Summary of the invention
At above situation, it is simple to the purpose of this invention is to provide a kind of composition, and have infection and antipyretic effect and can be used for the anti-infectives of treatment of eye disorders by what the extract combination of the effective ingredient of the effective ingredient of natural medicinal raw material or its medicinal raw material formed, make its active constituent content height, produce effects is fast, toxic and side effects is little, and can be made into multiple dosage forms such as oral, injection and eye external, to give full play to its pharmacological action.Further aim of the present invention is on this basis, can increase at least to select the types of drugs scope used as required for doctor and/or patient when infection, antipyretic-antalgic and ophthalmic.
The active drug composition of the said medicine of the present invention is fibrauretin and andrographolide or its soluble derivative separately, forms jointly with acceptable auxiliary element in the medicine.The part by weight of wherein said active drug composition is andrographolide/fibrauretin=1/0.1~10.
Andrographolide is the crystalline solid of a kind of colourless square, rectangle or prism-shaped, odorless, and bitter in the mouth, water insoluble and be dissolved in the ethanol that boils, slightly be dissolved in methanol.This composition can be by the mode of chemosynthesis, or is obtained after chemical modification is transformed by other related compounds, also can be obtained through ethanol extraction, separation by natural medicinal raw material Herba Andrographis.On this basis, even can also be to the total ethanol extract that obtains by Herba Andrographis without separation, and directly use this total ethanol extract of the Herba Andrographis raw material that contains said significant proportion amount andrographolide to replace.When using its soluble derivative, can use it to use the soluble derivative commonly used of the appropriate format that allows in potassium salt commonly used, sodium salt or the other medicines preparation for preparing by the volume of production and marketing lactone according to a conventional method.When its total ethanol extract of direct use, this extract normally comprises this lactone at interior total andrographolides, therefore as long as other wherein contained composition does not produce interference or adverse effect to the effect of active drug composition in the said medicine of the present invention, and according to the related request of different preparations and the scope of permission, allow it to exist simultaneously with appropriate format and/or ratio, and and do not require and its separation must be removed, to reduce cost.
The fibrauretin of said active drug composition is xanchromatic acicular crystal, and the water part omitted is molten and be soluble in hot water, and slightly soluble in ethanol or the chloroform is dissolved in ether hardly.What the present invention relates to is this composition in the medicine, except that can adopting existing commercially available fibrauretin finished product, also can transform obtain through chemical modification by chemosynthesis or by related compound, can also by natural medicinal raw materials such as the Caulis Fibraureae that contains the fibrauretin composition through the acid extraction of conventional form, alkalizing to separate through steps such as alcohol extraction, acidify and crystallizes with the crude extract that obtains of saltouing obtains again.When other composition except that fibrauretin contained in the total extract not to medicine of the present invention in the effect of said effective ingredient produce to disturb or adverse effect, requirement and allowed band according to different preparations, equally also can directly use this Caulis Fibraureae raw material total extract that contains said significant proportion amount fibrauretin, and and do not require that other composition beyond these active drug compositions must will separate and remove, allow it to exist simultaneously with suitable form and/or ratio.When using the soluble derivative of fibrauretin, can according to a conventional method it be prepared into corresponding salt, as having the soluble derivative of other appropriate format that allows in commercially available fibrauretin hydrochlorate or the other medicines preparation.
Medicine of the present invention by after the desired mixed, according to required dosage form, respectively by different dosage form preparation requirement, standard separately, and regulations such as processing, detection method, is made the medicine of corresponding dosage forms with above-mentioned active drug composition in the preparation.As: after adding suitable adjuvant, can be made into solid oral dosage forms such as corresponding tablet, pill, capsule, granule, or the medicine of liquid oral dosage forms such as syrup, oral liquid; By the requirement and the processing method of injection routine, can be made into the medicine of corresponding intramuscular injection or intravenous form.By the preparation requirement of eye medicinal dosage form, can also make corresponding external ophthalmic preparation.
Below example by the specific embodiment again foregoing of the present invention is described in further detail.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.Do not breaking away under the above-mentioned technological thought situation of the present invention, various replacements or change according to ordinary skill knowledge and customary means are made include within the scope of the invention.
The specific embodiment
Embodiment 1
Get the fibrauretin 100g and the andrographolide fine powder 50g that meet medicinal standard, mix with starch 50g even after, granulate with 10% starch slurry in the usual way, oven dry, adding 0.5g magnesium stearate mixing is pressed into 1000, obtains the tablet oral drugs.
Embodiment 2
Get the fibrauretin 25g that meets medicinal standard, andrographolidum Natrii Bisulfis 25g, with the dissolving of 200ml water for injection, add 0.1% active carbon and stirred 5 minutes, filter, water for injection adds to 1000ml, 10% sodium hydroxide solution is regulated pH value to 4.0-6.0, filters fill, sterilization promptly gets injection medicine.
Embodiment 3
Get the fibrauretin 2g that meets medicinal standard, andrographolidum Natrii Bisulfis 2g with the dissolving of 200ml water for injection, filters, and adds the injection water to 1000ml, and 10% sodium hydroxide solution is regulated pH value to 4.0-6.0, filter, and fill, sterilization promptly obtains the eye drop medicine.
Each test that becomes of medicine system with embodiment 1 composition form has been carried out following correlation test with sample.
Toxicity test
Acute toxicity test: get 20 of mices, by 15g/kg (be equivalent to clinical maximum administration 1000 times) dosage, gastric infusion was observed 8 days continuously, none animal dead.Fail to record LD
50
Long term toxicity test: get 20 of rat, by 1.5g/kg (be equivalent to clinical maximum administration 100 times) dosage, gastric infusion was observed 30 days continuously, organ no abnormality seens such as the heart, liver, spleen, lung, kidney.Result of the test shows that medicine of the present invention does not have overt toxicity.
Pharmacodynamics test
1. bacteriostatic test
Said medicine of the present invention is respectively 1/1 (sample 1), 1/2 (sample 2), 1/4 sample 3 by andrographolide/fibrauretin respectively) and the trial drug sample of four kinds of different proportion forms of 1/8 (sample 4), carried out external antibacterial activity contrast test respectively with the andrographolide and the fibrauretin of equivalent.The wherein andrographolide of medicine and fibrauretin in contrast, adopt fibrauretin sheet and andrographolide sheet respectively, the trial drug sample of the same concentration that dissolving or suspendible are mixed with behind porphyrize by andrographolide with identical active drug total amount and fibrauretin.The correlation test result of minimum inhibitory concentration (MIC) is as shown in table 1.
The test of table 1 antibacterial activity in vitro
Test strain | MIC(mg/ml) |
Sample 1 | Sample 2 | Sample 3 | Sample 4 | The fibrauretin matched group | The andrographolide matched group |
The bloodthirsty influenza bacterium of golden staphylococci bacillus cloacae Pseudomonas aeruginosa aerobacteria proteus pneumonia streptococcus Klebsiella pneumoniae hemolytic streptococcus | 0.40 1.20 2.40 1.20 1.20 0.80 2.40 0.80 2.40 | 0.42 1.26 1.26 1.26 0.84 0.84 1.26 0.84 2.52 | 0.38 1.14 1.14 1.14 0.76 0.76 1.14 0.76 1.90 | 0.41 1.23 2.46 1.23 1.23 0.82 2.46 0.82 2.05 | 0.60 1.50 3.00 1.50 1.50 1.20 3.00 1.20 3.00 | 0.58 1.45 2.90 1.16 1.45 1.16 2.90 1.46 2.90 |
The result of table 1 shows, the drug test sample of being made up of two kinds of active component of different proportion form of the present invention all demonstrates common Gram-positive and negative pathogenic bacterium and to have certain antibacterial vigor.Sample 1, sample 2, sample 3,4 pairs of golden staphylococcis of sample, bacillus cloacae, bacillus pyocyaneus, aerobacteria, Bacillus proteus, Ke Shi pneumobacillus etc. all have antibiotic preferably vigor.Antibacterial vigor to streptococcus pneumoniae, Hemolytic streptococcus is stronger.Bloodthirsty influenza bacterium also there is certain antibacterial vigor.The result of table 1 also shows, four groups of different proportion forms what the present invention relates to is that the antibacterial activity in vitro of drug test sample is basic identical, but wherein sample 2 and 3 liang of groups of sample are better than 4 groups in sample 1 and sample slightly.And the antibacterial vigor of sample 1, sample 2, sample 3, sample 4 all is better than the fibrauretin and the andrographolide reference substance group of single component, shows that the medicine of composition form of the present invention has potentiation aspect antibacterial action.Correlation test research also shows, as Gram-positive, negative bacillus such as Gram-positives such as Diplococcus pneumoniae, meningococcus, negative cocci and dysentery bacterium, Bacillus typhi, Salmonella paratyphi, vibrio cholera, diphtheria corynebacterium, bordetella pertussis, and pathogenic microorganism such as leptospira, Candida albicans, epidermophyton also has bacteriostasis to said medicine of the present invention to other.
2. antiinflammatory test
Adopt carrageenin to cause the rat paw edema method said medicine of the present invention, get 70 of rats, be divided into 1 group in same sample, 2 groups in sample, 3 groups in sample, 4 groups in sample, blank group, fibrauretin matched group and andrographolide matched group, matched group is irritated the isometric distilled water of stomach, all the other each groups are pressed 300mg/ml and are irritated stomach, after the administration 1 hour, the right sufficient plantar subcutaneous injection 1% carrageenin 0.1ml of each Mus causes inflammation, measure after the administration 1,2,3,5h about sufficient volume, calculate swelling degree and inhibitory rate of intumesce.Result of the test is as shown in table 2.
The result of table 2 shows, the drug test sample of the present invention of two kinds of active component of different proportion form, and on Carrageenan causes sufficient the swelling of rat all can obviously suppress rat paw edema.The result of table 2 shows that also the test specimen medicine of four groups of different proportion forms is basic identical to the inhibitory action of rat paw edema, but wherein sample 2 and 3 liang of groups of sample are better than 4 groups in sample 1 and sample slightly.And the inhibitory action of sample 1, sample 2, sample 3,4 pairs of rat paw edemas of sample all is better than the fibrauretin and the andrographolide reference substance group of single component, shows that the medicine of composition form of the present invention has potentiation on antiphlogistic effects.
Table 2 on Carrageenan cause rat paw edema influence (x ± s, n=10)
Group | Swelling degree (ml) |
1h after the administration | 2h after the administration | 3h after the administration | 5h after the administration |
Blank group sample 1 sample 2 samples 3 samples 4 fibrauretine control group andrographolide control groups | 0.265±0.062 0.226±0.047
* 0.197±0.044
* 0.183±0.069
* 0.206±0.039
* 0.246±0.041
* 0.256±0.049
| 0.401±0.088 0.319±0.080
* 0.245±0.047
** 0.228±0.071
** 0.267±0.059
** 0.359±0.082
* 0.389±0.077
| 0.548±0.119 0.409±0.097 0.339±0.046
** 0.284±0.094 0.368±0.084 0.469±0.091 0.509±0.089
| 0.722±0.123 0.560±0.117
** 0.481±0.111
** 0.433±0.110
** 0.519±0.112
** 0.590±0.102
* 0.660±0.100
* |
Annotate: compare with the blank group,
*: P<0.05,
*: P<0.01
3. separate heat test
Said medicine of the present invention is caused the influence test that rat temperature raises to dry yeast.Get rat and regularly measure the anus temperature, continuous 3 days, measure twice of anus temperature morning on the 4th, interval 1h, average as the body temperature before the administration, body temperature is changed the rat that is no more than 0.5 ℃ be divided into 5 groups at random, 10/group, be divided into as the test of pesticide effectiveness 11 group in said sample, 2 groups in sample, 3 groups in sample, 4 groups in sample, blank group, fibrauretin matched group and andrographolide matched group, matched group is irritated the isometric distilled water of stomach, all the other each group press 300mg/ml filling stomach, first administration after 6 hours with the method rechallenge.Cervical region subcutaneous injection 20% dry yeast liquid 1ml/100g pyrogenicity immediately after the first administration.Each group respectively at the injection yeast after 0.5,1,2,4,6,8,10,12h surveys the anus temperature once.Result of the test is as shown in table 3.
Table 3 pair dry yeast cause the influence that rat temperature raises (x ± s, n=10)
Group | | Matched group | Sample 1 | Sample 2 | Sample 3 | Sample 4 | The fibrauretin matched group | The andrographolide matched group |
Different time body temperature drop-out value after the administration (℃) | 0.5h 1h 2h 4h 6h 8h 10h 12h | -0.42±0.29 -0.28±0.45 0.28±0.52 1.20±0.53 1.81±0.43 1.62±0.41 1.32±0.51 0.73±0.38 | -0.53±0.47 -0.40±0.23 0.10±0.37 0.80±0.37 1.21±0.42
* 10.9±0.58
* 0.85±0.50 0.53±0.46
| -0.52±0.25 -0.41±0.29 -0.04±0.24 0.62±0.20
** 1.10±0.32
** 1.20±0.46
* 0.79±0.35
* 0.39±0.33
| -0.96±0.12 -1.00±0.37
** -0.67±0.47
** 0.35±0.42
* 0.72±0.29
** 0.76±0.26
* 0.51±0.29
* 0.29±0.22
* | -0.48±0.38 -0.60±0.33 -0.17±0.20 0.65±0.32
* 1.14±0.63
** 1.26±0.25
** 0.95±0.28 0.42±0.25
** | -0.50±0.42 -0.42±0.21 0.18±0.33 1.08±0.32 1.51±0.48
* 1.49±0.52
* 1.15±0.54 0.63±0.56
| -0.58±0.49 -0.43±0.25 0.15±0.37 0.90±0.37 1.42±0.47
* 1.22±0.51
* 0.95±0.52 0.58±0.36
|
Annotate: compare with the blank group,
*P<0.05,
*P<0.01
The result of table 3 shows, the drug test sample of the present invention of two kinds of active component of different proportion form causes the rat temperature rising to dry yeast and all can obviously suppress.The result of table 3 also shows, it is basic identical that the test specimen medicine of four groups of different proportion forms causes the inhibitory action that rat temperature raises to dry yeast, but wherein sample 2 and 3 liang of groups of sample are better than 4 groups in sample 1 and sample slightly.And sample 1, sample 2, sample 3,4 pairs of dry yeast of sample cause fibrauretin and andrographolide reference substance group that inhibitory action that rat temperature raises all is better than single component, show that medicine of the present invention has potentiation separating on the thermal effect.
4. antivirus test
Said medicine sample 2 of the present invention is respectively 10mg/ml (1 group) by total medicament contg, 5mg/ml (2 groups) 2.5ma/ml (3 groups), 1.25mg/ml (4 groups) different dilution factors are four test group, the antivirus test of being done by virazole control drug, fibrauretin control drug, andrographolide control drug and the blank group of the preparation of same dilution factor respectively, result of the test sees Table 4.
Table 4 antivirus test result
Test specimen | Medicine of the present invention | The fibrauretin control drug | The andrographolide control drug | The virazole control drug | The blank group |
Virus | ADV
3 | ADV
7 | RSV | ADV
3 | ADV
7 | RSV | ADV
3 | ADV
7 | RSV | ADV
3 | ADV
7 | RSV | ADV
3 | ADV
7 | RSV |
3 group of 4 papova 3+ group of 2 groups of medicine groups of 1 group of the present invention groups of cells- | - - - - 3+ - | - - - - 3+ - | - - - - 3+ - | 1+ 1+ 2+ 3+ 3+ - | 1+ 2+ 2+ 3+ 3+ - | 1+ 2+ 3+ 3+ 3+ - | - - - 1+ 3+ - | - - 1+ 1+ 3+ - | - - 1+ 2+ 3+ - | - - 1+ 3+ 3+ - | - 1+ 1+ 3+ 3+ - | - - 2+ 3+ 3+ - | 3+ 3+ 3+ 3+ 3+ - | 3+ 3+ 3+ 3+ 3+ - | 3+ 3+ 3+ 3+ 3+ - |
Annotate: no pathological changes in "-" expression cell in the table: how much "+" represents the interior lesion degree of cell
The result of the test of table 4 shows, each dilution group of drug sample of the present invention all has in various degree inhibitory action to three kinds of viruses, to ADV
3, ADV
7All be better than the control drug virazole with the inhibitory action of RSV, to ADV
3, ADV
7All be better than the fibrauretin and the andrographolide control drug of single component with the inhibitory action of RSV, show that medicine of the present invention has potentiation on antiviral effect.
5. analgesic test
Adopt 4 groups in 3 groups in 2 groups in 1 group in sample, sample, sample, sample, blank group, fibrauretin matched group and andrographolide matched group with the bacteriostatic test same form.Wherein matched group isometric(al) distilled water is irritated stomach, and all the other each groups are pressed 300mg/ml and irritated stomach.Equal administration every day of each treated animal 1 time, continuous 3d, 1h after the last administration, lumbar injection 0.6% acetum 0.2ml/ only carry out the influence test of the mouse writhing reaction that acid is brought out to ester, and mice turns round the body number of times in the record 30min.Result of the test is as shown in table 5.
Table 5 analgesic test result
Group | The blank group | Medicine group of the present invention | The fibrauretin matched group | The andrographolide matched group |
1 group | 2 groups | 3 groups | 4 groups |
Turn round the body number of times (inferior/30min) | 56.8±7.3 | 20.4±14.4 | 19.2±11.3 | 19.6±10.4 | 21.4±12.1 | 45.2±12.8 | 29.4±11.6 |
Suppression ratio (%) | - | 64.8
** | 66.3
** | 65.5
** | 62.3
** | 20.6 | 48.2
* |
Annotate: compare with the blank group,
*P<0.05,
*P<0.01
Result by table 5 shows, the drug test sample of the present invention of two kinds of active component of different proportion form, and the mouse writhing reaction that acid is brought out to ester all can obviously suppress.The inhibitory action of the test specimen medicine mouse writhing that acid is brought out to the ester reaction of four groups of different proportion forms is basic identical.The inhibitory action of the mouse writhing reaction that sample 1, sample 2, sample 3,4 pairs of ester acid of sample are brought out all is better than the fibrauretin and the andrographolide reference substance group of single component, shows that medicine of the present invention can produce potentiation.
6. medicament for the eyes test
The trial drug group is the ophthalmic preparation of the foregoing description 3, and it contains andrographolide 0.2%, fibrauretin 0.2%.The control drug group is respectively and contains " viral quiet eye drop " (People's Hospital, Sichuan Prov.) and the chemicals " ancitabine eye drop " (Hualian Pharmaceutical Co., Ltd., Shanghai) that refining andrographolide 0.5% is used for the treatment of herpes simplex keratitis.The result of the clinical observation on the therapeutic effect test that common multiple ophthalmic is carried out is as shown in table 6.
The clinical observation on the therapeutic effect of the common ophthalmic of table 6
| The medicament for the eyes that medicine of the present invention is made | The quiet eye drop of virus | The ancitabine eye drop |
Trachoma | Conjunctivitis | Keratitis | Epidemic hemorrhagic conjunctivitis | Trachoma | Conjunctivitis | Keratitis | Epidemic hemorrhagic conjunctivitis | Trachoma | Conjunctivitis | Keratitis | Epidemic hemorrhagic conjunctivitis |
The total routine number of recovery from illness enabledisable | 25 11 0 36 | 26 10 0 36 | 49 12 0 61 | 31 22 1 54 | 10 15 2 27 | 12 13 1 26 | 57 15 1 73 | 11 23 2 36 | - - - - | - - - - | 24 32 4 60 | - - - - |
The clinical efficacy result of table 5 shows that control drug " viral quiet eye drop " is the ophthalmic preparation that cures mainly herpes simplex keratitis, and indication and effect are single.Because various inflammation may be bacterial infection, viral infection or mixed infection respectively in the different courses of disease, this requires to select medicine at different situations, thereby has limited its performance clinical treatment effect; " ancitabine eye drop " can be used for treating herpes simplex keratitis for herpes simplex virus is had inhibiting chemicals.Because the corneal epithelial metabolism is influential, often take place clinically thus that the corneal epithelium infringement is prolonged does not heal, cause healing time obviously to prolong.The medicament for the eyes of medicine composition form of the present invention is owing to combine andrographolide antiviral and the antimicrobial advantage of fibrauretin, can have heat-clearing and toxic substances removing, the effect of anti-inflammation, thereby can be used for the different ophthalmic diseasess of antibacterial and viral infection, as trachoma, epidemic hemorrhagic conjunctivitis, and various antibacterials and/or the viral conjunctivitis that causes and/or the treatment of keratitis.
Above-mentioned relevant pharmacodynamics test and clinical research result show, and medicine of the present invention has infection, analgesia, many-sided effect such as analgesic; Clinical common pathogenic microbes such as various bacteria, virus had obvious and stronger inhibitory action; Also all can produce the inside and outside section various disease conditions that causes thus and to take stopgap measures preferably and effect a permanent cure effect; Obviously be better than single component in the said medicine component at aspects such as antiinflammatory, antibacterial, analgesic, antiviral, have obvious synergistic, potentiation; Ophthalmic diseases obviously is better than the single component preparation clinically.Simultaneously, said medicine decapacitation of the present invention increases the doctor when being used for the treatment of infection and antipyretic analgesic outside more alternative variety range and the leeway, also simple because of its composition, effective ingredient is concentrated, and can be made into the several formulations form, thereby the use amount of medicine is subtracted down relatively, reduce waste natural medicinal raw material, and make the scope of its occupation mode and/or patient's object more extensive, help promoting the use of on a large scale.