CN103751098A - Recombinant human interferon alpha 2a vaginal expansion suppository and preparation and detection methods thereof - Google Patents

Abstract

The invention relates to a recombinant human interferon alpha 2a vaginal expansion suppository and preparation and detection methods thereof. The expansion suppository comprises a medicated mucous layer, a substrate layer and an expansion carrier; the medicated mucous layer consists of recombinant human interferon alpha 2a and a protective agent, wherein the protective agent improves the stability of recombinant human interferon alpha 2a; when in local application, the medicated mucous layer can form mucosa by which the medicated mucous layer can be attached on vaginal skin to promote skin absorption so as to further improve the bioavailability of the suppository. The expansion suppository is characterized in that a substrate further comprises a specific hardening agent. Due to the adoption of six unique leading technologies, the recombinant human interferon alpha 2a vaginal expansion suppository has the beneficial effects of preventing outflow of liquid medicine, being high in stability and lasting in curative effect, and preventing superinfection, etc.

Description

Recombinant human interferon-alpha 2 а vagina expansible plugs and preparation method thereof and detection method

Technical field

The invention belongs to vaginal suppository field, particularly a kind of recombinant human interferon-alpha 2 а vagina expansible plugs and preparation method thereof and detection method.

Background technology

Recombinant human interferon-alpha 2 а, are the multi-functional and highly active albumen that induces that a class is produced by leukocyte and lymphocyte, meet light and moist lability; Being mainly used in clinically the treatment to cervicitis, is mainly to rely on its broad-spectrum disease resistance toxic action and the immunoloregulation function to body.The formulation development of recombinant human interferon-alpha 2 а is always very undesirable, has affected the application of recombinant human interferon-alpha 2 а, and the preparation that goes on the market at present mainly comprises tablet, suppository; But tablet is filled in after vagina, recombinant human interferon-alpha 2 а are difficult to stripping, be difficult to play a role, and tablet are bad to human vagina's adaptability; And there is slow, the difficult absorption of disintegrate in general suppository, use is inconvenient, onset is slow, the shortcoming of pollution clothes, the impact of position when therapeutic effect is used by patient, patient's posterior fornix facies ventralis affected part is difficult to touch medicine.A kind of effervescent tablet that recombinant human interferon-alpha 2 а make as active component of take is disclosed in Chinese patent CN1970079A; Although can make medicine fully contact performance drug effect with focus, it can not solve stability and the adaptability problem of preparation to human body of recombinant human interferon-alpha 2 а; And prior art rarely has the research to recombinant human interferon-alpha 2 а suppositorys, simultaneously because recombinant human interferon-alpha 2 а natures are unstable, make after suppository poor stability; And common recombinant human interferon-alpha 2 а suppositorys also exist not easy to clean, easy pollution clothes; Active component and people's contact level are little, cause absorbing insufficient, do not bring into play the technical problems such as optimum curative effect of prescription.

Summary of the invention

In order to solve the problems of the technologies described above, increase the stability of recombinant human interferon-alpha 2 а, give full play to curative effect, the present invention has studied a kind of recombinant human interferon-alpha 2 а vagina expansible plugs and preparation method thereof; A kind of science detection method that can control above-mentioned recombinant human interferon-alpha 2 а vagina expansible plug product qualities is also provided simultaneously.

The concrete technical scheme of the present invention is as follows:

The invention provides a kind of recombinant human interferon-alpha 2 а vagina expansible plugs, described expansible plug comprises recombinant human interferon-alpha 2 а, substrate and expandable expansion carrier; Described substrate is coated in expansion carrier surface, and the medicated layer that described recombinant human interferon-alpha 2 а form is coated in stromal surface, and the swell value of described expansion carrier after saturated water suction is greater than 1.1, and the parts by weight of described each composition of expansible plug are:

Recombinant human interferon-alpha 2 а 0.03-0.75

Substrate 80-190

Expansion carrier 50-400

Described substrate comprises one or more in Acrawax, natural acid ester, lipoidis substrate, water-soluble base, gel-type vehicle, hydrogenated oil and fat, glyceride, distillate oil or colloidal compound;

Described Acrawax comprises one or more in mixed fatty glycerides, propylene glycol stearate, fixed oil, semi-synthetic fatty acid glyceride, semi-synthetic cocos nucifera oil ester, semi-synthetic Petiolus Trachycarpi grease or semi-synthetic Fructus Litseae ester;

Described natural acid ester comprises one or more in oleum sapii, Oleum Linderae, Fructus Foeniculi fat, methyl oleate or cocoa butter;

Described lipoidis substrate comprises lanoline or lanonol;

Described water-soluble base comprises one or more in PEG400, polyethylene glycol 1500, Macrogol 4000, polyethylene glycol 6000, glycerin gelatine, polysorbate60, polysorbate65, polyoxyethylene monostearate or poloxamer;

Described gel-type vehicle comprises one or more in sodium carboxymethyl cellulose, hypromellose, carbomer, alginic acid or sodium alginate;

Described hydrogenated oil and fat comprises one or more in hydrogenated vegetable oil, hydrogenated groundnut, castor oil hydrogenated or cotmar;

Described glyceride comprises one or more in tripalmitin, glycerol tristearate, Gan You behenic acid ester or lauric acid triglyceride;

Described distillate oil comprises fractional distillation Petiolus Trachycarpi oil or fractionated coconut oil;

Described colloidal compound comprises one or more in arabic gum, gelatin or pectin.

The specific activity of described recombinant human interferon-alpha 2 а is every 1mg albumen containing 100,000,000 ius, 1.0 * 10 ⁸iU.

The present invention has adopted original creation " intravaginal sticking type medicine-feeding technology ", and this technology by adding expansion carrier to realize in suppository.Ordinary suppository is due to the relation of gravity, no matter user be standing, sitting posture or lie under formula position, always have vagina can not or the less pastille substrate that touches, after described expansion carrier expands, pastille substrate can be 360 ° and contact with vaginal walls, fully administration, 6 times of administration enlarged areas, medicine directly attaches to lesions position, makes the position that originally cannot contact medicine also can obtain effectively attaching treatment; And due to the buffer action that stype plays in the middle of vagina, make no longer to contact with each other between wound surface, thereby avoid superinfection, it is shown effect no longer repeatedly.In addition, " anti-side leakage key " design concept has been applied in the introducing of expansion carrier, because the expansion of expansion carrier tail end (non-pastille substrate) can be close to vaginal wall, and then prevent the outflow of medicine and substrate, reduce the pollution of medicated clothing and keep the valid density of medicine.And by " inner core coefficient of expansion control technology ", the swell value of carrier after saturated water suction that can make to expand is greater than 1.1.

Further, the parts by weight of described each composition of expansible plug are:

Recombinant human interferon-alpha 2 а 0.06-0.5

Substrate 100-170

Expansion carrier 80-250

Described substrate is the mixture of Acrawax and colloidal compound, and the ratio of weight and number of described Acrawax, colloidal compound is 6-9:1.After Acrawax in described substrate mixes with colloidal compound, can increase the fusing point of mixture, and then improve the stability of suppository, can promote the mixing of substrate and active component simultaneously, improve the cohesion of suppository.

Further, described Acrawax substrate is semi-synthetic Fructus Litseae ester, and described colloidal compound substrate is arabic gum, and the parts by weight of described each composition of expansible plug are:

Preferably, described substrate of the present invention also comprises that weight portion is the sclerosing agent of 1-2.5 part, and described sclerosing agent comprises one or more in behenic acid, hexadecanol or microwax.Described sclerosing agent comprises one or more in behenic acid, hexadecanol or microwax.

Described sclerosing agent can improve the molding of suppository; Selected specific sclerosing agent also has further prolong drug to vagina action time, makes " prolonged drug layering release ", within one day, is administered once, and drug effect reaches 24 hours, improves patient compliance.

Further preferably; recombinant human interferon-alpha 2 а vagina expansible plugs of the present invention; the medicated layer of described hypothallus outside is pastille mucous layer; described pastille mucous layer comprises that weight portion is that recombinant human interferon-alpha 2 а and the weight portion of 0.03-0.75 part is the protective agent of 5-20 part, and described protective agent comprises one or more in acetyl tributyl citrate or acetyltriethyl citrate.

Described protective agent can improve the stability of recombinant human interferon-alpha 2 а; Because the consumption of recombinant human interferon-alpha 2 а is very little, so we make pastille mucous layer by recombinant human interferon-alpha 2 а and carry out administration together with protective agent, can improve the bioavailability of medicine; Meanwhile, acetyl tributyl citrate or acetyltriethyl citrate are plasticizers, during local application, can form mucosa, and in use, pastille mucous layer just can be adsorbed on vagina skin, promote the absorption of skin, further improve the bioavailability of medicine.

Further preferably, described pastille mucous layer is discontinuous lamellar distribution.

Plasticization due to protective agent acetyl tributyl citrate or acetyltriethyl citrate; can cause the poor ductility of pastille mucous layer; and being designed to be discontinuous lamellar, pastille mucous layer distributes; can overcome well this shortcoming; improve the ductility of pastille mucous layer, further improve the bioavailability of medicine.

The swell value of expansion carrier of the present invention after saturated water suction is greater than 1.5, and described swell value is swell value radially; Described expansion carrier includes but not limited to cotton sliver, also can comprise the sterilizable material that other can expand, as non-woven fabrics, elastic fiber etc.

Substrate of the present invention is the front end 1/5-4/5 place that Semi surrounding type is coated in described expansion carrier, according to " human engineering suppository " shape, is designed to the shapes such as duckbill, spherical, avette, bullet shaped, torpedo, cylindrical, taper shape or clavate.

The rear end of expansion carrier of the present invention is connected with bracing wire, stype after expansion, by the rear end bracing wire of tractive expansion carrier, the dead bark that be adsorbed on the virus being killed on expansion carrier, toxin, comes off is also pulled out external in the lump, change dressings at every turn and just equal to do once thoroughly clean to vagina, fundamentally prevention of inflammation recurs again, and this expansible plug belongs to " integrated dosage form is cleaned in treatment ".

In order to reach recombinant human interferon-alpha 2 а vagina expansible plugs of the present invention, meet the effect that water can expand rapidly, after described expansion carrier water suction, the maximum water absorption of every is not less than 1.5 milliliters.Suitable water absorption, can guarantee suitable swell value, and too low or too high water absorption all can cause bad impact.When water absorption is too low, can not guarantee that recombinant human interferon-alpha 2 а vagina expansible plugs of the present invention meet the feature that water expand and make to expand insufficient; When water absorption is too high, expansion carrier is understood too much absorption ingredient and is limited its diffusion.Therefore, water absorption is too low or too high, is not the desired expansion carrier of the present invention.

The present invention provides the preparation method of recombinant human interferon-alpha 2 а vagina expansible plugs on the other hand, and described method comprises the steps:

1) preparation of substrate:

A. substrate is placed in to water-bath heating and melting, obtains fused mass;

B. by the fused mass of sclerosing agent and a step, mix homogeneously, makes substrate;

2) substrate is poured in bolt mould, inserted expansion carrier, cooling and shaping, makes substrate bolt core;

3) preparation of pastille mucous membrane agent:

Recombinant human interferon-alpha 2 а are mixed homogeneously with protective agent, make pastille mucous membrane agent;

4) by the pastille mucous membrane agent in step 3) with discontinuous Sprayable spray attachment in step 2) on the substrate bolt core that makes, on substrate bolt core, form one deck pastille mucous layer, make described recombinant human interferon-alpha 2 а vagina expansible plugs.

The preparation method of recombinant human interferon-alpha 2 а vagina expansible plugs of the present invention has adopted " the integrally formed technology of the disposable fill of stype ".

The present invention also provides the detection method of described recombinant human interferon-alpha 2 а vagina expansible plugs on the other hand, and described detection method comprises swell value assay method, weight differential detection method, assay etc.

Wherein said swell value assay method comprises:

A. along radially the choosing a bit or some points of described recombinant human interferon-alpha 2 а vagina expansible plug end faces, measure the initial length H of described expansion carrier;

B. after the saturated water suction of described recombinant human interferon-alpha 2 а vagina expansible plug, be determined at the rear length h of selected location place's expansion carrier expansion in a step;

While c. calculating described swell value, according to I formula, calculate;

$p=\frac{h}{H}---\left(I\right)$

Wherein, p represent axial swell value, h represent to expand after length, H represent initial length; Or swell value assay method is as follows:

A. along described recombinant human interferon-alpha 2 а vagina expansible plugs axially choose a place or some positions, measure the initial diameter R of described expansion carrier;

B. after the saturated water suction of described recombinant human interferon-alpha 2 а vagina expansible plug, be determined at the rear diameter r of selected location place's expansion carrier expansion in a step;

While c. calculating described swell value, by II formula, calculate;

$P=\frac{r}{R}---\left(\mathrm{II}\right)$

Wherein, P represent swell value, r radially represent to expand after diameter, R represent initial diameter; Described weight differential assay method comprises:

A. get described recombinant human interferon-alpha 2 а vagina expansible plugs, take weight M;

B. get the expansion carrier in the described recombinant human interferon-alpha 2 а vagina expansible plugs that scrape substrate, dry, take weight m, according to the weight X of substrate as described in calculating as III formula:

X=M-m

（Ⅲ）。

Further, in the detection method of described recombinant human interferon-alpha 2 а vagina expansible plugs, melt and become after time limit detection, then carry out following at least one detection method:

1) swell value assay method is as follows:

A. first along radially the choosing a bit or some points of described recombinant human interferon-alpha 2 а vagina expansible plug end faces, measure after the initial length H of described expansion carrier, then the different angles of rolling measure several times, Hi averages;

B. described recombinant human interferon-alpha 2 а vagina expansible plugs melt after the detection of change time limit, are determined at the rear length h of selected location place's expansion carrier expansion in a step; The different angles of rolling are again measured several times, ask the rear average length hi that expands;

While c. calculating described swell value, according to I formula, calculate;

$\mathrm{pi}=\frac{\mathrm{hi}}{\mathrm{Hi}}---\left(I\right)$

Wherein, pi represent axial swell value, hi represent to expand after average length, Hi represent average initial length;

Or described swell value assay method is as follows:

A. first an angle along described recombinant human interferon-alpha 2 а vagina expansible plugs axially choose a place or some positions, measure after the initial diameter R of described expansion carrier, then the different angles of rolling measure several times, Ri averages;

B. described recombinant human interferon-alpha 2 а vagina expansible plugs melt after the detection of change time limit, are determined at the rear diameter r of selected location place's expansion carrier expansion in a step; The different angles of rolling are again measured several times, ask the rear average diameter ri that expands;

While c. calculating described swell value, by II formula, calculate;

$P_i=\frac{\mathrm{ri}}{\mathrm{Ri}}---\left(\mathrm{II}\right)$

Wherein, Pi represent swell value, ri radially represent to expand after average diameter, Ri represent average initial diameter;

2) weight differential assay method:

A. get described recombinant human interferon-alpha 2 а vagina expansible plugs, take weight M;

B. scrape substrate and pastille mucous layer, expansion carrier is placed in to 20-90 ℃ of organic solvent, take out, in 50-150 ℃ of dry 1-10h, take weight m, according to the gross weight X of substrate as described in calculating as III formula and pastille mucous layer:

X=M-m?（Ⅲ）

Wherein, described organic solvent comprises one or more in ethanol, methanol or isopropyl alcohol.

Further, in the detection method of described recombinant human interferon-alpha 2 а vagina expansible plugs, described expansion carrier is cotton sliver, and described detection comprises following at least one method:

1) swell value assay method:

Get 3 of described recombinant human interferon-alpha 2 а vagina expansible plugs, with slide gauge, survey its afterbody cotton sliver diameter, roll approximately 90 ° and survey once again, survey twice for every, obtain 2 meansigma methods Ri of every mensuration; Above-mentioned 3 bolts are used for melting and become after the time limit measures and finish, take out immediately residue cotton sliver, treat disconnected of water, all gently be placed on glass plate, with slide gauge, measure two ends and middle three positions of each cotton sliver, roll after approximately 90 ° and measure three positions again, each cotton sliver obtains six data altogether, obtains 6 meansigma methods r of mensuration _i, calculate the swell value P of every _i, the swell value of three bolts all should be greater than 1.5;

2) weight differential assay method:

Get 10 of described recombinant human interferon-alpha 2 а vagina expansible plugs, the accurately weighed weight of difference, gently scrape substrate and pastille mucous layer (must not lose cotton sliver), cotton sliver is placed in to the 200-400ml ethanol of 50-80 ℃, preferably, cotton sliver is placed in to the 300ml ethanol of 60-70 ℃, and 80khz frequency ultrasonic cleaning 5 minutes, the remaining stromatolysis in cotton sliver surface is removed, taking out cotton sliver firmly extracts, with filter paper, inhale 3 times again, in 105 ℃, be dried 2 hours, take out, room temperature was placed after 1 hour, the accurately weighed cotton sliver weight of difference, obtain every containing substrate and pastille mucous layer gross weight and average substrate and pastille mucous layer gross weight with respect to total number, and the two is compared, what exceed average weight ± 10% must not be more than 2, and must not there is 1 times of 1 overrun.

The assay method of the swell value of recombinant human interferon-alpha 2 а vagina expansible plugs provided by the present invention adopts repeatedly measuring method of multi-angle, has improved the accuracy of measuring, and has increased the operability of measuring, and method is easier to judgement; When carrying out the standard detection recombinant human interferon-alpha 2 а vagina expansible plug of the present invention of state-promulgated pharmacopoeia appendix weight differential, substrate and pastille mucous layer need be scraped off, the scraping degree of grasping due to operator is different, cause every scraping degree difference larger, so the present invention adopts the substrate on dissolution with solvents expansible plug surface, through verification experimental verification, when adopting hot ethanol to clean the substrate on expansible plug surface, result is better, repeatability is higher, and described weight differential assay method is more convenient stable.

It is high that recombinant human interferon-alpha 2 а vagina expansible plugs provided by the present invention have stability, the beneficial effects such as good effect, expansion carrier in described recombinant human interferon-alpha 2 а vagina expansible plugs add " the disposable fill of stype is integrally formed " technology of having applied original creation, " control of the inner core coefficient of expansion " technology, " human engineering bolt type " technology, " treatment is clean integrated " dosage form, " administration of intravaginal sticking type " technology, " anti-side leakage key design " six technology, this expansion carrier can make the pastille mucous layer of recombinant human interferon-alpha 2 а vagina expansible plugs fully contact with vaginal walls, and prevent medicinal liquid outflow, also there is the effect that cleaning vagina prevents superinfection, described protective agent has improved the stability of recombinant human interferon-alpha 2 а, described protective agent mixes the pastille mucous layer making and just can be adsorbed on vagina skin with recombinant human interferon-alpha 2 а, promote the absorption of skin, further improves the bioavailability of medicine.

Accompanying drawing explanation

Fig. 1 is the structural representation of recombinant human interferon-alpha 2 а vagina expansible plugs of the present invention, and 1 is pastille mucous layer, and 2 is substrate, and 3 is expansion carrier, and 4 is bracing wire;

Fig. 2 and Fig. 3 swell value assay method schematic diagram.

The specific embodiment

Embodiment 1

Preparation method:

1) preparation of substrate:

Semi-synthetic Fructus Litseae ester and arabic gum are mixed, be placed in water-bath heating and melting, obtain fused mass;

2) substrate is poured in bolt mould, inserted cotton sliver, cooling and shaping, makes 100 pieces of conical substrate bolt cores;

3) by recombinant human interferon-alpha 2 а with discontinuous Sprayable spray attachment in step 2) on the substrate bolt core that makes, on substrate bolt core, form one deck medicated layer, make described recombinant human interferon-alpha 2 а vagina expansible plugs, every piece of heavily about 3.3g.

Embodiment 2

Preparation method:

1) preparation of substrate:

A. semi-synthetic Petiolus Trachycarpi grease and propylene glycol stearate are mixed, be placed in water-bath heating and melting, obtain fused mass;

B. by the fused mass of hexadecanol and a step, mix homogeneously, makes substrate;

2) substrate is poured in bolt mould, inserted cotton sliver, cooling and shaping, makes 100 pieces of cylindrical substrate bolt cores;

3) by recombinant human interferon-alpha 2 а with discontinuous Sprayable spray attachment in step 2) on the substrate bolt core that makes, on substrate bolt core, form one deck medicated layer, make described recombinant human interferon-alpha 2 а vagina expansible plugs, every piece of heavily about 3.3g.

Embodiment 3

Preparation method: make 100 pieces of clavate suppositorys by embodiment 2 methods, every piece of heavily about 4.9g.

Embodiment 4

Preparation method:

1) preparation of substrate:

A. PEG400 and lanoline are mixed, be placed in water-bath heating and melting, obtain fused mass;

B. by the fused mass of microwax and a step, mix homogeneously, makes substrate;

2) substrate is poured in bolt mould, inserted cotton sliver, cooling and shaping, makes 100 pieces of sphere matrix bolt cores;

3) preparation of pastille mucous membrane agent:

Recombinant human interferon-alpha 2 а are mixed homogeneously with acetyl tributyl citrate, make pastille mucous membrane agent;

4) by the pastille mucous membrane agent in step 3) with discontinuous Sprayable spray attachment in step 2) on the substrate bolt core that makes, on substrate bolt core, form one deck pastille mucous layer, make described recombinant human interferon-alpha 2 а vagina expansible plugs, every piece of heavily about 4.9g.

Embodiment 5

Preparation method: make 100 pieces of avette suppositorys by embodiment 4 methods, every piece heavily approximately 4.9 _g.

Embodiment 6

Preparation method: make 100 pieces of bullet shaped suppositorys by embodiment 4 methods, every piece heavily approximately 2.7 _g.

Embodiment 7

Preparation method: make 100 pieces of duckbill suppositorys by embodiment 4 methods, every piece heavily approximately 3.8 _g.

Embodiment 8

Preparation method: make 100 pieces of torpedo suppositorys by embodiment 4 methods, every piece of heavily about 3.4g.

Test example 1 stability test

1. accelerated test

Get recombinant human interferon-alpha 2 а vagina expansible plugs described in embodiment 2-8 and commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat), all at 25 ℃ ± 2 ℃ of constant temperature, relative humidity is to place 6 months under 65% ± 5% constant humidity condition, duration of test 1 month, 2 months, 3 months, sample respectively once 6 the end of month, press the regulation in Chinese Pharmacopoeia, detect the character of expansible plug, labelled amount (%) containing recombinant human interferon-alpha 2 а, melt and become the time limit, the accelerated test of the recombinant human interferon-alpha 2 а vagina expansible plugs described in embodiment 2-5 the results are shown in Table 1, the accelerated test of the recombinant human interferon-alpha 2 а vagina expansible plugs described in embodiment 6-8 and commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat) the results are shown in Table 2.

The accelerated test result of recombinant human interferon-alpha 2 а vagina expansible plugs described in table 1 embodiment 2-5

Recombinant human interferon-alpha 2 а vagina expansible plugs and commercially available recombinant human interferon-alpha 2 а bolts (trade name: accelerated test result difficult to understand putting down) described in table 2 embodiment 6-8

Recombinant human interferon-alpha described in embodiment of the present invention 2-8 2 а vagina expansible plugs as can be seen from the table, under the environment of constant temperature high humidity, place after 6 months, the color of described expansible plug does not change, described expansible plug smooth surface, do not occur speckle, the phenomenon of breaking, in described expansible plug there is not obvious variation in the labelled amount of recombinant human interferon-alpha 2 а, melts and become yet significantly prolongation of generation of time limit; And commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat) are placed after 1 month under high humidity, it is coarse that suppository surface just becomes, and places after 6 months, and suppository breaks, and it is more and more darker that color becomes; And described recombinant human interferon-alpha 2 а content significantly decline, melt and become time limit significant prolongation; Show that recombinant human interferon-alpha 2 а vagina expansible plugs of the present invention compare stability with commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat) and significantly improve.

2. long term test

Get recombinant human interferon-alpha 2 а vagina expansible plugs described in embodiment 2-8 and commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat), at 25 ℃ ± 2 ℃ of temperature, relative humidity is to place 36 months under 60% ± 10% condition, sampling in every 3 months once, respectively at 0 month, 3 months, 6 months, 9 months, sampling in 12 months, detect the character of expansible plug, contain the labelled amount (%) of recombinant human interferon-alpha 2 а and melt and become the time limit, after 12 months, still need to continue to investigate index of correlation, respectively at 18 months, 24 months, sampling in 36 months detects, and testing result and comparison in 0 month, the long-term test results of the recombinant human interferon-alpha 2 а vagina expansible plugs described in embodiment 2-3 is in Table 3, the long-term test results of the recombinant human interferon-alpha 2 а vagina expansible plugs described in embodiment 4-6 is in Table 4, embodiment 7-8 and commercially available recombinant human interferon-alpha 2 а bolts (trade name: long-term test results difficult to understand putting down) is in Table 5.

The long-term test results of recombinant human interferon-alpha 2 а vagina expansible plugs described in table 3 embodiment 2-3

The long-term test results of recombinant human interferon-alpha 2 а vagina expansible plugs described in table 4 embodiment 4-6

Recombinant human interferon-alpha 2 а vagina expansible plugs and commercially available recombinant human interferon-alpha 2 а bolts (trade name: long-term test results difficult to understand putting down) described in table 5 embodiment 7-8

Recombinant human interferon-alpha described in embodiment of the present invention 2-8 2 а vagina expansible plugs as can be seen from the table, in the long term test of 36 months, the color of described expansible plug does not change, described suppository smooth surface, there is not speckle, the phenomenon of breaking, in described expansible plug there is not obvious variation in the labelled amount of recombinant human interferon-alpha 2 а, melts and become yet significantly prolongation of generation of time limit; And commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat) were placed after 3 months, it is coarse that suppository surface just becomes, and places after 36 months, and suppository breaks, and it is more and more darker that color becomes; And the content of described recombinant human interferon-alpha 2 а significantly declines, melt and become time limit significant prolongation; Show that recombinant human interferon-alpha 2 а vagina expansible plugs of the present invention compare stability with commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat) and significantly improve.

Test example 2 swell value determination tests

Method one

Get 3 of described recombinant human interferon-alpha 2 а vagina expansible plugs, with slide gauge, survey its afterbody cotton sliver diameter, roll approximately 90 ° and survey once again, survey twice for every, obtain 2 meansigma methods Ri of every mensuration; Above-mentioned 3 bolts are used for melting and become after the time limit measures and finish, take out immediately residue cotton sliver, treat disconnected of water, all gently be placed on glass plate, with slide gauge, measure two ends and middle three positions of each cotton sliver, roll after approximately 90 ° and measure three positions again, each cotton sliver obtains six data altogether, obtains 6 meansigma methods r of mensuration _i, calculate the swell value P of every _i, the swell value of three bolts is all greater than 1.5.

As shown in table 6 according to the swell value of recombinant human interferon-alpha 2 а vagina expansible plugs described in above-mentioned swell value assay method mensuration embodiment 1-8.

The swell value of table 6 embodiment 1-8 recombinant human interferon-alpha 2 а vagina expansible plugs

Embodiment	Average diameter (mm) before expanding	Average diameter (mm) after expanding	Swell value (doubly)	Result
					Embodiment
1	12.19	21.20	1.74	Qualified
					Embodiment
2	11.93	20.09	1.69	Qualified
					Embodiment 3	12.30	19.97	1.62	Qualified
Embodiment 4	12.28	22.50	1.83	Qualified
					Embodiment 5	12.13	22.45	1.85	Qualified
Embodiment 6	12.01	22.36	1.86	Qualified
					Embodiment 7	12.13	22.45	1.85	Qualified

Embodiment 8

12.34

23.41

1.90

Qualified

Method two

As shown in Figures 2 and 3, the detection method of vagina expansible plug:

1) swell value assay method:

A. first along described vagina expansible plug end face, radially choose C1, C2, tri-points of C3, with slide gauge, measure after initial length H1, H2, the H3 of described expansion carrier, the 90 ° of measurements of rolling again, radially choose C1, C4, tri-points of C5 along described vagina expansible plug end face are measured after initial length H4, H5, the H6 of described expansion carrier with slide gauge, ask the meansigma methods Hi of H1, H2, H3, H4, H5 and H6;

B. described vagina expansible plug melts after the detection of change time limit, with slide gauge, is determined at selected C1, C2 in a step, C3 tri-expansions of position expansion carriers rear length h1, h2, h3; Roll again 90 °, with slide gauge, be determined at selected C1, C4 in a step, C5 tri-expansions of position expansion carriers rear length h4, h5, h6, ask the average length of h1, h2, h3, h4, h5 and h6, be the rear average length hi that expands;

While c. calculating described swell value, according to I formula, calculate;

$\mathrm{pi}=\frac{\mathrm{hi}}{\mathrm{Hi}}---\left(I\right)$

Wherein, pi represent axial swell value, hi represent to expand after average length, Hi represent average initial length;

Or, swell value assay method:

D. first an angle, along described vagina expansible plug, axially choose A1, A2 and tri-points of A3, with slide gauge, measure after initial diameter R1, the R2 and R3 of described expansion carrier, roll again 90 °, along described vagina expansible plug, axially choose B1, B2 and tri-points of B3, with slide gauge, measure after initial diameter R4, the R5 and R6 of described expansion carrier, ask the meansigma methods Ri of R1, R2, R3, R4, R5 and six initial diameters of R6;

E. described vagina expansible plug melts and becomes after the time limit detects, and after being determined in d step selected A1, A2 and A3 position expansion carrier and expanding, diameter is respectively r1, r2 and r3; Roll again 90 ° measure in d step selected B1, B2 and B3 position expansion carrier and expand after diameter be respectively r4, r5 and r6, ask the meansigma methods of r1, r2, r3, r4, r5 and r6, average diameter ri afterwards must expand;

While f. calculating described swell value, by II formula, calculate;

$P_i=\frac{\mathrm{ri}}{\mathrm{Ri}}---\left(\mathrm{II}\right)$

Wherein, Pi represent swell value, ri radially represent to expand after average diameter, Ri represent to put down

Equal initial diameter.

Test example 3 weight differential determination tests

Get 10 of described recombinant human interferon-alpha 2 а vagina expansible plugs, the accurately weighed weight of difference, gently scrape substrate and pastille mucous layer (must not lose cotton sliver), cotton sliver is placed in to the 300ml ethanol of 60-70 ℃, and 80khz frequency ultrasonic cleaning 5 minutes, the remaining stromatolysis in cotton sliver surface is removed, taking out cotton sliver firmly extracts, with filter paper, inhale 3 times again, in 105 ℃, be dried 2 hours, take out, room temperature was placed after 1 hour, the accurately weighed cotton sliver weight of difference, obtain every containing substrate and pastille mucous layer gross weight and average substrate and pastille mucous layer gross weight with respect to total number, and the two is compared, what exceed average weight ± 10% must not be more than 2, and must not there is 1 times of 1 overrun.

Repeat aforesaid operations, detect the recombinant human interferon-alpha 2 а vagina expansible plugs described in embodiment 1-8, the equal conformance with standard of its weight differential.

Test example 4 clinical trials

One, research method

(1) research approach is formulated with reference to relevant " clinical verification of the department of obstetrics and gynecology urogenital tract infection local application " content of Ministry of Health of the People's Republic of China's new drug clinical guidance principle.

(2) diagnostic criteria

1, cervicitis symptom: leucorrhoea grow in quantity, be purulence or milky, be in mucus shape or leucorrhea and accompany the blood streak, or haemorrhagia sub coitu, companion's lower abdomen is uncomfortable, pruritus vulvae, and waist and sacrum pain, increases the weight of to accompany lumbago menstrual period;

2, gynecologial examination: cervix uteri congestion and edema, or have erosion in various degree, and there is purulent secretion White House neck tube to discharge, while touching cervix uteri, can there is pain; The performance such as cervix uteri has hypertrophy, polyp, body of gland cyst, turn up, or see that cervix uteri mouth has purulent secretion, palpation cervix uteri is harder, or has contact bleeding;

3, lab testing: cervical scraping smear shows Pasteur's II level.

(3) test case standard

Include case history standard in: in 21-50 year, meet the married woman of bacteriodiagnosis.

(4) Therapeutic Method

1, treatment group: the recombinant human interferon-alpha 2 а vagina expansible plugs described in the use embodiment of the present invention 7 are treated, and one of every day, 7 days are a course for the treatment of.

2, matched group: using commercially available recombinant human interferon-alpha 2 а bolts (trade name: difficult to understand flat) treatment, one of every day, 7 days is a course for the treatment of.

(5) observation item

Therapeutic is observed

(1) symptom: the amount of secretions, color, state;

(2) gynecologial examination: comprise vaginal secretions inspection and cervix uteri inspection;

(3) lab testing: comprise papanicolaou test.

(6) curative effect determinate standard

1, recovery from illness: transference cure, gynecologial examination and lab testing are normal;

2, effective: symptom obviously alleviates, gynecologial examination and lab testing are clearly better;

3, effective: symptom alleviates to some extent, gynecologial examination and lab testing take a turn for the better;

4, invalid: unchanged before and after treatment.

Two, therapeutic outcome

(1) two group of therapeutic effect to cervicitis is relatively in Table 7;

The therapeutic effect comparison of table 7 liang group to cervicitis

Group	Case load	Disease more	Effective	Effectively	Invalid	Effective percentage (%)
							Treatment group	35	26	3	5	1	97.1
Matched group	25	11	3	5	6	76.0

Two groups of curative effect comparison treatment groups are better than matched group.

Three, conclusion

Recombinant human interferon-alpha 2 а vagina expansible plugs reach 97.1% to the effective percentage of cervicitis, and curative effect is better than matched group; In its curative effect process of checking, do not find toxic and side effects, safe and reliable, evident in efficacy.

Claims (11)

1. recombinant human interferon-alpha 2 а vagina expansible plugs, is characterized in that, described expansible plug comprises recombinant human interferon-alpha 2 а, substrate and expandable expansion carrier; Described substrate is coated in expansion carrier surface, and the medicated layer that described recombinant human interferon-alpha 2 а form is coated in stromal surface, and the swell value of described expansion carrier after saturated water suction is greater than 1.1, and the parts by weight of described each composition of expansible plug are:

Recombinant human interferon-alpha 2 а 0.03-0.75

Substrate 80-190

Expansion carrier 50-400

Described substrate comprises one or more in Acrawax, natural acid ester, lipoidis substrate, water-soluble base, gel-type vehicle, hydrogenated oil and fat, glyceride, distillate oil or colloidal compound;

Described Acrawax comprises one or more in mixed fatty glycerides, propylene glycol stearate, fixed oil, semi-synthetic fatty acid glyceride, semi-synthetic cocos nucifera oil ester, semi-synthetic Petiolus Trachycarpi grease or semi-synthetic Fructus Litseae ester;

Described natural acid ester comprises one or more in oleum sapii, Oleum Linderae, Fructus Foeniculi fat, methyl oleate or cocoa butter;

Described lipoidis substrate comprises lanoline or lanonol;

Described water-soluble base comprises one or more in PEG400, polyethylene glycol 1500, Macrogol 4000, polyethylene glycol 6000, glycerin gelatine, polysorbate60, polysorbate65, polyoxyethylene monostearate or poloxamer;

Described gel-type vehicle comprises one or more in sodium carboxymethyl cellulose, hypromellose, carbomer, alginic acid or sodium alginate;

Described hydrogenated oil and fat comprises one or more in hydrogenated vegetable oil, hydrogenated groundnut, castor oil hydrogenated or cotmar;

Described glyceride comprises one or more in tripalmitin, glycerol tristearate, Gan You behenic acid ester or lauric acid triglyceride;

Described distillate oil comprises fractional distillation Petiolus Trachycarpi oil or fractionated coconut oil;

Described colloidal compound comprises one or more in arabic gum, gelatin or pectin.

2. recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 1, is characterized in that, the parts by weight of described each composition of expansible plug are:

Recombinant human interferon-alpha 2 а 0.06-0.5

Substrate 100-170

Expansion carrier 80-250

Described substrate is the mixture of Acrawax and colloidal compound, and the ratio of weight and number of described Acrawax, colloidal compound is 6-9:1.

3. recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 2, is characterized in that, described Acrawax substrate is semi-synthetic Fructus Litseae ester, and described colloidal compound substrate is arabic gum, and the parts by weight of described each composition of expansible plug are:

4. recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 1, is characterized in that, described substrate also comprises that weight portion is the sclerosing agent of 1-2.5 part, and described sclerosing agent comprises one or more in behenic acid, hexadecanol or microwax.

5. recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 4; it is characterized in that; the medicated layer of described hypothallus outside is pastille mucous layer; described pastille mucous layer comprises that weight portion is that recombinant human interferon-alpha 2 а and the weight portion of 0.03-0.75 part is the protective agent of 5-20 part, and described protective agent comprises one or more in acetyl tributyl citrate or acetyltriethyl citrate.

6. recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 5, is characterized in that, described pastille mucous layer is discontinuous lamellar and distributes.

7. the recombinant human interferon-alpha 2 а vagina expansible plugs as described in as arbitrary in claim 1-6, is characterized in that, the swell value of described expansion carrier after saturated water suction is greater than 1.5, and described swell value is swell value radially; Described expansion carrier is cotton sliver; Described substrate is the front end 1/5-4/5 place that Semi surrounding type is coated in described expansion carrier, and the rear end of described expansion carrier is connected with bracing wire; After described expansion carrier water suction, the maximum water absorption of every is not less than 1.5 milliliters.

8. the preparation method of recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 6, is characterized in that, described method comprises the steps:

1) preparation of substrate:

A. substrate is placed in to water-bath heating and melting, obtains fused mass;

B. by the fused mass of sclerosing agent and a step, mix homogeneously, makes substrate;

2) substrate is poured in bolt mould, inserted expansion carrier, cooling and shaping, makes substrate bolt core;

3) preparation of pastille mucous membrane agent:

Recombinant human interferon-alpha 2 а are mixed homogeneously with protective agent, make pastille mucous membrane agent;

4) by the pastille mucous membrane agent in step 3) with discontinuous Sprayable spray attachment in step 2) on the substrate bolt core that makes, on substrate bolt core, form one deck pastille mucous layer, make described recombinant human interferon-alpha 2 а vagina expansible plugs.

9. as the detection method of the recombinant human interferon-alpha 2 а vagina expansible plugs of claim 1-6 or 8 as described in arbitrary, it is characterized in that, described detection method comprises following at least one method:

1) swell value assay method is as follows:

A. along radially the choosing a bit or some points of described recombinant human interferon-alpha 2 а vagina expansible plug end faces, measure the initial length H of described expansion carrier;

B. after the saturated water suction of described recombinant human interferon-alpha 2 а vagina expansible plug, be determined at the rear length h of selected location place's expansion carrier expansion in a step;

While c. calculating described swell value, according to I formula, calculate;

$p=\frac{h}{H}---\left(I\right)$

Wherein, p represent axial swell value, h represent to expand after length, H represent initial length; Or swell value assay method is as follows:

A. along described recombinant human interferon-alpha 2 а vagina expansible plugs axially choose a place or some positions, measure the initial diameter R of described expansion carrier;

B. after the saturated water suction of described recombinant human interferon-alpha 2 а vagina expansible plug, be determined at the rear diameter r of selected location place's expansion carrier expansion in a step;

While c. calculating described swell value, by II formula, calculate;

$P=\frac{r}{R}---\left(\mathrm{II}\right)$

Wherein, P represent swell value, r radially represent to expand after diameter, R represent initial diameter;

2) weight differential assay method:

A. get described recombinant human interferon-alpha 2 а vagina expansible plugs, take weight M;

B. scrape substrate and pastille mucous layer, expansion carrier is placed in to 20-90 ℃ of organic solvent, take out, in 50-150 ℃ of dry 1-10h, take weight m, according to the gross weight X of substrate as described in calculating as III formula and pastille mucous layer:

$X=M-m---\left(\mathrm{III}\right).$

10. the detection method of recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 9, is characterized in that, melts and becomes after time limit detection, then carry out following at least one detection method:

1) swell value assay method is as follows:

A. first along radially the choosing a bit or some points of described recombinant human interferon-alpha 2 а vagina expansible plug end faces, measure after the initial length H of described expansion carrier, then the different angles of rolling measure several times, Hi averages;

B. described recombinant human interferon-alpha 2 а vagina expansible plugs melt after the detection of change time limit, are determined at the rear length h of selected location place's expansion carrier expansion in a step; The different angles of rolling are again measured several times, ask the rear average length hi that expands;

While c. calculating described swell value, according to I formula, calculate;

$\mathrm{pi}=\frac{\mathrm{hi}}{\mathrm{Hi}}---\left(I\right)$

Wherein, pi represent axial swell value, hi represent to expand after average length, Hi represent average initial length;

Or swell value assay method is as follows:

A. first an angle along described recombinant human interferon-alpha 2 а vagina expansible plugs axially choose a place or some positions, measure after the initial diameter R of described expansion carrier, then the different angles of rolling measure several times, Ri averages;

B. described recombinant human interferon-alpha 2 а vagina expansible plugs melt after the detection of change time limit, are determined at the rear diameter r of selected location place's expansion carrier expansion in a step; The different angles of rolling are again measured several times, ask the rear average diameter ri that expands;

While c. calculating described swell value, by II formula, calculate;

$P_i=\frac{\mathrm{ri}}{\mathrm{Ri}}---\left(\mathrm{II}\right)$

Wherein, Pi represent swell value, ri radially represent to expand after average diameter, Ri represent average initial diameter;

2) weight differential assay method:

A. get described recombinant human interferon-alpha 2 а vagina expansible plugs, take weight M;

B. scrape substrate and pastille mucous layer, expansion carrier is placed in to 20-90 ℃ of organic solvent, take out, in 50-150 ℃ of dry 1-10h, take weight m, according to the gross weight X of substrate and pastille mucous layer as described in the calculating of III formula

X=M-m?（Ⅲ）

Wherein, described organic solvent comprises one or more in ethanol, methanol or isopropyl alcohol.

The detection method of 11. recombinant human interferon-alpha 2 а vagina expansible plugs as claimed in claim 10, is characterized in that, described expansion carrier is cotton sliver, and described detection comprises following at least one method:

1) swell value assay method:

Get 3 of described recombinant human interferon-alpha 2 а vagina expansible plugs, with slide gauge, survey its afterbody cotton sliver diameter, roll approximately 90 ° and survey once again, survey twice for every, obtain 2 meansigma methods Ri of every mensuration; Above-mentioned 3 bolts are used for melting and become after the time limit measures and finish, take out immediately residue cotton sliver, treat disconnected of water, all gently be placed on glass plate, with slide gauge, measure two ends and middle three positions of each cotton sliver, roll after approximately 90 ° and measure three positions again, each cotton sliver obtains six data altogether, obtains 6 meansigma methods r of mensuration _i, calculate the swell value P of every _i, the swell value of three bolts all should be greater than 1.5;

2) weight differential assay method:

Get 10 of described recombinant human interferon-alpha 2 а vagina expansible plugs, the accurately weighed weight of difference, gently scrape substrate and pastille mucous layer (must not lose cotton sliver), cotton sliver is placed in to the 200-400ml ethanol of 50-80 ℃, preferably, cotton sliver is placed in to the 300ml ethanol of 60-70 ℃, and 80khz frequency ultrasonic cleaning 5 minutes, the remaining stromatolysis in cotton sliver surface is removed, taking out cotton sliver firmly extracts, with filter paper, inhale 3 times again, in 105 ℃, be dried 2 hours, take out, room temperature was placed after 1 hour, the accurately weighed cotton sliver weight of difference, obtain every containing substrate and pastille mucous layer gross weight and average substrate and pastille mucous layer gross weight with respect to total number, and the two is compared, what exceed average weight ± 10% must not be more than 2, and must not there is 1 times of 1 overrun.

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