performed according to the method of Kita et al. (Nichi Pharmacology, Vol. 71, pp. 195-210, 1975). Using a thermostat for breeding, change the breeding environment temperature of ddY male mice from 10:00 am to 5:00 pm alternately between 4 ℃ and 24 ℃ every hour, then from 5:00 pm to 10:00 am the next morning Was maintained at 4 ° C., water and feed were freely fed, reared for 4 days, and subjected to repeated cold stress before being subjected to the experiment. The pain threshold was measured by the modified Randall-Selitto method (tail pressure method: Nihon Pharmacology, 72, 573-584, 1976) before and 30 minutes after administration of the test drug. That is, using a Randall-Selitto type analgesic effect measuring device with a pressure bar modified for the mouse tail, pressure stimulation was applied to the site 1.5 cm from the mouse ridge to the tip side at a rate of 16 g / sec. The analgesic coefficient was calculated by dividing the “pressurized weight (g) after administration of test drug” by the “pressurized weight (g) before administration of test drug”. Therefore, the analgesic coefficient is 1.0 when the test drug has no effect, and the analgesic coefficient increases as 1.1, 1.2, and 1.3 as the effect increases. Elution test of (1) above was conducted using 4 tablets containing 16.0 neurotropin units of rabbit inflammatory skin extract 1 immunized with vaccinia virus, and 20 mL per kg body weight of the mouse was used as the test drug using the sample solution prepared by concentration. And the analgesic efficacy was measured.
[0022]
FIG. 1 shows an example of the results of measuring the ultraviolet absorbance and analgesic coefficient of the eluate over time for the ultraviolet absorption measurement (2) and (3) analgesic efficacy test.
[0023]
(4) Ultraviolet Absorbing Substance Measurement Figure 1 shows the results of HPLC using the octadecylsilylated silica gel column (NUCLEOSIL 5C18, Chemco, 4.0 mm ID × 250 mm, 5 μm) for the sample solution used in the ultraviolet absorption measurement. It was shown in 2. The elution behavior of UV absorbing substances such as urocanic acid, uracil, hypoxanthine, xanthine and thymine showed good agreement with the change in the absorbance measured value at 270 nm of the eluate. In addition, the first solution (pH 1.2) and the second solution (pH 6.) described in the general test method and disintegration test method of the Japanese Pharmacopoeia, which is instructed to conduct dissolution tests on non-water elution solutions, ie, coating preparations. In the case of using 8) and McIlvaine buffer (pH 3.0 to 5.0), it was confirmed that the elution concentration of each UV-absorbing substance and the measured absorbance were similarly correlated.
[0024]
FIG. 2 shows an example of the results obtained by measuring the concentration of the ultraviolet absorbing substance and the ultraviolet absorbance in the eluate over time for the measurement of the ultraviolet absorbing substance in (4) above.
[0025]
【The invention's effect】
As is clear from the results shown in FIG. 1, in the elution of vaccinia virus inoculated rabbit inflammatory skin extract from tablets into aqueous solution, the UV absorbance of the eluate and the increase in analgesic coefficient over time show a good correlation. It was. In the case of the tablet used in the above test, both the ultraviolet absorbance and the analgesic coefficient reach a plateau after 20 minutes. Therefore, the tablet containing the vaccinia virus-inoculated rabbit inflammatory skin extract is about 20 minutes in water. It was confirmed that elution of the active ingredient was completed. In addition, as shown in FIG. 2, there is a good correlation between the absorbance in the eluate and the concentration in the eluate of UV-absorbing substances such as urocanic acid, uracil, hypoxanthine, xanthine and thymine. It became clear that the elution of the rabbit skin extract from the tablet into the aqueous solution can be confirmed not only by measuring the absorbance but also by using each ultraviolet absorption substance as an index.
[0026]
If the amount of active ingredient in rabbit skin inoculated with vaccinia virus is defined by the titer (analgesic efficacy, etc.), animals must be used to measure the amount of active ingredient in the eluate, and Requires a complicated operation such as concentration to obtain a sample that can be administered to animals. On the other hand, the operation of the test method of the present invention for measuring ultraviolet light absorption and ultraviolet light absorbing substances such as urocanic acid, uracil, hypoxanthine, xanthine, and thymine is carried out by separating a small amount of the eluate and using a spectrophotometer or It is extremely simple and can be carried out in a short time only by using HPLC. Thus, the present invention provides a dissolution test method that accurately reflects the dissolution of an active ingredient and can be easily measured with respect to a solid preparation for internal use such as a vaccinia virus inoculated rabbit inflammation skin extract tablet. The method of the present invention is novel and highly useful as a substitute for a quantitative test based on a titer that requires an animal and is complicated to operate.
[Brief description of the drawings]
FIG. 1 shows the results of measuring the ultraviolet absorbance and analgesic coefficient of an eluate over time in an elution test of a tablet containing vaccinia virus-inoculated rabbit inflammation skin extract as an active ingredient. .
[Fig. 2] Fig. 2 shows the results of measurement of the concentration of the UV-absorbing substance in the eluate and the UV-absorbance over time in the dissolution test of a tablet containing a rabbit skin inoculated with vaccinia virus as an active ingredient. Is shown.

Claims (4)

When elution of an active ingredient from a solid preparation containing a vaccinia virus-inoculated rabbit inflammatory skin extract as an active ingredient into an aqueous solution was conducted using the Japanese Pharmacopoeia paddle method, the elution solution at an elution time of 0 to 10 minutes A test method characterized by measuring and confirming urocanic acid, uracil, hypoxanthine, xanthine or thymine . Dissolution test method of claim 1, wherein the solid preparation is a tablet. In the method for producing a solid preparation containing an active ingredient from the inflamed skin tissue of a rabbit inoculated with vaccinia virus and producing a solid preparation containing the active ingredient, a necessary amount of the active ingredient is released from the solid preparation in the digestive tract In order to confirm this, when a dissolution test was performed by the Japanese Pharmacopoeia paddle method, urocanic acid, uracil, hypoxanthine, xanthine or thymine in the eluate at an elution time of 0 to 10 minutes was measured and the aqueous solution was removed from the solid preparation. A method for producing a solid preparation, which comprises confirming elution of an active ingredient in The method for producing a solid preparation according to claim 3 , wherein the solid preparation is a tablet.

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