CN101631777A

CN101631777A - Triol form of rosuvastatin

Info

Publication number: CN101631777A
Application number: CN200880008162A
Authority: CN
Inventors: V·尼达姆-希尔德谢姆; A·巴拉诺夫; I·韦恩伯格
Original assignee: Teva Pharmaceutical Industries Ltd
Current assignee: Teva Pharmaceutical Industries Ltd
Priority date: 2007-03-13
Filing date: 2008-03-13
Publication date: 2010-01-20

Abstract

Provided is a rosuvastatin triol and its use as a reference standard for analysis of rosuvastatin.

Description

The triol form of superstatin

The cross reference of related application

60/918,466 the rights and interests that the application requires to submit on March 13rd, 2007 U.S. Provisional Application is submitted to number on March 15th, 60/906,914 and 2007, they open incorporated herein in full by reference.

Invention field

The present invention relates to superstatin triol and conduct thereof and analyze the purposes of the reference standard of superstatin.

Background of invention

Statins (Statins) is at present obtainable being used for to reduce the most effective medicine in the treatment of low-density lipoprotein (LDL) granule density in the blood flow the patient who is in risk of cardiovascular diseases.Therefore, adopt statins treatment hypercholesterolemia, hyperlipoproteinemia and atherosclerosis.High-level LDL in the blood flow has related to the formation of the coronary artery pathological changes that hinders blood flow and may destroy and promote thrombosis.Goodman and Gilman, therapeutic pharmacological basis (The Pharmacological Basis of Therapeutics), 879 pages (the 9th edition, 1996).

Statins suppresses people's cholesterol biosynthesizing by competitive inhibition 3-hydroxy-3-methyl-glutaryl--coenzyme A (" HMG-CoA ") reductase enzyme.The HMG-CoA reductase enzyme promotes that HMG is converted into mevalonic acid (mevalonate), and this is the step of decision speed in the cholesterol biosynthesizing.The minimizing that cholesterol generates makes the amount of ldl receptor increase, thereby correspondingly reduces LDL particulate concentration in the blood flow.The decline of LDL level reduces the risk of coronary artery disease in the blood flow.J.A.M.A.1984，251，351-74。

The available statins comprises lovastatin, Simvastatin, Pravastatin, fluvastatin, Cerivastatin and atorvastatin at present.Give lovastatin (being disclosed in U.S. Patent number 4,231,938) and Simvastatin (being disclosed in U.S. Patent number 4,444,784) with lactone form.After the absorption, lactonic ring is opened in liver through chemistry or enzymic hydrolysis, and generates activity hydroxy acid.

Give Pravastatin (being disclosed in U.S. Patent number 4,346,227) as sodium salt.Also fluvastatin that gives as sodium salt (being disclosed in U.S. Patent number 4,739,073) and Cerivastatin (are disclosed in U.S. Patent number 5,006,530 and 5,177,080) be the synthetic compound fully, its part-structure is different from this class fungi derivative that contains the hexahydro naphthalene ring.Give atorvastatin and two kinds of new " super Statins ", superstatin and pitavastatin as calcium salt.

The ZD-4522 of Shionogi exploitation (two (+) 7-[4-(4-fluoro phenyl)-6-sec.-propyl-2-(N-methyl-N-methyl sulphonyl aminopyrimidine)-5-yl]-(3R, 5S)-and dihydroxyl-(E)-6-heptenoic acid) calcium is the HMG-CoA reductase inhibitor, every day is oral once with treatment hyperlipidaemia (Ann Rep, Shionogi, 1996; Direct communication (Direct communications), Shionogi, on February 8th, 1999 and on February 25th, 2000).ZD-4522 has following chemical formula:

ZD-4522 is with trade(brand)name

Listing is with treatment Mammals, for example people.According to

The producer, it gives with the about 40mg of the about 5mg-of per daily dose.More sharply do not reduce the patient of LDL-C or the patient who myopathy is had predisposing factor at needs, recommend 5mg dosage, and the recommended dose of general patient is 10mg, have remarkable hypercholesterolemia and significantly the lipid target (＞190mg/dL) patient's dosage is 20mg, and is 40mg to the patient's that do not respond than low dosage dosage.

U.S. Patent number 5,260,440 disclose superstatins, its calcium salt (2: 1) and lactone form thereof and they have been proposed claim.The method of being somebody's turn to do ' 440 patents is by under refluxing; in acetonitrile; make 4-(4-fluoro phenyl)-6-sec.-propyl-2-(N-methyl-N-methyl sulphonyl amino)-5-pyrimidine formaldehyde with (3R)-the inferior phosphoranyl methyl caproate reaction of 3-(t-butyldimethylsilyl oxygen base)-5-oxo-6-triphenyl, the preparation superstatin.Use hydrogen fluoride cracking silyl again, use the sodium borohydride (NaBH in the tetrahydrofuran (THF) (THF) subsequently ₄) and diethyl methoxyl group borane reduction, obtain the methyl esters of superstatin.

Under room temperature, in the ethanol,, remove ethanol subsequently and add ether then, obtain the sodium salt of superstatin with this ester of sodium hydroxide (NaOH) hydrolysis.Again sodium salt is converted into calcium salt.Make sodium salt be dissolved in water and remain under the nitrogen atmosphere.In this solution, add calcium chloride then, generate ZD-4522 (2: 1) precipitation.The intermediates preparation that is disclosed in ' 440 patents is incorporated herein by reference.

The mixture of reaction product seldom is that to be enough to meet the simplification compound of pharmaceutical standards pure.As a rule, the auxiliary reagent that can have reaction by-product and by product and be used to react.In some step that will be contained in during superstatin in the product mixtures is processed as active pharmaceutical ingredient (" API "), must typically analyze through HPLC or GC, analyze the purity of superstatin, whether be suitable for continuing processing or finally be suitable for use in medicament production to determine it.Superstatin needn't be definitely pure.Absolute purity is unapproachable theoretic ideal value.For guaranteeing not reduce the security of the clinical use of prepared API, would rather set purity rubric because of the existence of impurity.In the U.S., food and drug control rules (Food and Drug Administrationguidelines) are recommended, and the applicant limits some impurity below 0.1%.

In general, differentiate byproduct, by product and auxiliary reagent (being referred to as " impurity ") through spectrum and other physical method, thereby impurity is relevant with the peak position (or the spot on the TLC plate) in the chromatography.(953 pages of Strobel) (Strobel, H.A.; Heineman, W.R., chemical apparatus: be winding method, 3 editions. (Wiley; Sons:New York 1989)).Afterwards, can differentiate impurity, to post and after testing measure impurity routinely between several minutes of being called between the device wash-out specific components " retention time " injecting sample by the position in the chromatography.According to the condition of instrument and other many factors, this every day time period difference.For reducing the influence of this class variation to accurate discriminating impurity, the professional adopts " relative retention time " (" RRT ") to differentiate impurity (922 pages of Strobel).The RRT of impurity is the retention time of its retention time divided by some reference mark.In theory, itself can be used as reference mark superstatin, but as practical material, it is present in inundatory ratio and is tending towards being full of in the mixture of post, produce irreproducible retention time, that is, corresponding to the maximum peak of superstatin be tending towards vacillating (wander) (diagram of Figure 24 .8 (b) viewed this unsymmetrical peak when comprising column overload that Strobel is 879 pages).Therefore, want to select selective compound sometimes, it is with obvious enough detectable amount and fully be low to moderate the amount adding that is not full of post or be present in the mixture, and with this compound as reference mark.

The compound that is relative pure state can be used as " reference standard " (" reference mark " is similar to reference standard, but it is used for qualitative analysis), to quantize the amount of the compound in the unknown mixture.When compound is used as " external standard ", with the compound solution of constructed analysis as the concentration known of unknown mixture.(924 pages of Strobel, 549 pages of Snyder) (Snyder, L.R.; Kirkland, the modern liquid chromatography (LC) introduction of J.J., 2 editions. (John Wiley; Sons:New York 1979)).Can determine the amount of the compound in the mixture by the size of relatively detector response.Also referring to USP6,333,198, incorporated herein by reference.

If pre-determined " response factor " of the difference of the detector sensitivity that compensates two kinds of compounds, also can adopt the reference standard compound to quantize the amount (894 pages of Strobel) of another kind of compound in the mixture.For this reason, can directly the reference standard compound be added to mixture, in this case, it is called as " interior mark " (925 pages of Strobel, 552 pages of Snyder).

When the technology that is called " standard adding " (wherein preparing at least two kinds of samples by adding the known and different interior marks of measuring) by employing when unknown mixture contains some reference standard compound, even the reference standard compound also can be used as interior mark (Strobel 391-393 page or leaf, 571,572 pages of Snyder).Can determine to come from the detector response of original reference standard compound in mixture than (for example, Strobel, 392 pages of Figure 11 .4) by the extrapolation detector response figure extremely zero with respect to the amount that adds to the reference standard compound in each sample.

The invention provides and can be used as quantification and differentiate superstatin and the reference standard of impurity and the compound of reference mark that is present in each batch superstatin.

Summary of the invention

In one embodiment, the invention provides superstatin triol with following structure:

X wherein is hydrogen, C ₁-C ₄Alkyl or alkali or alkaline earth metal cation, prerequisite are that when X was alkaline-earth metal, bimolecular superstatin appeared in face of the metallic cation.

In another embodiment, the superstatin triol that is sour form provided by the invention has following structure:

In another embodiment, the invention provides the superstatin triol that is ester-formin with following structure:

R wherein is C ₁-C ₄Alkyl ester.

In one embodiment, the invention provides the superstatin triol that is salt form with following structure:

M wherein is alkali or alkaline earth metal cation, and prerequisite is that when X was alkaline-earth metal, bimolecular superstatin appeared in face of the metallic cation.

In one embodiment, the invention provides the superstatin triol that is lactone form with following structure:

In another embodiment, the invention provides and do not contain corresponding superstatin glycol form substantially, be through separating or the triol of the various above forms of purified form.

In another embodiment, the invention provides preparation superstatin triol C ₁-C ₄The method of ester, it comprises makes superstatin C ₁-C ₄Ester and borine methyl-sulfide complex solution merge in the organic solvent that is suitable for, and obtain reaction mixture, and the reaction mixture and the NaOH aqueous solution that are generated are merged, and add hydrogen peroxide (H ₂O ₂) and reclaim three alcohol esters.

In another embodiment, the invention provides preparation superstatin triol C ₁-C ₄The method of ester, it comprises oxidation superstatin glycol C ₁-C ₄Ester obtains having superstatin three alcohol esters of hydroxyl on 7.

In another embodiment, the invention provides a kind of method, this method comprises makes superstatin C ₁-C ₄Ester and the borine solution in organic solvent merges, and obtains reaction mixture, the aqueous solution of the reaction mixture that obtains and mineral alkali is merged and adding superoxide and reclaim three alcohol esters.

In one embodiment, the invention provides by measuring the amount of the superstatin calcitriol in each batch ZD-4522, selection has the ZD-4522 batch of the triol of desired level, and preparation has the medicinal compositions of the ZD-4522 batch through selecting, and reduces the method that is present in the impurity level in the ZD-4522.

In another embodiment, the invention provides the method that reduces the amount that is present in the superstatin triol calcium in the mixture that comprises superstatin glycol calcium and superstatin triol calcium, this method comprises measures each batch superstatin glycol C ₁-C ₄Superstatin triol C in the ester ₁-C ₄The amount of ester selects to contain triol C ₁-C ₄The superstatin glycol C of ester ₁-C ₄Ester batch and preparation contain superstatin glycol C through selecting ₁-C ₄The medicinal compositions of the superstatin glycol calcium of ester batch.

In another embodiment, the invention provides the method for determining the impurity level in superstatin ester (preferred tertiary butyl ester) sample, this method comprises by GC or HPLC to be measured corresponding to the area under the peak of superstatin three alcohol esters in the reference standard of superstatin three alcohol esters (preferred tertiary butyl ester) that comprise known quantity; Measure corresponding to the area under the peak of superstatin three alcohol esters in the sample that comprises superstatin three pure and mild superstatin diol esters (preferred tertiary butyl ester) by GC or HPLC; Compare the area of reference standard and the area of specimen with passing through, determine the amount of superstatin three alcohol esters in the sample.

In another embodiment, the invention provides the method for determining the impurity level in the ZD-4522 sample, it comprises through GC or HPLC measures corresponding to the area under the peak of the superstatin triol calcium in the reference standard of the superstatin triol calcium that is comprising known quantity; Measure corresponding to the area under the peak of the superstatin triol calcium in the sample that is comprising superstatin three pure and mild superstatin glycol calcium salts through GC or HPLC; Determine the amount of the triol calcium in the sample with the area of area by reference standard relatively and specimen.

In one embodiment, the invention provides the method for the relative retention time (RRT) of determining the impurity in superstatin diol ester (preferred tertiary butyl ester) sample, it comprises through GC or HPLC measures relative retention time (RRT) corresponding to three alcohol esters of the superstatin in the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin diol ester and superstatin three alcohol esters, to obtain having the GC or the HPLC chromatography of retention time; Compare the relative retention time (RRT) of reference mark and the relative retention time (RRT) of specimen with passing through, determine the relative retention time (RRT) of three alcohol esters in the sample.

In another embodiment, the invention provides the method for the relative retention time (RRT) of determining the impurity in the superstatin glycol calcium sample, it comprises through GC or HPLC measures relative retention time (RRT) corresponding to the superstatin triol calcium in the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin two pure and mild superstatin triol calcium salts, to obtain having the GC or the HPLC chromatography of retention time; Compare the relative retention time (RRT) of reference mark and the relative retention time (RRT) of specimen with passing through, determine the relative retention time (RRT) of the triol calcium in the sample.

In one embodiment, the invention provides the method that preparation has the superstatin triol acid of following structure:

This method comprises the ester of the following structure of hydrolysis

With the ester conversion that makes with acid through hydrolysis, R wherein is C ₁-C ₄Group.

In one embodiment, the invention provides the method that preparation has superstatin three alcoholic lactones of following structure:

This method comprises the ester of the following structure of hydrolysis

With make ester be converted into lactone through hydrolysis, R wherein is C ₁-C ₄Ester.

This method comprises that hydrolysis has the lactone of following structure

With make lactone be converted into salt through hydrolysis, M wherein is basic metal or alkaline-earth metal, prerequisite is, if metallic cation is an alkaline-earth metal, each positively charged ion is existed the superstatin of two molecules.

In one embodiment, the invention provides the method that preparation has superstatin three alkoxide of following structure:

This method comprises the acid that has following structure with the alkali contact,

Prerequisite is, if metallic cation is an alkaline-earth metal, each positively charged ion is existed the superstatin of two molecules.

The summary of figure

Fig. 1 is the NMR of TBRE (superstatin tertiary butyl ester) triol.

Fig. 2 is the HPLC chromatography of explanation superstatin triol calcium as reference standard (comprising reference mark).

Detailed Description Of The Invention

As used herein, term " glycol " refers to be present in two hydroxyls on the rosuvastatin. The glycol rosuvastatin and the rosuvastatin synonym that are used for this paper.

As used herein, term " does not substantially contain " and refers to the meter based on area percentage HPLC, tool Have to be less than about 30% respective compound (for example, glycol or diastereoisomer), more preferably few In about 20%, even more preferably less than about 10% with most preferably be less than about 5%.

As used herein, term " three alcoholic lactones " refers to the lactone of rosuvastatin triol.

As used herein, term " reference standard " refers to not only can be used for quantitative analysis but also can qualitative branch Analyse the compound of active pharmaceutical ingredient. For example, the retention time of the compound among the HPLC relates to Set relative retention time, thereby make qualitative analysis become possibility. Before injecting the HPLC post Solution in compound concentration relate to the comparison of the area under each peak in the HPLC chromatography, Thereby so that quantitative analysis becomes possibility.

In qualitative analysis, adopt " reference mark ", according to them, for example, at chromatography or thin Analyse layer by layer the position on (TLC) plate, each component of discriminating mixture (Strobel page or leaf 921,922, 953). For this reason, if this compound is present in the mixture, then it also optionally must add To mixture. " reference mark " only is used for qualitative analysis, and reference standard can be used to quantitatively or Qualitative analysis, or the two. Therefore, reference mark is the subset of reference standard, and is included in In the definition of reference standard.

Although briefly described those knowledge in the reference standard field, this with regard to this point Those skilled in the art still understand, and detector response may be, for example, and the peak of resulting chromatography Height or complete peak area for example, detect by UV or refractive index, derive from HPLC system The eluent of system, perhaps, for example, flame ionization detection or thermal conductivity detect, and derive from the gas phase layer The eluent of analysing, perhaps, other detector response, for example, the spot on the fluorescence TLC plate The UV absorptance. The position of reference standard can be used to calculate rosuvastatin and other impurity mutually To retention time.

The invention provides the rosuvastatin triol with following structure:

X wherein is hydrogen, alkali or alkaline-earth metal or C₁-C ₄Alkyl. X is preferably hydrogen, and (that is, Luo Su cuts down His spit of fland triol acid), calcium (Ca²⁺) (that is, Luo Su cuts down for (that is, rosuvastatin triol calcium) or the tert-butyl group His spit of fland triol tert-butyl ester (" TBRE ")).

The invention provides the rosuvastatin triol with following structure:

X wherein is hydrogen, alkali or alkaline-earth metal or C₁-C ₄Alkyl. X is preferably hydrogen, and (that is, Luo Su cuts down His spit of fland triol acid), calcium (Ca²⁺) (that is, Luo Su cuts down for (that is, rosuvastatin triol calcium) or the tert-butyl group His spit of fland triol tert-butyl ester (" TBRE ")), the form that preferably is its separation.

The invention provides the rosuvastatin triol that is sour form with following structure:

The invention provides the rosuvastatin triol that is ester-formin with following structure

Wherein R is C₁-C ₄Alkyl. Preferably, R is the tert-butyl group or methyl. More preferably, R is The tert-butyl group.

The invention provides the rosuvastatin triol that is salt form with following structure:

M wherein is alkali metal or alkaline earth metal cation. Preferably, M is calcium. This area Those of ordinary skill can be appreciated that, when M is alkaline earth metal cation, for example during calcium, salt will be Half calcium salt (2: 1 ratios):

The present invention also provides the rosuvastatin triol that is lactone form with following structure:

The present invention also provides the above various forms of corresponding rosuvastatin glycol shape that substantially do not contain The rosuvastatin triol of formula. Therefore, the invention provides:

A) substantially do not contain rosuvastatin glycol C₁-C ₄The rosuvastatin triol C of ester₁-C ₄Ester. Substantially the rosuvastatin triol that does not contain the rosuvastatin glycol tert-butyl ester tert-butyl ester also is provided;

B) substantially do not contain the rosuvastatin triol acid of rosuvastatin two alkyd;

C) substantially do not contain rosuvastatin three alkoxide of rosuvastatin two alkoxide (preferred calcium salt) (preferred calcium salt); With

D) substantially do not contain rosuvastatin three alcoholic lactones of rosuvastatin diol lactone.

The present invention also provides above various forms of be racemic, (7S) and (7R) sieve of configuration The rosuvastatin triol. (7S) and (7R) configuration is diastereoisomer.

Particularly, the invention provides:

A) be racemic, (7S) and (7R) the rosuvastatin triol C of form₁-C ₄Ester, preferred The tert-butyl ester. In one embodiment, (7S) form does not contain (7R) form substantially. A reality Execute in the scheme, (7R) form does not contain (7S) form substantially.

B) be racemic, (7S) and (7R) the rosuvastatin triol acid of form. A reality Execute in the scheme, (7S) form does not contain (7R) form substantially. In one embodiment, (7R) form Substantially do not contain (7S) form.

C) be racemic, (7S) and (7R) rosuvastatin three alkoxide (for example calcium) of form. In the embodiment, (7S) form does not contain (7R) form substantially. In one embodiment, (7R) Form does not contain (7S) form substantially.

D) be racemic, (7S) and (7R) rosuvastatin three alcoholic lactones of form. At one In the embodiment, (7S) form does not contain (7R) form substantially. In one embodiment, (7R) Form does not contain (7S) form substantially.

The present invention also provides the method for preparing rosuvastatin three alcohol esters. Can cut down by oxidation Luo Su He is spit of fland C₁-C ₄Ester, the especially tert-butyl ester prepare three alcohol esters. Can be by making rosuvastatin C₁-C ₄Ester, the especially tert-butyl ester and borine (for example, BH₃，B ₂H ₆) chemical combination, carry out the oxidation of ester. Can Adopt borine and various monoalkyl (C₁-C ₈)-and dialkyl group (C₁-C ₈The complex compound of)-borine. Excellent Selection of land is closed solution and the esterification of borine dimethyl sulfide complex compound in the organic solvent that is suitable for. Can Stirred reaction mixture. Make inorganic base, the aqueous solution and the reactant mixture of preferred NaOH close again And, add subsequently H₂O ₂(preferred about 30% in water). Preferably dropwise add H₂O ₂。H ₂O ₂Temperature during the adding preferably keeps below about 50 ℃.

Except H₂O ₂Outside, also can adopt other oxidising agent. For example, can adopt other any Peroxide comprises tert-butyl hydroperoxide (TBHP) and six hydrations of single peroxide phthalic acid magnesium Thing (MMPP).

Inorganic base preferred as alkali alkali, more preferably hydroxide bases, for example NaOH, KOH and LiOH. Adoptable another kind of alkali is NH₄OH。

Separable organic phase, and water and/or salt water washing, to remove easily mixed water-soluble accessory substance, for example the borine accessory substance (for example: H₃BO ₃). It also can wash through sodium sulfite, removes The hydrogen peroxide of amount. Then, concentrate organic phase, obtain residue. Can pass through pressure decreased To being lower than 1 atmospheric pressure, for example be less than about 100mmHg, concentrate.

After the reaction, if need, can add precipitating reagent, for example ammonium chloride or another kind of salt, with From reactant mixture, be settled out impurity. Available ammonium chloride is removed byproduct of reaction H₃BO ₃ Generation For adopting ammonium chloride, can adopt acid, for example among acetic acid or the HCl and alkaline mixt. Can be through water H is removed in washing₃BO ₃。

Then purifying rosuvastatin three alcohol esters and through chromatography from corresponding rosuvastatin glycol Ester separates.

The invention provides rosuvastatin three alcohol esters of the form that is its separation.

Can again three alcohol esters be converted into corresponding acid, salt or lactone.

The ion gun that can be suitable for through hydrolysis and the adding of ester makes three alcohol esters be converted into three alkoxide. For Obtain calcium salt, can or adopt the combination of NaOH and calcium chloride, perhaps can adopt hydroxide Calcium.

Can be suspended in the mixture of mixed organic solvents and water by making ester, make the superstatin ester be converted into salt, and with alkali for example sodium hydroxide merge, obtain solution.Organic solvent can be C ₁-C ₄Alcohol, preferred alcohol.Evaporate organic solvent down in decompression then, add calcium chloride subsequently, produce the calcium precipitation of triol.Can pass through routine techniques, for example filter, reclaim precipitation.

The invention provides superstatin three alkoxide that are its isolating form.

For obtaining the superstatin triol acid, make salt and acid, for example hydrochloric acid or sulfuric acid merge.In one embodiment, make superstatin triol calcium be suspended in organic solvent, for example in the methylene dichloride, to wherein adding moisture HCl.For example by separating organic phase, reaction mixture separates the superstatin triol acid, subsequently, for example, removes organic solvent by reduction vaporization then.

Also can be by after the ester hydrolysis, acidified reaction mixture and do not add calcium chloride and obtain acid.Can adopt mineral acid, for example HCl and H ₂SO ₄

The invention provides the superstatin triol acid that is its isolating form.

Then, facilitating under the condition that lactonizes, can obtain the rosuvastatin statin lactone by acid.In one embodiment, make superstatin triol calcium be dissolved in organic solvent, for example in the acetonitrile, to wherein adding moisture HCl.The restir reaction mixture.For example, can pass through vapourisation under reduced pressure, remove organic solvent and water, obtain lactone.

As mentioned above, these compounds, promptly superstatin triol acid, salt, lactone and ester can be used as reference mark/standard.Fig. 2 illustrates that The compounds of this invention can be used as reference standard, is present in the amount of the impurity in the superstatin composition with quantification and evaluation.On post, superstatin triol calcium is near superstatin glycol calcium, but not with the peak overlapping of superstatin.Because this makes quantification easier, the peak is not overlapping to be ideal.

The invention provides superstatin three alcoholic lactones that are its isolating form.

The invention provides the method that reduces the amount that is present in the superstatin triol calcium in the mixture that comprises ZD-4522 and superstatin triol calcium, it comprises the amount of the superstatin calcitriol of measurement in each batch superstatin glycol calcium, selection have desired level triol the superstatin glycol batch and preparation have the medicinal compositions of selected superstatin glycol batch.Usually the salt that is not calcium also can be used for present method.

The invention provides the method that reduces the amount that is present in the superstatin triol calcium in the mixture that comprises superstatin glycol calcium and superstatin triol calcium, it comprises that measurement is at each batch superstatin glycol C ₁-C ₄Superstatin triol C in the ester ₁-C ₄The amount of ester selects to contain triol C ₁-C ₄The superstatin glycol C of ester ₁-C ₄Ester batch and preparation contain selected superstatin glycol C ₁-C ₄The medicinal compositions of the superstatin glycol calcium of ester batch.Described ester preferred tertiary butyl ester.

The invention provides the method that reduces the amount that is present in the superstatin triol calcium in the mixture that comprises superstatin glycol calcium and superstatin triol calcium, it comprises the amount of superstatin three alcoholic lactones of measurement in each batch superstatin diol lactone, select to have desired level three alcoholic lactones the superstatin diol lactone batch and preparation have the medicinal compositions of the superstatin glycol calcium of selected superstatin diol lactone batch.

The invention provides the method that reduces the amount that is present in the superstatin triol calcium in the mixture that comprises superstatin glycol calcium and superstatin triol calcium, it comprises the amount of the superstatin triol acid of measurement in each batch superstatin two alkyd, select to have desired level triol acid superstatin two alkyd batch and preparation have the medicinal compositions of the superstatin glycol calcium of selected superstatin two alkyd batch.

The invention provides the method for the amount of determining the impurity in superstatin diol ester (preferred tertiary butyl ester) sample, it comprises through GC or HPLC measures corresponding to the area under the peak of superstatin three alcohol esters in the reference standard of superstatin three alcohol esters (preferred tertiary butyl ester) that comprising known quantity; Measure corresponding to the area under the peak of superstatin three alcohol esters in the specimen that is comprising superstatin three pure and mild superstatin diol esters (preferred tertiary butyl ester) through GC or HPLC; Compare the area of reference standard and the area of specimen with passing through, determine the amount of three alcohol esters in the sample.

The invention provides the method for the amount of determining the impurity in the ZD-4522 sample, it comprises through GC or HPLC measures corresponding to the area under the peak of the superstatin triol calcium in the reference standard of the superstatin triol calcium that is comprising known quantity; Measure corresponding to the area under the peak of the superstatin triol calcium in the specimen that is comprising superstatin three pure and mild superstatin glycol calcium salts through GC or HPLC; Compare the area of reference standard and the area of specimen with passing through, determine the amount of the triol calcium in the sample.Usually the salt that is not calcium also can be used for present method.

The invention provides the method for the amount of determining the impurity in the superstatin acid sample, it comprises through GC or HPLC measures corresponding to the area under the peak of the superstatin triol acid in the reference standard of the superstatin triol acid that is comprising known quantity; Measure corresponding to the area under the peak of the superstatin triol acid in the specimen that is comprising superstatin acid and superstatin two alkyd through GC or HPLC; Compare the area of reference standard and the area of specimen with passing through, determine the amount of the triol acid in the sample.

The invention provides the method for the amount of determining the impurity in the rosuvastatin statin lactone sample, it comprises through GC or HPLC measures corresponding to the area under the peak of superstatin three alcoholic lactones in the reference standard of superstatin three alcoholic lactones that comprising known quantity; Measure corresponding to the area under the peak of superstatin three alcoholic lactones in the specimen that is comprising superstatin three alcoholic lactones and superstatin diol lactone through GC or HPLC; Compare the area of reference standard and the area of specimen with passing through, determine the amount of three alcoholic lactones in the sample.

The invention provides the method for the relative retention time (RRT) of determining the impurity in superstatin diol ester (preferred tertiary butyl ester) sample, it comprises through GC or HPLC measures relative retention time (RRT) corresponding to three alcohol esters of the superstatin in the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin ester and superstatin three alcohol esters, obtain having the HPLC or the GC chromatography of retention time; Compare the relative retention time (RRT) of reference mark and the relative retention time (RRT) of specimen with passing through, determine the relative retention time (RRT) of three alcohol esters in the sample.Superstatin diol ester and superstatin three alcohol ester preferred tertiary butyl esters.

Therefore, in another embodiment, the invention provides the method for the relative retention time (RRT) of determining the impurity in superstatin glycol calcium sample, it comprises through GC or HPLC measures relative retention time (RRT) corresponding to the superstatin triol calcium in the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin two pure and mild superstatin triol calcium salts, obtain HPLC chromatography with retention time; Compare the relative retention time (RRT) of reference mark and the relative retention time (RRT) of specimen with passing through, determine the relative retention time (RRT) of the triol calcium in the sample.Usually the salt that is not calcium also can be used for present method.

Therefore, in another embodiment, the invention provides the method for the relative retention time (RRT) of determining the impurity in the superstatin two alkyd samples, it comprises through GC or HPLC measures relative retention time (RRT) corresponding to the superstatin triol acid in the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin two alkyd and superstatin triol acid, obtain having the HPLC chromatography of retention time; Compare the relative retention time (RRT) of reference mark and the relative retention time (RRT) of specimen with passing through, determine the relative retention time (RRT) of the triol acid in the sample.

Therefore, in another embodiment, the invention provides the method for the relative retention time (RRT) of determining the impurity in the superstatin diol lactone sample, it comprises through GC or HPLC measures relative retention time (RRT) corresponding to three alcoholic lactones of the superstatin in the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin two alkyd and superstatin triol acid, obtain having the HPLC chromatography of retention time; Compare the relative retention time (RRT) of reference mark and the relative retention time (RRT) of specimen with passing through, determine the relative retention time (RRT) of the triol acid in the sample.

Describe the present invention with reference to some embodiment preferred, from the thinking to this specification sheets, those skilled in the art can understand other embodiment.Following examples of method by describing preparation of compositions of the present invention and use in detail further limit the present invention.Those skilled in the art can understand, can carry out many modifications to raw material and method under not departing from the scope of the present invention.

Embodiment

Synthesizing of 1: three alcohol ester of embodiment

In inert atmosphere, TBRE (10g) is mixed with THF (56ml) solution of 1M borine methyl-sulfide complex compound.Under 20 ℃, 3h stirs the mixture.Water (5ml) solution that slowly adds NaOH (74g).Dropwise add H ₂O ₂(30% in water, 15ml), so that the temperature of mixture keeps below 50 ℃.0.5h stirs the mixture.Add dense ammonium chloride solution (150ml), and filter out precipitation.Separate each phase, with dense sodium sulfite solution (40ml), the mixed solution of water (100ml) and salt solution (100ml) is used salt solution (150ml) at last more earlier, the washing organic phase.Then, under reduced pressure remove organic solvent, obtain containing the semi-solid residue of triol, unreacted TBRE and some impurity.

Separate triol through the twice chromatographic partition method.At first, adopt in water from 40% to 45%EtOH gradient liquid, the RP-18 post (

The C-18 reversed-phase column) separates on.Employing is at CH ₂Cl ₂In from 0% to 1%EtOH gradient liquid as elutriant, common silicagel column (

The positive disposable column) carrying out the second time on separates.Purity 93%.MS(ES+)：M+H＝556?M+Na+＝578

Produce by Teledyne Isco company (Nebraska).

Synthesizing of embodiment 2:Rosu-Ca-triol

The 2g raw material of being made up of TBRE and 2.85% triol-ester (3.7mmole carboxyl) is suspended in EtOH (10mL)/water (6mL) mixture.Under the room temperature, dropwise add saturated NaOH solution (0.35g, 4.1mmole) and the 2h that stirs the mixture.Concentrated solution under the vacuum is removed EtOH.Under 40 ℃, by under agitation adding CaCl ₂(0.41g 3.7mmole), is settled out calcium salt from this aqueous solution.Under the room temperature, continue to stir 1h, precipitate after filtration, through water washing and drying.Raw material contains 2.82%Rosu-Ca-three pure and mild 95%Rosu-Ca.

Embodiment 3: superstatin triol acid synthetic

Rosu triol Ca (0.5g) is suspended in the methylene dichloride (5mL), and (1N is in water, 1mL) to add HCl.Stir after 15 minutes, separate each phase, the vacuum concentration organic phase is mainly comprised the resistates of product.Can be further purified through column chromatography (silica gel, the methylene chloride-methanol mixture is as elutriant), obtain pure Rosu triol acid.

Embodiment 4: superstatin three alcoholic lactones synthetic

Make Rosu triol Ca (4g) be dissolved in acetonitrile (40mL), and (1N is in water, 40mL) to add HCl.Stir the mixture under the room temperature all night.Remove acetonitrile and water through underpressure distillation.Can be further purified the resistates that contains product through flash chromatography method (silica gel, hexane-ethyl acetate mixture is as elutriant), obtain pure rosu three alcoholic lactones.

Embodiment 5: superstatin triol calcium synthetic

Pure triol-the ester of 2g (3.7mmol) is suspended in the mixed solution of EtOH (10mL)/water (6mL).Under the room temperature, dropwise add saturated NaOH solution (0.35g, 4.1mmole), and the 2h that stirs the mixture.Vacuum concentrated solution is removed EtOH.Under 40 ℃, add CaCl while stirring ₂(0.41g, aqueous solution 3.7mmole) precipitate calcium salt from the aqueous solution.Under room temperature, continue to stir 1h, precipitation after filtration, through water washing and drying, generate the Rosu-Ca-triol.

The MS condition

Instrument: Bruker Esquir 6000

The source: just/the negative ESI that switches

Aimed quality: 556Da

Compound stability: 50%

Trap drives: 100%

The ends of the earth (Octopole) RF:195.3Vpp

Capillary outlet: 111.8V

Dry gas flow velocity: 10L/min

Spraying gun: 60psig

Dry gas temperature: 365 ℃

V?Cap： 4000V

The HPLC method of the Impurity Distribution of TBRE

Post: C18

Moving phase: the gradient of elutriant A and elutriant B

Gradient: time (min) elutriant A (%) elutriant B (%)

0 100 0

2 84 16

23 84 16

36 10 90

40 10 90

Elutriant A:60%0.005M ammonium acetate 40% acetonitrile: ethanol=2: 3

Elutriant B:100% acetonitrile: ethanol=1: 4

UV detects: 243nm

Working time: 55min

Flow velocity: 0.6mL/min

Inject volume: 5 μ L

Column temperature: 5 ℃

Emission limit set: be less than 0.02%

Sample formulation: 0.5mg/mL

The RT of TBRE: about 24.5min

The RRT of triol-TBRE impurity is corresponding to 0.6 of TBRE main peak.

The HPLC method of ROSU Impurity Distribution

Post: C18

Moving phase: the gradient of elutriant A, elutriant B and elutriant C

Gradient: time (min) elutriant A (%) elutriant B (%) elutriant C (%)

0 100 0 0

15 0 100 0

20 0 93 7

30 0 78 22

40 0 5 95

45 0 5 95

Elutriant A: 60%0.05% Glacial acetic acid pH, 3.5 35% acetonitriles, 5% ethanol that contains ammonium hydroxide

Elutriant B: 55%0.05% Glacial acetic acid pH, 3.5 45% acetonitriles that contain ammonium hydroxide

Elutriant C:100% ethanol

UV detects: 243nm

Working time: 45min

Flow velocity: 0.5mL/min

Inject volume: 10 μ L

Column temperature: 20 ℃

Emission limit set: less than 0.02%

Sample formulation: 0.2mg/mL

The RT of ROSU: about 19min

The RRT of triol-ROSU impurity is corresponding to 0.7 of ROSU main peak.

Claims

1. superstatin triol with following structure:

X wherein is hydrogen, alkali or alkaline-earth metal or C ₁-C ₄Alkyl.

2. the superstatin triol of claim 1, wherein said superstatin triol is separated.

3. claim 1 or 2 superstatin triol, superstatin triol does not wherein contain corresponding superstatin glycol substantially.

4. be the superstatin triol of the claim 1 of sour form, it has following structure:

5. the superstatin triol of claim 4, wherein said superstatin triol is separated.

6. claim 4 or 5 superstatin triol, wherein said superstatin triol does not contain corresponding superstatin glycol substantially.

7. be the superstatin triol of the claim 1 of ester-formin, it has following structure:

R wherein represents C _1-4Alkyl.

8. the superstatin triol of claim 7, wherein said superstatin three alcohol esters are separated.

9. claim 7 or 8 superstatin triol, wherein said superstatin triol ester group does not originally contain corresponding superstatin diol ester.

10. claim 7,8 or 9 superstatin triol, ester wherein is the tert-butyl ester.

11. be the superstatin triol of the claim 1 of salt form, it has following structure:

M wherein represents alkali or alkaline earth metal cation.

12. the superstatin triol of claim 11, metallic cation wherein are the alkaline-earth metal that there are two superstatin molecules in each positively charged ion.

13. the superstatin triol of claim 12, metallic cation wherein is Ca2+.

14. claim 11,12 or 13 superstatin triol, wherein said superstatin triol is separated.

15. claim 11,12,13 or 14 superstatin triol, wherein said superstatin triol does not contain corresponding superstatin glycol substantially.

16. be the superstatin triol of the claim 1 of lactone form, it has following structure:

17. the superstatin triol of claim 16, wherein said superstatin three alcoholic lactones are separated.

18. the superstatin triol of claim 16 or 17, wherein said superstatin triol lactone does not contain corresponding superstatin glycol substantially.

19. each superstatin triol among the claim 1-18, wherein said superstatin triol is selected from: the superstatin triol that is (7S) form; The superstatin triol that is (7R) form; With racemic superstatin triol.

20. a method for preparing each triol among claim 7-10 or the 18-19, it comprises oxidation superstatin glycol C ₁-C ₄Ester obtains having at 7 bit strips superstatin three alcohol esters of hydroxyl.

21. comprising, the method for claim 20, wherein said method make superstatin C ₁-C ₄Ester and the borine solution in organic solvent merges, and obtains reaction mixture, the reaction mixture that obtains and the aqueous solution of mineral alkali is merged, and add superoxide and recovery three alcohol esters.

22. the method for claim 20 or 21, wherein said borine are the borane complexes of methyl-sulfide.

23. the method for claim 22, wherein said borine are monoalkyl-or dialkyl group-borines.

24. the method for claim 21 or 22, superoxide wherein is H ₂O ₂

25. the method for claim 21 or 22, superoxide wherein are tert-butyl hydroperoxide (TBHP) or single peroxide phthalic acid magnesium hexahydrate (MMPP).

26. the method for claim 0, alkali wherein is mineral alkali.

27. the method for claim 26, mineral alkali wherein is an alkali metal base.

28. the method for claim 27, alkali wherein is hydroxide bases.

29. the method for claim 28, hydroxide bases wherein are NaOH, KOH or LiOH.

30. the method for claim 21, alkali wherein is NH ₄OH.

31. each method among the claim 0-30, organic solvent wherein is C ₃-C ₈Ether.

32. the method for claim 31, organic solvent wherein is a tetrahydrofuran (THF).

33. a method for preparing three alkoxide of claim 11, it comprises makes three alcohol esters with following formula:

R wherein is C ₁-C ₄Ester is suspended in the mixture of water and inorganic solvent, and this suspension and alkali and ion source are merged.

34. the method for claim 33, organic solvent wherein is C ₁-C ₄Alcohol.

35. the method for claim 33, organic solvent wherein is an ethanol.

36. claim 33,34 or 35 method, ion source wherein is a calcium chloride.

37. a method for preparing the triol acid of claim 4, it comprises makes superstatin three alkoxide with following formula:

Contact with acid, M wherein is alkali or alkaline-earth metal.

38. the method for claim 37, acid wherein are hydrochloric acid or sulfuric acid.

39. the method for claim 37 or 38, wherein superstatin three alkoxide and organic solvent merge.

40. the method for claim 39, organic solvent wherein is a methylene dichloride.

41. a method for preparing the superstatin triol acid of claim 4, it comprises the ester that has following formula with acid hydrolysis:

R wherein is C ₁-C ₄Ester.

42. the method for claim 41, acid wherein are HCl or H ₂SO ₄

43. a method for preparing each lactone among the claim 16-19, it comprises that hydrolysis has three alcohol esters of following formula:

R wherein is C ₁-C ₄Ester, and make ester be converted into lactone through hydrolysis.

44. method for preparing each superstatin three alkoxide among claim 11-15 with following structure or the 18-19:

It comprises that hydrolysis has the lactone of following structure

And making lactone be converted into salt through hydrolysis, M wherein is basic metal or alkaline-earth metal.

45. method for preparing each superstatin three alkoxide among the claim 16-20 with following structure:

It comprises makes the acid with following structure

Contact with alkali.

46. a minimizing is present in the method for the amount of the impurity in the medicinal compositions of ZD-4522, it comprises the amount of measuring the superstatin triol calcium in each batch superstatin glycol calcium, selection have desired level superstatin triol calcium superstatin glycol calcium batch, and preparation has the medicinal compositions of selected superstatin glycol batch.

47. a minimizing is present in the method for the amount of superstatin three alcohol esters in the mixture that comprises superstatin diol ester and superstatin three alcohol esters, it comprises that measurement is at superstatin glycol C ₁-C ₄Superstatin triol C in each batch of ester ₁-C ₄The amount of ester selects to contain triol C ₁-C ₄The superstatin glycol C of ester ₁-C ₄Ester batch and preparation contain selected superstatin glycol C ₁-C ₄The superstatin glycol calcium medicinal compositions of ester batch.

48. the method for the impurity level in the definite superstatin diol ester sample, it comprise through GC or HPLC measure in the reference standard of superstatin three alcohol esters that comprising known quantity corresponding to the area under the peak of superstatin three alcohol esters; Measure corresponding to the area under the peak of three alcohol esters of the superstatin in the sample that is comprising superstatin three pure and mild superstatin diol esters through HPLC or GC; Determine the amount of three alcohol esters in the sample with the area of area by reference standard relatively and specimen.

49. the method for claim 48, three alcohol esters wherein are tert-butyl esters.

50. the method for the impurity level in the definite ZD-4522 sample, it comprises through GC or HPLC measures corresponding to the area under the peak of the superstatin triol calcium in the reference standard of the superstatin triol calcium that comprises known quantity; Measure corresponding to the area under the peak of the superstatin triol calcium in the sample that comprises superstatin three pure and mild superstatin glycol calcium salts through GC or HPLC; Determine the amount of the triol calcium in the sample with the area of area by reference standard relatively and specimen.

51. the method for the relative retention time (RRT) of impurity in the definite superstatin diol ester sample, it comprises the relative retention time (RRT) of superstatin three alcohol esters in GC or HPLC measure corresponding to the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin diol ester and superstatin three alcohol esters, obtain having the GC or the HPLC chromatography of retention time; With by the relative retention time (RRT) of the reference mark relative retention time (RRT) with specimen is compared, determine the relative retention time (RRT) of three alcohol esters in the sample.

52. the method for claim 51, three alcohol esters wherein are tert-butyl esters.

53. the method for the relative retention time (RRT) of impurity in the definite superstatin glycol calcium sample, it comprises through GC or HPLC measures relative retention time (RRT) corresponding to the superstatin triol calcium in the reference mark sample; Carry out GC or HPLC with the specimen that comprises superstatin two pure and mild superstatin triol calcium salts, obtain having the HPLC chromatography of retention time; With by the relative retention time (RRT) of the reference mark relative retention time (RRT) with specimen is compared, determine the relative retention time (RRT) of triol calcium in the sample.

54. as the purposes of reference standard or reference mark, it is used for determining the purity of superstatin acid, superstatin ester, superstatin salt (preferred calcium salt) and rosuvastatin statin lactone according to each compound among the claim 1-19.