CN1360567A - Sphingoid base devivs. and uses thereof - Google Patents
Sphingoid base devivs. and uses thereof Download PDFInfo
- Publication number
- CN1360567A CN1360567A CN00807004A CN00807004A CN1360567A CN 1360567 A CN1360567 A CN 1360567A CN 00807004 A CN00807004 A CN 00807004A CN 00807004 A CN00807004 A CN 00807004A CN 1360567 A CN1360567 A CN 1360567A
- Authority
- CN
- China
- Prior art keywords
- acid
- sphingosine base
- sphingosine
- base derivative
- hydrophilic
- 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.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C215/00—Compounds containing amino and hydroxy groups bound to the same carbon skeleton
- C07C215/02—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton
- C07C215/22—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being unsaturated
- C07C215/24—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being unsaturated and acyclic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/68—Sphingolipids, e.g. ceramides, cerebrosides, gangliosides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
Landscapes
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Biophysics (AREA)
- Rheumatology (AREA)
- Pain & Pain Management (AREA)
- Molecular Biology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Cosmetics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The present invention discloses sphingoid base derivatives which are salts of a sphingoid base. These salts have a substantially increased solubility in an aqueous environment and display an increased efficacy in compositions for topical use.
Description
Invention field
The present invention relates to the topical application field, particularly the topical application of sphingol alkali (sphingoidbase) derivative.
Background of invention
Sphingol alkali such as sphingosine are known is skin cell differentiation and propagation effective effector, by disturb basic biological chemistry cell processes carry out (Hannun, Y.A. and Bell, R.M. (1989), Science 243,500-507).For example, the activity of free sphingosine arrestin kinase c, thereby in signal transduction and cell fission adjusting, play a crucial role (Hannun, people such as Y.A. (1986), J.Biol.Chem.261,12604-12609).In the adjusting that acts on epidermal cell proliferation of free sphingosine is important factor, in case the speed that statocyte is lost by skin surface (Downing, D.T. (1992) J.Lipid Res., 33,301-313).In addition, by the agency of other biological activity of sphingol alkali, for example anti-microbial activity (Bibel, people such as D.E.. (1992), J.Invest.Dermatol.98,269-273).
Because they are to the effect and the anti-microbial activity of skin cell differentiation and propagation, sphingol alkali can be used as activeconstituents and is included in the various cosmetic compositions.For example, introduced various unusual and disease, for example xeroderma, xeroderma and the psoriasis that sphingosine is suitable for treating relevant skin.Sphingosine also can protect skin to resist various deleterious or undesirable effects, for example UV light and skin aging effect.Especially, sphingol alkali the office of being included in treat in the composition as anti-inflammatory agent or antiseptic-germicide (WO98/49999).
The shortcoming of sphingol alkali is their insoluble in aqueous environment.This phenomenon has hindered the use of these compounds at aqueous compositions.For example, in order to show effective anti-microbial activity, it is important making sphingol alkali be dissolved in aqueous compositions.
Invention is described
The invention discloses the derivative of sphingol alkali, they have the solubleness in water of remarkable rising than its free alkali counterpart.Thereby these sphingol alkali derivants show the effect of improving astoundingly when it is prepared into aqueous composition matter,
It is the salt of sphingol alkali that sphingol alkali of the present invention is derived.
According to the present invention, the negative ion source of the salt of sphingol alkali is from any suitable acid.In that respect, suitable acid is defined as a kind of acid, and it is mixed together with sphingol alkali in appropriate solvent, produces a kind of salt that has the solubleness of rising at water-bearing media, compares with the solubleness of sphingol alkali itself.
In an embodiment of the present invention, this acid is a kind of the acid that can have curative effect in topical application.
In an embodiment of the present invention, this acid is a kind of hydrophilic acid that can discharge sphingol alkali to the water of cosmetic or pharmaceutical composition.
Preferably, this acid is hydrophilic organic acid, for example Alpha-hydroxy paraffinic acid, beta-hydroxy paraffinic acid, α, beta-dihydroxyl paraffinic acid, chain docosandioic acid or mineral acid.The preferred example of hydrophilic organic acid is lactic acid, oxyacetic acid (glycolic acid), oxysuccinic acid, pyruvic acid, succsinic acid, fumaric acid, citric acid, xitix, glyconic acid and/or Pyrrolidonecarboxylic acid (pyrrolidone carboxylic acid).The example of preferred mineral acid is hydrochloric acid, nitric acid and/or phosphoric acid.
In another embodiment of the present invention, this acid is the lipophilic organic acid, like this so that improved the curative effect of lipophilic acid and sphingol alkali with the combination of sphingol alkali.
Sphingol alkali of the present invention can be prepared as follows.Dissolving sphingol alkali wherein adds the suitable acid of monovalent at least in suitable organic solvent.Usually, the affiliation that adds of acid causes about at least 3 units of pH decline.The end value that is appreciated that pH can depend on the acid of employing.The dissolving of sphingol alkali in organic solvent preferably produces in the temperature that raises, for example 50 ℃ to 70 ℃.During cooling mixture, the sphingol alkali salt can precipitate.By filtering and can choosing wantonly, preferably, from reaction mixture, reclaim the crystalline deposit thing with the used identical solvent of preparation salt with solvent wash.
The organic solvent that is fit to is final product wherein preferably, and promptly the sphingol alkali salt is insoluble solvent.The organic solvent that is fit to for example is ethanol or methyl iso-butyl ketone (MIBK).
In an embodiment of the present invention, the sphingol alkali salt is as the starting compound of another sphingol alkali salt preparation.
Sphingol alkali salt of the present invention preparation before they use is necessary, and for example they are included in the topical composition.The inclusion that additionally contains free sphingol alkali in the anionic topical composition of one or more acid of above definition can not cause that solubleness raises and/or curative effect increases.
Sphingol alkali salt of the present invention is preferably the salt of sphingol alkali sphingosine, D-sphinganine or phytosphingosine.More preferably, the sphingol alkali salt is a phytosphingosine salt.
In an embodiment of the present invention, obtain phytosphingosine through microbial fermentation.For example, phytosphingosine by Pichia ciferrii deutero-tetra-acetylated-phytosphingosine (TAPS), obtain by the deacetylation reaction that is fit to.Deacetylation can be chemical, for example passes through with the catalytic hydrolysis of potassium hydroxide base, perhaps enzymolysis.After the basic hydrolysis of TAPS, but the phytosphingosine of purifying gained.Such purifying can be undertaken by the known any method of those skilled in the art.Yeast-deutero-phytosphingosine is that human skin is equal to, and it is in the news and has the three-dimensional chemical configuration same as the Mammals phytosphingosine, that is, and and the red configuration of D-D-.
The solubleness that sphingol alkali salt of the present invention has in aqueous environment is significantly higher than the solubleness of free sphingol alkali.Show astoundingly further that by the present invention compare with free sphingol alkali, the sphingol alkali salt has the curative effect of increase, in addition also like this in the environment that free sphingol alkali also is solubilized form.Free sphingol alkali can be solubilized form when additional water-bearing media that has an organic solvent and surface active cpd.
The composition that comprises the sphingol alkali derivant according to the present invention is applicable to topical application, and topical application here is understood as cosmetic and/or the dermatological applications on the last leather lining that is included in skin, hair and mouth, nose, eye, urogenital tract or the like.
Sphingol alkali derivant of the present invention preferably mixes in the topical composition with 0.001 to 5 weight % concentration, preferred 0.005 to 5 weight %, more preferably 0.01 to 2.5 weight %, 0.02 to 1 weight % most preferably, preferred especially 0.02 to 0.5 weight %.
According to not wishing and/or unusual symptom that the topical composition that the present invention includes the sphingol alkali derivant is specially adapted to that various parts relevant with inflammation and/or microorganism active take place.
The topical composition that comprises the sphingol alkali derivant of the present invention can be favourable application, it is local that example undesirable and/or abnormal symptom takes place is eczema, psoriasis, atopic dermatitis, acne, seborrheic dermatitis, mouth and/or lip infection, mycosis, various other skin infections disease or vaginal infection.The topical composition that comprises described sphingol alkali derivant further acts on wound healing valuably and for example burns under the situation, and the normalizing of skin flora.
Because their anti-microbial activity, sphingol alkali derivant of the present invention make up and dermatological compositions in can also play sanitas, to reduce and/or to replace existing Chemical Preservative.
PS. Lactated preparation
The mixture of 50 gram phytosphingosines and 500ml dehydrated alcohol stirs and is heated to 65 ℃.Then with the paper filter filtered while hot this almost clear soln to 1 rise in the 3-neck flask.
(L)-when lactic acid (25.7g) stirs by partly adding in the filtrate, so that pH is reduced to 5.3 from 9.9, simultaneous temperature rises to 71 ℃ from 66 ℃.Mixture stirs and cooling.Begin crystallization in the time of about 45 ℃, continue cooling 3/4 hour to 21 ℃ simultaneously.
Leach precipitation, filter cake is with 150ml ethanol displacement (filter fast and replace, amount to 2 minutes).
Wet cake (110.8g) vacuum-drying produces the product of 51.2 grams.The NMR purity assay is 99.3%.
PS. the preparation of oxyacetate
The mixture of 50 gram phytosphingosines and 500ml dehydrated alcohol stirs and is heated to 65 ℃.Then with the paper filter filtered while hot this almost clear soln to 1 rise in the 3-neck flask.Hot ethanol flushing with 20ml.Filtrate is heated to 65 ℃ again.
By partly adding in the filtrate, so that pH is reduced to 5.6 from 9.9, simultaneous temperature rose to 68 ℃ from 65 ℃ when hydroxyethanoic acid (13.4g) stirred.Mixture stirs and cooling.Begin crystallization in the time of about 66 ℃, cooling simultaneously continues 20 minutes to 25 ℃.
Leach precipitation, filter cake is with 150ml ethanol displacement (filter fast and replace, amount to 3 minutes).
Wet cake (87g) vacuum-drying is spent the night and is produced the product of 56.6 grams.The NMR purity assay is 98.6%.
PS. the preparation of hydrochloride
The mixture of 50 gram phytosphingosines and 500ml dehydrated alcohol stirs and is heated to 65 ℃.Then with the paper filter filtered while hot this almost clear soln to 1 rise in the 3-neck flask.Hot ethanol flushing with 20ml.
Add when hydrochloric acid (36%, approximately 13ml) stirs in the filtrate, about 0 so that pH is reduced to from 10.3, simultaneous temperature rises to 50 ℃ from 45 ℃.Mixture stirs and cooling.After adding crystal seed, begin crystallization and continue to cool off 0.5 hour to 10 ℃ 34 ℃ the time.
Leach precipitation, filter cake is with the cold ethanol displacement of 100ml (slowly filter and replace, amount to 3/4 hour).
Wet cake (272g) vacuum-drying produces the product of 48.0 grams.The NMR purity assay is 96.7%.
PS. the preparation of pyroglutamate
The suspension of 25 gram phytosphingosine, 200ml methyl iso-butyl ketone (MIBK) (MIK) and 2ml water stirs and is heated to 66 ℃.
Then add 12 gram DL-Pyrrolidonecarboxylic acids, pH is become 5.8 from 9.4.
Obtain the glassy precipitate thing.Begin crystalline 1ml sample 45 ℃ of employings through scraping.This is used for giving mixture as crystal seed in further cooling period.
Next step, mixture further is cooled to 17 ℃ and filter with glass G3 filter, with the fresh MIK of 50ml (filtering fast) flushing/displacement.Wet cake (57g) vacuum-drying produces 34.3 gram products.
PS. the preparation of Citrate trianion
The suspension of 25 gram phytosphingosine, 200ml methyl iso-butyl ketone (MIBK) (MIK) and 1ml water stirs and is heated to 72 ℃.
Next step adds 18 gram citric acid monohydrate compounds, and pH is become 1.8 from 9.4.
Obtain throw out.Next step mixture is cooled to 14 ℃ and filter with glass G3 filter, with the fresh MIK of 50ml (filtering fast) flushing/displacement.Wet cake (84g) vacuum-drying produces 39.7 gram products.The NMR purity assay is 96.4%.
PS is to the zymic anti-microbial activity
Adopt two kinds of different yeast strains: yeast saccharomyces cerevisiae ATCC 9763 and Candida albicans ATCC 10231.Perhaps carry out all cultivations at 30 ℃ (yeast saccharomyces cerevisiaes) or 37 ℃ (Candida albicanss).Two primary yeast bacterial strains YEPD2% substratum growth (20g/l glucose, the 10g/l peptone, the 20g/l yeast extract, pH=6.0).The culture liquid of growing, by centrifugal collection 50 μ l cells in culture, with 1ml sterile buffer (10mM HEPES (pH=7.2 uses NaOH)+20g/l glucose) washing, centrifugal and in the sterile buffer of 0.5ml resuspending.
At a kind of solvent system, it is made up of the aqueous glycerin solution of 1 volume fraction ethanol, 2 volume fraction polysorbas20s and 17 volume fractions 50%, the reserve liquid of preparation phytosphingosine (PS) 10mg/ml.With this order, in the composition adding phytosphingosine with this solvent system, effectively vibrate solution after each the adding.When all solvents all add, mixture heating up to 40 ℃ reaches 15 to 30 minutes.Aqueous ethanolic solution 5% prepares the dilution (if desired) of reserve liquid.Prepared all solution, and remain on room temperature in 24 hours before using.
Adopt LIVE/DEAD
(USA) the research phytosphingosine is to the antifungic action of this two primary yeast for Molecular ProbesInc., Oregon for yeast life-span test kit L-7009.This test kit adopts two kinds of different fluorescence dyes, FUN-1
TMAnd Calcofluor
TMWhite M2R, with live and dead cell between show difference, this can adopt the fluorescence microscope with suitable spectral filter.
For this analysis, the fluorescence dye of following amount adds in the sterile buffer suspension of 0.5ml yeast cell, is prepared as previously mentioned: 1 μ l FUN-1
TMWith 2.5 μ lCalcofluor
TMWhite M2R.After the mixing, these suspension were cultivated 30 minutes.The suitable diluent that then adds 50 μ l phytosphingosine reserve liquids is to obtain the ultimate density as Fig. 1 a and 1b demonstration.After the mixing, cultivate these suspension, that live and mark time to time change dead cell.
0lympus BHB fluorescent microscope is used for microscopic examination, and it has two kinds of different filtering apparatus:
1. dichroic mirror blueness (B) excites filter IF490, emission filter 0530 (cell of living shows orange particle in cell, and dead cell equably green/orange).
2. dichroic mirror pansy (V) excites filter U95-B93, emission filter Y455 (cell of living shows the blue cell wall, and dead cell does not show).
The result is shown in Fig. 1 a and 1b.It clearly illustrates that two primary yeast bacterial strains are all killed by phytosphingosine (PS) with the dose-dependently form.
Embodiment 7
PS is to the antifungic action of hungry cell
In its natural living environment, most of time microorganism cellss are starvations.Whether this anti-microbial effect of pointing out us to study PS also obviously suppresses hungry cell.
For this purpose, revise the method that embodiment 6 introduces (all methods and condition be identical unless stated otherwise) a little.By centrifugal overnight culture collecting cell by Candida albicans ATCC 10231.Adopt two kinds of different damping fluids to wash and the resuspension cell: as 10mM HEPES (pH=7.2 uses the NaOH)+20g/l glucose that uses among the embodiment 6, and the same buffer that does not contain glucose.
Two kinds of cell suspending liquids contain and do not conform to glucose (each is 2.5ml), cultivate 10 minutes at 37 ℃.Then add the suitable dilution of 125 μ l phytosphingosine reserve liquids, obtain ultimate density shown in Figure 2.After the mixing, further cultivate these suspension, and, adopt 100 μ l samples at the time point that shows.Cell is by centrifugal collection, and cell is resuspending in containing 100 μ l damping fluids of glucose, and the concentration of fluorescence dye is identical with embodiment 6.This analysis of mixtures was cultivated more than 10 minutes so that dye distribution is gone into cell, as embodiment 6 described definite work with quantity dead cell.
As shown in Figure 2, hungry cell is more responsive than the cell of heavy meal to the antifungic action of PS.In fact, after the initial lag phase, 250mg/l PS proof kill on the hungry cell the same with 500mg/l effective.Lag phase is general owing to exist the cell endogenetic energy to stock, and this shows that again PS is effective especially to starvation.
Embodiment 8
The anti-microbial effect of PS in the cosmetic formulations in the agar diffusion test
The overnight culture (BHI substratum (Difco), 37 ℃ of cultivations) for preparing streptococcus aureus ATCC 14458 with the method that is similar to embodiment 6 introductions.
For diffusion test prepares agar plate, the BHI substratum of 300ml and 1% agar and 15% glycerine that add melt and are cooled to 50 ℃.The overnight culture that then adds aseptic glucose solution of 6ml (50%w/v) and 6ml microorganism.Petri diss is packed into the nutrient agar of 12.5ml, allows substratum solidify.
The preparation of test prepares as shown in Figure 6, and fat is octyl group dodecyl lactate mutually.It contains 1,2 or 5g/l PS.
For sample is added to test board, stainless steel ring (6mm internal diameter) is inserted empty sterile petri dish.In this ring, put into 2 round scraps of paper (6mm diameter) to cover the bottom, the test preparation of 50 μ l is inserted on the filter disc.Ring is placed on the surface of the agar plate that contains microorganism, and agar plate to allow the diffusion of testing liquid, is withdrawn ring 5 ℃ of storages afterwards in the period that shows in table.Subsequently, agar plate is cultivated (37 ℃) in the temperature that is fit to microorganism growth.After microorganism is grown in the non-inhibitory area of agar plate fully, measure the inhibition degree, as non-growth (or reducing growth) district, it extends two right angle orientation by using the zone.
The anti-microbial effect of PS in table 1. cosmetic formulations
| ??[PS](g/l) | | Spread 5 ℃ 14 days | ||||
| ??1 | ????- | ????- | ????- | ????- | ????- | ????- |
| ??2 | ????7 | ????6 | ????5 | ????7/14 | ????7/13 | ????5/11 |
| ??5 | ????5 | ????6.5 | ????7 | ????7.5/14 | ????9.5/16 | ????9/15 |
Two accompanying drawings given here, first refers to non-vitellarium, and second finger reduces the vitellarium.
As shown in Table 1, there is PS dose-dependently growth-inhibiting.
Embodiment 9
The antifungic action of PS and its part derivative
Have been found that the part Phytosphingosine derivate has the solubleness of improvement in Aquo System.This anti-microbial activity of pointing out our anti-microbial activity with them and PS itself relatively.Study following derivative: the oxyacetic acid of phytosphingosine, lactic acid and hydrochloride.Prepare the reserve liquid of phytosphingosine salt as embodiment 6 with regard to the method for phytosphingosine introduction, and adopt same implementation condition.
All three kinds of salt of finding test have the stronger anti-mycotic activity that specific ionization alkali has.This is owing to the negatively charged ion that exists in the salts solution, and (Fig. 3 a) owing to contain the no effect of blank group of lactic acid salt, hydrochloride or hydroxyl acetate of appropriate amount.In Fig. 3 b, the effectiveness that can find out hydrochloride is more than 2.5 times of effectiveness of PS alkali.
Embodiment 10
PS and the antifungic action of part derivative in solvent-free system thereof
In deionized water, prepare solution with the method (embodiment 6) that is similar to the solvent system introduction, comprise last heating steps.
Can find that in Fig. 4 a the shortage of solvent during specimen preparation almost eliminated the antifungic action of free PS alkali, and the effectiveness of PS salt does not reduce.In fact, the effectiveness of finding PS salt is at solvent-free system even can be higher, shown in Fig. 4 b and 4c.
Embodiment 11
PS is to the anti-microbial effect of bacterium
Adopt two kinds of different bacterial isolateses: streptococcus aureus ATCC 14458 and intestinal bacteria 421. to carry out all cultivations at 37 ℃.The method that two kinds of bacteriums such as embodiment 8 introduce is grown, by centrifugal collection 50 μ l cultures, with the washing of 1ml aseptic deionized water, and in the 0.5ml aseptic deionized water resuspension.
Preparation 10mg/ml PS reserve liquid as preceding introduction.The dilution (if desired) for preparing this reserve liquid with 5% aqueous ethanolic solution.Before using, prepare all solution in 24 hours, remain on room temperature.
Adopt LIVE/DEAD
BacLight
TM(USA) the research phytosphingosine is to the anti-microbial effect of these two kinds of bacteriums for MolecularProbes Inc., Oregon for bacterium life-span test kit L-7012.This test kit adopts two kinds of different fluorescence dyes, SYTO9 dyestuff and propidium iodide, with live and dead cell between show difference, this can adopt the fluorescence microscope with suitable spectral filter.
The solution of the dyestuff that test kit provides just mixed in 1: 1 before use, and the fluorescence dye mixture of 1.5 μ l adds in the bacterial suspension of 0.5ml.Subsequently, add the suitable dilution of the phytosphingosine reserve liquid of 50 μ l, to obtain the ultimate density that Fig. 5 shows.After the mixing, cultivate these suspension, that live and mark time to time change dead cell.
Adopt Olympus BHB fluorescent microscope to carry out microscopic inspection, it has following filtering apparatus: dichroic mirror blueness (B) excites filter IF490, emission filter 0530 (cell of living is green, and dead cell is orange/yellow).
Find that streptococcus aureus is killed strongly.Yet this effect does not quantize, because the dead cell attribution can not detect for dissolving: the measurement with fluorescence dye needs dead cell to keep structural integrity.Therefore, lethal effect obviously only reduces viable cell quantity strongly.
Bacillus coli cells is also effectively killed, and dead cell can be easy to quantize (Fig. 5) in this organism.Its showed cell is killed in the dose-dependently mode by PS, and this bacterium is more responsive than fungi to this compound.
Embodiment 12
Antibiotic and the antifungic action of PS derivative in diffusion test
Specify the organic overnight culture of test such as the method preparation that embodiment 6 and 8 introduces, what sample such as embodiment 8 introduced is applied on the test board.Sample is by being prepared into the suitable dilution thing as the reserve liquid of introducing preparation in the past.Agar plate spends the night 5 ℃ of storages, to spread in testing liquid, withdraws the point sample ring afterwards.Subsequently, agar plate is cultivated (37 ℃) in the temperature that is fit to microorganism growth.After growing fully in the non-inhibitory area of agar plate, measure the inhibition degree in microorganism, as non-growth (or reducing growth) district, it extends (in mm) by using the zone two right angle orientation.
Antibacterium and the antifungic action of table 2.PS-derivative in agar diffusion test
| Derivative | ??(g/l) | Streptococcus aureus ATCC 14456 | Candida albicans ATCC 10231 | ||||
| The PS-lactic acid salt | ????10 | ????12 | ??13 | ????13 | ????9 * | ????9.5 * | ????10.5 * |
| ????5 | ????12 | ??10.5 | ????12 | ????8 * | ????9 * | ????9 * | |
| ????2 | ????8 | ??8.5 | ????9 | ????± | ????5 * | ????5 * | |
| ????1 | ????6 | ??5 | ????5 | ????4 * | ????6 * | ????4 * | |
| The PS-hydroxyl acetate | ????10 | ????11.5 | ??9.5 | ????11 | ????7.5 | ????5 * | ????6 |
| ????5 | ????9 | ??9 | ????9 | ????3 | ????3 | ????3 | |
| ????2 | ????7 | ??6 | ????5 | ????- | ????- | ????- | |
| ????1 | ????3 | ??4.5 | ????3 | ????- | ????- | ????- | |
| The PS-hydrochloride | ????10 | ????12.5 | ??11 | ????11 | ????7 | ????6 * | ????7 |
| ????5 | ????9 | ??9 | ????9 | ????2 | ????3 | ????3 | |
| ????2 | ????6 | ??7 | ????5 | ????- | ????- | ????- | |
| ????1 | ????5 | ??5 | ????5 | ????- | ????- | ????- | |
* the data that are marked with asterisk refer to suppress the vitellarium; Unmarked data refer to not have the vitellarium.
Table 2 shows that the PS derivative has fully antibacterium and fungi activity in diffusion test, and this effect has tangible dose-dependently.
Claims (12)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP99200700.5 | 1999-03-09 | ||
| EP99200700 | 1999-03-09 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1360567A true CN1360567A (en) | 2002-07-24 |
Family
ID=8239966
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN00807004A Pending CN1360567A (en) | 1999-03-09 | 2000-03-09 | Sphingoid base devivs. and uses thereof |
Country Status (6)
| Country | Link |
|---|---|
| EP (1) | EP1159256A1 (en) |
| JP (1) | JP2002539110A (en) |
| KR (1) | KR20010108331A (en) |
| CN (1) | CN1360567A (en) |
| BR (1) | BR0009265A (en) |
| WO (1) | WO2000053568A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN119390593A (en) * | 2024-09-26 | 2025-02-07 | 优然创新(北京)技术研发中心有限公司 | A sphingosine supramolecular body and its preparation method and application |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002060405A1 (en) * | 2001-01-29 | 2002-08-08 | Cosmoferm B.V. | Veterinary dermatologic composition comprising a sphingoid base and/or a sphingoid base derivative |
| EP1287815A1 (en) * | 2001-08-31 | 2003-03-05 | Cosmoferm B.V. | Use of a sphingoid base for inhibiting ceramidase activity |
| EP1291340B1 (en) * | 2001-09-05 | 2006-03-15 | Charmzone Co., Ltd | Phytosphingosine derivatives with antitumor activity |
| FR2836630B1 (en) * | 2002-03-01 | 2004-07-09 | Lvmh Rech | COSMETIC USE OF PHYTOSPHINGOSINE AS A SLIMMING AGENT AND COSMETIC COMPOSITIONS CONTAINING PHYTOSPHINGOSINE |
| JP2005531542A (en) * | 2002-05-02 | 2005-10-20 | ドゥーサン コーポレイション | Anticancer agent composition containing N, N-dimethylphytosphingosine |
| DE10255554A1 (en) | 2002-11-28 | 2004-06-17 | Goldschmidt Ag | Water-based emulsifier wax gels |
| FR2855049B1 (en) * | 2003-05-19 | 2006-07-21 | Oreal | COMPOSITION COMPRISING A 6-HYDROXY SPHINGENINE CERAMIDE PRECURSOR AND A 6-HYDROXYLASE PATH ACTIVATOR FOR USE IN ENHANCING THE BARRIER FUNCTION OF THE SKIN |
| FR2855048B1 (en) * | 2003-05-19 | 2006-07-21 | Oreal | COMPOSITION COMPRISING A PHYTOSPHINGOSIN-BASED CERAMIDE PRECURSOR AND AN ACTIVATOR OF THE 4-HYDROXYLASE PATHWAY, USE FOR ENHANCING THE BARRIER FUNCTION OF THE SKIN |
| KR101288776B1 (en) | 2011-12-07 | 2013-07-22 | 가톨릭대학교 산학협력단 | Novel phytosphingosine derivatives and cosmetic composition for preventing and improving inflammatory skin diseases and hyperkeratotic disorders |
| GB2564797B (en) * | 2016-04-04 | 2020-01-15 | Ocusoft Inc | Compositions, kits and methods for maintaining eyelid hygiene |
| CN120957965A (en) | 2023-03-27 | 2025-11-14 | 柯泰亚生物科技(上海)有限公司 | Production of sphingosine base salts |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IT1235162B (en) * | 1988-12-02 | 1992-06-22 | Fidia Farmaceutici | LYSOSPHINGOLIPID DERIVATIVES |
| US5190876A (en) * | 1988-12-27 | 1993-03-02 | Emory University | Method of modifying cellular differentiation and function and compositions therefor |
| JP3594962B2 (en) * | 1992-04-03 | 2004-12-02 | コスモフェルム ベースローテン フェンノートシャップ | Selective N-acylation of amino alcohols |
| DE69419601T2 (en) * | 1993-03-17 | 1999-12-02 | Kao Corp., Tokio/Tokyo | AMINE DERIVATIVES AND DERMATOLOGICAL PREPARATIONS CONTAINING THEM |
| GB2323594A (en) * | 1997-03-25 | 1998-09-30 | Victor Martin | 2-amino-alkanoic acid derivatives, 2-amino alcohols and diamines |
| KR100541271B1 (en) * | 1997-05-02 | 2006-04-21 | 코스모페름 베.파우 | Antimicrobial compositions for topical use |
-
2000
- 2000-03-09 EP EP00907673A patent/EP1159256A1/en not_active Withdrawn
- 2000-03-09 BR BR0009265-7A patent/BR0009265A/en not_active IP Right Cessation
- 2000-03-09 JP JP2000604009A patent/JP2002539110A/en active Pending
- 2000-03-09 WO PCT/EP2000/002191 patent/WO2000053568A1/en not_active Ceased
- 2000-03-09 KR KR1020017011411A patent/KR20010108331A/en not_active Withdrawn
- 2000-03-09 CN CN00807004A patent/CN1360567A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN119390593A (en) * | 2024-09-26 | 2025-02-07 | 优然创新(北京)技术研发中心有限公司 | A sphingosine supramolecular body and its preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2002539110A (en) | 2002-11-19 |
| KR20010108331A (en) | 2001-12-07 |
| WO2000053568A1 (en) | 2000-09-14 |
| EP1159256A1 (en) | 2001-12-05 |
| BR0009265A (en) | 2001-11-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1360567A (en) | Sphingoid base devivs. and uses thereof | |
| DE2552638B2 (en) | Thienamycin, process for its preparation and drugs containing thienamycin | |
| Abraham et al. | Biofilm inhibition and bacterial eradication by C-dots derived from polyethyleneimine-citric acid | |
| Hussein et al. | Antimicrobial, antibiofilm, and antioxidant activity of chitosan nanoparticles synthesized by E. coli | |
| Li et al. | Green synthesis of antibacterial LFL-ZnO using L. plantarum fermentation liquid assisted by ultrasound-microwave | |
| Irzoqy et al. | Inhibitory effect of Saccharomyces cerevisiae against Propionibacterium acnes | |
| CN110227167A (en) | Chitosan graft lipid nanometer capsule contains rheum emodin and is inhibiting the purposes in pathogen biofilm | |
| CN113288880B (en) | Tea saponin/chitosan coated dihydromyricetin liposome and preparation method and application thereof | |
| CN102816717B (en) | Staphylococcus cohnii and method for preparing 5-aminolevulinic acid by using staphylococcus cohnii | |
| CN117431062B (en) | Preparation method and application of green luminescent amino acid-derived antibacterial carbon dots | |
| US4062943A (en) | Antibiotic produced from the microorganism (Pseudomonas lindbergii), its preparation and method of use | |
| RU2181144C1 (en) | Method to release dermatophytes out of clinical material | |
| WO2014115934A1 (en) | Glycerol derivative of hydroxy fatty acid and preparation method therefor | |
| US12178828B1 (en) | Method for making nano-sized chitosan/VS2 nanocomposites | |
| RU2237709C2 (en) | Microbiological medium for isolation of trichophyton verrucosum | |
| CN120420455B (en) | A baicalin and boric acid antimicrobial peptide assembly, its preparation method and application | |
| CN1662240A (en) | Phenyloxazolidinone Derivatives | |
| DE2652677A1 (en) | ANTIBIOTICA 890A LOW 1 AND 890A LOW 3 | |
| BR102024001280A2 (en) | Nanoparticles with bactericidal activity, method of preparation and use of nanoparticles. | |
| Ossman et al. | Isolation and diagnosis of Pseudomonas aeruginosa using traditional and Virtek-2 techniques | |
| Munir et al. | Comparative analysis of antimicrobial properties of maggot (Musca domestica) crude extract, maggot chitosan and chitosan nanoparticles | |
| CN1331836C (en) | C60 trans-succinate with biologic activity and its synthesis | |
| CN117567359A (en) | Pyridine-olefine acid type ionic liquid and preparation method and application thereof | |
| Goren et al. | The microbiome in hair disorders | |
| CN118419965A (en) | A kind of zinc oxide nano flower and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |