CN100339069C - Formulation comprising thymol useful in treatment of drug resistant bacterial infections - Google Patents
Formulation comprising thymol useful in treatment of drug resistant bacterial infections Download PDFInfo
- Publication number
- CN100339069C CN100339069C CNB008194823A CN00819482A CN100339069C CN 100339069 C CN100339069 C CN 100339069C CN B008194823 A CNB008194823 A CN B008194823A CN 00819482 A CN00819482 A CN 00819482A CN 100339069 C CN100339069 C CN 100339069C
- Authority
- CN
- China
- Prior art keywords
- thymol
- preparation
- menthae
- oleum menthae
- obtains
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
The present invention relates to a new preparation for treating drug-resistant bacterial infections, which contains an effective amount of 'thymol' obtained from a plant named Trychyspermum ammi, mint oil compositions containing proper quantities of monoterpene obtained from field mint and mentha viridis, and conventional additive agents. The present invention also provides a method for preparing the preparation by mixing the components and a method for treating the drug resistance of patients by the administration of a therapeutically effective amount of the preparation.
Description
Invention field
The present invention relates to can be used for treat the cooperative compositions of the novelty of drug resistant bacterial infections, described compositions comprises the thymol of effective dose, the field Herba Menthae of proper proportion (Mentha arvensis) and Herba Menthae Rotundifoliae (Mentha spicata) blend of essential oils or its monoterpene component and conventional additive.Described compositions can be used for treating drug resistance intestinal portion and systemic infection.The preparation that can strengthen the thymol action activity comprises the combination of thymol and described quintessence oil, and this is a kind of carvone, 1 that contains, the combination of the rare mixture of 8-limonene and menthol.The present invention also provide produce described method for compositions and use the thymol that obtains from plant Trychyspermum ammi (Ajwain) seed as be used to control drug tolerant bacteria the 4th generation antibiotic formulations method.More properly, the present invention relates to from the purposes of plant Trychyspermum ammi (Ajwain) seed distilled oil isolated compound " thymol ", be used to kill antibiotic antibacterial of the third generation and multidrug-resisting (mdr) microbial pathogens that tolerance is generally used, thus can be used as based on plant the 4th generation the medical herbs antibiotic formulations.
Background of invention
Infected by microbes is harm humans health and the main cause that causes a large amount of people's death in the whole world and misery.In addition, infecting thoroughly influences human efficiency, makes various metabolic functions and system lose function, for example digests, breathing, urinary system, circulation, nervous system and skin.Cause the successive pain of people then, up to the invasive organism of curing the patient fully.Bacterial infection brings serious threat for the people's of each age group health.Since AlexanderFleming since 20th century, found penicillin the forties, the medical worker has used antibiotic and antimicrobial drug to eliminate infectious factors and cure diseases.But, infective micro-organisms always can be resisted every kind of new medicine, produces the medicine/antibiotic drug resistance in using.
The appearance of multidrug-resisting bacterial strain has become the realistic problem in the medical domain.The main cause that drug resistance produces is the generation of random mutation.Sudden change can occur in the gene of being responsible for giving to the sensitivity of medicine.Owing to lack the antibiotics with mode of action relatively, we can find what we can do when the medical science disaster is coming.Now this is when utilizing modern target class screening implement to disclose dark plant molecular miracle of hiding, and has the advanced person's of novel binding mode medicine of new generation and antibiotic with exploitation.
By the structural modification of existing medicine, developed the antimicrobial that upgrades, develop the higher generation medicine that has spectrum activity and strengthen effect.The representative instance of higher generation antimicrobial family is penicillins and cephalosporins.They come by the chemical modification development of basic beta-lactam nucleus.Cephalosporin has represented a big class grandson of cefalotin for medicine.Similarly, since Lesher with work together and developed nalidixic acid as the antimicrobial quinolone medicine of the first generation in 1962, quinolinones family has also developed into the third generation.In fact, in the seventies in 20th century, it is found that the application of nalidixic acid in the treatment systemic infection is limited, the improvement to quinolones subsequently is also unimportant, as the acid of quinoline, pipemidic acid and cinoxacin.The eighties in 20th century is early stage, and the development of fluoro-carbostyril class is a breakthrough.What at first occur is norfloxacin, and it is to have 6-fluorine and a piperazine substituted second filial generation quinolinones of 7-by Wolfson and Hooper exploitation.It has strengthened the activity to Gram-negative and gram-positive bacterium, and difference is blue pus organism (Pseudomonas aeruginosa) and staphylococcus aureus (Staphylococcus aureus) for example.Then developed sisters' drug molecule, for example ciprofloxacin, enoxacin, ofloxacin and pefloxacin etc.Almost simultaneously, developed third generation medicine, for example lomefloxacin, fleroxacin, temafloxacin, tosufloxacin are compared with the second filial generation quinolones that has a fluorine in basic quinolinones 6-position, and the third generation has another one or a plurality of fluoro substituents.
Generally the using continuously of these quinolinoness/fluoro-carbostyril medicine causes the antibacterial that is used to resist insensitive gradually, and pathogen needs these chemical compounds of increasing dosage thus.Owing to be synthetic, the use of these medicines causes the side effect that forces.And then, even pathogen is also becoming drug resistance to these chemical compounds of high dose more, make to be worse off.In order to reverse this problem, the applicant has developed to use from known plants isolated compound thymol and has killed drug tolerant bacteria and check the new method of bacterial resistance sexual developments with other available antibiotic combinations.
Consider the most fatal bacterial infection, pulmonary tuberculosis is the No.1 killer in the world, seizes every year to surpass 3,000,000 life in the world.More thrilling is that the M ﹠ M of this disease in the HIV positive individuals is higher.Phthisical resurrection and the sickness rate in the positive crowd of human immunodeficiency virus thereof are all paid close attention to pressing for the more advanced medicine of new generation of exploitation in developing country and industrialized country.
Present common recognition has reflected that repeatedly the common antibiotics that is used for the treatment of diseases such as pulmonary tuberculosis is invalid.These problems are now in developing country's ratio that takes advantage.Adopt the novel drugs combination of the bioactive molecule of natural origins such as plant to need system development, otherwise the result to be doomed in the new millennium to the mankind be destructive and out of control.In the most promising progress in drug development field is to find new molecule or available novel application of compound, and they have known safety, without any side effect.The combination of this class active biomolecule and other antibiotic can be killed drug tolerant bacteria, checks simultaneously that infectious microorganism is chemical sproof to further develop.
Thymol generally is used for various herbal medicinal products, from collutory and intestinal illness to skin infection.Parent oil and thymol itself treated general disease as ancient family's prescription component, has been found that child and adult effectively same.These purposes have been used to the Indian subcontinent, and cross over the continent by the tradition and the formal and informal communication of medical herbs knowledge.
Here need specifically to quote to establish the relevant or prior art of some of uniqueness of the present invention and comprise following different purposes.
1, be used for the treatment of viral infection, comprise compositions (Pruthi etc., 1999 of HIV; United States Patent (USP) 5,980,903:1999 November 9) and correlation technique (Badaway1998; United States Patent (USP) 5,801,153; In JIUYUE, 1998; Rohatgi 1996; United States Patent (USP) 5,529,778; In June, 1996; Hozumi etc., 1995; United States Patent (USP) 5,411,733; May nineteen ninety-five).
2, (Beck 1991 to can be used for the analgesic composition of respite arthritic symptom; United States Patent (USP) 5,073,366; On December 17th, 1991) and correlation technique (Arbir etc., 1981; United States Patent (USP) 4,307,109; In December, 1981; Dubash etc., 1983; United States Patent (USP) 4,383,986; May nineteen eighty-three; Seth 1985; United States Patent (USP) 4,540,572; In JIUYUE, 1985; Geria 1987: United States Patent (USP) 4,702,916; In October, 1987; Grohe 1989; United States Patent (USP) 4,844,902; In July, 1989; Bisset etc., 1989: United States Patent (USP) 4,847,071; In July, 1989; Deckner etc., 1989; United States Patent (USP) 4,863,725; In July, 1989).
3, collutory (Smigel etc., 1999: United States Patent (USP) 4,925,655; May 15 nineteen ninety) and correlation technique (Fleischman 1936; United States Patent (USP) 2,035,267; In March, 1936; Poetschke 1937; United States Patent (USP) 2,094,671; October nineteen thirty-seven; Welsh etc., 1970; United States Patent (USP) 3,518,343; In June, 1970; Breece etc., 1974; United States Patent (USP) 3,821,117; In June, 1974; Cheng 1976; United States Patent (USP) 3,936,385; In February, 1976).
Can find out obviously that from the list of references and the literature search of being quoted thymol and compositions thereof the purposes in drug tolerant bacteria/microorganism treatment is not all mentioned or described in known invention.But meanwhile, it behaves used is set, therefore need not toxicology test.
Therefore, press for the serious means with life-threatening bacterial infection of effective and cheap treatment, comprise TB, side effect is few or do not have.The present invention is devoted to this pressing for, provide known herb ingredients native compound and with other antibiotic/compound compositions, be used for control of bacterial infection, comprise pulmonary tuberculosis.Determination of biological activity method based on concrete target can limit from the new purposes of plant Trychyspermum ammi (Ajwain) the isolating plant compound of seed distilled oil " thymol ", in order to kill tolerance general third generation antibiotic antibacterial and multidrug-resisting (mdr) microbial pathogens.The character of thymol and former conventional use and show about the available information that the mankind use, as based on plant the 4th generation the medical herbs antibiotic formulations, it can be used safely.Therefore, the applicant is devoted to strengthen the activity of thymol effect, has developed novel preparation, comprises the combination of thymol and the suitable oil from Herba Menthae (field Herba Menthae and Herba Menthae Rotundifoliae)/monoterpene combination, wherein contain carvone, 1, the rare combination of 8-limonene and menthol.
The applicant has been found that brand-new and far-reaching application, is used for helping human health nursing comprehensively, to infected by microbes due to the anti-drug resistance pathogen.Common available antibiotic unscrupulous and excessive use has continued to cause the single and multidrug-resisting of appearance in infection.The applicant the classification of bacterial drug resistance as foundation to the human health classification of risks, this is well beyond traditional taxonomy category.The present invention is specifically related to continue the drug tolerant bacteria that existence catastrophic event afterwards produces by described antibiotic.In experiment, the applicant uses escherichia coli (Escherichia coli) and mycobacteria as model system, to monitor to the development of quinolinones and fluoroquinolone Drug tolerance and the appearance of mdr bacterial strain, find the new purposes of plant molecular (thymol), in order to the drug tolerant bacteria of new generation that kills the mdr bacterial strain and under the existing of these medicines, evolve.This mode is a kind of discovery of uniqueness, has great practicality at field of medicaments.
Goal of the invention
Main purpose of the present invention provides the novel formulation that can be used for treating drug resistant bacterial infections.
Another object of the present invention provides the novel formulation that can be used for treating bacterial infection, comprise effective dose from the deutero-thymol of plant Trychyspermum ammi (Ajwain) with from the combination of field Herba Menthae and deutero-aromatic oil of Herba Menthae Rotundifoliae or monoterpene.
Also have a purpose to provide the preparation method of novel formulation of the present invention.
Another object of the present invention provides the antibacterial agent that can be used for treating bacterial infection, comprises thymol.
Also have another purpose to provide the method for use thymol control drug tolerant bacteria.
Summary of the invention
The invention provides novel preparation, suitable Oleum menthae combination and the conventional additive that comprise the thymol that obtains from plant Trychyspermum ammi (Ajwain) of effective dose, obtain from field Herba Menthae and Herba Menthae Rotundifoliae.The present invention also provides the described preparation method that can be used for treating the novel formulation of drug resistant bacterial infections.And then, the invention provides the antibacterial agent that can be used for controlling drug tolerant bacteria, comprise the thymol of effective dose.The present invention also provides the method for using thymol control drug tolerant bacteria.
Detailed description of the invention
The invention provides the collaborative preparation of the novelty that can be used for treating drug resistant bacterial infections, described preparation comprise effective dose " thymol " that obtain from plant Trychyspermum ammi (Ajwain), contain the Oleum menthae combination and the conventional additive of an amount of or specific monoterpene that obtains from field Herba Menthae and Herba Menthae Rotundifoliae.
In one embodiment, the content of thymol is in 100 to 500mg or 20 to 50%w/w scopes.
In another embodiment, the content of Oleum menthae combination is about 0.1 to 0.5mg.
In another embodiment, additive is selected from the citric acid of about 2-10mg, the calcium carbonate of 100-200mg, the magnesium hydroxide gel of 50-100mg, the lactose of 200-600mg, 0.1 to 1% Mel, sodium glutamate and the sodium buffer of 200mg.In another embodiment, if necessary, Oleum menthae is diluted to 10 times.
In another embodiment, Oleum menthae comprise about 6 to 25% 1,8-limonene, about menthol of 0.50 to 2.50%, about carvone of 64.0 to 76%.
Another embodiment comprises Mel is diluted to 10 times.
In another embodiment, said preparation effectively resists the antibacterial that is selected from Mycobacterium or Escherichia.
In another embodiment, said preparation effectively resists drug tolerant bacteria, the medicine that is tolerated is selected from bromination 3,8-diaminourea-5-ethyl-6-phenylphenanthridineand , isoniazid, chloromycetin, tetracycline, rifampicin, nalidixic acid, oxolinic acid, sparfloxacin, ciprofloxacin and lomefloxacin.
Novel collaborative preparation can be used for treating drug resistant bacterial infections, the Oleum menthae that described preparation comprises " thymol " that obtain from plant Trychyspermum ammi of effective dose, obtain from field Herba Menthae and Herba Menthae Rotundifoliae hybrid and the additive of routine.
It should be noted that the surprising and unexpected antibacterial property of collaborative preparation performance of the present invention.Each composition of said preparation does not have the character of the compositions that can be used for treating drug resistant bacterial infections.The Oleum menthae that said preparation comprises " thymol " that obtain from plant Trychyspermum ammi (Ajwain) of effective dose, obtain from field Herba Menthae and Herba Menthae Rotundifoliae hybrid and the additive of routine show surprising and beyond thought antibacterial property.
Preparation of the present invention can derive from and be various physical form, for example tablet, syrup, powder, injection etc., and this is known in the art.
In order to prepare the preparation of various physical form, with the neccessary composition of said preparation, chemical compound thymol and to have an aromatic oil combination of required monoterpene combination mixed with the additive of above-mentioned routine just, for example Mel, sodium glutamate, citric acid etc. are to improve clinical efficacy.It is also to be noted that, the amount of each composition of preparation mentioned in this article only be the amount of exemplary, suitable each composition with different, determined by those skilled in the art easily.Content ratio in the preparation of the present invention is not crucial, has nothing in common with each other yet.Yes adopts the thymol of aforementioned proportion and the combination of specific Oleum menthae and obtain for best result.The optimum content of preparation composition will be different because of the medication of preparation.
The present invention also provides the preparation method of the preparation that can be used for treating drug resistant bacterial infections, and described method comprises the Oleum menthae combination step mixed with conventional additive that obtains with the thymol that obtains from plant Trychyspermum ammi of effective dose, from field Herba Menthae and Herba Menthae Rotundifoliae.
In one embodiment, the content of thymol is in 100 to 500mg or 20 to 50%w/w scopes.
In another embodiment, the content of Oleum menthae is about 0.1 to 0.5mg.
In another embodiment, additive is selected from the citric acid of about 2-10mg, the calcium carbonate of 100-200mg, the magnesium hydroxide gel of 50-100mg, the lactose of 200-600mg, 0.1 to 1% Mel, sodium glutamate and the sodium buffer of 200mg.
In another embodiment, if necessary, Oleum menthae is diluted to 10 times.
In another embodiment, Oleum menthae comprise about 6.8 to 23.2% 1,8-limonene, 0.66 to 2.50% menthol, 64.0 to 76.1% carvone.
Another embodiment comprises Mel is diluted to 10 times.
In further embodiment, thymol and Oleum menthae are dispersed in 0.1 to 1% Mel, obtain syrup.
In another embodiment, thymol and Oleum menthae and 2 to 10mg citric acids is mixed, be dissolved in the buffer that contains sodium glutamate, obtain injection.
In another embodiment, said preparation effectively resists the antibacterial that is selected from Mycobacterium or Escherichia.
In another embodiment, said preparation effectively resists drug tolerant bacteria, the medicine that is tolerated is selected from and comprises bromination 3,8-diaminourea-5-ethyl-6-phenylphenanthridineand , isoniazid, chloromycetin, tetracycline, rifampicin, nalidixic acid, oxolinic acid, sparfloxacin, ciprofloxacin and lomefloxacin.
And then the present invention also provides the Therapeutic Method of human drug resistant bacterial infections, comprises novel formulation with the treatment effective dose when needed to the step of curee's administration.
In one embodiment, said preparation is by oral or subcutaneous route administration.
In another embodiment, said preparation is dissolved in the 5ml 0.05M sodium buffer (pH 7.0) that contains the 200mg sodium glutamate, be used for subcutaneous injection.
In further embodiment, said preparation is effectively to bacterial-infection resisting, for example intestinal and systemic infection.
In another embodiment, treatment comprises the administration of said preparation, be used for the bacterial infection that causes by drug tolerant bacteria, the medicine that is tolerated is selected from bromination 3,8-diaminourea-5-ethyl-6-phenylphenanthridineand , isoniazid, chloromycetin, tetracycline, rifampicin, nalidixic acid, oxolinic acid, sparfloxacin, ciprofloxacin and lomefloxacin.
In one embodiment, said preparation is used for the treatment of the infection that is caused by the multidrug-resisting antibacterial, and this multidrug-resisting antibacterial is selected from Mycobacterium or Escherichia.
In another embodiment, said preparation is used to kill drug tolerant bacteria, the pharmaceutical pack that is tolerated contains bromination 3,8-diaminourea-5-ethyl-6-phenylphenanthridineand , isoniazid, chloromycetin, tetracycline, rifampicin, nalidixic acid, oxolinic acid, sparfloxacin, ciprofloxacin and lomefloxacin.
The present invention is the result by the predetermined process of the test of activity specific biological assessment algoscopy.The purpose of research be determine and estimate from the plant compound thymol of " Ajwain " oil as antibiotic of new generation and exploitation strengthened active, particularly kill the potentiality of the active medical herbs antibiotic formulations of drug tolerant bacteria.Test is devoted to find the purposes of this novelty, we observe for the first time, and the coli strain that tolerates nalidixic acid (a kind of wide spectrum quinolone medicine) owing to the sudden change of gyrA gene is responsive more to Trychyspermum ammi oil and key component " thymol " thereof on the contrary.
The following example is described the present invention in detail, and they should not be interpreted as the restriction to the scope of the invention or category.
Embodiment 1
Utilize disk diffusion measurement method to measure the antibacterial activity of the no thymol part of Trychyspermum ammi oil, thymol and oil, represent (table 1) with the growth inhibited district that on the lawn of endurance strain, produces.
Table 1: by the inhibition zone (mm) of disk diffusion measurement method mensuration
| Coli strain | Trychyspermum ammi oil | The oily part of no thymol | Thymol | ||
| 50mg/ml | 50mg/ml | 10mg/ml | 20mg/ml | 50mg/ml | |
| CA8000 | 4.5 | - | 1.0 | 2.0 | 5.0 |
| DH5α | 5.0 | - | 1.0 | 2.0 | 5.5 |
| ET8000 | 5.5 | - | 1.0 | 3.0 | 6.0 |
Here, CA8000 is colibacillary wild-type strain, and DH5 α and ET8000 concealment gyr sudden change that is to say that coding is responsible for the gene of the lax DNA gyrase Gyr A subunit of DNA and has been modified.Because the DNA gyrase has been changed, these mutant tolerance quinolinoness.By shifting the gene complementation effect of the plasmid of cloning gyr A, confirm specific sudden change.Just as shown in Table, thymol to kill the district more under the situation of gyr mutants which had of tolerance said medicine.In the no thymol part of oil, do not observe activity.
Embodiment 2
Poisonous agar method is measured nalidixic acid toleration coli strain to different antibiotic sensitivity
By that introduced and spontaneous gene mutation, the independent mutation body of split tolerance nalidixic acid in escherichia coli in the wild type CA8000 bacterial strain.Test their sensitive mode then, because the drug resistance of the second filial generation and third generation medicine is also only caused by the sudden change in the gyrA gene to nalidixic acid and other quinolinoness of new generation and fluoroquinolone medicine.
Following table 2 shows the crossing drug resistant sexual norm of this class mutant.
Table 2: escherichia coli Nal
RThe crossing drug resistant sexual norm of mutant
| Colibacillary mutant (Nal R) bacterial strain | Growth in poisonous agar culture medium wherein contains | ||||
| Nalidixic acid | Oxolinic acid | Sparfloxacin | Ciprofloxacin | Lomefloxacin | |
| 50μg/ml | 5μg/ml | 5μg/ml | 5μg/ml | 5μg/ml | |
| CAN 101 | + | - | + | - | - |
| CAN 102 | + | - | + | - | - |
| CAN 103 | + | - | + | - | - |
| CAN 104 | + | - | + | - | - |
| CAN 105 | + | - | + | - | - |
| CAN 106 | + | + | + | - | - |
| CAN 107 | + | + | + | - | - |
| CAN 108 | + | + | + | - | - |
| CAN 109 | + | + | + | - | - |
| CAN 110 | + | + | + | - | - |
| CAN 111 | + | + | + | - | - |
| CAN 112 | + | - | + | - | - |
| CAN 113 | + | + | + | - | - |
| CAN 114 | + | + | + | - | - |
| CAN 115 | + | + | + | - | - |
| CAN 116 | + | + | + | - | - |
| CAN 117 | + | + | + | - | - |
| CAN 118 | + | + | + | - | - |
| CAN 119 | + | - | + | - | - |
| CAN 120 | + | - | + | - | - |
| CAN 121 | + | + | + | - | - |
| CAN 122 | + | + | + | - | - |
| CAN 123 | + | + | + | - | - |
| CAN 124 | + | + | + | - | - |
| CAN 125 | + | + | + | - | - |
| CAN 126 | + | + | + | - | - |
| CA8000(WT) | - | - | - | - | - |
+=growth,-=not growth, the WT=wild type
The isolating nalidixic acid toleration strain growth of institute is as above shown described in the presence of different antibiotic.These mutants show positive growth in the presence of high concentration naphthyridines keto acid.Some also tolerates oxolinic acid, all tolerates sparfloxacin, but neither one tolerance second filial generation medicine ciprofloxacin and third generation medicine lomefloxacin.Just as was expected to these antibiotic is responsive for contrast wild-type strain CA8000.
Embodiment 3
Thymol antagonism escherichia coli Nal
RThe active biological assessment of mutant
Test said mutation body is further confirmed above-mentioned hypothesis, promptly colibacillary Nal to the sensitivity of thymol
RTherefore mutant can effectively be killed by thymol on the contrary, and its purposes as bouvardin control drug tolerant bacteria has been described.Thymol clearly shows all Nal of antagonism in these algoscopys
RNo matter the activity of mutant is single or multidrug-resisting (table 3).
Table 3: the active disk diffusion measurement method that is used to measure thymol antagonism nalidixic acid toleration escherichia coli mutant
| Colibacillary mutant (Nal R) bacterial strain | Thymol (the mm number of inhibition zone) | ||
| 50 μ g/ dish | 100 μ g/ dish | 250 μ g/ dish | |
| CAN 101 | Tr. | 1.5 | 4.0 |
| CAN 102 | Tr. | 2.0 | 7.0 |
| CAN 103 | Tr. | 2.0 | 4.0 |
| CAN 104 | Tr. | 2.0 | 6.0 |
| CAN 105 | Tr. | 2.0 | 5.0 |
| CAN 106 | 1 | 2.0 | 4.0 |
| CAN 107 | 1 | 3.0 | 5.0 |
| CAN 108 | Tr. | 2.0 | 5.0 |
| CAN 109 | 1 | 3.0 | 6.0 |
| CAN 110 | 1 | 3.0 | 6.0 |
| CAN 111 | 1 | 3.0 | 5.0 |
| CAN 112 | 1 | 3.0 | 5.0 |
| CAN 113 | Tr. | 2.0 | 5.0 |
| CAN 114 | 1 | 3.0 | 6.0 |
| CAN 115 | 1 | 3.0 | 5.0 |
| CAN 116 | 1 | 3.0 | 5.0 |
| CAN 117 | 1 | 3.0 | 5.0 |
| CAN 118 | 1 | 3.0 | 5.0 |
| CAN 119 | 1 | 3.0 | 6.0 |
| CAN 120 | 1 | 3.0 | 6.0 |
| CAN 121 | 1 | 3.0 | 5.0 |
| CAN 122 | Tr. | 2.0 | 5.0 |
| CAN 123 | 1 | 3.0 | 5.0 |
| CAN 124 | 1 | 3.0 | 5.0 |
| CAN 125 | 1 | 3.0 | 5.0 |
| CAN 126 | 1 | 2 | 5.0 |
| CA 8000(WT) | 1 | 2 | 4 |
Tr=trace activity (＜0.5mm) .WT=wild type
In fact, the chemical compound thymol can kill all nalidixic acid toleration escherichia coli, and efficient is higher than wild-type strain CA8000.
Embodiment 4
The separation and the characterized of the escherichia coli mutant of tolerance lomefloxacin (third generation fluoroquinolone medicine)
In growth medium, in the presence of lomefloxacin, after selecting, separate inductive mutant.According to the growth of poisonous agar method research lomefloxacin toleration coli strain in the presence of different antibiotic, to measure cross resistance.The expection of lomefloxacin (third generation) toleration bacterial strain also tolerates the first generation and second filial generation antibiotic (table 4).
Table 4: the sensitive mode of lomefloxacin toleration Bacillus coli cells
| Colibacillary mutant (Lom R) bacterial strain | Growth in poisonous agar culture medium wherein contains | |||||
| Lomefloxacin | Nalidixic acid | Oxolinic acid | Aorfloxacin | Ciprofloxacin | Sparfloxacin | |
| 1.5μg/ml | 50μg/ml | 5μg/ml | 20μg/ml | 2μg/ml | 20μg/ml | |
| CAL 101 | + | + | + | + | + | + |
| CAL 102 | + | + | + | + | + | + |
| CAL 103 | + | + | + | + | + | + |
| CAL 104 | + | + | + | + | + | + |
| CAL 105 | + | + | + | + | + | + |
| CAL 106 | + | + | + | + | + | + |
| CAL 107 | + | + | + | + | + | + |
| CAL 108 | + | + | + | + | + | + |
| CA8000(WT) | - | - | - | - | - | - |
+=growth,-=not growth, the WT=wild type
Embodiment 5
Thymol antagonism escherichia coli Lom
RThe active biological assessment of mutant
Tolerate the sensitivity of the anti-thymol of mutant of lomefloxacin by the test of disk diffusion measurement method.Can find out obviously that from table 5 thymol effectively kills colibacillary all Lom
RMutant cells shows that it can directly be used as the medicine of anti-drug resistance antibacterial of new generation.
Table 5: the active disk diffusion measurement method that is used to measure thymol antagonism lomefloxacin toleration escherichia coli mutant
| Colibacillary mutant (Lom R) bacterial strain | Thymol (the mm number of inhibition zone) | |
| 50 μ g/ dish | 100 μ g/ dish | |
| CAL 101 | 2.0 | 4.0 |
| CAL 102 | 3.0 | 5.0 |
| CAL 103 | 2.0 | 3.0 |
| CAL 104 | 1.0 | 1.5 |
| CAL 105 | 2.0 | 4.0 |
| CAL 106 | 2.0 | 4.0 |
| CAL 107 | 2.0 | 4.0 |
| CAL 108 | 3.0 | 5.0 |
| CA8000 | 3.0 | 5.0 |
Tr=trace activity (＜0.5mm), the WT=wild type
Embodiment 6
The activity rating of thymol antagonism nalidixic acid toleration smegma mycobacteria (Mycobacteriumsmegmatis) mutant
In case find challenging thymol antagonism forms in escherichia coli as antibacterial agent various levels and the chemical sproof susceptibility result of kind, we have also designed the mycobacteria test.We adopt the smegma mycobacteria, and this is the quick growth model system that is used to screen antituberculotics.At wild-type strain MC
2The a series of drug resistance mutants of middle separation.At first test the sensitivity of nalidixic acid toleration bacterial strain to thymol.Wild type smegma mycobacterium strain is grown in the culture medium that contains 50 μ g/ml nalidixic acids, separates the nalidixic acid mutant.Can find out obviously that from table 6 thymol can effectively kill the Nal of smegma mycobacteria
RBacterial strain has been established the effectively serviceability of anti-bacterial drug of its conduct, even the antagonism mycobacteria.
Table 6: the active disk diffusion measurement method that is used to measure thymol antagonism nalidixic acid toleration smegma mycobacteria mutant
| Mutant (the Nal of smegma mycobacteria R) bacterial strain | Nalidixic acid | Thymol (the mm number of inhibition zone) | ||
| 50 μ g/ dish | 50 μ g/ dish | 100 μ g/ dish | 250 μ g/ dish | |
| MSN 101 | - | Tr. | 3 | 5 |
| MSN 102 | - | Tr. | 2 | 5 |
| MSN 103 | - | 1 | 2 | 5 |
| MSN 104 | - | 1 | 2 | 5 |
| MSN 105 | - | - | - | 2 |
| MSN 106 | - | 1 | 3 | 5 |
| MSN 107 | - | 1 | 3 | 6 |
| MSN 108 | - | 1 | 3 | 5 |
| MSN 109 | - | 1 | 2 | 5 |
| MSN 110 | - | 1 | 2 | 5 |
| MSN 111 | - | 1 | 2 | 5 |
| MSN 112 | - | 1 | 3 | 5 |
| MSN 113 | - | Tr. | 2 | 3 |
| MSN 114 | - | Tr. | 1.5 | 3 |
| MSN 115 | - | Tr. | 2 | 3 |
| MC -(WT) | - | 1 | 2 | 4 |
Tr=trace activity (＜0.5mm), the WT=wild type
Embodiment 7
Thymol antagonism smegma mycobacteria Lom
RThe active biological assessment of mutant
Make the smegma mycobacterium growth in the culture medium that contains 20 μ g/ml lomefloxacins (third generation fluoroquinolone antibiotics), separate the lomefloxacin mutant.These mutant tolerances are up to the lomefloxacin of 60 μ g/ml.What point out here is that to appear at after the sudden change that drug resistance level in the mycobacteria cell compares with escherichia coli be very high, can kill this class Lom
RThe medicine of mutant will be need badly with valuable.What is interesting is, also find these mutant tolerance second filial generation antibiotic, as ciprofloxacin.Check then they to thymol, be the sensitivity of The compounds of this invention, just as shown in table 7, thymol effectively kills these multidrug-resisting sexual cells: the smegma mycobacteria, thereby confirmed thymol can serve as the 4th the generation/this inference of medical herbs antibiotic of new generation.
Table 7: the active disk diffusion measurement method that is used to measure thymol antagonism lomefloxacin toleration smegma mycobacteria mutant
| Mutant (the Lom of smegma mycobacteria R) bacterial strain | Lomefloxacin | Thymol (the mm number of inhibition zone) | |
| 200 μ g/ dish | 100 μ g/ dish | 150 μ g/ dish | |
| MSL 101 | - | 1 | 3 |
| MSL 102 | - | 1 | 3 |
| MSL 103 | - | 1 | 3 |
| MSL 104 | - | 1 | 3 |
| MSL 105 | - | 1 | 3 |
| MSL 106 | - | Tr. | 3 |
| MSL 107 | - | 1 | 3 |
| MSL 108 | - | 1 | 3 |
| MSL 109 | - | 1 | 2.5 |
| MSL 110 | - | 1 | 3 |
| MSL 111 | - | 1 | 3 |
| MSL 112 | - | 1 | 3 |
Tr=trace activity (＜0.5mm),-=activity, WT=wild type do not had
Embodiment 8:
The activity of the bacteria variants of the another kind of line antitubercular agent isoniazid of thymol antagonism tolerance
Table 8: isoniazid toleration escherichia coli mutant cells is to the sensitivity of thymol
| Coli strain | Thymol (the mm number of inhibition zone) |
| 100 μ g/ dish | |
| CA8000 | 6 |
| CAO3 | 2 |
| CAO3 (revertant) | 8 |
Isoniazid is that another kind is widely used in the phthisical medicine of control, and owing to not existing of oxyR system, antibacterial is responsive to this medicine.The antibiotic (1000 μ g/ml) of wild-type e. coli bacterial strain tolerance high concentration.Use this bacterial strain of N ' N-methyl nitrosoguanidine mutation, in meat soup, to be separated in killed sensitive strain (CAO3) under the 250 μ g/ml concentration.And then, detect spontaneous mutation from the sensitive mutant culture that tolerates 1000 μ g/ml isoniazids.Test the sensitivity of these cultures to 100 μ g/ dish thymol by disk diffusion measurement method, it is more responsive observing isoniazid toleration revertant bacterial strain (CAO3 revertant).
Embodiment 9
Thymol antagonism bromination 3, the activity of 8-diaminourea-5-ethyl-6-phenylphenanthridineand toleration (mdr) mutant
Table 9
| The smegma mycobacterium strain | Thymol (100 μ g/ dish) |
| Wild type | 8mm |
| Bromination 3,8-diaminourea-5-ethyl-6-phenylphenanthridineand toleration bacterial strain | 11mm |
Reach 12 μ g brominations 3 by the normal sensitivity wild-type strain of continuous enrichment, 8-diaminourea-5-ethyl-6-phenylphenanthridineand /ml meat soup, the mutant of separation smegma mycobacteria.This mutant is in this concentration bromination 3, well-grown under 8-diaminourea-5-ethyl-6-phenylphenanthridineand , and wild type is killed under 8-diaminourea-5-ethyl-6-phenylphenanthridineand /ml meat soup in 3 μ g brominations 3.And then, find this bromination 3,8-diaminourea-5-ethyl-6-phenylphenanthridineand toleration strains is multidrug-resisting (mdr), as chloromycetin (20 μ g/ml), tetracycline (10 μ g/ml) and rifampicin (40 μ g/ml).
Embodiment 10
The The compounds of this invention thymol under different pH to the activity of coli strain CA8000
Table 10
| Concentration | PH (the mm number of inhibition zone) | |||
| 4.00 | 7.00 | 9.00 | 1M NaOH | |
| 12.5 μ g/ dish | - | - | - | - |
| 25 μ g/ dish | Trace | 1.00 | - | - |
| 50 μ g/ dish | 2.00 | 2.00 | 1.00 | 1.00 |
| 100 μ g/ dish | 4.00 | 6.00 | 3.00 | 2.00 |
Last table clearly illustrates that the activity of The compounds of this invention is maximum under neutral pH (7.00).
Embodiment 11
The preparation of the Synergistic antimicrobial composition of the thymol of antagonism multidrug-resisting antibacterial
We observe from research, and thymol is the antibacterial of a kind of effective antagonism multidrug-resisting antibacterial.Further observe, can improve this effect in conjunction with Oleum menthae from field Herba Menthae and Herba Menthae Rotundifoliae.In different growth stage, the combination of oil contains 1,8-limonene (6.8-23.2%), menthol (0.66-2.45%), carvone (64.0-76.1%) and without the part of differentiating, and aromatic oil amounts to 100%.When using thymol concentration is that 0.1% line of oils is fashionable, and the antibacterial activity of thymol improves 45%.In addition, oil also increases the carvone fragrance and the menthol abnormal smells from the patient of compositions.In view of this, we have prepared following different compositions.
Table 11: be used to measure thymol and nalidixic acid synergistic disk diffusion measurement method to Bacillus coli cells
| Nalidixic acid concentration (μ g/ dish) | Inhibition zone (mm), disk contains | Thymol concentration (μ g/ dish) | ||
| Nalidixic acid | Combination (Nal+ thymol) | Thymol | ||
| 0.25 | 3 | 4 | Tr. | 12.5 |
| 0.50 | 6 | 12 | 2 | 25.0 |
| 1.00 | 9 | 10 | 3 | 50.0 |
| 1.50 | 11 | 10 | 5 | 75.0 |
| 2.00 | 12 | 10 | 6 | 100.0 |
Tr=trace activity (＜0.5mm), the WT=wild type
Table 12: be used to measure thymol and Herba Menthae hybrid oil synergistic disk diffusion measurement method to Bacillus coli cells
| Herba Menthae hybrid oil (μ g/ dish) | Inhibition zone (mm), disk contains | Thymol concentration (μ g/ dish) | ||
| Herba Menthae hybrid oil | Combination (oil+thymol) | Thymol | ||
| 50 | 0 | 1.0 | 3.0 | 50 |
| 5 | 0 | 5.0 | 3.0 | 50 |
| 0.5 | 0 | 2.5 | 3.0 | 50 |
Tr=trace activity (＜0.5mm), the WT=wild type
Compositions 1
1, thymol 100-500mg (20～50%w/w)
2, from aromatic oil 0.1～0.5mg of hybrid
3, citric acid 2～10mg
4, calcium carbonate 100～200mg
5, magnesium hydroxide gel 50～100mg
6, lactose 200～600mg
7, gross weight 500～1000mg
Each composition is suitably mixed, pulverize, be packaged in the commercially available capsule.
Compositions 2
With 0 to 10 times of aromatic oil dilution.
1, the thymol 50～300mg (10～60%) in the aromatic oil combination
These preparations are immersed in the 250mg lactose ball.
Compositions 3
Mel is diluted 0 to 10 times in sterile distilled water, adds following ingredients:
1, thymol 50～300mg (10～60%)
2, the aromatic oil from Herba Menthae makes up (0.1～1.0%).
Compositions 4
1, thymol 100～500mg
2, citric acid 2～10mg
These two kinds of components are dissolved in the 5ml 0.05M sodium buffer (pH 7.0) that contains the 200mg sodium glutamate.
Claims (13)
1, a kind of preparation that can be used for treating drug resistant bacterial infections, described preparation comprises the thymol that dosage is 100～500mg, the dosage that obtains from the hybrid of field Herba Menthae and Herba Menthae Rotundifoliae is Oleum menthae and the conventional additives of 0.1～0.5mg, described Oleum menthae necessarily comprise 6～25% 1,8-limonene, 0.50～2.50% menthol and 64.0～76% carvone.
2, preparation as claimed in claim 1, wherein thymol obtains from plant Trachyspermum ammi.
3, preparation as claimed in claim 1, wherein said additive are selected from the citric acid of 2～10mg, the calcium carbonate of 100～200mg, the magnesium hydroxide gel of 50～100mg, the lactose of 200～600mg, 0.1～1% Mel, sodium glutamate and the sodium buffer of 200mg.
4, preparation as claimed in claim 1 wherein is diluted to Oleum menthae 10 times.
5, preparation as claimed in claim 3 wherein is diluted to Mel 10 times.
6, a kind of preparation method that can be used for treating the preparation of drug resistant bacterial infections as claimed in claim 1, it is that the thymol of 100～500mg, the dosage that obtains from the hybrid of field Herba Menthae and Herba Menthae Rotundifoliae are Oleum menthae and the mixed step of conventional additives of 0.1～0.5mg that described method comprises dosage, described Oleum menthae comprise 6～25% 1,8-limonene, 0.50～2.50% menthol, 64.0～76% carvone.
7, method as claimed in claim 6, wherein said thymol obtains from plant Trachyspermum ammi.
8, method as claimed in claim 6, wherein said additive are selected from the citric acid of 2～10mg, the calcium carbonate of 100～200mg, the magnesium hydroxide gel of 50～100mg, the lactose of 200～600mg, 0.1～1% Mel, sodium glutamate and the sodium buffer of 200mg.
9, method as claimed in claim 6 wherein is diluted to Oleum menthae 10 times.
10, method as claimed in claim 8 wherein is diluted to Mel 10 times.
11, method as claimed in claim 6 wherein is dispersed in thymol and Oleum menthae in 0.1～1% the Mel, obtains syrup.
12, method as claimed in claim 6, wherein that the citric acid of thymol and Oleum menthae and 2～10mg is mixed, and be dissolved in the buffer that contains sodium glutamate, obtain injection.
13, contain thymol and the preparation of the Oleum menthae that obtains is used for preparing the treatment toleration bacterial infection and the effective purposes of the medicine of control of bacterial infection from the hybrid of field Herba Menthae and Herba Menthae Rotundifoliae, wherein said Oleum menthae comprise 6～25% 1,8-limonene, 0.50～2.50% menthol, 64.0～76% carvone.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/IN2000/000033 WO2001072317A1 (en) | 2000-03-28 | 2000-03-28 | Formulation comprising thymol useful in the treatment of drug resistant bacterial infections |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1452491A CN1452491A (en) | 2003-10-29 |
| CN100339069C true CN100339069C (en) | 2007-09-26 |
Family
ID=11076234
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB008194823A Expired - Fee Related CN100339069C (en) | 2000-03-28 | 2000-03-28 | Formulation comprising thymol useful in treatment of drug resistant bacterial infections |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN100339069C (en) |
| BR (1) | BR0017198A (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104803853B (en) * | 2014-01-29 | 2017-02-22 | 中国科学院海洋研究所 | Anti-Vibrio vulnificus monoterpene derivative, and preparation method and application thereof |
| CN108567814A (en) * | 2018-04-27 | 2018-09-25 | 江西中医药大学 | A kind of Herba Moslae volatile oil is for sterilizing and/or antibacterial, antimicrobial agent application and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996037210A2 (en) * | 1995-05-26 | 1996-11-28 | Ropapharm B.V. I.O. | Pharmaceutical compositions, based on etheric oils obtained from plants for use in the human and veterinary medical field |
-
2000
- 2000-03-28 BR BR0017198-0A patent/BR0017198A/en not_active Application Discontinuation
- 2000-03-28 CN CNB008194823A patent/CN100339069C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996037210A2 (en) * | 1995-05-26 | 1996-11-28 | Ropapharm B.V. I.O. | Pharmaceutical compositions, based on etheric oils obtained from plants for use in the human and veterinary medical field |
Also Published As
| Publication number | Publication date |
|---|---|
| BR0017198A (en) | 2003-04-08 |
| CN1452491A (en) | 2003-10-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gillespie et al. | Fluoroquinolones: a new treatment for tuberculosis? | |
| US6514541B2 (en) | Formulation comprising thymol useful in the treatment of drug resistant bacterial infections | |
| AU785159B2 (en) | Inhibition of pathogens by bacillus coagulans strains | |
| US11253556B2 (en) | Lactobacillus fermentum GKF3, composition comprising the strain and method for improving psychataxia using the same | |
| HUT61464A (en) | Preventive and curative pharmaceutical composition for curing diarrhoea and pseudomembranous colitis caused by clostridium difficile | |
| CN1649513A (en) | Bioavailability/bioefficacy enhancing activity of cuminum cyminum and extracts and fractions thereof | |
| CN111705042B (en) | Pasteurella phage vB _ PmuP _ PS02, phage composition and application thereof | |
| Gallani et al. | Motile Aeromonas septicemia in tambaqui Colossoma macropomum: Pathogenicity, lethality and new insights for control and disinfection in aquaculture | |
| Owhe-Ureghe et al. | Antibacterial activity of garlic and lime on isolates of extracted carious teeth | |
| CN100339069C (en) | Formulation comprising thymol useful in treatment of drug resistant bacterial infections | |
| KR100884681B1 (en) | The preparation method of lactic acid fermentation broths using rice washouts | |
| KR101871342B1 (en) | Novel Salmonella specific bacteriophage ST1 and antibacterial composition comprising the same | |
| CN1874785A (en) | Immunotherapic agent which is used for the combined treatment of tuberculosis together with other pharmaceuticals | |
| US6824795B2 (en) | Formulation comprising thymol useful in the treatment of drug resistant bacterial infections | |
| CN105831391B (en) | Traditional Chinese medicine micro-ecological granules for preventing and treating newcastle disease, feed and application thereof | |
| KR101840306B1 (en) | Composition for preventing or treating hyperlipidemia | |
| Dabanka et al. | Antibacterial Activity of Phyllanthus amarus (Schum and Thonn) Extract Against Salmonella typhi Causative Agent of Typhoid Fever | |
| RU2245719C2 (en) | Thymol-containing composition useful in treatment of patients suffering from drug-resistant bacterial infections | |
| Dangana et al. | Preliminary evaluation of antibacterial potential of four common Nigerian plants against isolates of Staphylococcus aureus and Escherichia coli | |
| CN1045895C (en) | Compound immune antibiotic | |
| CN1210030C (en) | Antimicrobial composition and mthod for producing same | |
| ES2820312B2 (en) | MYCOBACTERIUM CHELONAE STRAIN AND PHARMACEUTICAL, VETERINARY AND FOOD COMPOSITIONS CONTAINING IT FOR THE PREVENTION AND / OR CONTROL OF TUBERCULOSIS | |
| Teo | Antimicrobial study of pyroligneous extract from Rhizophora apiculate against urinary tract pathogens | |
| CN107320471A (en) | One kind treats seromycin pharmaceutical composition lungy and its application | |
| Gupta et al. | Resistance, Phylogenetic Analysis of Pathogenic Escherichia coli in Human Isolate and Their Sensitivity with Plant Extracts |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070926 Termination date: 20130328 |