RU2115415C1 - Solid dosaged form for oral using showing antiviral activity (variants) - Google Patents

Abstract

FIELD: pharmaceutical industry. SUBSTANCE: dosaged form is made as a tablet or granule. Form has 10-carboxymethyl-9-acridanone ammonium or N-methylglucaminic salt as an active component and special additions. Dosaged form is covered by protective acid-resistant coating made of shellac or acetylphthalyl cellulose. Tablets and granules are used for prophylaxis and treatment of viral diseases. EFFECT: improved quality of form, enhanced antiviral activity. 2 cl, 11 tbl

Description

 The invention relates to the pharmaceutical industry and is intended for the prevention and treatment of diseases of a viral nature, such as hepatitis, encephalitis, chlamydia, herpes, as well as HIV-related immunodeficiency states.

 In connection with the wide spread of viral infections, the need for long courses of treatment, as well as the acute need for the prevention of viral infections, it is advisable to create an oral dosage form.

 Oral dosage forms based on substituted acridanones, namely 10-carboxymethyl-2-chloro-9-acridanone, are known for the treatment of viral diseases (US Pat. No. 3,681360, C 07 D 37/20, application. 04.04.71, publ. 08/01/72).

в количестве 42-79%;
традиционные связующие добавки: крахмал, масло какао, стеарат кальция или магния, лактоза, желатин, дизамещенный фосфат кальция, тальк в количестве суммарно 58-21%.The prototype has the following composition:
as the active substance is 10-carboxymethyl-2-chloro-9-acridanone of structural formula (I)

in the amount of 42-79%;
traditional binders: starch, cocoa butter, calcium or magnesium stearate, lactose, gelatin, disubstituted calcium phosphate, talc in the amount of 58-21% in total.

 The active substance is mixed with inactive binding agents and the resulting granulate is compressed in the form of tablets containing 250 or 500 mg of the active substance.

 The oral medicinal product described above practically does not exhibit biological activity due to low bioavailability.

 The applicant has experimentally found that when a tablet enters the stomach, the active substance is released in the form of acridonoacetic acid, which is not absorbed in the stomach and excreted unchanged with the contents of the intestine, so the drug mentioned above does not have bioavailability.

 To create a bioavailable oral medicinal product with high therapeutic efficacy, the active substance must be in a tablet in the form of a pharmacologically acceptable salt of acridonoacetic acid, and such a tablet should disintegrate, bypassing the stomach, in the intestine.

 When a tablet containing the active substance in the form of a pharmacologically acceptable salt is introduced into the stomach, and if it disintegrates in the stomach, where the pH of the gastric juice is 1-2, the active substance will be released in the form of insoluble acridonoacetic acid, which, as described above, does not It is bioavailable and does not have a therapeutic antiviral effect.

 In the event of a tablet disintegration in the intestine, where, as you know, the alkaline medium, the active substance is released in the form of a salt that has bioavailability, is well absorbed and has a high biological effect.

 The introduction into the intestine of the active substance in the form of a pharmacologically acceptable salt leads to the rapid absorption of the substance and the manifestation of a high therapeutic effect.

 With the introduction of the active substance in the form of acridonoacetic acid into the intestine, and not its pharmacological salt, low (8-10 times less) therapeutic efficacy of the drug was experimentally established compared to the claimed drug. This is due to the insufficiently high intestinal pH for the formation of salt from acridonoacetic acid and the limited residence time of the tablet in the body, during which the drug does not have time to absorb and have a high therapeutic effect.

 The objective of the invention is the creation of an oral drug with high bioavailability and exhibiting high antiviral efficacy.

где
R = NH $\genfrac{}{}{0ex}{}{\text{+}}{\text{4}}$ , N⁺ - метилглюкамин,
а на ядро таблетки дополнительно нанесено защитное кислотоустойчивое покрытие, выбранное из группы: шеллак, ацетилфтолилцеллюлоза при следующих соотношениях компонентов, в мас. ч.:
Указанная соль 10-карбоксиметил-9-акриданона - 269-900
Целевые добавки - 50-225
Шеллак или ацетилфталилцеллюлоза - 35-50
Второй вариант лекарственного средства представляет собой гранулы, предназначенные для детей, содержащие активное вещество и целевые добавки, нанесенные на инертную частицу, при этом в качестве активного вещества в него включена соль 10-карбоксиметил-9-акридона структурной формулы

где
R = NH $\genfrac{}{}{0ex}{}{\text{+}}{\text{4}}$ , N⁺ - метилглюкамин,
а на гранулу дополнительно нанесено защитное кислотоустойчивое покрытие, выбранное из группы: шеллак, ацетилфтолилцеллюлоза при следующем соотношении компонентов, в мас. ч.:
Указанная соль 10-карбоксиметил-9-акриданона - 187-195
Инертная частица - 380-400
Целевые добавки - 10-15
Шеллак или ацетилфталилцеллюлоза - 40- 45
Обе заявляемые лекарственные формы направлены на решение одной и той же задачи - повышения биодоступности и увеличения противовирусной активности препарата и основаны на едином изобретательском замысле, заключающемся в том, что в композиции в качестве активного вещества используется 10-карбоксиметил-9-акриданон в виде фармакологически приемлемой соли, обеспечивающей быстрое проникновение активного вещества в органы и ткани организма. На таблетку или гранулу нанесено кислотоустойчивое защитное покрытие, способствующее высвобождению активного вещества в кишечнике.The problem is solved by the fact that two variants of the oral drug are claimed. The first variant of the medicinal product is a tabletted medicinal product for adults containing the active substance and target additives, while the 10-carboxymethyl-9-acridanone salt of the structural formula is included as an active substance in it

Where
R = NH $\genfrac{}{}{0ex}{}{\text{+}}{\text{4}}$ , N ⁺ - methylglucamine,
and on the core of the tablet is additionally applied a protective acid-resistant coating selected from the group: shellac, cellulose acetate in the following ratios of components, in wt. hours:
The specified salt of 10-carboxymethyl-9-acridanone - 269-900
Target Supplements - 50-225
Shellac or Acetylphthalyl Cellulose - 35-50
The second version of the drug is granules intended for children, containing the active substance and the target additives, applied to an inert particle, while the active substance contains 10-carboxymethyl-9-acridone salt of the structural formula

Where
R = NH $\genfrac{}{}{0ex}{}{\text{+}}{\text{4}}$ , N ⁺ - methylglucamine,
and the granule is additionally coated with a protective acid-resistant coating selected from the group: shellac, cellulose acetate in the following ratio of components, in wt. hours:
The specified salt of 10-carboxymethyl-9-acridanone - 187-195
Inert particle - 380-400
Targeted Supplements - 10-15
Shellac or acetylphthalyl cellulose - 40-45
Both of the claimed dosage forms are aimed at solving the same problem - increasing the bioavailability and increasing the antiviral activity of the drug and are based on a single inventive concept, namely that 10-carboxymethyl-9-acridanone is used in the composition as a pharmacologically acceptable salt, providing rapid penetration of the active substance into the organs and tissues of the body. An acid-resistant protective coating is applied to the tablet or granule, which promotes the release of the active substance in the intestine.

обладающим широким спектром биологической активности: интерфероногенной, противовирусной, в том числе антиВИЧ.The solution to this problem is illustrated by experimental data on the structure, method of preparation and biological activity of the active substance of the claimed dosage forms. The active substance of the claimed dosage forms relates to salts of derivatives of acridanones, for example, N-methylglucamine salt of acridonoacetic acid, of the General formula

having a wide range of biological activity: interferonogenic, antiviral, including anti-HIV.

Example 1. 99.6 g of N-methyl-N- (α, D-glucopyranosylamine) / N-methylglucamine / are dissolved in 200 ml of distilled water and 125 g of finely ground 2- (acridon-9-one-10) are added in portions with stirring. -yl) acetic acid, stirring is continued at room temperature until completely dissolved and 1000 ml of ethyl alcohol are added. The precipitate formed is filtered off, washed with a filter of 100 ml of ethanol and dried at 60 ^° C. for 1 hour. 224 g (100%) of N-methyl-N- (α, D-glucopyranosyl) -ammonium-2 are obtained in the form of yellow crystals - (acridon-9-one-10-yl) acetate / salt of N-methylglucamine and acridonoacetic acid /, which after recrystallization from lower alcohols has a melting point of 129-131 ^o C.

Elemental analysis of C ₂₂ H ₂₆ N ₂ O ₈ :
Calculated,%: C 58.74, H 5.83, N 6.73
Found,%: C 58.52, H 5.64, N 6.81.

 Physico-chemical properties of the active substance of the claimed dosage forms are presented in table. one.

 Mutagenic, terratogenic, embryotoxic and allergenic effects of N-methyl-N- (α, D-glucopyranosyl) -ammonium-2- (acridon-9-one-10-yl) acetate does not have.

 The study of the biomedical properties of N-methyl-N- (α, D-glucopyranosyl) -ammonium-2- (acridon-9-one-10-yl) acetate was carried out in an animal experiment, and a wide spectrum of biological activity of the claimed substance was revealed combined with its low toxicity, good tolerance and high therapeutic efficacy index.

 Experience A. Acute toxicity of the active substance was studied on outbred white mice of the crystalline line weighing 18-20 g.

The drug was administered intravenously and intramuscularly in various doses from 2000 mg / kg and below with a double step. The observation period is 14 days. The lethal dose (LD ₅₀ ) was calculated according to the Kerber method. Data on the study of acute toxicity are presented in table. 2.

 It is known that the basis of the antiviral activity of any chemical compound can be the development by the body of a universal antiviral protein - interferon. The applicant has established that the claimed compound also possesses this property.

 Experience B. Interferonogenic activity.

 The interferon-inducing activity of the active substance was studied in mice and in cell culture of animal and human origin.

10-12 g CBA mice were injected once subcutaneously with the active substance at a dose of 200 mg / kg and tiloron at a dose of 50 mg / kg. At certain time intervals, blood titers of interferon were determined by the standard method on homologous L-929 cells grown in 96-well plates in a CO ₂ incubator. Encephalomyocarditis virus served as a test virus. Comparative data on the interferon-inducing activity of the claimed compounds and tilorone are given in table. 3.

 As can be seen from the data presented, the claimed compound induces the production of early (2-8 h) interferon in high titers. The peak production of interferon in this case exceeds the level of interferon in the serum of mice in response to the introduction of tilorone 1000 times. High interferon-inducing activity of the claimed compounds is also confirmed in cell culture.

Spleen lymphocytes and peripheral blood lymphocytes were isolated from CBA mice and induced by the claimed compound and tilorone. Landing lymphocyte concentration was 5 • 10 ⁶ cells / ml. We studied the dynamics of the accumulation of interferon in the culture fluid of lymphocytes grown on plastic panels in the presence of CO ₂ .

 Interferon titers in splenocytes and blood cells after administration of the claimed compounds and tilorone are presented in table. 4.

 The results presented in table. 4 demonstrate that the claimed compound induces significantly higher levels of interferon in a mixed culture of spleen and peripheral blood lymphocytes than tiloron.

Confirmed interferonogenic activity of the claimed compounds when acting in human cell culture. To this end, human peripheral blood lymphocytes were obtained by dividing the leukocyte mass of donated blood. Landing lymphocyte concentration was 2 • 10 ⁶ cells / ml. Cells were grown in 24-well plastic panels. Lymphocyte induction was carried out by the claimed compound and tilorone at a concentration of 600 and 200 mg / ml, respectively. Interferon in the culture fluid was determined by titration on diploid human M-19 cells. The test virus was the vesicular stomatitis virus. Interferon samples were neutralized with standard antisera to α-9 and β-9 of human interferon, and international standards of human interferon served as a control.

 The induction data of interferon in human lymphocytes are given in table. 5.

 As follows from the data table. 5, the claimed compound induces the production of interferon in lymphocytic cultures of human peripheral blood 90 times higher than tiloron.

 Experience B. Antiviral activity of the claimed compounds.

 1B. The high activity of N-methyl-N- (α, D-glucopyranosyl) ammonium-2- (acridon-9-one-10-yl) acetate was established during experimental infection of mice infected with tick-borne encephalitis virus (TBE).

 Currently, only an inactivated vaccine is used to combat this infection, which has very low efficiency and requires annual vaccinations due to a rapid decrease in resistance to this infection. Practical medicine has no other means of protection against CE.

To establish the effectiveness of the claimed compounds in relation to TBE, the TBE virus strain ABSETTAR was used. The virus titer with intracerebral infection was 7.0-8.0 lg LD ₅₀ , with subcutaneous infection - 5.0 lg LD ₅₀ . The experiment was carried out on nonlinear white mice weighing 12-14 g. The animals were infected by subcutaneous injection of the TBE virus. The inventive compound was dissolved in saline and was administered subcutaneously to the experimental group of mice. The control group of mice was injected subcutaneously with saline. Observation of experimental animals was carried out for 21 days. The criteria for evaluating the effectiveness of the claimed compounds were the survival and average life expectancy of animals of the experimental group compared with the control. The results of the experiment are presented in table. 6.

 The data presented indicate a pronounced protective effect of the drug. In addition, specially conducted studies showed that on days 4 and 7 after infection, the TBE virus was not detected in the brains of mice receiving the claimed compound, while the virus was reproduced in high titers in the control group of animals.

 2B. The activity of the claimed compounds in relation to the human immunodeficiency virus (HIV). Currently, the only known drug approved for use in medical practice as a tool for the treatment of acquired immunodeficiency syndrome (AIDS). This is the nucleoside nature of azidothymidine (retrovir, zidovudine), the significant disadvantages of which are high toxicity and the rapid formation of resistance to it of virus strains. In addition, the drug does not provide survival, but only prolongs the life expectancy of AIDS patients.

To assess the effectiveness of the claimed compounds for HIV reproduction used a culture of monocyte cells of the I-937 line. Cells were grown in test tubes on RPMI-1640 medium with 20% fetal calf serum at a final concentration of 0.5-0.7 • 10 ⁶ cells / ml. Before the preparations were added to the support medium, the cell suspension was infected with a culture fluid concentrate, feeding the HIV-producing cells obtained from the NTA-41 line. Then the cells were incubated at 37 ^o C for 3-4 days, followed by a change in nutrient medium. On days 5 and 7 after infection, the presence of a virus-specific antigen in the indirect immunofluorescence reaction using HIV immunoglobulins was determined.

 The results of evaluating the inhibitory effect of the claimed compound on the expression of HIV antigens in HIV-inducing monocyte cells of the I-937 line, in comparison with azidothymidine, are presented in table. 7.

 Thus, the claimed compound in concentrations almost 1000 times lower than azidothymidine, has an equal effect on the expression of HIV antigens in cell culture of monocytes.

Ammonium (NH) has similar properties. $\genfrac{}{}{0ex}{}{\text{+}}{\text{4}}$ ) acridonoacetic acid salt.

 The therapeutic efficacy of the drug was studied on tick-borne encephalitis models - a viral disease in white mice. Three groups of animals were involved in the experiment.

 The first group - infected white mice were administered orally the active substance in the form of 10-carboxymethyl-9-acridanone, described in the prototype, and in the form of N-methylglucamine salt shown in the claims.

 In the second group of infected animals, the same compounds and in the same doses were injected directly into the intestine, bypassing the stomach.

 The third group of infected mice did not receive treatment (placebo).

 As a result of the experiment, the survival rate of infected \\ mice and the protective effect of the drug were taken into account.

 As follows from the table. 8, with the introduction of the drug, intradermally in the form of salt, the effectiveness of treatment increases by 8-10 times compared with the prototype, which indicates a high bioavailability of the drug. Due to the fact that the claimed drug is oral, it is necessary to apply a protective acid-resistant coating, which ensures drug delivery and its release in the intestine.

 Medical and biological tests have established that therapeutic therapeutic doses of 10-carboxymethyl-9-acridanone salts are 269-900 mg per person per day with a single application. In accordance with this dose, the applicant has developed the optimal composition of the drug in the form of tablets and granules.

 Below are the experimental data on the selection of the optimal composition of the core tablets, table. nine.

 The ratio of the active substance to the binder additives was established experimentally. The criteria for assessing the selected ratios were bioavailability indicators and qualitative technological characteristics of the tablet core: compressibility, strength, hygroscopicity.

 As can be seen from the table. 9, the optimal options for the composition of the tablet core are options I and IV, providing maximum bioavailability of the active substance and high quality technological characteristics of the tablet core.

 Here are ways to get the tablet core.

 Example 2. In a glass dissolve 550 wt.h. ammonium salt of 10-carboxymethyl-9-acridanone (active substance) in 100 parts by weight distilled water, 75 parts by weight lactose, 2 parts by weight Tween-80 and 13 parts by weight calcium stearate (binders). The resulting paste is dried at room temperature for 18 hours in a layer with a thickness of not more than 0.5 cm. The dried granulite is triturated, sieved through a sieve and fed to a tablet machine mixer, where tablets weighing 640 mg are pressed. Each tablet contains 500 mg of active substance.

 Example 3. In the glass make 45 wt.h. distilled water, 445 parts by weight the active substance in the form of N-methylglucamine salt of 10-carboxymethyl-9-acridanone, 13 parts by weight tween-80 (binder). The resulting paste is thoroughly mixed and dried, as in example 2. In the mixer of the tablet machine load the dried and sifted paste and 12 wt.h. talc, after which tablets weighing 500 mg are pressed. Each tablet contains 250 mg of active substance.

 The tablets thus obtained are a semi-finished product (core) and are coated on top with an acid-resistant coating that protects the tablets from disintegration in the gastric juice for at least one hour (passage through the stomach) and rapid disintegration in the intestine for 5-10 minutes. These properties of the dosage form depend on the thickness of the layer of acid-resistant coating, which we established experimentally and is regulated by the amount of the corresponding ingredient supplied to the apparatus.

 In the table. 10 provides the rationale for choosing the amount of protective acid-resistant coating applied to the core of the tablet.

The selection criteria were two indicators:
preservation time of the tablet in the gastric juice;
tablet disintegration time in an alkaline environment (intestines).

 As follows from the table. 10, the preservation of tablets in gastric juice for 60-73 minutes and their rapid disintegration in alkaline media simulating the intestinal environment is ensured in options III and IV, when the amount of protective acid-resistant coating is 35-50 parts by weight, which is 5- 7% by weight of tablet cores.

 With an increase in the amount of protective coating (option I), a long disintegration of the tablet in the intestine takes place, which leads to a decrease in the effectiveness of the drug.

 With a decrease in the amount of protective coating (option II), the tablet disintegrates in the stomach and, as shown above, the drug completely loses its effectiveness.

 Here is an example of applying a protective acid-resistant coating on the tablet core: 350 wt. including cores of tablets obtained by the method described in examples 2 and 3, are placed in a fluidized bed installation, where 20 wt. including AFC or shellac in the form of a 1% solution in a suitable solvent (e.g. alcohol, chloroform, acetone). In this case, the surface of the tablet cores is uniformly coated with a protective acid-resistant shell. After supplying the entire amount of AFC, air is supplied to remove the solvent and fix the protective shell on the surface of the tablet core for at least 1.5-2 hours.

 Here is an example embodiment of the invention based on various compositions.

I composition
The active substance in the form of the ammonium salt of 10-carboxymethyl-9-acridanone - 269 mg
Targeted Supplements - 231 mg
Shellac or Acetylphthalyl Cellulose - 35 mg
Total: - 535 mg
II composition
The active substance in the form of N-methylglucamine salt of 10-carboxymethyl-9-acridanone - 445 mg
Targeted Supplements - 55 mg
Shellac or Acetylphthalyl Cellulose - 35 mg
Total: - 535 mg
III composition
The active substance in the form of the ammonium salt of 10-carboxymethyl-9-acridanone - 538 mg
Targeted Supplements - 102 mg
Acetylphthalyl cellulose - 45 mg
Total: - 685 mg
IV composition
The active substance in the form of N-methylglucamine salt of 10-carboxymethyl-9-acridanone - 890 mg
Targeted Supplements - 55 mg
Acetylphthalyl cellulose - 47 mg
Total: - 992 mg
Considering the possibility of using 10-carboxymethyl-9-acridanone salt as an antiviral agent in children and the impossibility of crushing tablets coated with a protective acid-resistant coating, a similar oral dosage form in the form of granules for pediatrics was developed.

 As mentioned above, an effective therapeutic dose of 10-carboxymethyl-9-acridanone salt of 269-900 mg per person per day was experimentally established, which amounts to 3.8 - 12.8 mg per 1 kg of body weight. For ease of dosage of the number of granules at one time for children, the applicants have chosen the content of the active substance in one granule of 6-7 mg, corresponding to the daily requirement per 1 kg of body weight of the child.

Upon receipt of the granules retained all the basic principles of creating an effective oral drug, namely:
to ensure maximum bioavailability, 10-carboxymethyl-9-acridanone is administered as a pharmacologically acceptable salt;
each granule is coated with a protective acid-resistant membrane, which contributes to the release of the active substance in the intestine.

 As a basis for the granules, standardly produced inert spherical sugar particles calibrated on a sieve with a pore diameter of 2-3 mm were used.

 The optimum ratios of the components to obtain granules have been experimentally established. The main criteria were: the specific content of the active substance in one granule, which should be 6-7 mg, technological indicators of the quality of the granulate (moisture, adhesion), the shelf life of the granules in the gastric juice. The experimental data are given in table. eleven.

 As follows from the table. 11, the optimal options for producing granules are the ratio of the components shown in option III and IV. The amount of protective acid-resistant coating is 40-45 wt. hours, which corresponds to 5-7% by weight of the granules, the same as for the above tablets. Exceeding this content of a protective acid-resistant coating in the granule leads to an increase in the disintegration time of the granule in the intestine (options II, IV, V of table 11), which negatively affects the effectiveness of the drug.

 We give an example of the preparation of granules of an oral drug for pediatrics.

 390 parts by weight of inert spherical sugar particles calibrated on a sieve with a pore diameter of 2-3 mm are placed in a fluidized bed apparatus into which an aqueous solution consisting of a mixture of 10 parts by weight is sprayed. methyl cellulose (binders) and 180 parts by weight of active substance. After spraying the specified solution, the layer of the obtained granules is dried for 1.5-2 hours, then the application of the solution continues with 40 parts by weight AFC or shellac in the form of a 5% solution in acetone or other easily volatile solution (chloroform, alcohol), after which air is blown for 1.5-2 hours to fix the protective layer and completely remove the solvent.

 Thus, the claimed oral medicines are highly effective drugs for the prevention and treatment of a wide range of diseases of viral etiology in adults and children.

Claims (1)

1. A solid oral dosage form having antiviral activity, containing as active substance a pharmaceutically acceptable salt of 10-carboxymethyl-9-acridanone and target additives, characterized in that it is made in the form of a tablet containing a pharmaceutically acceptable salt of 10-carboxymethyl 9-acridanone of the general formula

where R is NH $\genfrac{}{}{0ex}{}{\text{+}}{\text{4}}$ , N ⁺ -methylglucamine,
with a protective acid-resistant coating of shellac or cellulose acetate, the components are taken in the following ratio, wt. hours:
The specified salt of 10-carboxymethyl-9-acridanone - 269 - 900
Target additives - 50 - 225
Shellac or cellulose acetate-35-50
2. A solid oral dosage form having antiviral activity, containing as an active substance a pharmaceutically acceptable salt of 10-carboxymethyl-9-acridanone and target additives, characterized in that it is made in the form of granules with a protective acid-resistant coating of shellac or cellulose acetate, containing a pharmaceutically acceptable salt of 10-carboxymethyl-9-acridanone of the general formula

where R is NH $\genfrac{}{}{0ex}{}{\text{+}}{\text{4}}$ N ⁺ methylglucamine,
applied together with the target additives to an inert particle, while the components are taken in the following ratio, parts by weight:
The specified salt of 10-carboxymethyl-9-acridanone - 187 - 195
Inert particle - 380 - 400
Target Supplements - 10 - 15
Shellac or acetylphthalyl cellulose - 40 - 45sh

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