CN102716115A - Pharmaceutical composition - Google Patents
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Abstract
The invention belongs to the technical field of medicines and provides a pharmaceutical composition. The pharmaceutical composition comprises germacrone, furanodiene, curdione, beta-elemene, curcumol, curzerene and borneol. The components of the pharmaceutical composition are definite, and the quality can be controlled. The pharmaceutical composition can be used for preparing vaginal or rectal preparations and has a good effect of treating HR-HPV (high risk human papillomavirus) and/or LR-HPV (low risk human papillomavirus) infection and cervical diseases and can be used widely clinically.
Description
Technical field
The invention belongs to medical technical field, be specifically related to a kind of pharmaceutical composition that contains 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, beta-elemene, curcumenol, curzerene, Borneolum Syntheticum.
Background technology
Human papillomavirus (HPV) is that a kind of papillary tumor vacuolating virus A that belongs to papovaviridae belongs to, and is spherical DNA viruses, can cause the squamous epithelial cancer propagation of human body skin mucosa.Isolated kind more than 130 at present, different hypotypes causes the different clinical performance, invades different tissue sites.By sending out classification carcinous, can be divided into low risk HPV and high-risk HPV.The HPV resistance is strong, can tolerate dry and long preservation, but heating or handle deactivation through formalin, so but high-temperature sterilization and 2% glutaraldehyde sterilization deactivation.HPV mainly comprises 13 kinds of hypotypes, and the generation that becomes (CIN) with cervical cancer, cervix uteri squamous epithelial cancer tumor is closely related.Cervix uteri persistent infection HPV 8-10 can develop into cervical cancer; And " cervical cancer " is the second largest gynecological cancer after breast carcinoma, global new cases 460,000/year, and the Asia accounts for half the; 80% occurs in developing country, dies from nearly 200,000 people of patient of cervical cancer every year.China's new cases 130,000/year are died from cervical cancer 4-5 ten thousand/year, and M & M all presents increase trend." cervical cancer " is the clear and definite cancer of preventing and treating of present unique cause of disease, and removing HPV is the determiner of the blocking-up course of disease, prevention cervical cancer.
In recent ten years, be that the Chinese medicine of representative is widely used in the treatment that female genital tract HPV infects with BAOFUKANG SHUAN (main component is Oleum Curcumae and Borneolum Syntheticum), and obtained better curative effect; Therefore, on this basis to the new pharmaceutical composition of active component exploitation come prophylactic treatment HPV infect with and the diseases such as optimum or malignant change of bringing out, particularly infect significant to HR-HPV to cervical cancer.
Rhizoma Curcumae is the dry rhizome of zingiberaceous plant Rhizoma Curcumae Curcuma phaeocaulis Valeton, Guangxi zedoary C.kwangsiensis S.G.Lee et C.F.Liang or RADIX CURCUMAE C.wenyujin Y.H.Chen et C.Ling.The latter practises title " warm Rhizoma Curcumae ".Rhizoma Curcumae contains volatile oil 1.5%～2%, and is the highest with curzerenone content, is curcumenol, Rhizoma Curcumae enol, curcumadiol etc. secondly.Still contain the Rhizoma Curcumae polysaccharide in addition.Guangxi zedoary contains volatile oil 1%-1.2%, contain 20 in the oil surplus kind of composition, content is up to Camphora, about 17.7%, eucalyptol is about 7.5%, and contains zingiberene, curcumenol, ar-curcumene, fragrant zingiberone, curdione etc.RADIX CURCUMAE contains volatile oil 1.4%-2.0%, contain 20 in the oil surplus kind of main constituent, be mainly curcumenol, curdione, gima ethylenic, 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-etc., nearly report therefrom tell α-, β-, δ-elemene, wherein beta-elemene is main anti-tumor active ingredient.
Compare the Rhizoma Curcumae in above-mentioned three places of production " comparisons of 3 kind Rhizoma Curcumae volatile oil chemical constituents " (Chinese herbal medicine, the 36th the 12nd phase of volume, 1785～1787), though point out the different cultivars Rhizoma Curcumae a lot of total compositions is arranged, and the mass fraction difference is bigger; The amount significant difference of composition such as curzerenone, curdione in the Rhizoma Curcumae of the different places of production.And the notable difference of the main component of Rhizoma Curcumae and content, meeting directly influence the curative effect of aromatic turmeric oil preparation.
CN102058568 discloses the application of 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-in preparation treatment HPV infection medicine; CN102091059 discloses the application of curdione in preparation treatment HPV infection medicine; CN102091064 discloses the application of furanodiene in preparation treatment HPV infection medicine; CN102091065 discloses the application of curcumenol in preparation treatment HPV infection medicine.Mentioned component all is proved to be HPV is infected effectively; But because the HPV hypotype is numerous; Each hypotype viral phenotype, growth characteristics, pathogenic, everyways such as the sensitivity of medicine and clinical prognosis there are differences; Thereby single component can not be effective to all hypotypes, so the preparation of single component is unfavorable for clinical use.
Summary of the invention
For solving existing problem in the above-mentioned prior art, the invention provides a kind of new pharmaceutical composition and preparation thereof.
The purpose of this invention is to provide a kind of pharmaceutical composition, said composition has good therapeutical effect to HR-HPV (high-risk human mammilla papillomavirus) and/or LR-HPV (high-risk human mammilla papillomavirus) infection.
The purpose of this invention is to provide a kind of pharmaceutical composition, said composition is effective to vaginitis.
The purpose of this invention is to provide a kind of preparation that contains this pharmaceutical composition.
The purpose of this invention is to provide a kind of application of preparation in anti-HPV virus that contains this pharmaceutical composition.
The purpose of this invention is to provide a kind of application of preparation in the treatment vaginitis that contains this pharmaceutical composition.
Particularly, the invention provides:
A kind of pharmaceutical composition, its characteristic pharmaceutical composition comprises 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, beta-elemene, curcumenol, curzerene, Borneolum Syntheticum.
Above-mentioned described pharmaceutical composition comprises the composition of following weight portion:
3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-5-16 weight portion;
Furanodiene 10-40 weight portion;
Curdione 5-25 weight portion;
Beta-elemene 4-11 weight portion;
Curcumenol 1-10 weight portion;
Curzerene 8-22 weight portion;
Borneolum Syntheticum 30-90 weight portion.
Above-mentioned described Borneolum Syntheticum is selected from natural Broneolum Syntheticum and/or synthetic borneol.
Above-mentioned described preparation of pharmaceutical compositions becomes vagina administration preparation or rectally preparation.
Said preparation comprises suppository, ointment, capsule, effervescent tablet, gel, lotion, membrane or foam.
Above-mentioned described pharmaceutical composition prevents and/or treats the application in the medicine of human papilloma virus infection in preparation.
Wherein human papilloma virus infection comprises high-risk human mammilla papillomavirus infection and/or low risk human papilloma virus infection.
Above-mentioned described pharmaceutical composition prevents and/or treats the application in the medicine of vaginitis, cervical erosion, cervical cancer in preparation.
The present invention compared with prior art has the following advantages and good effect:
1, the bright described pharmaceutical composition definite ingredients of we is quality controllable;
2, pharmaceutical composition of the present invention does not receive the restriction in the places of origin of raw materials, is fit to the big production of industry;
3, pharmaceutical composition of the present invention all has good therapeutical effect to HR-HPV and/or LR-HPV, cervical disease etc., is suitable for wide clinical application.
Pharmacological experimental example
Below description through the specific embodiment the present invention is described further; But this is not to be limitation of the present invention; Those skilled in the art are according to basic thought of the present invention; Can make various modifications or improvement, but only otherwise break away from basic thought of the present invention, all within scope of the present invention.
HPV nucleic acid amplification (PCR) fluorescence detection reagent kit is available from the PiJi Biology Engineering Co., Ltd., Shenzhen City.
Oleum Curcumae is available from the magnificent spice in Ji'an, Jiangxi refinery, and detecting 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-content according to the requirement of 2010 editions first one of Chinese Pharmacopoeia is 12.3%.
3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, beta-elemene, curcumenol, curzerene are available from the suitable vigorous biological company limited in Shanghai.
Borneolum Syntheticum is available from Tianjin Li Lang Chemical Industry Science Co., Ltd.
Test Example 1: the effect of anti-HPV
Test reagent: HPV nucleic acid amplification (PCR) fluorescence detection reagent kit (comprising DNA extraction liquid 1, DNA extraction liquid 2, PCR reactant liquor, Taq enzyme, UNG).Other reagent are analytical pure.
Trial drug:
Test 1 group: 88mg Oleum Curcumae and 75mg Borneolum Syntheticum.
Test 2 groups: 5mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+40mg furanodiene+8mg curdione+10mg beta-elemene+4mg curcumenol+21mg curzerene+75mg Borneolum Syntheticum.
Test 3 groups: 8mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+32mg furanodiene+15mg curdione+8mg beta-elemene+7mg curcumenol+18mg curzerene+75mg Borneolum Syntheticum.
Test 4 groups: 13mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+36mg furanodiene+12mg curdione+7mg beta-elemene+6mg curcumenol+14mg curzerene+75mg Borneolum Syntheticum.
Test 5 groups: 14mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+28mg furanodiene+20mg curdione+8mg beta-elemene+2mg curcumenol+16mg curzerene+75mg Borneolum Syntheticum.
Test 6 groups: 10mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+20mg furanodiene+25mg curdione+11mg beta-elemene+10mg curcumenol+12mg curzerene+75mg Borneolum Syntheticum.
Test method:
HPV infected specimen preparation: make a definite diagnosis HPV from hospital outpatient and infect the BIAO and BEN of taking (the HPV type is 16,52 mixed types, belongs to the mucosa low risk), focus in 1 centrifuge tube; The 30mg that weighs uses glass homogenizer homogenate, is adding the 5ml normal saline; Be made into the stripped suspension of 6mg/ml, subsequent use.
Sample treatment: draw 25 μ l sample liquid and 25 μ l BIAO and BEN suspensions to the 15ml centrifuge tube with liquid-transfering gun; Jolting is even, and sample and BIAO and BEN can fully be acted on, and puts in 37 ℃ of water baths and cultivates;, respectively get 1 group and carry out following test after 1 day, 3 days, 5 days, 7 days at the sample treatment BIAO and BEN.
The extraction of HPV-DNA: from water bath, take out BIAO and BEN suspension, extract DNA by the test kit step: add 50 μ l normal saline, add 100 μ l DNA extraction liquid behind the mixing again through sample treatment; Jolting is even, the centrifugal 10min of 1200r/min, abandoning supernatant; Add 25 μ l DNA extraction liquid 2 again, abundant mixing, 100 ℃ of boiling water boil 10min; The centrifugal 10min of 1200r/min, supernatant is the HPV-DNA template very.
DNA cloning: get the PCR reactant liquor 37.6 μ l in the detection kit, Taq archaeal dna polymerase 0.4 μ l, UNG0.03 μ l are adding HPV-DNA template 2 μ l in the PCR reaction tube, and button strict control lid places quantitative PCR appearance cocycle amplification.HPV-DNA carries out cyclic amplification through high-temperature denatured, process annealing and extension, and cyclic program is set to: 37 ℃, and 5min; 94 ℃, 1min; 95 ℃, 5sec; 60 ℃, 30sec circulates 40 times.
Amount standard curve: detect quantitatively with fluorescent probe,, set up the quantitative positive criteria article of HPV-DNA of 4 series concentration, be followed successively by 5 * 10 according to the detection kit requirement
7/ ml, 5 * 10
6/ ml, 5 * 10
5/ ml, 5 * 10
4/ ml, (its detection sensitivity is 1 * 10 through the PCR detection
3/ ml, the result is judged to be feminine gender less than this concentration person).Natural logrithm with initial copy number is an abscissa, and circulation fault value is a vertical coordinate, and the regression straight line that obtains is a standard curve, in view of the above the amplification times of sample is carried out quantitatively.
Result of the test: see table 1.
Table 1: the DNA exercising result that the different tests medicine infects HPV
Annotate :-represent negative.
The HPV infected specimen changed into be diagnosed as HPV from hospital outpatient and infect the isolated preparation take (the HPV type is the mixed type of HPV1 and HPV12; Belong to the skin low risk); Make an experiment according to above-mentioned identical condition and method, it is all negative to test 2～6 groups of results that cultivate 1 day, 3 days, 5 days, 7 days.
The HPV infected specimen changed into be diagnosed as HPV from hospital outpatient and infect the isolated preparation take (the HPV type is the mixed type of HPV8 and HPV36; Belong to the skin high-risk-type); Make an experiment according to above-mentioned identical condition and method, it is all negative to test 2～6 groups of results that cultivate 1 day, 3 days, 5 days, 7 days.
The HPV infected specimen changed into be diagnosed as HPV from hospital outpatient and infect the isolated preparation take (the HPV type is the mixed type of HPV11 and HPV53; Belong to the mucosa low risk); Make an experiment according to above-mentioned identical condition and method, it is all negative to test 2～6 groups of results that cultivate 1 day, 3 days, 5 days, 7 days.
Conclusion (of pressure testing): above-mentioned result of the test shows that pharmaceutical composition according to the invention has the good restraining effect to the DNA that different subtype HPV infects.
Test Example 2: to the inhibiting influence of human cervical carcinoma cell
Test material
(1) oncocyte: human cervical carcinoma Hela cell system, available from the Xiangya Medical College, Zhongnan Univ cell centre.
(2) trial drug:
Test 1 group: BAOFUKANG SHUAN, Hainan Bikai Pharmaceutical Co., Ltd produces, and every heavy 1.74g wherein contains Oleum Curcumae 88mg, and Borneolum Syntheticum 75mg, all the other compositions are substrate; BAOFUKANG SHUAN is dissolved in the RPMI1640 culture fluid of 44ml serum-free, its concentration is 3.70 * 10
3Mg/l (active component), treat solution clarification after, using the aperture is the filter filtration sterilization of 0.22 μ m, 4 ℃ keep in Dark Place subsequent use after the packing.
Test 2 groups: 5mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+40mg furanodiene+8mg curdione+10mg beta-elemene+4mg curcumenol+21mg curzerene+75mg Borneolum Syntheticum prepares suppository according to embodiment 1 said method, every heavy 1.74g; Suppository is dissolved in the RPMI1640 culture fluid of 44ml serum-free, its concentration is 3.70 * 10
3Mg/l (active component), treat solution clarification after, using the aperture is the filter filtration sterilization of 0.22 μ m, 4 ℃ keep in Dark Place subsequent use after the packing.
Test 3 groups: 8mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+32mg furanodiene+15mg curdione+8mg beta-elemene+7mg curcumenol+18mg curzerene+75mg Borneolum Syntheticum, every heavy 1.74g; Suppository is dissolved in the RPMI1640 culture fluid of 44ml serum-free, its concentration is 3.70 * 10
3Mg/l (active component), treat solution clarification after, using the aperture is the filter filtration sterilization of 0.22 μ m, 4 ℃ keep in Dark Place subsequent use after the packing.
Test 4 groups: 13mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+36mg furanodiene+12mg curdione+7mg beta-elemene+6mg curcumenol+14mg curzerene+75mg Borneolum Syntheticum, every heavy 1.74g; Suppository is dissolved in the RPMI1640 culture fluid of 44ml serum-free, its concentration is 3.70 * 10
3Mg/l (active component), treat solution clarification after, using the aperture is the filter filtration sterilization of 0.22 μ m, 4 ℃ keep in Dark Place subsequent use after the packing.
Test 5 groups: 14mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+28mg furanodiene+20mg curdione+8mg beta-elemene+2mg curcumenol+16mg curzerene+75mg Borneolum Syntheticum, every heavy 1.74g; The suppository bolt is dissolved in the RPMI1640 culture fluid of 44ml serum-free, its concentration is 3.70 * 10
3Mg/l (active component), treat solution clarification after, using the aperture is the filter filtration sterilization of 0.22 μ m, 4 ℃ keep in Dark Place subsequent use after the packing.
Test 6 groups: 10mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+20mg furanodiene+25mg curdione+11mg beta-elemene+10mg curcumenol+12mg curzerene+75mg Borneolum Syntheticum, every heavy 1.74g; Suppository is dissolved in the RPMI1640 culture fluid of 44ml serum-free, its concentration is 3.70 * 10
3Mg/l (active component), treat solution clarification after, using the aperture is the filter filtration sterilization of 0.22 μ m, 4 ℃ keep in Dark Place subsequent use after the packing.
(3) reagent: super NBCS, Hangzhou Ilex purpurea Hassk.[I.chinensis Sims Company products; RPMI1640, U.S. Gibco Company products, four formylmerphalan azoles blue (MTT), trypsin are U.S. Sigma Company products; Dimethyl sulfoxide (DMSO) is homemade analytical pure.
(4) key instrument: ELX800 type enzyme-linked immunosorbent assay instrument, U.S. BIO-TEK company produces; Epics XL flow cytometer, U.S. Beckmen Coulter product.
Test method
(1) cell culture: the human cervical carcinoma Hela cell puts 37 ℃, 5% CO with the RPMI1640 culture fluid that contains 10% super NBCS
2Cultivate in the incubator.The trophophase cell of taking the logarithm is used for experiment.
(2) MTT measures: with 1 * 10
5/ ml inoculating cell is in 96 well culture plates, and every hole 100 μ l are divided into administration group and matched group behind the adherent 8h, establishes 6 multiple holes for every group.Add above-mentioned each the test group medicine of 20 μ l in the administration group; Matched group only adds the equivalent culture fluid.Leave heart 10min with per minute 800 after continuing to cultivate 48h; Absorb culture fluid, add serum-free medium 90 μ l and 10 μ l MTT (5mg/ml), hatch the centrifugal supernatant that goes behind the 6h for 37 ℃; Add DMSO 150 μ l, vibration 10min is after enzyme-linked immunosorbent assay instrument is surveyed the A570 value.Repeat 3 plates, every plate all has and only adds the zeroing hole that culture fluid does not add cell.Cell proliferation inhibition rate=(1-experimental group A
570Value/control group A
570Value) * 100%.
(3) flow cytometry analysis: 0.25% trypsinization collect matched group and test group effect 48h cell each 1 * 10
6Individual, add cold PBS washing 2 times, 70% cold ethanol is fixed, 4 ℃ of preservations.The centrifugal ethanol of abandoning during mensuration, PBS washing 2 times, 37 ℃ of digestion of RNA enzyme (0.1mg/ml) 30min; (propidium iodide PI) goes up the machine test behind dyeing (50mg/ml) 15min to propidium iodide, prints dna content distribution prescription figure; Automatically simulate cell cycle each the time phase ratio
Calculate proliferation index: PI=(S+G2/M)/(G0/G1+S+G2/M) * 100% with formula.
Statistical disposition
The gained experimental result is handled through the SPSS statistical package, relatively adopts the t check between group.
Result of the test
1, to the influence of Hela cell proliferation, sees table 2
Each test group of table 2 is to the influence of Hela cell proliferation
| Test group | A 570Value | Suppression ratio (%) |
| Matched group (culture fluid) | 0.384±0.0656 | - |
| Test 1 group | 0.354±0.0727 | 7.81 |
| Test 2 groups | 0.263±0.0513**# | 31.51 |
| Test 3 groups | 0.220±0.0517**# | 42.70 |
| Test 4 groups | 0.193±0.0396**# | 49.73 |
| Test 5 groups | 0.155±0.0283**# | 59.63 |
| Test 6 groups | 0.130±0.0218**# | 66.15 |
Annotate: compare: compare * * P＜0.01 with matched group, compare #P＜0.05 for 1 group with test with matched group.
Experiment conclusion: presentation of results pharmaceutical composition according to the invention can obviously suppress the propagation of Hela cell, and is superior to the combination of Oleum Curcumae and Borneolum Syntheticum.Its suppression ratio reaches and reaches as high as 66.15%.
2, to the Hela influence of cell generation cycle, see table 3.
Table 3: to the Hela influence of cell generation cycle
Experiment conclusion: the flow cytometer testing result shows that pharmaceutical composition effect 48h of the present invention can make Hela cell proliferation index (PI) descend, and S phase cell proportion is reduced, and G0/G1 phase cell proportion increases, and more Hela cell is blocked the phase in G0/G1.
Test Example 3: to pressing down the tumor experiment in the body of mouse cervical cancer model
Test material
(1) oncocyte: human cervical carcinoma Hela cell system.
(2) experimental animal: the female healthy mice of Kunming kind, 2-3 monthly age, body weight (25 ± 3) g.
(3) trial drug:
1 group of administration: 88mg Oleum Curcumae and 75mg Borneolum Syntheticum.
2 groups of administrations: 10mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+20mg furanodiene+25mg curdione+11mg beta-elemene+10mg curcumenol+12mg curzerene+75mg Borneolum Syntheticum.
(4) reagent: super NBCS, Hangzhou Ilex purpurea Hassk.[I.chinensis Sims Company products; Trypsin, Beijing Geyuantianrun Biotechnology Co., Ltd..
The foundation of mouse cervical cancer model:
The Hela cell is at 37 ℃, 5%CO
2, containing in the DMEM culture fluid of 10% hyclone and cultivate, the cell that will be in exponential phase is used 0.25% trypsinization, processes 5 * 10 with aseptic PBS liquid is resuspended
7The single cell suspension of/ml, every mice be the subcutaneous vaccination 0.2ml of nape portion cell suspension in the right side, inoculates 30 altogether, observes each injection point every day and have or not red and swollen ulceration.
Press down the tumor experiment
30 mices are divided into 3 groups at random, and 10 every group, administration group and blank group are established in experiment.The administration group is respectively at the medicine (preparing with normal saline) of inoculating cell suspension 2 all pneumoretroperitoneum injection 100mg/kg dosage, every day 1 time, continuous 4 weeks; And the isopyknic normal saline of matched group lumbar injection.Observe the gross tumor volume and the speed of growth in the experiment, experiment is carried out the back cervical vertebra dislocation of 4 weeks and is put to death mice, and it is heavy with tumor to weigh in.
The result
1, whole 30 mices all become to live in experimentation, and each cell suspension inoculation is put not swollen ulceration of show and bleeding.
2, at the 14th day of cell suspension inoculation, each inoculation point was all seen tumor nodule, diameter＞0.5cm.
3, mice carries out perusal to the tumor tissue under peeling off in the mice body after giving medicine, is nodositas; Canescence; Quality is hard partially, and there is pseudocapsule on the surface, and easy and surrounding tissue is peeled off; Tangible necrosis region appears in two most of visible central authorities in medication group tumor transverse section, the then rare necrosis region of matched group.
4, weighing administration group and control group mice body weight, and tumor body weight, the result sees table 4.
Table 4: mice average weight and tumor are heavy
Annotate: compare * * P＜0.01, * P＜0.05 with matched group; Compare #P＜0.05 for 1 group with administration.
The result: the body weight of administration group mice is apparently higher than mice in control group, and P＜0.05; Relatively the mice body weight of two administration groups does not have obvious difference, but tumor heavy be that 2 groups of administrations (giving pharmaceutical composition of the present invention) significantly are lower than 1 group of administration, P＜0.05.Explain that Oleum Curcumae+Borneolum Syntheticum and pharmaceutical composition of the present invention are all inhibited to cervical cancer.
Test Example 4: in-vitro antibacterial test
Trial drug: 10mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+20mg furanodiene+25mg curdione+11mg beta-elemene+10mg curcumenol+12mg curzerene+75mg Borneolum Syntheticum.
Method: adopt trace continuously doubling dilution measure the minimal inhibitory concentration (MIC) of pharmaceutical composition of the present invention to the mark bacterial strain of ETEC, staphylococcus aureus, Pseudomonas aeruginosa, staphylococcus epidermidis, proteus vulgaris, streptococcus faecalis, gonococcus and ETEC, staphylococcus aureus, Pseudomonas aeruginosa, staphylococcus epidermidis, proteus vulgaris, streptococcus faecalis, gonococcus clinical separation strain, adopt dull and stereotyped infection protocol to measure the MBC (MBC) of pharmaceutical composition of the present invention to above-mentioned antibacterial.ETEC, staphylococcus aureus, Pseudomonas aeruginosa, staphylococcus epidermidis, proteus vulgaris, vagina Gartner bacterium are inoculated in MH meat soup, put common incubator and cultivate 24h for 37 ℃; Streptococcus faecalis is inoculated in TPY meat soup (anaerobe nutrient broth), puts the anaerobism incubator and cultivates 48h for 37 ℃; Gonococcus is inoculated in the MH meat soup of antiperspirant 5% calf serum, puts 5%CO
2Cultivation property is cultivated 48h for 37 ℃.
Table 5 pharmaceutical composition of the present invention is to the MIC and the MBC of reference culture
Can know by last table; The mark bacterial strain of the ETEC that pharmaceutical composition of the present invention is selected for use test, staphylococcus aureus, Pseudomonas aeruginosa, staphylococcus epidermidis, proteus vulgaris, streptococcus faecalis, gonococcus all has than obvious suppression and deactivation with vagina Gartner bacterium clinical the separation with reference to strain, and its MBC is 2～4 times of MIC.
Table 6 pharmaceutical composition of the present invention is to MIC, the MIC of 405 strain clinical isolates strains
50, MIC
99And MBC (the mg compositions/ml)
| Bacterial strain | The strain number | MIC | MIC 50 | MIC 99 | MBC |
| Staphylococcus aureus | 100 | 0.39～1.56 | 1.0526 | 1.3084 | 0.78～6.25 |
| Escherichia coli | 120 | 6.25～50 | 18.9163 | 23.1657 | 25～100 |
| Staphylococcus epidermidis | 50 | 0.39～3.13 | 0.7232 | 1.5016 | 1.56～6.25 |
| Bacillus proteus | 100 | 12.5～50 | 26.125 | 34.0726 | 25～100 |
| Gonococcus | 24 | 0.195～0.39 | 0.2438 | 0.2751 | 0.195～0.78 |
| Vagina Gartner bacterium | 11 | 0.39～1.56 | 0.7800 | 0.9146 | 0.85～1.65 |
Visible by last table, the 405 strain clinical isolates strains that pharmaceutical composition of the present invention is selected for use test all have certain inhibition and deactivation, and its MBC is 2～4 times of MIC.
Test Example 5: external antifungal test
Trial drug: 10mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+20mg furanodiene+25mg curdione+11mg beta-elemene+10mg curcumenol+12mg curzerene+75mg Borneolum Syntheticum.
Method: adopt the continuous doubling dilution of trace to measure pharmaceutical composition of the present invention, adopt dull and stereotyped infection protocol to measure pharmaceutical composition of the present invention to oidiomycetic MBC to oidiomycetic MIC.Adopt the continuous doubling dilution of trace to measure the MIC of pharmaceutical composition of the present invention, adopt dull and stereotyped infection protocol to measure the MBC of pharmaceutical composition of the present invention mycete to mycete.
Table 7 pharmaceutical composition of the present invention is to the MIC and the MBC of test strain
Can know by last table; Mark bacterial strain or the clinical separation of the Candida albicans that pharmaceutical composition of the present invention is selected for use test, Candida parapsilosis, monilia guilliermondii, candida parakrusei, Oidium tropicale, penicillium, Aspergillus flavus all have than obvious suppression and deactivation with reference to strain, and its MBC is 2～4 times of MIC.
Table 8 pharmaceutical composition of the present invention is to MIC, the MIC of the strain of Candida albicans clinical isolates
50, MIC
99And MBC (the mg compositions/ml)
| The strain number | MIC | MIC 50 | MIC 99 | MBC |
| 50 | 1.56～6.25 | 0.0991 | 0.1243 | 6.25～12.5 |
Visible by last table, the 50 strain Candida albicans clinical isolates strains that pharmaceutical composition of the present invention is selected for use discoloration test have certain inhibition and deactivation.
Test Example 6: anti-trichomonal vaginitis test
Trial drug: 10mg 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-+20mg furanodiene+25mg curdione+11mg beta-elemene+10mg curcumenol+12mg curzerene+75mg Borneolum Syntheticum.
Method: adopt extracorporeal culture-ing to measure the minimum parasite killing concentration of pharmaceutical composition of the present invention to trichomonal vaginitis.Trichomonal vaginitis is cultivated 48h for 37 ℃ in the CPLM culture medium of improvement, and the infusorian motion is active, well-grown, and natural mortality rate＜2%, it is 1.8～2.5 * 10 that test uses trichomonal vaginitis liquid to contain worm concentration
3/ ml.
The result shows that pharmaceutical composition of the present invention is external effective to killing trichomonal vaginitis.
Specific embodiment
Embodiment 1
Prescription: 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-15g, furanodiene 39g, curdione 24g, beta-elemene 9g, curcumenol 8g, Borneolum Syntheticum 75g.
Dosage form: suppository.
Method for preparing: with 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, beta-elemene, curcumenol and 75g polyoxyethylene sorbitan monoleate (Tween-80) mixing of recipe quantity, Borneolum Syntheticum 75g dissolves with adequate amount of ethanol, with above-mentioned solution mixing.
The Myrj 45 1491g of recipe quantity puts in the water-bath heating and makes fusing, adds above-mentioned medicinal liquid, stirs, and 1000 of suppositorys are processed in fill.
Embodiment 2-8
By following prescription preparation suppository, method for preparing is with embodiment 1.
| Embodiment | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)- | 5.5 | 6 | 9 | 10 | 11 | 13 | 14 |
| Furanodiene | 37 | 33 | 30 | 28 | 25 | 15 | 11 |
| Curdione | 6 | 8 | 9 | 15 | 18 | 20 | 23 |
| Beta-elemene | 10.5 | 10 | 9.5 | 8 | 6.5 | 5 | 4.5 |
| Curcumenol | 1.5 | 2 | 4 | 6 | 8 | 9 | 9.5 |
| Curzerene | 8.5 | 9 | 10 | 15 | 14 | 16 | 21 |
| Borneolum Syntheticum | 30 | 40 | 50 | 60 | 70 | 80 | 90 |
| Polyoxyethylene sorbitan monoleate | 55 | 60 | 65 | 70 | 75 | 80 | 85 |
| Myrj 45 | 1563 | 1574 | 1467 | 1572 | 1583 | 1543 | 1548 |
Embodiment 9
Prescription: 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-6.5g, furanodiene 32g, curdione 21g, beta-elemene 9.5g, curcumenol 2.5g, Borneolum Syntheticum 35g.
Dosage form: effervescent tablet.
Method for preparing:
1) get beta-schardinger dextrin-, it is an amount of to add water by 25ml/g, stirs it is dissolved entirely, and heating in case of necessity gets beta-schardinger dextrin-solution; The 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-of recipe quantity, furanodiene, curdione, beta-elemene, curcumenol, Borneolum Syntheticum slowly adds in the beta-schardinger dextrin-solution with dehydrated alcohol dilution back; Continue to be stirred to into homogeneous phase, place in the refrigerator and spend the night sucking filtration; With a small amount of petroleum ether 3 times, freezing, white powder.
2) with above-mentioned white powder and 35g Borneolum Syntheticum, citric acid 150g and lactose 160g mixing; Other gets sodium bicarbonate 120g and lactose 160g mixing, granulates as binding agent with the 5%PVP ethanol solution respectively, and 50 ℃ of dryings, granulate, PEG6000 is an amount of in adding, mixing, tabletting.
Embodiment 10
Prescription: 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-7.5g, furanodiene 31g, curdione 19g, beta-elemene 5.5g, curcumenol 3.5g, curzerene 11g, Borneolum Syntheticum 45g.
Dosage form: capsule
Method for preparing:
1) the capsule leather is equipped with: press gelatin: glycerol: the weight portion mixture of water=10: 3.7: 9.5, get gelatin, and the adding suitable quantity of water makes the gelatin imbibition.Other gets glycerol and remaining water and puts and be heated to 70 ℃ 2 in the glue pot, and mix homogeneously adds expansible gelatin, stir, and fusion, insulation, vacuum suction is removed bubble, considers, and adds the PEG400 of about 3% weight portion, and mixing is incubated 50 ℃ then, and is subsequent use.
2) content preparation: get the 300g PEG400 and be heated to 80 ℃, add Borneolum Syntheticum 45g, treat to dissolve fully back adding 150g Macrogol 4000 and be stirred to dissolving; When temperature is reduced to 40～50 ℃, add 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, beta-elemene, curcumenol, the curzerene of recipe quantity, stir; Be cooled to 25～30 ℃, cross colloid mill mill 2 times, each 5min; Process content, under the heat-retaining condition, process 1000 soft capsules.
Embodiment 11
Prescription: 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-8.5g, furanodiene 29g, curdione 21g, beta-elemene 4.5g, curcumenol 4.5g, curzerene 22g, Borneolum Syntheticum 55g.
Dosage form: membrane.
Method for preparing: get polyvinyl alcohol 800g and in 800ml water, soaked 24 hours, under 80 ℃ of temperature, dissolve in the water-bath; With 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, the beta-elemene of recipe quantity, curcumenol, curzerene, Borneolum Syntheticum are dissolved in the 1000ml ethanol (W/V=75%), and above-mentioned poly-vinyl alcohol solution is mixed, and stir; In above-mentioned mixed liquor, add antioxidant sodium pyrosulfite 10g, wetting agent glycerol 80g, plasticizer three triacetin 80g, stir; Deviate from bubble by conventional method, after the coating film forming, drying is cut, and promptly gets 1000 membrane products.
Embodiment 12
Prescription: 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-9.5g, furanodiene 26g, curdione 23g, beta-elemene 4.2g, curcumenol 5.5g, curzerene 19g, Borneolum Syntheticum 65g.
Dosage form: gel
Method for preparing: get carbomer 20g and swell among the propylene glycol 760g, leave standstill to swelling and fully regulate pH value to 4～7 in the back, and adding propylene glycol 600g, to process substrate subsequent use; With 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, the beta-elemene of recipe quantity, curcumenol, curzerene, Borneolum Syntheticum are dissolved in and get mixed solution in the 120g PEG400, stir, and with mixed liquor and above-mentioned substrate mix homogeneously, packing promptly gets.
Embodiment 13
Prescription: 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-10.5g, furanodiene 23g, curdione 24.5g, beta-elemene 4.6g, curcumenol 6.5g, curzerene 14g, Borneolum Syntheticum 70g.
Dosage form: lotion
Method for preparing: get Borneolum Syntheticum 70g, add an amount of ethanol and make its dissolving; With 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, the beta-elemene of recipe quantity, curcumenol, curzerene and azone 20g, 2500g Tween 80 mix homogeneously; Add ethanol to 2800g; Stir back adding water for injection to 10kg, stir packing; Get concentrated solution, 10 times of uses of thin up during use.
Embodiment 14
Prescription: 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-12.5g, furanodiene 17g, curdione 18.5g, beta-elemene 6.3g, curcumenol 7.5g, curzerene 16g, Borneolum Syntheticum 80g.
Dosage form: ointment
Method for preparing: after 80～100 ℃ of temperature controls were melted down, 115～120 ℃ of sterilizations of temperature control were cooled to 70～80 ℃ with sterilized vaseline with vaseline 800g; With lanoline 40g in 115～120 ℃ of sterilizations of temperature control; With 55～65 ℃ of extremely fusings of dimethyl sulfoxine 25g temperature control; Vaseline is put into material-compound tank, put into a half and stir; With Borneolum Syntheticum, dimethyl sulfoxine and lanoline congruent melting, filter and drop in the material-compound tank and stir again; With 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, the beta-elemene of recipe quantity, curcumenol, curzerene add in the material-compound tank then, stir.
Claims (8)
1. pharmaceutical composition, its characteristic pharmaceutical composition comprises 3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-, furanodiene, curdione, beta-elemene, curcumenol, curzerene, Borneolum Syntheticum.
2. pharmaceutical composition according to claim 1, wherein said pharmaceutical composition comprises the composition of following weight portion:
3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-5-16 weight portion;
Furanodiene 10-40 weight portion;
Curdione 5-25 weight portion;
Beta-elemene 4-11 weight portion;
Curcumenol 1-10 weight portion;
Curzerene 8-22 weight portion;
Borneolum Syntheticum 30-90 weight portion.
3. pharmaceutical composition according to claim 1 and 2, wherein said Borneolum Syntheticum is selected from natural Broneolum Syntheticum and/or synthetic borneol.
4. preparation of pharmaceutical compositions according to claim 1 and 2 becomes vagina administration preparation or rectally preparation.
5. preparation according to claim 4, wherein preparation comprises suppository, ointment, capsule, effervescent tablet, gel, lotion, membrane or foam.
6. pharmaceutical composition according to claim 1 and 2 prevents and/or treats the application in the medicine of human papilloma virus infection in preparation.
7. application according to claim 6, wherein human papilloma virus infection comprises high-risk human mammilla papillomavirus infection and/or low risk human papilloma virus infection.
8. pharmaceutical composition according to claim 1 and 2 prevents and/or treats the application in the medicine of vaginitis, cervical erosion, cervical cancer in preparation.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103611076A (en) * | 2013-11-27 | 2014-03-05 | 符耿哲 | Curcumin-containing pharmaceutical composition |
| CN106074703A (en) * | 2016-08-13 | 2016-11-09 | 广州婕熹卡生物科技有限公司 | A kind of pharmaceutical composition for preventing and treat human papilloma virus infection and cervical cancer |
| CN107952034A (en) * | 2017-11-30 | 2018-04-24 | 陈容 | A kind of medical composition and its use |
| CN108524917A (en) * | 2017-11-29 | 2018-09-14 | 吉林大学 | Application of the melittin in the infection for the treatment of high-risk HPV virus and uterine neck cancer drug |
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| CN101244237A (en) * | 2007-05-24 | 2008-08-20 | 海南碧凯药业有限公司 | Suppository and preparation thereof |
| CN101422585A (en) * | 2008-07-28 | 2009-05-06 | 海南碧凯药业有限公司 | Pharmaceutical use of medicine composition containing zedoary turmeric oil |
| CN101791392A (en) * | 2010-04-22 | 2010-08-04 | 海南碧凯药业有限公司 | Application of medicinal composition |
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| CN101244237A (en) * | 2007-05-24 | 2008-08-20 | 海南碧凯药业有限公司 | Suppository and preparation thereof |
| CN101422585A (en) * | 2008-07-28 | 2009-05-06 | 海南碧凯药业有限公司 | Pharmaceutical use of medicine composition containing zedoary turmeric oil |
| CN101791392A (en) * | 2010-04-22 | 2010-08-04 | 海南碧凯药业有限公司 | Application of medicinal composition |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103611076A (en) * | 2013-11-27 | 2014-03-05 | 符耿哲 | Curcumin-containing pharmaceutical composition |
| CN106074703A (en) * | 2016-08-13 | 2016-11-09 | 广州婕熹卡生物科技有限公司 | A kind of pharmaceutical composition for preventing and treat human papilloma virus infection and cervical cancer |
| CN108524917A (en) * | 2017-11-29 | 2018-09-14 | 吉林大学 | Application of the melittin in the infection for the treatment of high-risk HPV virus and uterine neck cancer drug |
| CN107952034A (en) * | 2017-11-30 | 2018-04-24 | 陈容 | A kind of medical composition and its use |
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