WO2017084612A1 - Interferon vaginal effervescent capsules and preparation method therefor - Google Patents

Abstract

Interferon vaginal effervescent capsules and a preparation method therefor. The interferon vaginal effervescent capsules consist of interferon freeze-dried powder, effervescent powder, and pharmaceutically acceptable accessories, the mass ratio of the interferon freeze-dried powder to the effervescent powder being 1:1.0~1.5, and preferably, 1:1.2. The vaginal effervescent capsules continuously release interferon for a long time at a low dosage, vaginal continuous low-dosage drug administration can be realized, local symptoms can be better removed, the treatment time can be effectively prolonged, and adverse effects of long-term medication are reduced; and meanwhile, during a continuous six-month acceleration test under the temperature of 40°C±2°C and a two-year long-term test under the temperature of 25°C±2°C of the vaginal effervescent capsules, both the valence (bioactivity) and the appearance thereof are not obviously changed, and good stability is expressed.

Description

Interferon vaginal effervescent capsule and preparation method thereof Technical field

The invention relates to the field of medicines, in particular to an interferon vaginal effervescent capsule and a preparation method thereof.

Background technique

Interferon has anti-pathogenic, anti-cell proliferation and immune regulation effects, so since its discovery, it has become an effective drug widely used in the treatment of viral diseases and certain tumors, such as treatment of viral hepatitis, herpes zoster , anti-poliovirus, coxsackie virus, flu virus, etc.

Chronic cervicitis and cervical erosion are common gynecological diseases and frequently-occurring diseases. About half of married women suffer from this disease. The cause of the disease is mostly caused by HPV-16 and HPV-18 infection, and it is a high risk factor for inducing cervical cancer. Since interferon is a biologically active substance with characteristics of being sensitive to light and heat and unstable in a liquid state, how to prepare it into a stable, uniform, efficient and convenient vaginal drug for treating gynecological viral infection becomes The goal of development.

"Research on the preparation technology of recombinant human interferon α-2b vaginal effervescent tablets" [Xu Xiao, Guan Jinghua. "Research on the preparation technology of recombinant human interferon α-2b vaginal effervescent tablets", preparation and technology [J], 2009, 6(14):65-67】Using acid-base mixed non-aqueous granules, micro-silica gel adsorption interferon stock solution is dried at room temperature, mixed with acid-base granules to prepare interferon vaginal effervescent tablets, and prescription for effervescent tablets is orthogonal. The design method was optimized. The results showed that the micro-powder silica gel adsorption interferon can better maintain the biological activity of interferon without loss, the acid-base mixed non-aqueous granule effervescence effect is better, and the tablet is more uniform.

“Development of recombinant human interferon α-2b vaginal effervescent tablets” [Lu Chunyan, Wang Ronghai. Development of recombinant human interferon α-2b vaginal effervescent tablets [J], Biology Journal, 2006, 23(3): 42 -43] A method for preparing a recombinant human interferon alpha-2b vaginal effervescent tablet is disclosed, which comprises: granulating and drying a recombinant human interferon α2b semi-finished product, citric acid, lactose and a binder PVP K ₃₀ solution, carbonic acid Sodium hydride is granulated with lactose and PVP K ₃₀ solution; after mixing the two granules, magnesium stearate is added to adjust the tablet weight and tableting.

"Study on the preparation technology of recombinant human α-2b interferon effervescent tablets" [Wang Yin. Research on preparation technology of recombinant human α-2b interferon effervescent tablets [J], Journal of Jiangxi College of Traditional Chinese Medicine, 2005, 17(4): 51-52] discloses a process for preparing a recombinant human α-2b interferon effervescent tablet by preparing recombinant human α-2b interferon semi-finished product with bovine serum albumin and citric acid to prepare recombinant human α-2b interferon The lyophilized powder was divided into two parts, one part was mixed with the recombinant human α-2b interferon lyophilized powder, and the other part was mixed with sodium hydrogencarbonate, and 5% PVP aqueous solution was used as the binder granulation, 30 After drying at °C, the whole granules are mixed with magnesium stearate and compressed, and the pressure range is controlled to be 1000-1500 kg.

When the vaginal effervescent tablet is applied topically, the medicine can be fully distributed in the parts of the cervix, sputum and the like which are difficult to reach, and the medicine can penetrate the vaginal wall and wrap around the cervix, and quickly and fully contact with the lesion, thereby enabling Improve the efficacy of the drug.

CN1569224A also discloses an interferon vaginal effervescent capsule and a preparation method thereof. The vaginal effervescent capsule is composed of an interferon, a water-soluble adjuvant, an effervescent disintegrant, an auxiliary disintegrant, and a binder. The capsule has the advantages of simple process, convenient administration, rapid disintegration and rapid effervescence covering the wound surface to release the drug, does not pollute the clothes, has uniformity and good stability, and has a good therapeutic effect on cervicitis and cervical erosion. The interferon vaginal effervescent capsule is administered vaginally and absorbed through the vaginal mucosal epithelium, and directly exerts an antiviral effect in the local part. The interferon which enters the body is partially hydrolyzed by protease, and the other part is excreted by the urine prototype to cervicitis and cervical erosion. Has a very good therapeutic effect.

However, as an interferon, it is a kind of highly active and multifunctional inducing protein with antiviral, antitumor and immunomodulatory functions. It is one of the earliest cytokines to enter clinical application. In vivo, interferon-based drugs work through immunoregulatory functions, so it is not possible to evaluate the efficacy of drugs based on their plasma drug concentrations. The diversity and complexity of the interferon effect make the relationship between dose and effect more complicated. Studies have shown that long-term continuous use of interferon can induce tolerance, intermittent administration can obtain satisfactory results, especially for chronic cervicitis, cervical erosion and other gynecological diseases, the best local treatment is sustained interferon absorption To eliminate local symptoms without causing adverse reactions due to systemic absorption. Unfortunately, the above vaginal effervescent tablets and effervescent capsules are not capable of achieving sustained release of interferon for a long period of time in a low dose. In addition, the above vaginal effervescent tablets can be stored for two years at a low temperature (2 to 8 ° C), thereby having a certain limiting effect in clinical use.

In view of this, the present invention has been specifically proposed.

Summary of the invention

The object of the present invention is to provide an interferon vaginal effervescent capsule which can continuously release interferon at a low dose for a long time, and can achieve sustained low dose administration of the vagina, thereby better eliminating local symptoms. Effectively extend treatment time and reduce adverse reactions of long-term medication.

In order to achieve the object of the present invention, the present invention adopts the following technical solutions:

An interferon vaginal effervescent capsule, wherein the interferon vaginal effervescent capsule is composed of interferon lyophilized powder, effervescent powder and pharmaceutically acceptable excipient, wherein the interferon lyophilized powder and The mass ratio of the effervescent powder is from 1:1.0 to 1.5, preferably 1:1.2.

For gynecological diseases such as chronic cervicitis and cervical erosion, the best topical treatment is sustained interferon absorption to eliminate local symptoms without causing adverse reactions due to systemic absorption. However, the prior art vaginal effervescent capsules and effervescent tablets are unable to achieve low-dose long-term sustained release of interferon. Moreover, the prior art Interferon vaginal effervescence can be preserved for two years at low temperature (2 ~ 8 ° C), which has a certain limiting effect in clinical use. The present inventors have surprisingly found that interferon is prepared as a lyophilized powder and then prepared into an interferon effervescent capsule with effervescent powder and a pharmaceutically acceptable excipient, which can not only continuously release interferon at a low dose for a long time, thereby achieving vaginal persistence. Low-dose administration to better eliminate local symptoms, prolong the treatment time, and reduce the adverse reactions of long-term medication; and the accelerated test at a temperature of 40 °C ± 2 °C for 6 months and two at 25 °C ± 2 °C In the long-term test, there was no significant change in potency (biological activity) and appearance, showing good stability.

Further, the interferon lyophilized powder is obtained by freeze-drying an interferon freeze-dried solution, which is an effervescent disintegrant binder composed of a carbonate and an acid substance. After the granulation, the binder is hydroxypropylmethylcellulose.

At present, in the preparation of interferon effervescent tablets and effervescent capsules containing carbonates and acid substances, although water-soluble binders should be used in terms of drug usability, in order to avoid carbonation during preparation Neutralization of salts and acids does not allow moisture to exist, so PVP, which is both soluble in water and soluble in anhydrous ethanol, is an ideal binder. In the prior art, a PVP or a mixed solution containing PVP is also basically used as a binder.

In the present invention, it has been unexpectedly found that an effervescent powder is prepared by using hydroxypropylmethylcellulose as a binder, and then interferon vaginal effervescent capsule is prepared by mixing with an interferon lyophilized powder and a pharmaceutically acceptable adjuvant. Low-dose continuous release of interferon for a long time, to achieve sustained low-dose administration of the vagina, thereby better eliminating local symptoms and effectively prolonging the treatment time. Probably because hydroxypropylmethylcellulose is a viscous polymer material, after the formulation is used as a binder, the contents of the capsule shell are melted and the contents are rapidly hydrated, and all the powders are bonded together to form a capsule-like semi-solid surface. The high-viscosity gel layer is formed, so that the preparation adheres to the vaginal surface for a long time, and the capsule-like semi-solid has a larger contact area with the vaginal mucosa than the capsule, so that the drug can be uniformly released. At the same time, hydroxypropylmethylcellulose may also be a retarder for the slow release of interferon, which plays a role in the sustained release of interferon for a long period of time.

The interferon of the present invention is natural or recombinant interferon alpha-2b, and analogs thereof.

Further, the interferon lyophilized stock solution is composed of an interferon alpha-2b stock solution, a protective agent, an excipient, a surfactant, and a buffer.

The protective agent is one or more of lactose, sucrose, glucose, sorbose or trehalose, preferably sucrose; the excipient is mannitol, starch, dextrin, glycerin, sorbitol, polyethyl One or more of diol, gelatin, sodium carboxymethylcellulose or hydroxypropylcellulose, preferably mannitol; the surfactant is polysorbate 80, sodium dodecyl sulfate or poly One or more of sorbate 20, preferably polysorbate 80; the buffer is a phosphate buffer, a boric acid buffer, a citrate buffer or an acetate buffer, preferably a phosphate buffer.

Preferably, the composition of the interferon lyophilized stock solution is as follows:

More preferably, the composition of the interferon lyophilized stock solution is as follows:

In the present invention, the percentage concentration is a mass percentage concentration.

Further, the mass ratio of the carbonate to the acid substance in the effervescent disintegrant is 1:2 to 4, preferably 1:3.

The carbonate is calcium carbonate, sodium carbonate, sodium hydrogencarbonate, potassium carbonate or potassium hydrogencarbonate, and the acid substance is citric acid, malic acid, tartaric acid, alginic acid or boric acid, preferably citric acid.

The pharmaceutically acceptable adjuvant is sodium carboxymethyl starch, hydroxypropyl cellulose, starch, dextrin, glycerin, mannitol, sorbitol, polyethylene glycol, gelatin, sodium carboxymethylcellulose or hydroxy One or more of propyl cellulose, preferably sodium carboxymethyl starch.

The object of the present invention is also to provide a preparation method of the interferon vaginal effervescent capsule, wherein the preparation process is stable, and the prepared interferon vaginal effervescent capsule has good release reproducibility between batches.

The preparation method of the interferon vaginal effervescent capsule of the present invention comprises the following steps:

1) Preparation of interferon lyophilized powder

Take the buffer, add the excipient, dissolve, filter, cool, add the protective agent and surfactant to obtain the diluent; take the interferon α-2b stock solution, and mix with the obtained diluent to obtain the interferon freeze-dried solution, and freeze Dry, that is, interferon lyophilized powder;

2) Preparation of effervescent powder

Mixing an effervescent disintegrant composed of a carbonate and an acid substance, and spraying the binder solution to obtain an effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, pharmaceutically acceptable excipients and effervescent powder are sequentially added, and evenly mixed, that is, interferon vaginal effervescent capsule is obtained.

Wherein, the buffer has a pH of 3.0 to 7.5.

The recombinant human interferon ɑ-2b is stable under acidic conditions. In the present invention, the pH of the buffer is controlled to be in an acidic environment of 3.0 to 7.5, and the biological activity of the recombinant human interferon ɑ-2b can be maintained. It is not damaged, and it does not change the acidic environment of the vagina, and does not cause irritation to the vaginal mucosa.

Compared with the prior art, the present invention has the following advantages:

(1) The interferon vaginal effervescent capsule of the present invention can continuously release interferon at a low dose for a long time, and can achieve sustained low-dose administration of the vagina, can better eliminate local symptoms, effectively prolong the treatment time, and reduce the poor long-term medication. reaction;

(2) The interferon vaginal effervescent capsule of the present invention has an increase in potency (biological activity) and appearance in an accelerated test at a temperature of 40 ° C ± 2 ° C for 6 months and a two-year long test at 25 ° C ± 2 ° C. No obvious change, showing good stability;

(3) The interferon vaginal effervescent capsule of the invention has beautiful appearance, is comfortable, convenient and clean and hygienic when used, and is dispersed in a powder form for the medicine, and the dosage is accurate;

(4) The interferon vaginal effervescent capsule prepared by the invention has good reproducibility and good preparation process, and is a safe, effective and convenient vaginal administration dosage form.

DRAWINGS

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing the release profile of interferon vaginal effervescent capsules of the present invention and prior art.

detailed description

The following is a specific embodiment of the invention, which is intended to further describe the invention and not to limit the invention.

Example 1

1) Interferon lyophilized powder

Take the appropriate amount of phosphate buffer, add mannitol, dissolve, filter, cool, add sucrose and polysorbate 80 to obtain a dilution; take interferon ɑ-2b stock solution, and mix with the resulting dilution to obtain interferon lyophilized stock solution, Freeze-dried to obtain interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; citric acid and calcium carbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 2

1) Interferon lyophilized powder

Take the appropriate amount of phosphate buffer pH 7.2, add mannitol, dissolve, filter, cool, add sucrose and polysorbate 80 to obtain a dilution; take interferon ɑ-2b stock solution, and mix with the resulting dilution to get interference The freeze-dried stock solution is lyophilized to obtain the interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; citric acid and calcium carbonate are mixed at a ratio of 10:20, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 3

1) Interferon lyophilized powder

Take the appropriate amount of phosphate buffer with a pH of 3.0, add mannitol, dissolve, filter, cool, add sucrose and polysorbate 80 to obtain a dilution; take the interferon ɑ-2b stock solution, and mix with the obtained dilution to obtain interference The freeze-dried stock solution is lyophilized to obtain the interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; citric acid and calcium carbonate are mixed at a ratio of 10:40, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 4

1) Interferon lyophilized powder

Take the appropriate amount of phosphate buffer pH 6.0, add starch, dissolve, filter, cool, add sucrose and Polysorbate 80, a dilution solution is obtained; the interferon ɑ-2b stock solution is mixed with the obtained diluent to obtain an interferon lyophilized stock solution, which is lyophilized to obtain an interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; citric acid and calcium carbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 5

1) Interferon lyophilized powder

Take the appropriate amount of phosphate buffer with a pH of 5.0, add dextrin, dissolve, filter, cool, add sucrose and polysorbate 80 to obtain a dilution solution; take the interferon ɑ-2b stock solution and mix with the obtained dilution to obtain interference The freeze-dried stock solution is lyophilized to obtain the interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; citric acid and calcium carbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 6

1) Interferon lyophilized powder

Take the appropriate amount of pH 7.0 phosphate buffer, add glycerin, dissolve, filter, cool, add sucrose and polysorbate 80 to obtain a dilution; take interferon ɑ-2b stock solution, and mix with the resulting dilution to obtain interferon The lyophilized stock solution is lyophilized and lyophilized to obtain an interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; citric acid and calcium carbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 7

1) Interferon lyophilized powder

Take the appropriate amount of phosphate buffer with a pH of 4.8, add sorbitol, dissolve, filter, cool, add sucrose and polysorbate 80 to obtain a dilution; take interferon ɑ-2b stock solution, and mix with the obtained dilution to obtain interference The freeze-dried stock solution is lyophilized to obtain the interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; citric acid and calcium carbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 8

1) Interferon lyophilized powder

Take the appropriate amount of phosphate buffer pH 7.2, add polyethylene glycol, dissolve, filter, cool, add trehalose and polysorbate 20 to obtain a dilution; take interferon ɑ-2b stock solution, mix with the resulting dilution The interferon lyophilized stock solution is obtained and lyophilized to obtain an interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; boric acid and potassium hydrogencarbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 9

1) Interferon lyophilized powder

Take appropriate amount of boric acid buffer, add gelatin, dissolve, filter, cool, add sorbose and sodium lauryl sulfate to obtain a diluent; take interferon ɑ-2b stock solution, and mix with the obtained dilution to obtain interferon freeze-dried The original solution is lyophilized to obtain an interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; the alginic acid and potassium carbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Add interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder in sequence, and mix evenly to obtain interferon vagina Effervescent capsules. The amount of each component is as follows:

Example 10

1) Interferon lyophilized powder

Take the appropriate amount of citrate buffer, add sodium carboxymethylcellulose, dissolve, filter, cool, add glucose and polysorbate 80 to obtain a dilution; take interferon ɑ-2b stock solution, and mix with the resulting dilution to get interference The freeze-dried stock solution is lyophilized to obtain the interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; tartaric acid and sodium hydrogencarbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, sodium carboxymethyl starch and effervescent powder were added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule was obtained. The amount of each component is as follows:

Example 11

1) Interferon lyophilized powder

Take the appropriate amount of acetic acid buffer, add hydroxypropyl cellulose, dissolve, filter, cool, add lactose and polysorbate 80 to obtain a dilution; take the interferon ɑ-2b stock solution, and mix with the obtained dilution to obtain interferon frozen The dried stock solution is lyophilized to obtain the interferon lyophilized powder. The specific proportions of the components are as follows:

2) Preparation of effervescent powder

After dissolving 15 g of hydroxypropylmethylcellulose with an appropriate amount of 70% medical alcohol, the volume is adjusted to 6 L to obtain a binder solution; malic acid and sodium carbonate are mixed at a ratio of 10:30, and the obtained binder solution is sprayed. Made of effervescent powder;

3) Interferon effervescent capsules

Interferon lyophilized powder, starch and effervescent powder are added in sequence, and evenly mixed, that is, interferon vaginal effervescent capsule is obtained. The amount of each component is as follows:

Comparative Example 1. Preparation of interferon vaginal effervescent capsule according to the method of CN1569224A

prescription:

Preparation Process:

Pour the interferon stock solution into 200 ml of phosphate buffer solution (0.2 mol/L, pH 7.2), mix well, add the sorbitol excipients in the prescription, stir and mix, vacuum dry at temperature of 20 degrees for more than 24 hours, remove The particles in the prescription are gradually mixed in an equal amount to form a fraction of the particles containing the interferon main drug.

Weigh the amount of malic acid, sodium bicarbonate and sodium carbonate according to the prescription, put it into the boiling granulator, ventilate the boiling material, set the inlet air temperature to 60 degrees, and after 10 minutes, press 0.5% hydroxypropyl in the prescription. A mixed solution of a methylcellulose ethanol solution and a 10% polyvinylpyrrolidone ethanol solution was spray granulated until uniform particles containing an effervescent component were obtained.

According to the prescription, pregelatinized starch and crospovidone which were dried at 70 degrees in an oven for 4 hours or more were weighed and uniformly mixed with the above two kinds of particles.

The final mixed granules were filled into capsules, and the interferon ɑ-2b titer was measured: 9.02 × 10 ⁵ IU / granule, moisture: 1.13%.

Test Example 1, release test

The interferon vaginal effervescent capsule (test article) prepared in Example 1 of the present invention and the interferon vaginal effervescent capsule (reference substance) prepared in Comparative Example 1 were respectively measured by the release method ("People's Republic of China Pharmacopoeia" 2010 edition Appendix XD first method), using the dissolution method ("People's Republic of China Pharmacopoeia" 2010 edition of Appendix XC) first method device, with phosphate buffer (pH = 7.4) 500mL as solvent, the rotation speed is 100r / min, respectively 5 mL of the solution was taken at 0.5, 1.0, 2.0, 3.0, 4.0, 6.0, 8.0, 10.0 and 12.0 h, filtered through a filter (pore size 0.45 μm), the primary filtrate was discarded, and the filtrate was taken as the test solution, and timely 5 mL of phosphate buffer (pH=7.4) was added, and the release amount of interferon ɑ-2b was measured by ELISA. The release curve is shown in Figure 1.

As can be seen from Figure 1, the interferon vaginal effervescent capsule (i.e., the test article) of the present invention has significant sustained release characteristics as compared with the interferon vaginal effervescent capsule (i.e., the control) prepared by the prior art method. .

The above-mentioned tests were also carried out on the interferon vaginal effervescent capsules prepared in other embodiments of the present invention, and compared with the above-mentioned reference materials. The method is the same as above, and the cumulative release results of interferon ɑ-2b are shown in Table 1 below:

Table 1. Cumulative release results of interferon ɑ-2b

	0.5h	1.0h	2.0h	3.0h	4.0h	6.0h	8.0h	10.0h	12.0h
Reference substance	52.3	75.1	83.5	87.8	88.6	-	-	-	-
Example 2	16.8	26.3	36.9	44.1	51.3	62.5	72.4	80.4	85.6
Example 3	17.5	26.9	37.2	45.9	52.2	62.3	72.8	80.9	85.3
Example 4	17.9	26.7	37.4	45.2	51.6	62.8	71.9	80.3	85.1

Example 5	18.1	25.8	37.5	44.7	52.3	62.3	71.5	80.7	85.0
Example 6	16.5	26.1	36.4	45.3	51.7	62.1	72.6	81.2	86.3
Example 7	17.4	26.3	36.8	45.1	51.9	61.9	72.3	81.0	86.7
Example 8	18.3	27.1	37.1	44.8	52.5	61.5	72.1	80.5	86.2
Example 9	17.2	26.7	37.8	44.3	52.3	62.3	71.6	80.4	85.9
Example 10	17.8	26.5	36.3	44.5	51.0	61.7	71.4	81.9	85.2
Example 11	18.0	27.3	36.0	45.9	52.8	61.2	72.3	81.3	85.7

As can be seen from the above test results, the interferon vaginal effervescent capsule of the present invention has a marked sustained release profile as compared with the interferon vaginal effervescent capsule (i.e., the control) prepared by the prior art method.

Test Example 2, Stability Test

The stability test of the interferon vaginal effervescent capsule of the present invention shows that the effervescent capsule of the present invention has an accelerated test at a temperature of 40 ° C ± 2 ° C for 6 months and a two-year long test at 25 ° C ± 2 ° C. There was no significant change in potency (biological activity) and appearance, as shown in Tables 2 and 3, which showed good stability.

Table 2, Interferon α-2b vaginal effervescent capsule stability investigation results accelerated test: temperature 40 ° C ± 2 ° C

Table 3, Interferon α-2b vaginal effervescent capsule stability investigation results Long-term test: temperature 25 ° C ± 2 ° C

Test Example 3, Clinical Trial

1.1 General Information

A total of 120 patients who were positive for HPV DNA typing in a clinic from January 2013 to December 2014 were selected. 60 patients with cervical HPV infection treated with interferon vaginal effervescent capsules prepared in Example 1 of the present invention were treated. In the group, 60 patients with cervicin HPV infection were treated with Xinfuning. There were no acute genital inflammation in the two groups. All patients underwent liquid-based thin-layer cytology (TCT) or colposcopy to exclude cervical cancer and high-grade cervical intraepithelial neoplasia. change. In the treatment group, there were 42 cases with cervical columnar epithelial displacement area <1/3, and 18 cases with cervical columnar epithelial translocation area >1/3, <2/3, age 20-50 years old, no birth 12 example. In the control group, 45 cases with cervical columnar epithelial displacement area <1/3, combined with cervical columnar epithelial displacement area >1/3, < 15 cases of 2/3, aged 20 to 50 years, 10 cases of non-fertility. There were no significant differences in age, pregnancy, abortion frequency, cervical columnar epithelial displacement area and typing between the two groups, which were comparable.

1.2 treatment methods

1.2.1 Method of placing drugs

Three days after the menstruation was cleaned, one of the interferon vaginal effervescent capsules prepared in Example 1 of the present invention and one capsule of Xin Funing were placed in the deep vagina of the treatment group and the control group, once a day, 10 days. 1 course of treatment.

1.2.2HPV detection

The human papillomavirus nucleic acid amplification typing test kit produced by Kaipu Biotechnology Co., Ltd. was used to detect different subtypes of HPV, and each test was performed before treatment and after 1, 2, and 3 treatments.

1.2.3 Effect evaluation

The cure was negative for HPV subtype detection, and improved to HPV for at least one subtype negative. Ineffective for HPV detection, none of the subtypes were negative or increased. Healing plus improvement is considered effective.

1.3 statistical methods

All data were tested by x ² .

2, the results

2.1 The end of the first course of observation

At the end of the first course of treatment, the HPV subtype in the treatment group was completely negative in 12 cases, and at least one subtype was negative in 9 cases, with a total effective rate of 35.00%. In the control group, HPV subtypes were completely negative in 10 cases, at least one subtype was negative in 6 cases, and the total effective rate was 26.67%. The difference between the two groups was statistically significant, and the x ² test was P < 0.05. The results are shown in Table 4.

2.2 The end of the second course of observation

At the end of the second course of treatment, the HPV subtype of the treatment group was completely negative in 22 cases, and at least one subtype was negative in 10 cases, with a total effective rate of 53.33%. In the control group, HPV subtypes were completely negative in 20 cases, at least one subtype was negative in 9 cases, and the total effective rate was 48.33%. The difference between the two groups was statistically significant, and the x ² test was P < 0.05. The results are shown in Table 4.

2.3 The end of the third course of observation

At the end of the third course of treatment, the HPV subtype of the treatment group was completely negative in 50 cases, at least one subtype was negative in 6 cases, the total effective rate was 93.33%; 4 cases were ineffective. In the control group, HPV subtype was completely negative in 37 cases, at least one subtype was negative in 10 cases, and the total effective rate was 78.33%. The difference between the two groups was statistically significant, and the x ² test was P < 0.05. The results are shown in Table 4.

Table 4

2.4 adverse reactions

No adverse reactions were observed in all patients during the course of treatment.

The above test was also carried out on the interferon vaginal effervescent capsules prepared in other embodiments of the present invention, and the results obtained were similar.

Claims (10)

1. An interferon vaginal effervescent capsule, characterized in that the interferon vaginal effervescent capsule is composed of interferon lyophilized powder, effervescent powder and pharmaceutically acceptable excipient, wherein the interferon is freeze-dried The mass ratio of the powder to the effervescent powder is from 1:1.0 to 1.5, preferably 1:1.2.
2. The interferon vaginal effervescent capsule according to claim 1, wherein the interferon lyophilized powder is obtained by freeze-drying an interferon lyophilized stock solution, wherein the effervescent powder is a carbonate The effervescent disintegrant composed of an acid substance is obtained by granulating with a binder, and the binder is hydroxypropylmethylcellulose.
3. The interferon vaginal effervescent capsule according to claim 2, wherein the interferon lyophilized stock solution consists of a stock solution of interferon alpha-2b, a protective agent, an excipient, a surfactant, and a buffer.
4. The interferon vaginal effervescent capsule according to claim 3, wherein the protective agent is one or more of lactose, sucrose, glucose, sorbose or trehalose, preferably sucrose; The agent is one or more of mannitol, starch, dextrin, glycerin, sorbitol, polyethylene glycol, gelatin, sodium carboxymethylcellulose or hydroxypropylcellulose, preferably mannitol; The surfactant is one or more of polysorbate 80, sodium dodecyl sulfate or polysorbate 20, preferably polysorbate 80; the buffer is phosphate buffer, boric acid buffer, lemon Acid buffer or acetate buffer, preferably phosphate buffer.
5. The interferon vaginal effervescent capsule according to claim 4, wherein the composition of the interferon lyophilized stock solution is as follows:

   

   Preferably, the composition of the interferon lyophilized stock solution is as follows:

   
6. The interferon vaginal effervescent capsule according to any one of claims 2 to 5, wherein the mass ratio of the carbonate to the acid substance in the effervescent disintegrant is 1:2 to 4, preferably 1:3.
7. The interferon vaginal effervescent capsule according to claim 6, wherein the carbonate is calcium carbonate, sodium carbonate, sodium hydrogencarbonate, potassium carbonate or potassium hydrogencarbonate, preferably calcium carbonate; The substance is citric acid, malic acid, tartaric acid, alginic acid or boric acid, preferably citric acid.
8. The interferon vaginal effervescent capsule according to any one of claims 1 to 7, wherein the pharmaceutically acceptable adjuvant is sodium carboxymethyl starch, hydroxypropyl cellulose, starch, dextrin, One or more of glycerin, mannitol, sorbitol, polyethylene glycol, gelatin, sodium carboxymethylcellulose or hydroxypropylcellulose, preferably sodium carboxymethyl starch.
9. The method for preparing an interferon vaginal effervescent capsule according to any one of claims 1 to 8, wherein the preparation method comprises the following steps:

   1) Preparation of interferon lyophilized powder

   Take the buffer, add the excipient, dissolve, filter, cool, add the protective agent and surfactant to obtain the diluent; take the interferon α-2b stock solution, and mix with the obtained diluent to obtain the interferon freeze-dried solution, and freeze Dry, that is, interferon lyophilized powder;

   2) Preparation of effervescent powder

   Mixing an effervescent disintegrant composed of a carbonate and an acid substance, and spraying the binder solution to obtain an effervescent powder;

   3) Interferon effervescent capsules

   Interferon lyophilized powder, pharmaceutically acceptable excipients and effervescent powder are sequentially added, and uniformly mixed to obtain interferon α-2b vaginal effervescent capsule.
10. The method according to claim 9, wherein the buffer has a pH of 3.0 to 7.5.

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