CN106866789A - A kind of method for isolating and purifying Daptomycin RS-8 impurity - Google Patents
A kind of method for isolating and purifying Daptomycin RS-8 impurity Download PDFInfo
- Publication number
- CN106866789A CN106866789A CN201510919777.2A CN201510919777A CN106866789A CN 106866789 A CN106866789 A CN 106866789A CN 201510919777 A CN201510919777 A CN 201510919777A CN 106866789 A CN106866789 A CN 106866789A
- Authority
- CN
- China
- Prior art keywords
- daptomycin
- impurity
- solution
- acid solution
- trifluoroacetic acid
- 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
Abstract
The invention discloses a kind of method for isolating and purifying Daptomycin RS-8 impurity, the method obtains RS-8 impurity crude liquids using Daptomycin filtrate after resin chromatography, again by preparative chromatography, the Daptomycin RS-8 impurity of high-purity of the chromatographic purity more than 90% is obtained.The method is simple and easy to apply, with low cost.
Description
Technical field
The present invention relates to pharmaceutical formulating art, more particularly to Daptomycin RS-8 impurity is isolated and purified, more particularly to one kind
The method that high-purity daptomycin RS-8 impurity is prepared using Daptomycin resin chromatography and preparative chromatography.
Background technology
Development and the abuse of antibiotic with antibiotic, pathogen be to the drug resistance of antibiotic today's society face it is most severe
Challenge.In addition to controlling abuse of antibiotics, a kind of effective antibiotic to antimicrobial agent into this problem of solution is found at present
Optimal path, vancomycin was once considered as the last line of defense to resisting gram-positive bacteria, but now world clinical
It was found that increasing resistance to this medicine bacterium.
Daptomycin (Daptomycin) is by Lilly (gift comes) company's original research, a ring of Cubist drugmakers exploitation
Lipopeptide antibiotic.Answer active demand of the patient to new resistance antibiotic, the end of the year 2003, FDA (FDA)
Ratifying injection Daptomycin (Daptomycin) (trade name cubicin) by quick trial program is used to treat blue by some leather
Concurrency skin and skin structure infection that family name positive sensitive strain causes, such as abscess, surgery cut infection and skin ulcer.Reach
The mechanism of action of Tobramycin is different from other antibiotic, and it is by upsetting transhipment of the cell membrane to amino acid, so as to hinder bacterium thin
The biosynthesis of cell wall peptide glycan, changes the property of cytoplasma membrane；In addition, it can also make it by destroying the cell membrane of bacterium
Content leaks and reaches sterilized purpose, therefore bacterium may be relatively difficult to Daptomycin generation drug resistance.
Although Daptomycin realizes industrialized production in the U.S., in Daptomycin product often include dehydration Daptomycin,
The impurity such as isomery Daptomycin, Daptomycin lactone hydrolysate, have a strong impact on product quality, are that this can be separately separated and is purified into
Impurity in Daptomycin simultaneously carries out research and is very important.
Existing Daptomycin impurity extracting method, such as the Daptomycin impurity ownership described by European patent 1586580A2,
It is not specifically related to Daptomycin impurity separation method.
The content of the invention
It is an object of the invention to provide a kind of easy to operate, with low cost and can quickly obtain high-purity daptomycin RS-8 impurity
Preparation method.
According to European patent " PROCESS FOR THE PURIFICATION OF DAPTOMYCIN " (EP 1 586 580
A2 method is detected to Daptomycin disclosed in), the Daptomycin impurity ownership with reference to described by European patent 1586580A2,
To the name of Daptomycin impurity ownership and positioning scenarios as shown in figure 1, DT represents Daptomycin (retention time 38.555min) in Fig. 1,
Daptomycin RS-8 impurity peaks are located at retention time 60.672min.
The method that the present invention isolates and purifies Daptomycin RS-8 impurity comprises the following steps：
1) by Daptomycin filtering fermentation liquor, filtrate obtains Daptomycin RS-8 impurity crude product solutions through resin chromatography；
2) Daptomycin RS-8 impurity crude product solution is obtained into chromatographic purity by preparative chromatography column separating purification>90% up to hold in the palm it is mould
Plain RS-8 impurity preparation solution；
3) preparation solution of Daptomycin RS-8 impurity is freezed, obtains Daptomycin RS-8 impurity.
Preferably, above-mentioned steps 1) in filtrate through FPDA13 resin chromatographies, obtain chromatographic purity>40% Daptomycin RS-8
Impurity crude product solution.Wherein after the upper FPDA13 resin chromatography posts of filtrate, first with 0.03N sodium chloride, 0.06N sodium acetates, acetic acid
The solution for adjusting pH6.5-7.0 rinses pillar, then adjusts the molten of pH6.5-7.0 with 0.3N sodium chloride, the sodium acetate of 0.06N, acetic acid
Liquid is eluted, and collects Daptomycin RS-8 impurity of the chromatographic purity more than 40%.
Above-mentioned steps 2) solution of Daptomycin RS-8 impurity is further separated using preparative chromatography piece-rate system, mobile phase uses second
The mixed liquor of nitrile and trifluoroacetic acid solution, specific chromatographic condition is：
Chromatographic column：LP-C8、250×21.2mm
Wavelength：214nm
Mobile phase：Acetonitrile, trifluoroacetic acid solution
Flow velocity：19mL/min
Column temperature：20~30 DEG C of room temperatures
Wherein, trifluoroacetic acid solution concentration is 0.01%-0.2%, preferably 0.01%-0.05%；
Mobile phase ratio is acetonitrile:Trifluoroacetic acid solution=20:80~60:40 (volume ratios, similarly hereinafter), preferably 20:80~40:60.
In the methods of the invention, after Daptomycin chromatographic solution is processed through preparative chromatography piece-rate system, the Daptomycin RS-8 for obtaining is miscellaneous
Matter its chromatographic purity is more than 95%.The method is simple and easy to apply, with low cost.
Brief description of the drawings
Fig. 1 is Daptomycin HPLC detection collection of illustrative plates, which show the ownership and positioning scenarios of each impurity of Daptomycin.
Specific embodiment
The present invention, the scope of but do not limit the invention in any way are further described by the following examples.
Embodiment 1
The upper FPDA13 resin chromatographies of filtrate after Daptomycin filtering fermentation liquor, with 0.03N sodium chloride, 0.06N sodium acetates, acetic acid
The solution 2BV for adjusting pH6.5-7.0 rinses pillar, and 0.3N sodium chloride, the sodium acetate of 0.06N, acetic acid adjust the solution of pH6.5-7.0
Wash-out, collects Daptomycin RS-8 impurity of the chromatographic purity more than 40%, and through preparative chromatography, (mobile phase is acetonitrile:0.01% trifluoro
Acetic acid=25:75, volume ratio) isolate and purify, and with efficient liquid phase tracing detection (testing conditions and European patent " PROCESS
FOR THE PURIFICATION OF DAPTOMYCIN " methods disclosed in (A2 of EP 1 586 580) are identical), collect
Chromatographic purity>90% Daptomycin RS-8 impurity preparation solutions.
Preparation solution vacuum freeze-drying, (testing conditions are special with Europe using high performance liquid chromatography for gained solid Daptomycin RS-8 impurity
Sharp " PROCESS FOR THE PURIFICATION OF DAPTOMYCIN " method disclosed in (A2 of EP 1 586 580)
It is identical) detection Daptomycin RS-8 impurity purity, its purity is 98%.
Embodiment 2
The upper FPDA13 resin chromatographies of filtrate after Daptomycin filtering fermentation liquor, 0.03N sodium chloride, 0.06N sodium acetates, acetic acid are adjusted
The solution 2BV for saving pH6.5-7.0 rinses pillar, and 0.3N sodium chloride, the sodium acetate of 0.06N, the solution of acetic acid regulation pH6.5-7.0 are washed
It is de-, Daptomycin RS-8 impurity of the chromatographic purity more than 40% is collected, through preparative chromatography, (mobile phase is acetonitrile:0.01% trifluoro second
Acid=30:70, volume ratio) isolate and purify, and with efficient liquid phase tracing detection (testing conditions and European patent " PROCESS FOR
Method disclosed in THE PURIFICATION OF DAPTOMYCIN " (A2 of EP 1 586 580) is identical), collect chromatogram
Purity>90% Daptomycin RS-8 impurity preparation solutions.
Preparation solution vacuum freeze-drying, (testing conditions are special with Europe using high performance liquid chromatography for gained solid Daptomycin RS-8 impurity
Sharp " PROCESS FOR THE PURIFICATION OF DAPTOMYCIN " method disclosed in (A2 of EP 1 586 580)
It is identical) detection Daptomycin RS-8 impurity purity, its purity is 99%.
Embodiment 3
The upper FPDA13 resin chromatographies of filtrate after Daptomycin filtering fermentation liquor, 0.03N sodium chloride, 0.06N sodium acetates, acetic acid are adjusted
The solution 2BV for saving pH6.5-7.0 rinses pillar, and 0.3N sodium chloride, the sodium acetate of 0.06N, the solution of acetic acid regulation pH6.5-7.0 are washed
It is de-, Daptomycin RS-8 impurity of the chromatographic purity more than 40% is collected, through preparative chromatography, (mobile phase is acetonitrile:0.01% trifluoro second
Acid=33:67, volume ratio) isolate and purify, and with efficient liquid phase tracing detection (testing conditions and European patent " PROCESS FOR
Method disclosed in THE PURIFICATION OF DAPTOMYCIN " (A2 of EP 1 586 580) is identical), collect chromatogram
Purity>90% Daptomycin RS-8 impurity preparation solutions.
Preparation solution vacuum freeze-drying, (testing conditions are special with Europe using high performance liquid chromatography for gained solid Daptomycin RS-8 impurity
Sharp " PROCESS FOR THE PURIFICATION OF DAPTOMYCIN " method disclosed in (A2 of EP 1 586 580)
It is identical) detection Daptomycin RS-8 impurity purity, its purity is 92%.
The present invention provides a kind of fast and convenient, with low cost preparation high-purity daptomycin RS-8 impurity using preparative chromatography technology
Technology.Wherein prepare acetonitrile and trifluoroacetic acid solution ratio and trifluoroacetic acid concentration even more high-purity in mobile phase mould up to support
Critical process control point prepared by plain RS-8 impurity.Although herein to being described this invention as described detailed, it is to be understood that,
On the basis of without prejudice to spirit of the invention and essence, those skilled in the art can be modified or change, these modifications or
Change within the scope of protection of present invention.
Claims (9)
1. a kind of method for isolating and purifying Daptomycin RS-8 impurity, comprises the following steps：
1) by Daptomycin filtering fermentation liquor, filtrate obtains Daptomycin RS-8 impurity crude product solutions through resin chromatography；
2) Daptomycin RS-8 impurity crude product solution is obtained into chromatographic purity by preparative chromatography column separating purification>90% up to hold in the palm it is mould
Plain RS-8 impurity preparation solution；
3) Daptomycin RS-8 impurity preparation solutions are freezed, obtains Daptomycin RS-8 impurity.
2. the method for claim 1, it is characterised in that step 1) in use FPDA13 resin chromatographies, obtain chromatographically pure
Degree>40% Daptomycin RS-8 impurity crude product solutions.
3. method as claimed in claim 2, it is characterised in that step 1) in after the upper FPDA13 resin chromatography posts of filtrate, first use
0.03N sodium chloride, 0.06N sodium acetates, acetic acid regulation pH6.5-7.0 solution rinse pillar, then with 0.3N sodium chloride,
The sodium acetate of 0.06N, acetic acid adjust the eluant solution of pH6.5-7.0, collect Daptomycin RS-8 of the chromatographic purity more than 40%
Impurity.
4. the method for claim 1, it is characterised in that step 2) preparative chromatography column separating purification in, mobile phase is used
The mixed liquor of acetonitrile and trifluoroacetic acid solution.
5. method as claimed in claim 4, it is characterised in that step 2) preparative chromatography column separating purification in, the trifluoro second
The concentration of acid solution is 0.01%~0.2%.
6. method as claimed in claim 5, it is characterised in that the concentration of the trifluoroacetic acid solution is 0.01%-0.05%.
7. method as claimed in claim 5, it is characterised in that step 2) preparative chromatography column separating purification in, the second of mobile phase
Nitrile:The volume ratio of trifluoroacetic acid solution is 20:80~60:40.
8. method as claimed in claim 7, it is characterised in that the acetonitrile of mobile phase:The volume ratio of trifluoroacetic acid solution is
20:80~40:60.
9. the method as described in claim 4~8 is any, it is characterised in that step 2) chromatographic condition of preparative chromatography is：
Chromatographic column：LP-C8、250×21.2mm；
Wavelength：214nm；
Mobile phase：The mixed liquor of acetonitrile and trifluoroacetic acid solution；
Flow velocity：19mL/min；
Column temperature：20~30 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510919777.2A CN106866789A (en) | 2015-12-11 | 2015-12-11 | A kind of method for isolating and purifying Daptomycin RS-8 impurity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510919777.2A CN106866789A (en) | 2015-12-11 | 2015-12-11 | A kind of method for isolating and purifying Daptomycin RS-8 impurity |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106866789A true CN106866789A (en) | 2017-06-20 |
Family
ID=59176954
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510919777.2A Pending CN106866789A (en) | 2015-12-11 | 2015-12-11 | A kind of method for isolating and purifying Daptomycin RS-8 impurity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106866789A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113929743A (en) * | 2020-06-29 | 2022-01-14 | 鲁南制药集团股份有限公司 | Method for preparing daptomycin impurity RS-1 and impurity RS-3 |
| CN114685610A (en) * | 2020-12-28 | 2022-07-01 | 杭州中美华东制药有限公司 | Lactone hydrolysate of daptomycin RS-8a impurity and preparation method thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1586580A2 (en) * | 2000-01-20 | 2005-10-19 | Cubist Pharmaceuticals, Inc. | Process for the purification of daptomycin |
| CN102492024A (en) * | 2011-12-09 | 2012-06-13 | 厦门大学 | Method for extracting daptomycin from fermentation broth |
| CN102675426B (en) * | 2012-04-26 | 2013-12-11 | 杭州华东医药集团生物工程研究所有限公司 | Extraction and purification method of daptomycin |
| CN103724400A (en) * | 2012-10-10 | 2014-04-16 | 北大方正集团有限公司 | Method for separating and purifying dehydrated daptomycin |
| CN103159829B (en) * | 2011-12-08 | 2014-11-05 | 北大方正集团有限公司 | Extraction method for daptomycin |
| CN104387444A (en) * | 2014-11-13 | 2015-03-04 | 北大医药重庆大新药业股份有限公司 | Method for preparing high-purity sample of impurity RS-2 in daptomycin |
| CN104650189A (en) * | 2013-11-19 | 2015-05-27 | 北大方正集团有限公司 | Preparation method of isomeric daptomycin |
| CN104846043A (en) * | 2014-02-17 | 2015-08-19 | 上海医药工业研究院 | Process for separating and purifying fidaxomicin from fermentation broth |
-
2015
- 2015-12-11 CN CN201510919777.2A patent/CN106866789A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1586580A2 (en) * | 2000-01-20 | 2005-10-19 | Cubist Pharmaceuticals, Inc. | Process for the purification of daptomycin |
| CN103159829B (en) * | 2011-12-08 | 2014-11-05 | 北大方正集团有限公司 | Extraction method for daptomycin |
| CN102492024A (en) * | 2011-12-09 | 2012-06-13 | 厦门大学 | Method for extracting daptomycin from fermentation broth |
| CN102675426B (en) * | 2012-04-26 | 2013-12-11 | 杭州华东医药集团生物工程研究所有限公司 | Extraction and purification method of daptomycin |
| CN103724400A (en) * | 2012-10-10 | 2014-04-16 | 北大方正集团有限公司 | Method for separating and purifying dehydrated daptomycin |
| CN104650189A (en) * | 2013-11-19 | 2015-05-27 | 北大方正集团有限公司 | Preparation method of isomeric daptomycin |
| CN104846043A (en) * | 2014-02-17 | 2015-08-19 | 上海医药工业研究院 | Process for separating and purifying fidaxomicin from fermentation broth |
| CN104387444A (en) * | 2014-11-13 | 2015-03-04 | 北大医药重庆大新药业股份有限公司 | Method for preparing high-purity sample of impurity RS-2 in daptomycin |
Non-Patent Citations (1)
| Title |
|---|
| 张定丰 等: "达托霉素在不同酸碱条件下产生的杂质及其结构研究", 《中国抗生素杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113929743A (en) * | 2020-06-29 | 2022-01-14 | 鲁南制药集团股份有限公司 | Method for preparing daptomycin impurity RS-1 and impurity RS-3 |
| CN114685610A (en) * | 2020-12-28 | 2022-07-01 | 杭州中美华东制药有限公司 | Lactone hydrolysate of daptomycin RS-8a impurity and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3064214B1 (en) | Separation and purification method for vancomycin hydrochloride of high purity | |
| CN101830970B (en) | Purification and preparation method of high-purity Daptomycin | |
| CN103159829B (en) | Extraction method for daptomycin | |
| CN101148410B (en) | Method for extracting high pure chicoric acid from Coneflower | |
| CN103724400B (en) | A kind of method of separation and purification dehydration daptomycin | |
| CN108373499B (en) | A kind of purifying and ionic control method of Teriparatide acetate | |
| CN102718839A (en) | Method for separating and purifying daptomycin | |
| CN103275152A (en) | Preparation method of high-purity fidaxomicin | |
| CN106866790A (en) | The preparation method of Daptomycin RS-5/6, RS-7 and RS-7a/7b impurity | |
| CN106866789A (en) | A kind of method for isolating and purifying Daptomycin RS-8 impurity | |
| CN101787071A (en) | Purification method of vapreotide | |
| CN102492024A (en) | Method for extracting daptomycin from fermentation broth | |
| CN101279979B (en) | Separation and purification method of cefamandole nafate and preparation of cefathiamidine freeze-dried injectable powder | |
| CN106866791A (en) | A kind of preparation method of high-purity daptomycin lactone hydrolysate | |
| CN101851277A (en) | Preparation method of Dalbavancin key intermediate A40926 BO component by purification | |
| CN102276696B (en) | Method for purifying daptomuycin | |
| CN102964430B (en) | Purification method of teicoplanin | |
| CN107382943B (en) | Method for subcritical water extraction of dihydroquercetin in sorghum bran | |
| CN106565820A (en) | Method for preparing high-purity sample of vancomycin hydrochloride impurities 3 and 8 | |
| CN105111285A (en) | Daptomycin extraction method | |
| CN105223296A (en) | The purification process of one class polypeptide | |
| CN107163102A (en) | A kind of method of hydrophilic polypeptides purifying | |
| CN103898182B (en) | The preparation method of beauvericin | |
| CN107778357A (en) | A kind of extraction of Pneumocandin B0, purification process | |
| CN104974041B (en) | High-purity chlorogenic acid and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170620 |