CN102552888B - Suppository for treating mammal endometritis - Google Patents

Abstract

The invention belongs to the technical field of medicines, and specifically relates to a suppository for treating mammal endometritis. The preparation disclosed in the invention comprises the following components of: by weight, 0.004-0.01% of staphylococcus lysozyme, 0.1-4% of lysozyme, 10-80% of semisynthetic fatty glyceride, 1-20% of poloxamer 188(F68), 1-20% of HPMC, 5-50% of sodium bicarbonate and 5-50% of sodium dihydrogen phosphate. After the preparation provided by the invention enters into mammal uterus through administration, the preparation can undergo effervescence decomposition in a body fluid. The medicine is mixed in foams and rapidly released. Contact area between the medicine and mucosal of uterus becomes larger and the medicine can penetrate into deep portions of mucosa wrinkles so as to prolong the action time between the medicine and the mucosa, raise the concentration of the medicine on local tissues, further enhance the curative effect and effectively kill various pathogens which can cause endometritis.

Description

A kind of suppository being used for the treatment of mammalian endometritis

Technical field

The present invention relates to a kind of pharmaceutical preparation, specifically, relate to a kind of suppository being used for the treatment of mammalian endometritis.

Background technology

Endometritis is the commonly encountered diseases of mammal as milch cow, pig etc.The cause of disease is complicated, and sickness rate is high.The sickness rate of cow endometritis is between 15%-20%, and the sickness rate of this disease of pig farm is about 30%.Endometritis can cause involution of uterus time lengthening, makes the estrus cycle disorderly.Inflammatory exudate can change the acid-base value of intrauterine environment, is unfavorable for the existence of sperm and ovum, and conception rate is declined, and causes infertile, simultaneously also can long-time stimulus uterus inwall sensor, by reflection, hormonal regulation is lacked of proper care, causes ovarian disease.In addition also can secondary mastitis etc.

Treat endometritis at present in the world and all use antibiotic, but there is many unfavorable factors in antibiotic therapy.As (1) easily makes antibacterial produce drug resistance; (2) single medicine antimicrobial spectrum is wideless; (3) easily antibiotic remains is caused; (4) other side effect.In recent years due to antibiotic a large amount of use, Resistant strain constantly occurs, especially staphylococcus aureus produces drug resistance to Multiple Classes of Antibiotics, and drug resistance is obvious ascendant trend, is difficult to treatment.American-European countries just dropped into the medicine of a large amount of human and material resources research substitute antibiotics but there is no definite result so far before more than 40 years.

Staphylococcus lysozyme is a kind of bacteriolysin of gram positive bacteria, antibacterial can be killed by cracking antibacterial rapidly, action time is short, antibacterial is not easy to produce drug resistance, bactericidal effect is good simultaneously, has stronger sterilizing ability equally to the staphylococcus aureus of methicillin-resistance (MRSA).

1991, the bovine mastitis that the Oldham ER etc. of Agricultural Research Institute of Princeton Cyanamid company causes with dissolving staphylococcal bacteria enzyme treatment drug-resistant S. aureus, demonstrated the effectiveness of staphylococcus lysozyme for this disease for the treatment of.Next year causes the immunoreation in body when they have studied again local use restructuring dissolving staphylococcal bacteria enzyme treatment bovine mastitis, think that restructuring staphylococcus lysozyme is effective and safe as injection in body, and propose staphylococcus lysozyme and can substitute traditional antibiotic.

Lysozyme is a kind of alkaline enzyme that can be hydrolyzed mucopolysaccharide in pathogenic bacterium.Mainly through the β-Isosorbide-5-Nitrae glycosidic bond between the-acetylmuramic acid in destruction cell wall and NAG, make the insoluble mucopolysaccharide of cell wall resolve into solubility glycopeptide, cause cell wall rupture content overflow and make bacterolysis.Lysozyme also directly can be combined with electronegative virus protein, forms double salt, make virally inactivated with DNA, RNA, apoprotein.The narrow spectrum cell wall acting on order geomicrobes of energy, has antibacterial action to gram positive bacteria, aerobic sporeformer, bacillus subtilis, Bacillus licheniformis etc.

Make compound enzyme after lysozyme and staphylococcus lysozyme being mixed by a certain percentage, there is wider fungicidal spectrum, press down bactericidal effect and be better than single enzyme.Chinese patent ZL03142299.3 (a kind of preparation for prevention and therapy cow endometritis and preparation method thereof), disclose with staphylococcus lysozyme, lysozyme, bacteriostatic peptide and the medically preparation made of acceptable carrier, can prevention and therapy cow endometritis.

Chinese patent ZL200510028617.5 (a kind of lysostaphin freeze dried powder for preventing and treating cattle endometritis), discloses and adopts lysostaphin freeze dried powder treatment cow endometritis and preparation method thereof.But freeze dried powder needs to redissolve with water before administration temporarily, administration is comparatively inconvenient.And with the administration of aqueous solution mode, medicinal liquid viscosity is little, not easily with Uterine mucosa effect, be namely excreted at short notice.

Summary of the invention

Technical problem to be solved by this invention is to provide a kind of biological antibiotic effervescent suppository for mammalian endometritis, to overcome the use inconvenience that prior art exists, repeatedly with medicine cause uterine involution and be affected in excessive flushing uterus in treatment, intrauterine drug distribution area is little, remaining time is short, the interaction of liquefying with mucosa is weak, is unfavorable for the shortcoming penetrating into Uterine mucosa.

Design of the present invention is such:

Type of effervescent suppository belongs to a kind of special form in suppository, is mixed by medicine and substrate, appropriate amount of auxiliary materials, surfactant and soda acid system (gas-producing disintegrant).Type of effervescent suppository meets body fluid or water, and bronsted lowry acids and bases bronsted lowry chemically reactive wherein produces carbon dioxide and makes the disintegrate of type of effervescent suppository effervescent, dissolves release, and forms foam with Action of Surfactant.Originally in suppository surface shallow-layer generation neutralization reaction, the carbon dioxide effusion of generation makes it form a lot of hole, is beneficial to moisture and gos deep into suppository internal layer, makes soda acid system continue reaction and produces great amount of carbon dioxide.Form certain pressure on the one hand, make other disintegrating agents expand and make suppository spalling, carbon dioxide effusion on the other hand, is conducive to moisture and continues to enter suppository inside, impels the disintegrate of whole suppository effervescent within a short period of time, dissolves release.

The substrate of suppository is divided into water solublity and oil-soluble.In Screening matrix, consider that water-less environment is conducive to the long-term maintenance of enzymatic activity, and water-soluble base more or less needs to add moisture in preparation process, the present invention have selected and keeps more favorably oil-soluble substrate for a long time to enzymatic activity.It is lower that semi-synthetic fatty acid glyceride has cost as the most frequently used suppository oil-soluble substrate, and the advantages such as fusing point is constant are therefore selected.

Find when screening compound enzyme type of effervescent suppository soda acid system, the activity of the acid-base pair enzyme that great majority are conventional has inhibitory action, may be that salting out or foaming process too acutely cause protein structure to change causing the inactivation of enzyme.And surfactant is also affect one of albumen qualitative factor.Tween 80 is the most frequently used surfactant of effervescent formulation, has consumption few, bubbles rapidly, the fine and smooth lasting advantage of foam.But find in the process of screening adjuvant, tween 80 can reduce the activity of compound enzyme.For keeping the activity of enzyme in effervescent process, ensure drug effect, the present invention has carried out investigating to disintegrate system and surfactant and has introduced the adjuvant of the current less application in effervescent formulation of HPMC and F68 these two kinds.

Hydroxypropyl emthylcellulose (HPMC) is cellulose derivative, and be applied to the history that food and medicament have more than 40 years, recording in American Pharmacopeia 19-23 version, is the pharmaceutical polymers with premium properties comparatively widely.HPMC is generally used as thickening agent, film coating agent, slow releasing preparation pore material, hydrophilic gel in a medicament, also can make solid dispersal agent material, with the bioavailability etc. of the stability and insoluble drug that improve medicine.In the production of effervescent formulation, HPMC is often used as Effervescent tablet disintegration agent to accelerate drug release, in type of effervescent suppository, then rarely have use.In the present invention, HPMC is except accelerating except the disintegrate release of suppository as co-disintegrant, it can also promote the absorption of medicine as surfactant, increase the volume of foam and fine and smooth degree, thus increase the contact area of itself and Uterine mucosa, and the fold depths making it more easily infiltrate ordinary preparation not easily to arrive.HPMC, as a kind of macromolecular material, has the effect of certain maintenance protein stabilization, contributes to the activity keeping enzyme.And the stickiness that it has can make effective ingredient adhere to uterus inwall within the longer time, avoid being excreted too quickly.

PLURONICS F87 (F68) is commonly used for the lubricant of Emulsion, ointment and suspensoid at pharmaceutical field, the solubilizing agent of tablet or capsule and dispersant, also can be used as the carrier of solid dispersion, and be applied to intravenous uniquely artificial emulsifying agent at present, mean molecule quantity is 7680-9510.F68, as a kind of surfactant, also demonstrates unique effect in cytoprotective.The HLB value of F68 is 29, has stronger surface-active action.In the present invention, it can produce more foam as surfactant, can not only increase the contact area of medicine and mucosa, and can strengthen preparation to endometrial infiltration, strengthens the performance of its bactericidal action, strengthens the stability of enzyme simultaneously.In addition F68 have one be shaped as gel do use, the gel formed with HPMC combined effect can extend the time that compound enzyme retains in uterus.

Technical scheme of the present invention is as follows:

A kind of suppository for the treatment of mammalian endometritis, its composition and weight percentage comprise, staphylococcus lysozyme 0.004 ~ 0.01%, lysozyme 0.1 ~ 4%, semi-synthetic fatty acid glyceride 10 ~ 80%, PLURONICS F87 (F68) 1 ~ 20%, HPMC1 ~ 20%, sodium bicarbonate 5 ~ 50%, sodium dihydrogen phosphate 5 ~ 50%.

The suppository of the treatment mammalian endometritis addressed, the composition of its optimization formula and percentage by weight comprise staphylococcus lysozyme 0.004 ~ 0.008%, lysozyme 2 ~ 4%, semi-synthetic fatty acid glyceride 20 ~ 70%, PLURONICS F87 (F68) 2 ~ 10%, HPMC2 ~ 10%, sodium bicarbonate 10 ~ 30%, sodium dihydrogen phosphate 10 ~ 30%.

Concrete method for making is as follows:

Take staphylococcus lysozyme, lysozyme, F68, HPMC, sodium bicarbonate, sodium dihydrogen phosphate according to the above ratio.All materials are all pulverized, and cross 80 mesh sieves, and mixing, joins in the semi-synthetic fatty acid glyceride of 50 DEG C of meltings, pour rapidly molding in suppository mould into.

Staphylococcus lysozyme and lysozyme compound are prepared into effervescent suppository, intrauterine administration.Not only convenient drug administration, and compound enzyme type of effervescent suppository can effervescent disintegrate under the effect of body fluid, and medicament mixed is discharged fast in foam.Compared with freeze dried powder and ordinary suppository, the contact area of type of effervescent suppository Chinese medicine and mucosa is larger, and the surfactant wherein added and macromolecular material contribute to the contact increasing medicine and mucosa, make medicine can infiltrate mucosa fold deep, prolong drug and mucosa action time, improve local organization drug level, and then strengthen therapeutic effect, effectively kill the various pathogen causing endometritis.In addition, the acidic ingredients that effervescent suppository contains, can form the slant acidity environment suppressing anaerobe growth, more be conducive to killing anaerobe.Form gel after HPMC and F68 disintegrate, alive the stablizing and extend effective ingredient retention time in uterus of enzyme can be maintained, play the effect of medicine to greatest extent, obtain better therapeutic effect.

Staphylococcus lysozyme suppository disclosed by the invention has following features:

1, release rapidly, and at 37 DEG C, 15min release completely.The bubble produced is tiny and lasting, medicaments uniformity is distributed in a large amount of foam break up very to be soon distributed in each gap of endometrium, makes medicine play curative effect rapidly.

2, overcome enzyme and meet conventional disintegrating agents and the deactivated situation of surfactant, bactericidal effect is strong.Said preparation has very strong bactericidal action to mammalian endometritis pathogenic bacteria.Experiment in vitro can effectively kill gram positive bacteria and negative bacterium.

3, good stability.This product preserves 2 years at 25 DEG C of room temperature, and biological activity does not change substantially.Lyophilized powder is better than in the storage and transport of preparation.

4, the present invention is biological preparation, and overcome traditional chemical class preparation and have zest to Uterine mucosa, antibacterial easily produces the shortcoming of drug resistance.Easily accept by body, have no side effect.

5, the mode of topical is adopted, convenient drug administration.Other dissolving is not needed, convenient drug administration compared with lyophilized powder.

Detailed description of the invention

Embodiment 1 compound enzyme suppository preparation method 1 (formula 1)

1, composition and proportioning

Staphylococcus lysozyme 0.007%, lysozyme 0.1%, F6820%, HPMC 10%, sodium bicarbonate 10%, sodium dihydrogen phosphate 30%, semi-synthetic fatty acid glyceride 29.893%.

2, prepare

Take staphylococcus lysozyme 0.007g, lysozyme 0.1g, F6820g, HPMC 10g, sodium bicarbonate 10g%, sodium dihydrogen phosphate 30g.Above raw material pulverizing crosses 80 mesh sieves, and mixing, with 29.893g melting semi-synthetic fatty acid glyceride Homogeneous phase mixing, pours rapidly molding in suppository moulds into.

Embodiment 2 compound enzyme suppository preparation method 2 (formula 2)

1, composition and proportioning

Staphylococcus lysozyme 0.004%, lysozyme 2%, F6810%, HPMC 20%, sodium bicarbonate 20%, sodium dihydrogen phosphate 10%, semi-synthetic fatty acid glyceride 37.996%.

2, prepare

Take staphylococcus lysozyme 0.004g, lysozyme 2g, F6810g, HPMC 20g, sodium bicarbonate 20g%, sodium dihydrogen phosphate 10g.Above raw material pulverizing crosses 80 mesh sieves, and mixing, with 37.996g melting semi-synthetic fatty acid glyceride Homogeneous phase mixing, pours rapidly molding in suppository moulds into.

Embodiment 3 compound enzyme suppository preparation method 3 (formula 3)

1, composition and proportioning

Staphylococcus lysozyme 0.01%, lysozyme 4%, F681%, HPMC 1%, sodium bicarbonate 30%, sodium dihydrogen phosphate 20%, semi-synthetic fatty acid glyceride 43.99%.

2, prepare

Take staphylococcus lysozyme 0.01g, lysozyme 4g, F681g, HPMC 1g, sodium bicarbonate 30g%, sodium dihydrogen phosphate 20g.Above raw material pulverizing crosses 80 mesh sieves, and mixing, with 43.99g melting semi-synthetic fatty acid glyceride Homogeneous phase mixing, pours rapidly molding in suppository moulds into.

The Pharmacodynamics in vitro experiment of embodiment 4 suppository

1, experiment equipment

Experimental strain: staphylococcus aureus (ATCC 6538); Escherichia coli (ATCC 11229); Phosphate buffered solution (PBS 0.03mol/L pH7.2)

2, experimental technique

Each one piece of the suppository of Example 1, embodiment 2, embodiment 3 preparation, in 37 DEG C of water-bath releases.Take out and discard overlayer matrix and foam after cooling, get lower aqueous solution and become series concentration with MH broth dilution, to be measured.

Experimental bacteria 24h slant culture PBS is washed down, makes the bacteria suspension of 0.5 Maxwell than turbid standard, with MH meat soup, above-mentioned bacteria suspension is carried out 1: 100 dilution.Suppository is discharged gained series concentration solution with 1: 1 ratio mix with bacteria suspension.Final often pipe dissolving staphylococcal bacteria enzyme concentration is 10,5,2.5,1.25,0.625,0.31,0.15U/mL, bacteria suspension concentration is 5 × 105CFU/mL.Put 37 DEG C to cultivate 18 ~ 24 hours, observe clarity.

3, experimental result

Table 1 each formula compound enzyme effervescent suppository In Vitro Bacteriostasis effect

As can be seen from Table 1, each formula compound enzyme effervescent suppository all effectively can suppress staphylococcus aureus and colibacillary growth.

The stability experiment of embodiment 5 compound enzyme suppository

1, experiment equipment

Staphylococcus lysozyme standard substance (90U/mL); Color source substrate KNR-PG, for subsequent use with 0.2mol/L Gly-NaOH buffer (pH10.0) 1: 5 (m/v) suspension; 0.2mol/L Gly-NaOH buffer (pH10.0) for subsequent use; 0.05mol/L Tris-HCL diluent (pH7.5);

2, experimental technique

Get compound enzyme effervescent suppository to discharge in 37 DEG C of water-baths.Take out and discard overlayer matrix and foam after cooling, get lower aqueous solution and be diluted to debita spissitudo with water, treat after filtration to add reagent and sample with table 2 order.

Table 2 compound enzyme activity determination method

The centrifugal 10min of 10000rpm, get supernatant 595nm and sentence No. 0 pipe for blank mensuration absorbance, the optical path of cuvette is 1cm.

According to the absorbance A value recorded and corresponding standard enzyme concentration C (U/ml), drawing standard curve: A=KC+B

Wherein K is the slope of standard curve, and B is the intercept of standard curve, and A is absorbance, and C is the concentration (U/ml) of enzyme.

3, experimental result

Not there is significant change in the enzymatic activity after compound enzyme effervescent suppository deposits 24 months at 25 DEG C as can be seen from Table 3

Result is determined in the enzyme work that compound enzyme type of effervescent suppository after 24 months deposited by table 325 DEG C

Embodiment 6 compound enzyme suppository is to the action effect of pig endometritis

1, trial drug

Compound enzyme suppository; Antibiotic.

2, case load and case selection standard

(1) acute endometritis case: 24, puerperal or 10 days post-abortions, clinical symptoms is that the rising of disease temperature of pig body subtracts food or do not eat, be reluctant to stand up, with micturition, frequently exert duty, in company with exerting the bronzing mucus or purulent secretion of blaming from intravaginal elution band stink, what have accompanies afterbirth fragment.

(2) chronic endometritis case: 24, case source is the same as with acute endometritis case identical pig farm; Fail to respond to any medical treatment and ill more than 20 days sows through antibiotic therapy or other drug.

3, test grouping

The sow 48 being suffered from endometritis is divided into 4 groups at random, often organizes 12 (6 acute, 6 chronic), wherein 3 test group (formula 1, formula 2, formula 3); 1 drug control group, matched group medicament selection antibiotic (penicillin 1,600,000 unit, streptomycin 5,000,000 units/head/time).

4, medication

Normal saline flushing uterus is first used to tested sow.Wherein test group will require that the compound enzyme type of effervescent suppository of dosage drops into intrauterine.The next day, offers medicine 1 time, and being used in conjunction is for 3 times 1 course for the treatment of; Drug control group is by penicillin 1,600,000 unit, streptomycin 5,000,000 unit, and normal saline dilution becomes solution to inject, medication every day 1 time. and being used in conjunction is for 3 times 1 course for the treatment of.

5, efficacy determination method and standard

The laggard line correlation inspection course for the treatment of.

(1) cure inflammation of uterus to disappear, intrauterine is discharged without rheuminess thing and pus, and recover of uterus is normal, and secretions transparent clear, oestruses on schedule, normal pregnancy.

(2) effectively inflammation of uterus is obviously alleviated, and uterine secretion thing is close to normal, and bacteria distribution qualification cross-reference, main or fraction of pathogens bacterium is eliminated.

(3) after invalid medication, inflammation of uterus has no light and slow, and uterine secretion thing has no improvement even to be worsened, and bacteria distribution shows there are other pathogenic bacterial infections.

6, experimental result

Table 4 compound enzyme effervescent suppository group compares with antibiotic group therapeutic effect

As can be seen from Table 4, the antibiotic therapy method more traditional by compound enzyme suppository in treatment pig endometritis effect is significantly improved, cure rate the former comparatively the latter improve nearly 20 to 30 percentage points.

Claims (1)

1. one kind is used for the treatment of the suppository of mammalian endometritis, be characterised in that, by weight percentage, its composition comprises, staphylococcus lysozyme 0.004 ~ 0.008%, lysozyme 2 ~ 4%, semi-synthetic fatty acid glyceride 20 ~ 70%, PLURONICS F87 (F68) 2 ~ 10%, HPMC2 ~ 10%, sodium bicarbonate 10 ~ 30%, sodium dihydrogen phosphate 10 ~ 30%.