CN1035434A - The preparation method that is used for the pharmaceutical preparation of prevention of STD and treatment urethral trichomoniasis - Google Patents

Abstract

A kind of preparation method that is used for the pharmaceutical preparation of individual prophylaxis sexually transmitted disease (STD) and treatment urethral trichomoniasis, this method comprises, with effective dose nitro-alpha-chloro cinnamic aldehyde and dimethyl sulfoxide are mixed into the cooperative action mixture of solution shape, and be diluted to predetermined concentration with medicinal diluent.Using molecular weight is that 400 Polyethylene Glycol is as medicinal diluent.

Description

The preparation method that is used for the pharmaceutical preparation of prevention of STD and treatment urethral trichomoniasis

The present invention relates to medical domain, more precisely, relate to the preparation method of the pharmaceutical preparation that is used for individual prophylaxis sexually transmitted disease (STD) and treatment urethral trichomoniasis.

The pharmaceutical preparation that makes by the method for recommending clinical practice in Dermatology and gynaecopathia.

Now, at the medicine that medically is used for the individual prophylaxis sexually transmitted disease (STD), well-known have a silvery agent, potassium permanganate preparation and mercurial, mercuric chloride solution for example, calomel Ointment, " д у ь л о з а н " ointment.Existing people proposes to use " О к с и р и р д о Λ " preparation to prevent syphilis and gonorrhea, and this preparation is to use 0.035%(weight) contain mercuric oxycyanide (О к с и ц и а и с т а я р т у т ь) and green soap is dissolved in glycerol (accounting for 50%(weight in glycerol)) in and make (М. Х. ъ e р г о ь ц "

" nineteen fifty-two,

).But this preparation does not have enough bactericidal activities for the pathogen of syphilis and gonorrhea, and it has stronger stimulation and can cause the desquamation and the dissolving of epithelium appearance mucous membrane of urethra, is unsafe clinically therefore.Because above-mentioned preparation lacks enough reliabilities, and that some mercurial can not guarantee is harmless, so these preparations all do not obtain practical application widely.

In recent years, the someone proposes chlorhexidine (Х Λ о р г e к с и д и н) (chlohexidine) solution with 0.05～0.1% as being used for the pharmaceutical preparation of prevention of STD.Test of carrying out in vitro and the test of carrying out on one's body animal show that all above-mentioned this preparation has enough bactericidal activities (в e с т н и к д e р м а т о

о г и и и в e н e р о

о г и и, 1978 the 6th phases; 49 pages).But,, find that it has stimulation and can make it become dry people's skin and sexual organ's mucosa when this preparation being applied to when clinical.Under working condition, the preparation of chlohexidine is very complicated, and requires a large amount of labour expenditures.

The external used medicine that is used for the treatment of urethral trichomoniasis has boric acid, silver nitrate, and zinc sulfate, copper sulfate, chloromycetin and acetarsol ( ) etc.The effect of these preparations is good inadequately, therefore also is not widely used.

Now, the most effective medicine that is used for the treatment of these diseases is the imidazoles preparation, for example fasigyne (

), metronidazole ( ),

But, these medicinal applications when clinical, find that infusorian produces more and more big Drug resistance to the effect of these medicines, therefore, this causes the recurrence (reaching 8.4%) in certain stage of the state of an illness and can cause the trichomoniasis sequela, these preparations can cause side effect, when hemopoietic system destroys and during the central nervous system activities morbidity, and the use that is under an embargo of these medicines.

Basic task of the present invention is the method that will work out a kind of like this pharmaceutical preparation of preparation, said this medicine medicine form easily is used for the individual prophylaxis of sexually transmitted disease (STD) and to the treatment of trichomoniasis, this medicine has very high curative effect and can not cause side effect when clinical practice.

For solving described task, proposed to be used for the preparation method of the pharmaceutical preparation of prevention of STD and treatment urethral trichomoniasis, according to the present invention, the preparation of this kind preparation be with effective dose to nitro-alpha-chloro cinnamic aldehyde

Be mixed into the cooperative action mixture of solution shape with dimethyl sulfoxide, and be diluted to predetermined concentration with medicinal diluent.

This kind method can guarantee to obtain the preparation of efficient prevention of STD of a kind of energy and treatment urethral trichomoniasis.

These have the various mechanism of action by the above-mentioned Chemical composition that active component mixes, these compositionss play the potentiation synergism on the whole, it can cause syphilis, the destruction fully of the protoplasm of the pathogen of gonorrhea and trichomoniasis and cell wall, and can guarantee that the preparation that is obtained has sterilization and kills the protozoacide ability.Under the effect of this preparation, as long as pathogen is decomposed.

The technology of this method is simple, and is convenient, and do not need a large amount of labour expenditures under the condition of industrial-scale production.All components all are harmless to the operator that produce.

Using molecular weight is that 400 Polyethylene Glycol is fit in the method for the invention as pharmaceutical diluents, uses above-mentionedly to nitro-alpha-chloro cinnamic aldehyde, and dimethyl sulfoxide and molecular weight are that 400 the suitable weight ratio of Polyethylene Glycol is 0.3: 5.0: 94.7.

The preparation that has this diluent does not have partial stimulation to skin and mucosa top layer.This diluent does not have toxicity, and is osmo active substance, thereby can impel active component to manifest maximum biological activity.But, also can use some other medicines diluent commonly used for this purpose, glycerol for example, dipropylene glycol, 1,2-propylene glycol.Press the preparation of the liquid medicine form of said component ratio preparation, when these medicines can have excellent contact with the trichomoniasis pathogen of infiltrating skin and mucosa, just have the reliability of height preventive effect and the therapeutic activity of anti-trichomonal.

Become aerocolloidal form when acting on the preparation of liquid condition with pressure for pharmaceutical preparation is formed in, preferably toward by 0.3%(weight) above-mentioned to nitro-alpha-chloro cinnamic aldehyde and 5%(weight) dimethyl sulfoxide and 10～30%(weight) molecular weight be to be added into 3.0～6.0%(weight in the solution formed of 400 Polyethylene Glycol) surfactant and water, make total amount reach 100%(weight), come the thoroughly saturated solution that is obtained that is contained in the aerosol container with propylene, the consumption of propylene is about 3～10% of solution weight.The selection of these material requirements is to obtain the rapid destructive foam of energy when using.

This method can guarantee to obtain a kind of like this preparation of medicament forms, and it can not only guarantee antiveneral ability in the case of necessary, and can guarantee individually the women of needs treatment urethral trichomoniasis is treated.

The preparation of being recommended is a kind of transparency liquid of viscosity, and its color has faint special odor from pale yellow to deep yellow, and it is stable when preserving in the place of lucifuge.

Said preparation can be sold with a kind of 5～10 milliliters form of the little phial of sealing or the form of pressurized package.

According to the present invention, we have carried out the preceding test of clinical practice with said preparation, comprise with test tube with experiment with syphilis rabbit being tested with the sterilization of research preparation and the activity of killing protozoa; The safety of research said preparation, comprising research its acute and chronic toxicity, partial stimulation, anaphylaxis, to immune influence, with and some aspect of pharmacological action.

We are that 0.075%, 0.15% and 0.3% preparation is studied to concentration.In preparation, the content to nitro-alpha-chloro cinnamic aldehyde with above-mentioned chemical formula is to measure like this.Promptly the content of dimethyl sulfoxide is immobilized (5%), and be 400 polyethyleneglycol content equivalent variation with molecular weight.Study the safety of preparation with 0.3% concentration.

In order to study the gonococcida ability of said preparation, isolate pure gonococcus culture on one's body and be inoculated on the large-area culture medium (" cotton-padded mattress ") from the gonorrhea patient.Chloride isosmotic solution with sterilization comes to wash circadian culture from cotton-padded mattress, obtains a kind of suspended substance from eluate, wherein contains 10 ⁹Individual microbial body (by " standard turbidity ").In the test tube of each sterilization, add 0.5 milliliter of suspension that is obtained respectively, respectively add the preparation of being studied of 0.5 milliliter of double strength again, in order that in each component the later acquisition of mixing needed (research) concentration through 5 minutes.This mixture was carried out centrifugalize 10～15 minutes (on the flush away gonococcus with preparation) with the rotating speed that per minute 1500～3000 changes.Remove supernatant, on precipitate, add 1 milliliter of sodium chloride isosmotic solution (reaching initial volume) again and carry out the second time centrifugal (washing once again).Remove supernatant, precipitate is inoculated on the tilting agar culture medium (handles the culture of each part gonococcal strain with the culture medium of 10 test tubes).The test tube that culture medium is housed in calorstat under 37 ℃ a kind of have in the atmosphere that improves carbonic acid gas content place.4 round the clock during in measured the propagation situation of a gonococcus culture in per 24 hours.

The suspension of the gonococcus culture of use equal volume (0.5 milliliter) and the mixed liquor of sodium chloride isosmotic solution are tested in contrast, once more this solution branch is installed in each test tube, identical as the situation in the test tube test, centrifugalize is inoculated into precipitation in the identical medium of composition.

Add caseinhydrolysate, do not have the yeast cells slurry of antiseptic and have serum of angle draught animal and lyophily medium to make meat peptone agar with underground mutton as culture medium.

Study of the effect of said preparation with following method to treponema pallidum and trichomonas vaginitis in the test tube test:

Obtain treponema pallidum on one's body from a kind of patient who has suffered from Te Zhong testis inflammation 8～10 days rabbit and suffered from infectiousness syphilis.The Buddhist nun is restrained tired (н и к о Λ о с) and № 8

The tissue culture of (ц в к и) bacterial strain is studied.Isolate the trichomonas vaginitis culture on one's body from the urethral trichomoniasis patient, and with synthetical culture medium ( ) cultivate.

The method of experiment comprises, suspension of equal volume treponema pallidum (or infusorian culture fluid) and the formulation soln that will study are mixed, and the concentration of adding preparation should be required twice, because its concentration will drop to 1/2 of original content after mixing.By the preparation of this mixture from the preparation of making, and in dark colour (to treponema pallidum and infusorian) or carry out microscopic examination on one contrasty (to trichomonas vaginitis) microscope microscope carrier mutually.According to the mobility of pathogen and its form effect and speed are estimated the effect of institute's test preparation.Can be used as the following several situations of having of sure evaluation: pathogen (refers in the moment of allocating preparation into) lose mobility at once, perhaps reached in about 3-5 minute concerning treponema and lost mobility in 5～10 minutes concerning infusorian.Can think that to the effect of institute's test preparation more definite results is meant that preparation can change the form of treponema and infusorian, promptly during above-mentioned, morbific pathogen loses its form, and (treponema pallidum attenuates, the wreath spread apart, present expansible individuality, part, perhaps dissolving fully; Trichomonas vaginitis loses flagellum, and is cracked, dissolving).

Other research approaches to the treponemicidal effect of institute's test preparation comprise in turn toward a suspension and a tested medicine that adds a treponema pallidum on glass; Tip with the pasteurization dropper carefully stirs the mixture, and covers and observes on visual microscopical dark microscope carrier immediately with a cover glass.This research makes after being contacted about 20～30 seconds with suspension by medicine can demonstrate this medicine to treponemal effect.In order to contrast, test in contrast with isopyknic treponema pallidum (20～30 to 80 thalline are arranged in the visual field) or the float of trichomonas vaginitis and the mixture of sodium chloride isosmotic solution with these experiments."

" on the microscopical dark microscope carrier and the microscope that produces by means of " Opton " company microscopic examination is carried out in the effect of said preparation, this kind microscope have dark microscope carrier ( ) and have condenser lens and the object lens (25 * 0.45 that differ; 40 * 0.60).

In order to study the preventive effect of said preparation, in the test of experimental syphilis, utilize that some are sexually matured, nutrition good, the variety classes of body weight between 2.5 to 4 kilograms and the male rabbit of hair color (Chilean rabbit, white, Lycoperdon polymorphum Vitt or silvery white).

In order to foreclose, before experimentizing, all rabbits are all observed more than one month earlier suffering from spontaneous spirochetotic rabbit.All these rabbits all used the complex to the syphilis seroreaction to carry out observation before experimentizing.

In the experimental syphilis test of rabbit, study the preventive effect of said preparation with two kinds of parallel experimental programs.

First kind of scheme is included in the go out treponemal effect of the mixture of subcutaneous injection treponema pallidum suspension and said preparation with the research said preparation.For this purpose, with known conventional method experiment is carried out subcutaneous test with rabbit, in the halves of the scrotum of each rabbit, each injects 0.75 milliliter the mixture of being formed by equal-volume by test preparation by treponema pallidum suspension and double strength (because the concentration of mixing afterwards them with suspension is all corresponding drop to original 1/2).

After stirring 10 minutes, immediately this mixture is injected in one group of rabbit body the said components contact and with the sterilization Glass rod.Said mixture and equal-volume sterilization sodium chloride isosmotic solution contact 10 minutes and in addition after centrifugal (for the flush away preparation), injecting another then and organize in rabbit body.

The purpose of carrying out these experiments is the probabilities in order to get rid of inactivation (precipitation) in the protein medium of said preparation in animal body and therefore to lose its sterilization effect.In the mode of centrifugal back injection treponema suspension, determined through it the treponema pallidum effect is just being reached enough treponemal effects of killing after 10 minutes when said preparation.For the rabbit that compares usefulness, with the scrotum position of the subcutaneous injection of the equal-volume mixture of the suspension of treponema pallidum and sodium chloride isosmotic solution rabbit.

The basic scheme of second kind of experiment comprises carries out such experiment, will make the condition of hare infection syphilis will approach the condition of human infection's syphilis under field conditions (factors) to a certain extent in this experiment as far as possible.For this reason, the treponema pallidum suspension that is dissolved in the sodium chloride isosmotic solution is embrocated from the skin galling of scrotum, so that rabbit infects.For this reason, in advance a cotton plug is dipped in ethanol, and then dip in ether, it comes the skin of wiping scrotum reuse.Then, left-handed finger farthest launches the here skin of scrotum, and simultaneously, the right hand in turn frays the skin of scrotum two halves with fine sandpaper.By about 1～1.5 centimetre of the area of galling place ², should note making the skin surface roughening that is rubbed to reach the local degree that small number of groups is knitted liquid that flows out during wiping.It is hemorrhage that attention should be avoided.Again prepare the suspension of syphilis substance then, its content is reached in the microscopical visual field be issued to 20～25 treponema pallidums, get 0.1 milliliter of galling place that is added to skin of scrotum then, and rubbed 1～1.5 minute with the glass scuppit of a sterilization.Then, be added with the infective agent place and be not less than 0.5 centimetre place and clamp leaving with two neaH anchor clamps, make two mutual decussations of clip, in order that make skin form wrinkle, can avoid like this treponema drying (in order to form best infectious condition-according to С. Т. the narration of П а в Λ о в).Two clips were clamped about 1～1.5 hour across, then clip is taken away.

After underproof rabbit infects processing through different interval (through 15 minutes, 1 hour, 2 hours and 3 hours), then to the rabbit disinfection.This each the infected position that comprises past skin of scrotum of disinfecting adds that 0.1 milliliter of underproof preparation also carefully rubbed 1～1.5 minute with the glass scuppit.The rabbit that is used for contrast is not then disinfected.

Laboratory animal was carried out a clinical observation weekly and used the complex to the syphilis seroreaction to carry out serum test in every month in 4-12 month, said serum reverse should have: the antigenic В а of cuorin and treponema с с e р м а н reaction, the precipitation of К а н and 3 а к с-В ц т e ъ с к и й, immunofluorescence reaction (Р И φ and Р И φ-200) and microprecipitation reaction (Р М П).

The evaluation rabbit is according to the following fact to the reliability of the preventive effect of syphilis, promptly do not occur morbific clinical picture and seroreaction on one's body and show as feminine gender, and transplant to the indication rabbit result that is negative from the lymph node of the rabbit Shen Shang popliteal portion of being tested in going down to posterity in succession according to secondary rabbit.Final result shows, has obtained definite results when inoculating to underproof and rabbit that go down to posterity with homologous treponema pallidum bacterial strain.

Employed in all experiments all is the suspension of freshly prepd н и к о Λ ъ с treponema pallidum bacterial strain.

Below provided experimental studies results to the sp act of said preparation.

Preparation is to the gonococcal sterilization functions of н e й с с e р: use from 7 aegs and 5 isolated on one's body 12 groups of pathogen bacterial strains of Aeg to experimentize, experimental result shows that said preparation has suppressed the vital functions of gonococcus culture fully.In all test tube, the neither one test tube demonstrates the gonococcus culture and obtains increasing, and increases significantly and observe culture in all contrast test tubes.

The treponemicidal activity of preparation: experiment shows, treponema pallidum moment is promptly lost vigor, simultaneously when preparation with can be in 3～4 minutes after the syphilis substance contacts syphilis substance destruction up to dissolving fully.Ruined treponemal discrete fragment has appearred in first minute after contact, the amorphous proteinic colony of the fine-grained form be made up of dissolved pathogen then appears in rectangular motionless " silk " and " band " (because spring is rolled up unfolded result) that has attenuated in the situation of most preparations.Under the influence of preparation, no matter be that the н и к о Λ ъ с bacterial strain or the treponema pallidum of No. 8 Ц К В И bacterial strains all are subjected to same destruction.The treponemicidal activity of preparation concerning all from the direct on one's body isolated pathogen of infectiousness patients with syphilis all without any the difference on the degree.The test that comes rabbit is done with syphilis with experiment has also obtained treponemicidal effect.In these experiments, 120 buck have been used altogether.To skin galling place at rabbit De testis scrotum position introduce behind the infected material 15 minutes to 3 hours during in said preparation Prevention Processing is carried out at the position of infection, the result, animal reaches 100% to the prevention of syphilis.Simultaneously, using treponema pallidum rabbit to be carried out the experiment of subcutaneous infection, also obtained to prevent the positive result of syphilis at the equal-volume mixture of the suspension of sodium chloride isosmotic solution and said preparation.

The result shows, the experiment rabbit is fully reliably to the prevention of syphilis, in these rabbits, both do not found ill clinical symptoms, seroreaction to syphilis be negative (animal was observed 4 to 12 months) again, and, in twice continuous passage, transplant to the indication rabbit result that is negative and (transplant to first generation indication rabbit from the experiment rabbit from experiment rabbit popliteal portion lymph node on one's body, transplant to second filial generation indication rabbit from first generation indication rabbit again), the seroreaction of syphilis also is negative.Experiment showed, that the homology bacterial strain with treponema pallidum inoculates,, still concerning the rabbit of going down to posterity, all obtained definite results no matter be to experiment rabbit itself.

Rabbit proves the prevention ability of syphilis under the situation of the experiment rabbit being carried out subcutaneous infection, as long as preparation contacts 10 minutes with treponema pallidum, just is enough to eliminate fully the vitality of syphilis substance.Show also that on the other hand preparation is stable equally in as the protein medium of rabbit organic organization.

Preparation is to the effect of trichomonal vaginitis (trichomonas Vaginales): use from suffering from 13 aegs and 3 totally 16 kinds of trichomonas vaginitis bacterial strains that the Aeg separates of urethral trichomoniasis on one's body, in test tube, experimentize, the result shows that preparation has very strong protozoa activity extremely (having carried out 256 the microscopic examination research to preparation).In first minute under the effect of preparation, trichomonas vaginitis has just lost the mobility and the flagellum that come off.In 2-3 minute, protozoa is just cracked fully afterwards, only in discrete microscopic examination to preparation, finds to have the last cracked infusorian that does not have flagellum of the discrete ability of losing vigor.

In order to study the safety of preparation, we have measured its acute and chronic toxicity, and it is to local excitation and its anaphylaxis and the immunization of skin and mucosa.Simultaneously, also studied the pharmacological action that preparation is applied to skin and is produced when making it enter mucosa.

Four kinds of animals have been carried out the determination of acute toxicity of preparation.Utilize following four kinds of animals to carry out four groups of experiments, they are: 150 average weights are the white mice of 18.5 grams.18 rat (165 gram), 18 rabbits (3000 gram) and 12 Cavia porcelluss (400 gram).When calculating, considered in the future it is applied to human situation (concerning average weight was 70 kilograms people, each kg body weight was 0.07 milliliter) by the dose of test preparation.

Consider the characteristics (wipe in skin and introducing vagina) of preparation when practical application, studied the toxicity of Ц и д п о Λ preparation, except carrying out subcutaneous injection (to rat, white mice) outside, also carry out injecting in the direct body-to female injection vagina, to male injection rectum (to rat, Cavia porcellus and rabbit).When using tested medicine, constantly the laboratory animal what happens is carried out perusal in round the clock at 14.

With two kinds of laboratory animals Ц и д и п о Λ preparation is carried out chronic toxicity research: 24 body weight are the rat of 170～200 grams and the Chinchilla rabbit that 24 body weight are 2.5～3.0 kilograms.Dispensing every day in during one month.Because consider the purpose that will be applied to preparation to prevent, so the dosage of the single dose in the current experiment when carrying out acute toxicity test is identical.

To throwing the preparation of above-mentioned therapeutic dose in one group of laboratory animal (each 6 of rabbit and rat) is during one month every day.Two kinds of animals (each 6) to other group experiment, throw the preparation of inferior toxicity dosage every day in a fortnight, promptly 10 times to therapeutic dose (0.7 milliliter/kilogram), during the 3rd week to one month finishing this, to the used dose of rabbit is 2.1 milliliters/kilogram, and the time rat be 3.5 milliliters/kilogram.The dosage of this back length is equivalent to 30 to 50 times of therapeutic dose.The rabbit of another group experiment and rat (each 6) as a comparison, they are not thrown preparation but throw the sodium chloride isosmotic solution.Approach situation as much as possible for the method for laboratory animal administered formulation to people's medication.To jenny is that the preparation of 1/3 dosage is injected vagina and rectum, and the while, the preparation of 2/3 dosage was added to the inside surface of shank and abdominal surface takes out on the skin of fine hair, and then rubbed 1 minute; To buck then is that 1/3 dosage is injected rectum, 2/3 dosage is added on the skin on the inside surface of scrotum, abdominal part and shank is also rubbed simultaneously.

Emphasis ground observation its total state and behavior in during observing animal whole, and the variation of mensuration material and the temperature of rectum.Studied of the influence of said preparation to following various organ functions, these internal organs comprise: central nervous system (Ц Н С), cardiovascular system, respiratory system, liver, kidney, endocrine gland, blood comprise the Biochemical Research of having carried out the heart, lung, liver, kidney and endocrine gland.When one month dispensing process finishes, carry out study of histological morphology with animal slaughtering and to following internal organs, these internal organs comprise: brain, spinal cord, heart, lung, blood vessel, liver, kidney, pancreas, gonad, blood and hemopoietic organ (spleen and lymph node).In addition, also to the mucosa obliterating of skin and vagina and rectum the position of preparation study, and measured the weight coefficient of internal organs.Jede Woche is measured the temperature of once intravital material and rectum.Other physiologic index is all noted respectively before experiment and after the experiment.Biochemical research is carried out three times altogether, promptly respectively before experiment, after 2 weeks and after 1 month (promptly when experiment finishes).

Chronic toxicity index to animal comprises: the state of appearance and behavior; The variation of body temperature and body substances; Physiological test (with " open place " method, lid Ke Senna anesthesia test, α KT test method(s)); The clinical research of urine; The end is the composition morphological indexes of blood slightly; The biochemical indicator of blood (total mensuration, direct and indirect bilirubin, kreatinin, carbamide, residual nitrogen, transaminase, alkalescence Activity, serum cholinesterase activity, alpha-amylase activity, total protein, Herba thymi vulgaris reagent reacting; Mensuration has the albumen of C-reaction); Measure trace element: potassium, sodium, calcium, chlorine; The pathomorphology research of the inner internal organs of animal.

Method according to " open place " has been studied neural function vigor, in 3 minutes time according to run, bathing and the number of times of standing write down motion vigor and the behavior reaction of rat.In addition, injected by 70.0 milligrams/kilogram dosage 0.7% lid Ke Saina ( ) solution and the lid Ke Saina anesthesia test of carrying out, estimated the functional status of central nervous system (Ц Н С) according to the time length of the clothes phase of diving.

By means of З Λ К А Р 087 galvo-recorder aroused in interest that has the output of two-way standard biological electric current, write down the electric vigor of heart, the speed of used paper-feeding mechanism is 50 mm/second.When handling electrocardiogram, measured systaltic frequency, the zigzag magnitude of voltage of G, R, S, T in addition, has also been write down in complete heartbeat circulation interval R-R, GRS and the gap of GRST.

After supply water (per 100 gram animal weight with 5 milliliters of sodium chloride isosmotic solution), the volume of the urine of arranging according to diel and the proportion of urine come the drain function and the concentrating function of evaluation of renal.According to the concentration of residual nitrogen, the concentration of urine and the denitrogen function of coming evaluation of renal according to the content of kreatinin in the serum of blood.

The blood sample of rat is to obtain from the tail end that breaks apart by chopping, and the blood sample of rabbit is to obtain from the vein at ear edge.The content of haemachrome is to utilize the reagent of one group of standard to carry out photometric according to unified hemoglobin cyaniding.It is photometric that erythrocytic content is that usefulness-56M instrument carries out: carry out the mensuration of erythrocyte absolute magnitude in advance in the darkroom of φ З К instrument with photometry, the optical density of 0.1 unit is equivalent to have 1,217 hundred ten thousand erythrocyte in 0.01 milliliter of blood.Erythrocytic sedimentation velocity, leukocytic total amount and leukocytic kind method is routinely measured.The content of the bilirubin in the blood, creatine liver, carbamide, transaminase, total protein and C-reactive protein uses one group of standard reagent to measure according to unified method.

Be the research Pathomorphology, internal's sample is to select from the animal (rat) that is breaked end or from the animal (rabbit) that is killed by secluding air.Sample specimen is kept in 10% the neutral formalin, and uses hematoxylin and eosin stain.Material and its weight coefficient of internal have been measured simultaneously.

Carried out the research of preparation with three groups of tests to the local irritant effect of skin and mucosa.35 Cavia porcelluss and 30 Chilean rabbits are used in first group of test, the preparation wiping are gone into to take out on that part of skin of fine hair, and splash in two the conjunctiva gap (in 10 days, dripping every day once to twice).

Having studied the local irritant effect of preparation to rabbit eyes mucosa in second group of test (8 rabbits), is 10 times of recommended density for the concentration of the used preparation of consideration of prevention.As first group of test, preparation is splashed into two, drip once or twice every day in 10 days, is spaced apart 2-3 hour.

Studied the local irritant effect of preparation in the 3rd group of test (when studying chronic poisoning), used condition approaches the condition of practical application as much as possible.Use rabbit (12) and white mice (12) to test, during one month in every day be added to preparation on the skin and injection vagina (to female) and rectum (to male).

Except every day the behavior of animal and reaction that preparation and skin and mucosa rose being observed, after off-test, take off and be subjected to that part of of preparation effect in skin and the mucosa, so that carry out histological research.

Come the allergy effect of preparation is studied with Cavia porcellus because this animal to study artificial irritated effect be a kind of optimal object (А а о А. д., 1970).To the animal of experimental group, six round the clock during in every day embrocate skin with preparation, perhaps inject in the body cavity (female injection vagina, male injection rectum) with the syringe holder preparation.The dosage of preparation equals to treat consumption (0.07 milligram/kilogram) or is equivalent to treat 10 times of consumptions and 15 times (0.7 and 1.1 milligram/kilogram).During 21～23 days, use distinguishable dosage, promptly equal to cause the total amount of irritated effect.24 Cavia porcelluss (6 every group, totally four groups, wherein one group is matched group) in experiment, have been used.About the judgement of irritated usage degree, can be according to the more observed features of visual energy (congested, as to play phlycten, skin and mucosa infiltration, not exclusively suitable animal behavior, abnormal gait ， Zhan Piao etc.).Also can be according to the immunologic assay of laboratory; The acidophil in the blood and the variation of basophilic leukocyte number slightly of the test of leukocytic agglomeration reaction, leukopenia, thrombocytopenia index and blood vessel end.

According to the above-mentioned result that research method obtained, the percentage rate of leukocyte agglomeration reaction increases more than 30%, at 1mm ³Leukocytic decreased number is 1000 in the blood, and the thrombocytopenia index increases more than 22%, no matter be the number to acidophil and basophilic leukocyte, still to by the caused irritated effect of preparation, all estimates.

In the research chronic poisoning, preparation and Main Ingredients and Appearance thereof have been studied to immune influence with 24 rabbits, (four treated animals, 6 every group, wherein a group is matched group).Immunologic research is carried out twice altogether; In when beginning, determine the original level of index earlier, then, after promptly finishing after through one month, measure the key component of inferior toxicity dosage particles with therapeutic dose and inferior toxicity dosage administered formulation.As follows to the test that the feature of T-and the B-immune factor and the non-special protection factor is done: the level of C, A and M immunoglobulin like protein in the quantitative assay serum (uses the radioimmunoassay method (to press Mancini; 1956)); the change of lymphocyte and mononuclear cell number; the alexinic titer of serum (colorimetry); have a liking for the titer (П а у Λ ь-Ъ у н н e Λ я reaction) of different in nature agglutinin; the C-reactive protein amount and the total protein concentration of serum, Pi is with the Histological research of popliteal portion lymph node.

Because preparation is used to part (skin and mucosa), therefore measure from skin and the probability of mucosa picked-up preparation and be determined at that the metabolic probability of preparation is very important in the human body.The content of preparation in blood is to measure with highly sensitive gas phase liquid chromatography, and the formulation concentrations that this method can be measured is 10 ^-6～10 ^-9Grams per milliliter.

The statistical disposition of data is carried out with the EC-1045 electronic computer.

Above-mentioned result of the test is as follows:

In giving the white mice abdomen ejection preparation with the condition of carrying out acute poisoning test under, the dosage of observing the lightest toxicity performance is 8.32 milligrams of/kilogram animal weight, and the dosage of the heaviest toxicity performance is 20.0 milligrams of/kilogram animals.The data that the acute poisoning institute is obtained is according to Ъ. М. and the method for (1980) such as Ш т а ъ с к и й is carried out statistical procedures.Under the situation of carrying out injecting in the abdomen to white mice, the half lethal dose of this preparation is 9.81 milligrams/kilogram (9.67～9.94), toxic grade partitioning according to С и д о р о в K.K, can put this preparation under relatively nontoxic one-level, this belongs to the toxic grade of minimum level (и з м e р о в etc., 1977).

The result of said preparation chronic poisoning research proves: in one month time, throw the preparation of treatment with dosage and inferior toxicity dosage every day, the internal organs of rabbit and rat and the functional status of human body are not caused injurious effects.During to off-test, the index level of being studied or match with raw data, perhaps consistent with the corresponding index of matched group, do not exceed Э а п а д н ю к И. П. wait the excursion of these type of animal physiologic index that (nineteen eighty-three) carried out.Absorbed the animal (rabbit and rat) of inferior toxicity dosage particles two group leader's phases, we observe, the function of liver there is the influence of harmful trend to a certain degree, is checked through bilirubin direct higher in the serum and unconjugated bilirubin content (P＞0.05) and high slightly transaminase activity (P＞0.05).The Pathomorphology of two types the test group and the internal organs of control animals studies show that, no matter be to use preparation itself, also is to use the main component of preparation, does not either way show obvious variation.

Result of study shows, when preparation rubs on the skin of the rabbit of experiment and rat and inject in the body cavity (vagina and rectum), local irritant effect do not take place.The morphology of being done in order to the rabbit and the part skin that takes off on one's body of rat of experiment studies show that, through embrocating this part skin of this preparation in one month, does not show any variation.The destruction that vagina (jenny dispenser place) mucosa and rectum (buck dispenser place) mucosa do not show histological structure yet.No matter be once a day or twice preparation splashed into the conjunctiva gaps of two of rabbit and Cavia porcelluss with the concentration of therapeutic dose, only bring short time (1～2 minute) to shed tears and inappreciable congested mocosa.

When the preparation that surpasses 10 times of therapeutic doses being dripped to the eye conjunctiva of rabbit, just manifest significantly mucous hyperemia, and this reaction continuity long period (reaching 1.5～3 hours).To studies show that of the eye tissue structure of experimental animal, no matter being therapeutic dose, used amount of formulation still is 10 times of this dosage, variation has all taken place in the eye functions feature.And the mucosa of eyes part does not demonstrate tangible disruptive features.

With such method to said preparation to studies show that of the local irritant effects of three types of animals, no matter be the skin that preparation acts on tested animal, still act on the mucosa of eyes, vagina and rectum, all do not produce partial stimulation.

Preparation studies show that the anaphylaxis effects of 24 Cavia porcelluss, uses preparation can not bring the reaction of animal whole body and the variation of blood aspect by therapeutic dose, tests the check that pair cell done with test tube and shows that cell does not change.Also show simultaneously, only used preparation reach therapeutic dose more than 10 times the time can cause to a certain extent that just the development of allergic phenomena feature, these features are, leukocytic agglomeration rises to original more than thousand times, at 1mm ³Blood in, the leukocyte number reduces by 1500, the platelet count purpose increases and reaches 70%, the content of acidophil is increased to more than original 4 times (P＜0.05).

The neither meeting of preparation therapeutic dose or even inferior toxicity dosage makes a difference to immune mechanism: promptly do not change the number of immunocyte, also do not change their function, particularly the comprehensive function of the immune body of main kind.Do not demonstrate the downtrod sign of the non-special protection factor of animal organism yet.The histological result of study of spleen and lymph node shows that not finding has reacting with lymphocytic infiltration and Cytoplasm of macrophage in spleen and lymph node, and this has just confirmed the result of immunological testing largely.

No matter be that preparation is embrocated on the skin of rabbit or on mucosa, it all is ingested.But than absorbing more slowly, measure also less by mucosa by skin.Also found out, though the picked-up of preparation by mucosa than skin carry out faster, capture process is still slow after all.

Experimental study according to the present invention shows that this preparation has very high activity especially for the pathogen of syphilis, gonorrhea and trichomoniasis.

About preparation studies show that to experimental animal human body adverse effect, result of study according to acute poisoning, this preparation can be listed relatively nontoxic first-class in, and it is used with therapeutic dose and inferior toxicity dosage for a long time, can not bring tangible illeffects to the organ and the system of laboratory animal.

Result of study before the clinical practice of preparation is shown, can recommend a kind of preparation of prevention of STD most effectively, the pharmaceutical preparation of this liquid state has following composition (weight %):

To nitro-alpha-chloro cinnamic aldehyde, molecular formula is:

Dimethyl sulfoxide 5.0

Polyethylene Glycol (molecular weight 400) adds to 100,0

For prevention of STD and treatment women urethral trichomoniasis, recommend a kind of preparation of the liquid condition in pressurized package with pressure, its composition (weight %) is:

To nitro-alpha-chloro cinnamic aldehyde, molecular formula is:

Dimethyl sulfoxide 5.0

Polyethylene Glycol (molecular weight 400) 10.0～30.0

Surfactant 3.0～6.0

Water is supplemented to 100.0 and add propylene and make it account for 3～10% of total amount

Preparation carried out research in some big Dermatology mechanisms of the Soviet Union as the clinical trial of individual prophylaxis sexually transmitted disease (STD) medicine.These researchs are to carry out in the medical station of some individual prophylaxis sexually transmitted disease (STD)s, and there, the staff of medical station gives the client and prevents to help, and also has the people of irregularity relation to provide pharmaceutical preparation to some.Result of study shows, accidental sexual relationship was once arranged and carried out treatment with this preparation and in the middle of the observed man, the situation of syphilis, gonorrhea and trichomoniasis do not take place to infect at those.Do not find that this preparation caused poisoning, local excitation and sign hypersensitive when acting on patient's human body yet.

According to the present invention, use this preparation that the patient of 150 urethral trichomoniasis is treated, be 5～7 days the course of treatment, twice of every day, each 1.5～10 milliliters, the result of treatment shows that this preparation has the very high medical effect that kills protozoa, comprising on the complete nosetiology with medicine on therapeutic effect.Observe in the supervision in long-term (3 months) to patient, do not find the situation of trichomoniasis recurrence, also do not find the side effect phenomenon.

Claims (4)

1, a kind of preparation method of pharmaceutical preparation that is used for individual prophylaxis sexually transmitted disease (STD) and treatment urethral trichomoniasis is characterized in that, with effective dose to nitro-alpha-chloro cinnamic aldehyde

Be mixed into solution shape cooperative action mixture with dimethyl sulfoxide, and be diluted to predetermined concentration with medicinal diluent.

2, the method for claim 1 is characterized in that, as medicinal diluent is that molecular weight is 400 Polyethylene Glycol.

3, method as claimed in claim 1 or 2 is characterized in that, above-mentioned used be that the weight ratio of 400 Polyethylene Glycol is 0.3: 5.0: 94.7 to nitro-alpha-chloro cinnamic aldehyde, dimethyl sulfoxide and molecular weight.

4, a kind of preparation method that is contained in the pharmaceutical preparation as claimed in claim 1 or 2 in the aerosol pressure packing with liquid form, it is characterized in that, toward a kind of by 3%(weight) above-mentioned to nitro-alpha-chloro cinnamic aldehyde, 5%(weight) dimethyl sulfoxide and 10～30%(weight) molecular weight is to add 3.0～6.0%(weight in the solution formed of 400 Polyethylene Glycol) and surfactant and water make it to reach 100%(weight), then with the abundant saturated solution that obtained that is contained in the pressurized package of the propylene that accounts for total solution weight 3～10%.