CN104402848B - Preparation method of impurity compound in ramelteon and prepared standard substance - Google Patents
Preparation method of impurity compound in ramelteon and prepared standard substance Download PDFInfo
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- CN104402848B CN104402848B CN201410481974.6A CN201410481974A CN104402848B CN 104402848 B CN104402848 B CN 104402848B CN 201410481974 A CN201410481974 A CN 201410481974A CN 104402848 B CN104402848 B CN 104402848B
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
Abstract
The invention relates to the pharmaceutical field, and in particular provides a preparation method of an impurity compound type shown as formula (I) or / and formula (II) in ramelteon raw material and the prepared standard substance containing the compound shown as formula (I) or / and (II). The methods includes the steps of: 1) preparing the ramelteon material containing the compounds shown as formula (I) and / or (II) by using a preparative chromatography method, wherein the chromatographic conditions are chromatographic column of kromasil C18 10 mu 21.2*250 mm, a mobile phase of acetonitrile-water (45:55), flow rate of 30ml / min, and detection wavelength of 225nm; and respectively collecting the peaks of the compounds shown as formula (I) (relative retention time of 0.5) and formula (II) (relative retention time of 1.5).
Description
Technical field
The present invention relates to pharmaceutical field is and in particular to a kind of preparation method of the impurity compound of ramelteon and preparation
Standard substance, the quality analysiss for medicine ramelteon and inspection.
Background technology
Ramelteon (Ramelteon), chemical name is：N- [2- [(8S) -1,6,7,8- tetrahydrochysenes -2H- indeno [5,4-b]
Furan -8- base] ethyl] propionic acid amide .), it is having difficulty in going to sleep insomnia and slow for treatment of developing of Japanese Wu Tian company (TAKEDA)
Property insomnia, recruit's medicine of short-term insomnia, it lists in the U.S. and Japan respectively at 2005 and priority in 2008.This medicine
For melatonin receptors agonist, the physiological action of the melatonin secreted by pinuies can be simulated, contribute to adjusting sleep cycle, change
Kind sleep quality.It is stronger to the affinity of melatonin receptors, to the no affinity such as aminobutyric acid receptors, opiate receptor, therefore
Drug dependence will not be produced；Gerontal patient's determined curative effect to patients with chronic insomnia, chronic primary insomnia disease, and something lost effect without issue
Should, untoward reaction is similar to placebo.
Continuous improvement drug safety being required with people, the requirement to drug quality is also lifted constantly, especially
It is to need further clear and definite for the impurity in pharmaceutical raw material and control, therefore, Quality Research miscellaneous in pharmaceutical raw material is become
The focus and emphasis of current those skilled in the art's research.
Ramelteon not yet at home and abroad records in pharmacopeia at present, and existing academic information also not yet reports this pharmaceutical raw material
In the preparation method of relative substance, the sale of corresponding impurity product is not had on market.
R.Scott et al. (Metabolism of Ramelteon in Human Liver Microsomes and
Correlation with the Effect of Fluvoxamine on Ramelteon Pharmacokinetics,
《Drug Metabolishm and Disposition》2010, vol 38, No.8,1381-1391) disclose formula (I) chemical combination
Thing is ramelteon metabolite in vivo, and finds that in practical study this compound is also a kind of catabolite,
The impurity of formula (I) compound equally can be produced in the preparation or storage of ramelteon product.CN101056867A discloses
The impurity of formula (II) compound can be produced in the preparation process of ramelteon.
And formula (I) and/or (II) compound are significant for the correlational study of impurity in ramelteon raw material, can be used for
The Qualitative and quantitative analysis of impurity in ramelteon production process, such that it is able to improve the quality standard of ramelteon, are the people
Masses'safety medication provides important directive significance.
But not yet there are at present any record of preparative separation method and the report of formula (I) and/or formula (II) compound, more not
There are preparation and the sale of the formula (I) using as analysis of control and/or formula (II) compound standard product.
Therefore, a kind of formula (I) and/or the preparation method of formula (II) compound are provided and corresponding standard substance are that have very much must
Want, this is conducive to promoting the raising of ramelteon drug standard, can ensure the medication of ramelteon medicine further
Safety.
Content of the invention
For case above, the invention provides impurity formula (I) compound or/and formula (II) compound in ramelteon
Preparation method, and the standard substance of impure formula (I) compound using the method preparation or/and formula (II) compound.
Formula (I) of the present invention or/and the preparative separation method of formula (II) compound, it includes：
1) the ramelteon raw material containing formula (I) and/or (II) compound is prepared using preparative chromatography, wherein
Described chromatographic condition is：
Chromatographic column：kromasil C18 10μ21.2×250mm；
Mobile phase：Acetonitrile: water=40: 60～50: 50 mixed solution；
Flow velocity：25ml/min～35ml/min；
Detection wavelength：210nm～300nm；
Collect the stream part at peak of formula (I) and formula (II) compound respectively, obtain final product.
In the inventive method, described " preparative hplc " refers to the preparative high performance liquid chromatography in described technical field.
As one of embodiment, step 1 described in the inventive method) in chromatographic column include but is not limited to kromasil
C18 10 μ 21.2 × 250mm chromatographic column；Those skilled in the art can be combined with the present invention and art technology general knowledge, preparation
Or the commercial chromatographic column same or like with chromatographic column property described in the inventive method is to substitute step 1 of the present invention) described in
Chromatographic column, such as including but not limited to Sepax HP-Cyano10 μ 21.2 × 250mm chromatographic column.
As one of embodiment, mobile phase of the present invention is acetonitrile: water=45: 55 mixed solution；
As one of embodiment, the flow velocity of mobile phase of the present invention is 30ml/min；
As one of embodiment, Detection wavelength of the present invention is 225nm；
Under above-mentioned chromatographic condition, the relative retention time of wherein said formula (I) compound is 0.5；Formula (II) compound
Relative retention time be 1.5.
In order to the object of the invention is better achieved, from the interference of other impurities in sample size and destruction sample, keep away simultaneously
Exempt to prepare time-consuming longer during sterling, relatively costly, preferably can adopt secondary preparation method as one of embodiment, the present invention
Preparation formula (I) and/or (II) compound, you can selectively, as one of embodiment, formula (I) of the present invention or/and formula
(II) the preparative separation method of compound further includes：
2-1) by step 1) in gained formula (I) compound according to step 1) described in chromatographic condition made again
Standby, then collect the stream part at peak of above-mentioned formula (I) compound, obtain final product；
As one of embodiment, the chromatographic condition that described (I) compound is prepared again is：
Chromatographic column：kromasil C18 10μ21.2×250mm
Mobile phase：Acetonitrile: water=45: 55 mixed solution
Flow velocity：30ml/min
Detection wavelength：225nm
Collect the stream part at peak of above-mentioned formula (I) compound, obtain final product；
As one of embodiment of the present invention, under above-mentioned chromatographic condition, formula (I) compound relative retention time is about 0.5.
As one of embodiment, the method for the invention further includes：
2-2) step is by step 1) in formula (II) compound of gained carry out chromatograph preparation method preparation, wherein, described system again
The chromatographic condition of standby formula (II) compound is：
Chromatographic column：YMC C18ODS-AQ 10μ10×250mm
Mobile phase：Acetonitrile: water=36: 64 mixed solution
Flow velocity：10ml/min
Detection wavelength：225nm
Collect the stream part at peak of formula (II) compound, obtain final product.
As one of embodiment of the present invention, formula (II) compound relative retention time is about 1.5.
As one of embodiment, step 2-2 described in the inventive method) in chromatographic column include but is not limited to YMC
C18ODS-AQ 10 μ 10 × 250mm chromatographic column；Those skilled in the art can be combined with the present invention and art technology general knowledge,
Preparation or the commercial chromatographic column same or like with chromatographic column property described in the inventive method are to substitute step 2-2 of the present invention)
Described in chromatographic column, such as including but not limited to Sepax C18-P 10 μ 10 × 250mm.
As one of embodiment, the preparative separation method of formula (I) of the present invention or/and formula (II) compound is further
Including：
3) by step 1) and/or step 2) gained contains formula (I) or the eluent of formula (II) compound, removes acetonitrile, water
Mutually use organic solvent extraction；Then remove organic solvent, gained solid organic solvent carries out purification and obtains final product.
As one of embodiment of the present invention, the described organic solvent for extracting aqueous phase includes but is not limited to dichloromethane
Alkane, normal hexane or ethyl acetate or their arbitrary composition；More preferably dichloromethane；
As one of embodiment of the present invention, gained solid is carried out purification organic solvent include but is not limited to acetonitrile,
Methanol, ethanol, benzyl alcohol or its aqueous solution；As one of embodiment of the present invention, preferably 90% acetonitrile solution.
As one of embodiment of the present invention, gained contains formula (I) or the eluent of formula (II), steams acetonitrile, and aqueous phase is used
Dichloromethane extraction；Then dichloromethane is evaporated, the products therefrom aqueous dissolution of 90% acetonitrile put at room temperature until
Separate out solid, filter, obtain final product.As one of embodiment, room temperature lower standing time including but not limited to about 12 can be placed in little
When.
In the preparation method of formula (I) of the present invention or/and formula (II) compound, described step 1) in containing formula (I) or/and
The source of the ramelteon raw material of formula (II) compound is not limited by any, can be real as long as containing in this ramelteon raw material
The amount containing formula (I) or/and formula (II) compound now separating preparation may be used to the preparation method of the present invention.
As one of embodiment, step 1 described in the inventive method) in the thunder containing formula (I) or/and formula (II) compound
U.S. replaces amine raw material, including but not limited to adopts and prepares with the following method：
A) by ramelteon raw material be exposed to intensity of illumination under conditions of 4500 ± 500lx certain time (generally 5～
30 days) so as to produce one or more described impurity；
B) ramelteon raw material is placed in and places certain time under hot conditionss and carry out high temperature so as to produce described
One or more impurity；And/or
C) it is exposed to intensity of illumination with pharmaceutic adjuvant after ramelteon raw material being mixed one under conditions of 4500 ± 500lx
Fix time (generally 5～30 days) so as to produce one or more described impurity.
As one of embodiment of the present invention, described step 1) in the thunder U.S.A containing formula (I) or/and formula (II) compound replace
In the preparation method of amine raw material, the temperature of high temperature can be by art technology any combination present invention and this area
General knowledge is determined, and as one of embodiment, the temperature of the described high temperature in described step b) is 80 DEG C～200 DEG C,
Preferably 100 DEG C～180 DEG C.
As one of embodiment of the present invention, described step 1) in the thunder U.S.A containing formula (I) or/and formula (II) compound replace
In the preparation method of amine raw material, the time of high temperature can be by art technology any combination present invention and this area
General knowledge is determined, and as one of embodiment, the high temperature time in described step b) is 30～240min, preferably 60～
180min.
As one of embodiment of the present invention, described step 1) in the thunder U.S.A containing formula (I) or/and formula (II) compound replace
In the preparation method of amine raw material, the adjuvant in step c) includes starch, Lactose, hydroxypropyl cellulose, magnesium stearate, hydroxypropyl first fibre
Dimension element, Copolyvidone, titanium dioxide, ferric oxide, Macrogol 8000, or their any mixture.
In the inventive method, under given conditions, described impurity formula can be improved with ramelteon after above-mentioned adjuvant is mixed
(I) and/or formula (II) compound content, improve yield, and impurity formula (I) and/or formula (II) compound will not be divided
From having adverse effect on.
In order to ensure to contain the ramelteon raw material energy of formula (I) or/and formula (II) compound used in the inventive method
The object of the invention is enough better achieved, before being prepared, can be using the conventional method in this area first to ramelteon raw material
In whether contain formula (I) or/and formula (II) compound and their content is detected.
As one of embodiment, the inventive method can including but not limited to adopt HPLC or LS-MS to containing formula (I)
And/or the ramelteon raw material of formula (II) compound is detected.
As one of embodiment, HPLC and/or LC-MS chromatographic condition of the present invention is：
Chromatographic column：Hypersil GOLD 5μ250×4.6mm
Eluant A：0.1% formic acid solution
Eluant B：Acetonitrile
Flow velocity：0.8ml/min
Sampling volume：20μL
Detector：288nm(UV)/MS.
In the detection method of the present invention, described impurity formula (I) compound and formula (II) compound are in HPLC or HPLC-MS
Under method, relative retention time is respectively 0.7 and 1.1.
The present invention still further provides the standard substance containing formula (I) compound using the inventive method preparation, described
Standard substance are off-white color material, and chromatographic purity is 60%～98%.
As one of embodiment, invention still further provides changing containing formula (I) using the inventive method preparation
The standard substance of compound, described standard substance are off-white color material, and chromatographic purity is 96.1～98.2%..
The present invention still further provides the standard substance containing formula (II) compound using the inventive method preparation, described
Standard substance are white crystal, and chromatographic purity is 85%～98%.
As one of embodiment, invention still further provides changing containing formula (I) using the inventive method preparation
The standard substance of compound, described standard substance are white crystal, and chromatographic purity is 96.5%～98.7%.
Preparation method of the present invention has that simple to operate, raw material is easy to get, with low cost, yield is suitable for, and can be simultaneously used for Lei Mei
Preparation for impurity formula (I) and formula (II) compound in amine raw material.
The impurity formula (I) of preparation method gained of the present invention, the standard substance of formula (II) compound can be used for ramelteon medicine
The analysis of impurity, detection in thing, solve and not yet have the formula (I) using as analysis of control and/or formula (II) compound at present
The technical barrier of standard substance, thus improve and ensured the drug safety of ramelteon medicine.
Brief description
Fig. 1：In embodiment 1, ramelteon destroys sample HPLC collection of illustrative plates；
Fig. 2：The UV collection of illustrative plates of ramelteon and target impurity in embodiment 1；
Fig. 3：Impurity formula (I) MS collection of illustrative plates in embodiment 1；
Fig. 4：Impurity formula (II) MS collection of illustrative plates in embodiment 1.
Specific embodiment
The present invention is expanded on further the present invention by following examples and experimental example, but the present invention is not limited to this.
Embodiment 1
Step one, ramelteon destroy the preparation of sample
Take 20 grams of ramelteon raw material (being prepared by Zhuhai United Laboratories Ltd, lot number is 140218), be placed in
Place 2 hours under the conditions of 160 DEG C so as to produce described impurity.
Step 2, HPLC measure ramelteon and destroy sample
High performance liquid chromatograph：Shimadzu 20A type HPLC (is equipped with DAD detector)；
Chromatographic column：Hypersil GOLD 5μ250×4.6mm；
Mobile phase A：0.1% formic acid solution；Mobile phase B：Acetonitrile；
Gradient：0min(A：90%th, B：10%), 5min (A：90%th, B：10%), 45min (A：40%th, B：60%),
50min(A：10%th, B：90%), 51min (A：90%th, B：10%), 60min (A：90%th, B：10%)；
Flow velocity：0.8ml/min；
Sampling volume：20μL；
Detection wavelength：288nm.
Take above-mentioned destruction sample appropriate, be configured to the solution containing ramelteon about 2mg/ml with 50% acetonitrile-water, sample introduction divides
Analysis, in result display sample, relative retention time is chromatographic peak content (area normalization method calculating) difference of 0.7 and 1.1 positions
For 13.1% and 3.4%, target impurity content is higher, can carry out subsequent experimental.
Step 3, LC-MS measure ramelteon and destroy sample
High performance liquid chromatograph：Agilent 1260 type HPLC (is equipped with the single level Four bar detector of Agilent 6120)；
Chromatographic column：Hypersil GOLD 5μ250×4.6mm；
Mobile phase A：0.1% formic acid solution；Mobile phase B：Acetonitrile；
Gradient：0min(A：90%th, B：10%), 5min (A：90%th, B：10%), 45min (A：40%th, B：60%),
50min(A：10%th, B：90%), 51min (A：90%th, B：10%), 60min (A：90%th, B：10%)；
Sampling volume：20μL；
Flow velocity：0.8ml/min, through being diverted into mass spectrum.
Mass Spectrometry Conditions are electron spray ionisation source cation (ESI+) detection mode；Nitrogen flow rate：10L/min；Nebulizer pressure
Power：35psig；Dry gas temperature：300℃；Collision induced dissociation constant：70.
Take above-mentioned destruction sample appropriate, be configured to the solution containing ramelteon about 2mg/ml with 50% acetonitrile-water, sample introduction divides
Analysis, in result display sample, relative retention time is the chromatograph peak molecular weight respectively 273 and 257 of 0.7 and 1.1 positions, with mesh
Mark impurity molecule amount is consistent, can carry out subsequent experimental.
The preparation of step 4, impurity formula (I) compound (referred to as " impurity 1 ") and formula (II) compound (referred to as " impurity 2 ")
Prepare for the first time
Instrument：Chinese nation science and technology NP2000, chromatographic column：Kromasil C18 10 μ 21.2 × 250mm, Detection wavelength is
225nm, flow velocity is 30ml/min, and mobile phase is acetonitrile-water (45: 55).Step one gained is destroyed sample 50% acetonitrile-water
Dissolving, injecting chromatograph, collecting relative retention time is 0.5 eluent corresponding with the peak at 1.5, two groups of eluents of gained
It is respectively placed in vacuum rotary steam in 50 DEG C of water-baths to steam acetonitrile in mobile phase, gained aqueous phase uses 100ml dichloromethane to extract respectively
Twice, then by dichloromethane it is evaporated, obtains grease, then carry out secondary preparation.
Prepare for second
The preparation of impurity 1
Instrument：Chinese nation science and technology NP2000, chromatographic column：Kromasil C18 10 μ 21.2 × 250mm, Detection wavelength is
225nm, flow velocity is 30ml/min, and mobile phase is acetonitrile-water (45: 55).First time is prepared gained grease (when relatively retaining
Between be 0.5 corresponding effluent) with 50% acetonitrile-water dissolving, injecting chromatograph, collection corresponding eluent, be placed in 50 DEG C of water
Vacuum rotary steam in bath, acetonitrile is steamed, and gained aqueous phase is extracted twice with 100ml dichloromethane, then dichloromethane is evaporated, gained
Product is dissolved with 90% acetonitrile-water, is placed in and places 12 hours under room temperature, obtains off-white powder after filtration.Chromatographically pure after measured
Spend for 98.2% (about 250mg), MS result display molecular weight is consistent with impurity 1.
The preparation of impurity 2
Instrument：Chinese nation science and technology NP2000, chromatographic column：YMC C18ODS-AQ 10 μ 10 × 250mm, Detection wavelength is
225nm, flow velocity is 10ml/min, and mobile phase is acetonitrile-water (36: 64).First time is prepared gained grease (when relatively retaining
Between be 1.5 corresponding effluents) with 50% acetonitrile-water dissolving, injecting chromatograph, collection corresponding eluent, be placed in 50 DEG C of water
Vacuum rotary steam in bath, acetonitrile is steamed, and gained aqueous phase is extracted twice with 100ml dichloromethane, then dichloromethane is evaporated, gained
Product is dissolved with 90% acetonitrile-water, is placed in and places 12 hours under room temperature, obtains white crystal after filtration.Chromatographic purity after measured
For 98.7% (about 150mg), MS result display molecular weight is consistent with impurity 2.
The Structural Identification of step 5, impurity 1 and impurity 2
LC-MS and nuclear magnetic resonance spectroscopy (1H-NMR, 13C-NMR) is adopted to carry out structural identification the impurity of gained, number
According to as follows：
The Structural Identification data of impurity 1：
MS(ESI)：274.2[M+H]+
1H-NMR (400MHz, CDCl3) δ (ppm)：1.12～1.15 (t, 3H), 1.63～1.73 (m, 1H), 1.91 (s,
2H, H2O), 2.15～2.24 (q, 2H；M, 1H), 2.39～2.44 (q, 1H), 2.83～2.90 (q, 1H), 3.17～3.24 (m,
1H), 3.25～3.30 (q, 2H), 3.32～3.42 (m, 2H), 4.64～4.71 (q, 1H), 4.72～4.79 (q, 1H), 5.74
(s, 1H), 6.80～6.82 (d, 1H), 7.57～7.59 (d, 1H)；13C-NMR (400MHz, CDCl3) δ (ppm)：9.78,
27.43,29.63,33.43,35.23,37.72,42.80,72.50,110.11,123.17,125.17,130.37,154.83,
166.49,174.01,203.64.
The Structural Identification data of impurity 2：
MS(ESI)：258.2[M+H]+
1H-NMR (400MHz, CDCl3) δ (ppm)：1.08～1.12 (t, 3H), 1.72～1.181 (m, 1H), 1.88～
1.95 (m, 1H), 2.10～2.16 (q, 2H), 2.17～2.20 (m, 1H), 2.34～2.43 (m, 1H), 2.88～2.95 (m,
1H), 3.00～3.08 (m, 1H), 3.36～3.40 (q, 2H), 3.42～3.49 (m, 1h), 5.48 (s, 1H), 6.76 (d, 1H),
7.11～7.14 (d, 1H), 7.30～7.32 (d, 1H), 7.60 (d, 1H)；13C-NMR (400MHz, CDCl3) δ (ppm)：
9.80,29.69,31.34,31.76,34.56,38.05,42.47,104.63,109.56,120.42,123.56,137.60,
138.63,145.06,154.72,173.67.
Embodiment 2
Step one, ramelteon destroy the preparation of sample
Take 10 grams of ramelteon raw material (being prepared by Zhuhai United Laboratories Ltd, lot number is 140218), be placed in
Place 3 hours under the conditions of 180 DEG C so as to produce described impurity.
Step 2, HPLC measure ramelteon and destroy sample
High performance liquid chromatograph：Shimadzu 20A type HPLC (is equipped with DAD detector)；
Chromatographic column：Hypersil GOLD 5μ250×4.6mm；
Mobile phase A：0.1% formic acid solution；Mobile phase B：Acetonitrile；
Gradient：0min(A：90%th, B：10%), 5min (A：90%th, B：10%), 45min (A：40%th, B：60%),
50min(A：10%th, B：90%), 51min (A：90%th, B：10%), 60min (A：90%th, B：10%)；
Flow velocity：0.8ml/min；
Sampling volume：20μL；
Detection wavelength：288nm.
Take above-mentioned destruction sample appropriate, be configured to the solution containing ramelteon about 2mg/ml with 50% acetonitrile-water, sample introduction divides
Analysis, in result display sample, relative retention time is chromatographic peak content (area normalization method calculating) difference of 0.7 and 1.1 positions
For 13.7% and 3.1%, target impurity content is higher, can carry out subsequent experimental.
Step 3, LC-MS measure ramelteon and destroy sample
High performance liquid chromatograph：Agilent 1260 type HPLC (is equipped with the single level Four bar detector of Agilent 6120)；
Chromatographic column：Hypersil GOLD 5μ250×4.6mm；
Mobile phase A：0.1% formic acid solution；Mobile phase B：Acetonitrile；
Gradient：0min(A：90%th, B：10%), 5min (A：90%th, B：10%), 45min (A：40%th, B：60%),
50min(A：10%th, B：90%), 51min (A：90%th, B：10%), 60min (A：90%th, B：10%)；
Sampling volume：20μL；
Flow velocity：0.8ml/min, through being diverted into mass spectrum.
Mass Spectrometry Conditions are electron spray ionisation source cation (ESI+) detection mode；Nitrogen flow rate：10L/min；Nebulizer pressure
Power：35psig；Dry gas temperature：300℃；Collision induced dissociation constant：70.
Take above-mentioned destruction sample appropriate, be configured to the solution containing ramelteon about 2mg/ml with 50% acetonitrile-water, sample introduction divides
Analysis, in result display sample, relative retention time is the chromatograph peak molecular weight respectively 273 and 257 of 0.7 and 1.1 positions, with mesh
Mark impurity molecule amount is consistent, can carry out subsequent experimental.
The preparation of step 4, impurity formula (I) compound (referred to as " impurity 1 ") and formula (II) compound (referred to as " impurity 2 ")
Prepare for the first time
Instrument：Chinese nation science and technology NP2000, chromatographic column：Sepax HP-Cyano 10 μ 21.2 × 250mm, Detection wavelength is
225nm, flow velocity is 35ml/min, and mobile phase is acetonitrile-water (40: 60).Step one gained is destroyed sample 50% acetonitrile-water
Dissolving, injecting chromatograph, collecting relative retention time is 0.5 eluent corresponding with the peak at 1.5, two groups of eluents of gained
It is respectively placed in vacuum rotary steam in 50 DEG C of water-baths to steam acetonitrile in mobile phase, gained aqueous phase uses 100ml ethyl acetate to extract respectively
Twice, then by ethyl acetate it is evaporated, obtains grease, then carry out secondary preparation.
Prepare for second
The preparation of impurity 1
Instrument：Chinese nation science and technology NP2000, chromatographic column：Sepax HP-Cyano 10 μ 21.2 × 250mm, Detection wavelength is
225nm, flow velocity is 35ml/min, and mobile phase is acetonitrile-water (40: 60).First time is prepared gained grease (when relatively retaining
Between be 0.5 corresponding effluent) with 50% acetonitrile-water dissolving, injecting chromatograph, collection corresponding eluent, be placed in 50 DEG C of water
Vacuum rotary steam in bath, acetonitrile is steamed, and gained aqueous phase is extracted twice with 100ml ethyl acetate, then ethyl acetate is evaporated, gained
Product is dissolved with 90% methanol-water, is placed in and places 12 hours under room temperature, obtains off-white powder after filtration.Chromatographically pure after measured
Spend for 96.1% (about 120mg), MS result display molecular weight is consistent with impurity 1.
The preparation of impurity 2
Instrument：Chinese nation science and technology NP2000, chromatographic column：Sepax C18-P 10 μ 10 × 250mm, Detection wavelength is 225nm,
Flow velocity is 10ml/min, and mobile phase is acetonitrile-water (36: 64).First time is prepared gained grease, and (relative retention time is
1.5 corresponding effluents) dissolved with 50% acetonitrile-water, injecting chromatograph, collect corresponding eluent, be placed in 50 DEG C of water-baths
Vacuum rotary steam, acetonitrile is steamed, and gained aqueous phase is extracted twice with 100ml ethyl acetate, then ethyl acetate is evaporated, products therefrom
Dissolved with 90% methanol-water, be placed in and place 12 hours under room temperature, after filtration, obtain white crystal.Chromatographic purity is after measured
96.5% (about 60mg), MS result display molecular weight is consistent with impurity 2.
Embodiment 3
Step one, ramelteon destroy the preparation of sample
Take 10 grams of ramelteon raw material (being prepared by Zhuhai United Laboratories Ltd, lot number is 140218), be placed in
Place 1.5 hours under the conditions of 140 DEG C so as to produce described impurity.
Step 2, HPLC measure ramelteon and destroy sample
High performance liquid chromatograph：Shimadzu 20A type HPLC (is equipped with DAD detector)；
Chromatographic column：Hypersil GOLD 5μ250×4.6mm；
Mobile phase A：0.1% formic acid solution；Mobile phase B：Acetonitrile；
Gradient：0min(A：90%th, B：10%), 5min (A：90%th, B：10%), 45min (A：40%th, B：60%),
50min(A：10%th, B：90%), 51min (A：90%th, B：10%), 60min (A：90%th, B：10%)；
Flow velocity：0.8ml/min；
Sampling volume：20μL；
Detection wavelength：288nm.
Take above-mentioned destruction sample appropriate, be configured to the solution containing ramelteon about 2mg/ml with 50% acetonitrile-water, sample introduction divides
Analysis, in result display sample, relative retention time is chromatographic peak content (area normalization method calculating) difference of 0.7 and 1.1 positions
For 13.0% and 3.5%, target impurity content is higher, can carry out subsequent experimental.
Step 3, LC-MS measure ramelteon and destroy sample
High performance liquid chromatograph：Agilent 1260 type HPLC (is equipped with the single level Four bar detector of Agilent 6120)；
Chromatographic column：Hypersil GOLD 5μ250×4.6mm；
Mobile phase A：0.1% formic acid solution；Mobile phase B：Acetonitrile；
Gradient：0min(A：90%th, B：10%), 5min (A：90%th, B：10%), 45min (A：40%th, B：60%),
50min(A：10%th, B：90%), 51min (A：90%th, B：10%), 60min (A：90%th, B：10%)；
Sampling volume：20μL；
Flow velocity：0.8ml/min, through being diverted into mass spectrum.
Mass Spectrometry Conditions are electron spray ionisation source cation (ESI+) detection mode；Nitrogen flow rate：10L/min；Nebulizer pressure
Power：35psig；Dry gas temperature：300℃；Collision induced dissociation constant：70.
Take above-mentioned destruction sample appropriate, be configured to the solution containing ramelteon about 2mg/ml with 50% acetonitrile-water, sample introduction divides
Analysis, in result display sample, relative retention time is the chromatograph peak molecular weight respectively 273 and 257 of 0.7 and 1.1 positions, with mesh
Mark impurity molecule amount is consistent, can carry out subsequent experimental.
The preparation of step 4, impurity formula (I) compound (referred to as " impurity 1 ") and formula (II) compound (referred to as " impurity 2 ")
Instrument：Chinese nation science and technology NP2000, chromatographic column：Kromasil C18 10 μ 21.2 × 250mm, Detection wavelength is
225nm, flow velocity is 30ml/min, and mobile phase is acetonitrile-water (45: 55).
Step one gained destruction sample is dissolved with 50% acetonitrile-water, injecting chromatograph, collecting relative retention time is
0.5 eluent corresponding with the peak at 1.5, two groups of eluents of gained are respectively placed in 50 DEG C of water-baths vacuum rotary steam by mobile phase
Middle acetonitrile steams, and gained aqueous phase is extracted twice with 100ml dichloromethane respectively, and dichloromethane is evaporated, and is vacuum dried at 80 DEG C
24 hours, obtain two kinds of yellowish-brown powder.Impurity 1 chromatographic purity is 59.8% (about 300mg) after measured, impurity 2 chromatographic purity
For 86.5% (about 150mg).
In MS result display above-mentioned substance, main component is consistent with target impurity molecular weight.
Claims (12)
1. the preparation method of formula (I) or/and formula (II) compound is it is characterised in that methods described includes：
1) the ramelteon raw material containing formula (I) and/or (II) compound is prepared using preparative chromatography, wherein said
Chromatographic condition is：
Chromatographic column：kromasil C18 10μm 21.2×250mm
Mobile phase：Acetonitrile:Water=40:60～50:50 mixed solution
Flow velocity：25～35ml/min
Detection wavelength：210nm～300nm
Collect the stream part at peak of formula (I) and/or formula (II) compound respectively, obtain final product；
Wherein, the described ramelteon raw material containing formula (I) or/and formula (II) compound is adopted and is prepared with the following method：
B) by ramelteon raw material be placed under the conditions of 80～200 DEG C place 30～240min time carry out high temperature so as to
Produce one or more described impurity.
2. method according to claim 1 is it is characterised in that methods described further includes：
2-1) by step 1) in gained formula (I) compound according to step 1) described in chromatographic condition be prepared again, so
Collect the stream part at peak of above-mentioned formula (I) compound afterwards, obtain final product.
3. method according to claim 1 is it is characterised in that methods described further includes：
2-2) step is by step 1) in the formula II compound of gained carry out chromatograph preparation method preparation again, wherein prepare formula (II)
The chromatographic condition of compound is：
Chromatographic column：YMC C18ODS-AQ 10μm 10×250mm
Mobile phase：Acetonitrile:Water=36:64 mixed solution
Flow velocity：10ml/min
Detection wavelength：225nm
Collect the stream part at peak of formula (II) compound, obtain final product.
4. method according to claim 1 and 2 is it is characterised in that described step 1) in mobile phase be acetonitrile:Water=45:
55 mixed solution.
5. method according to claim 1 and 2 is it is characterised in that described step 1) in the flow velocity of mobile phase be 30ml/
min.
6. method according to claim 1 and 2 is it is characterised in that described step 1) in Detection wavelength be 225nm.
7. method according to claim 1 and 2 is it is characterised in that methods described also includes：
3) by step 1) and/or step 2-1) gained contains the eluent of formula (I) compound, or step 1) and/or step 2-2)
Eluent containing formula (II) compound, removes acetonitrile, aqueous phase organic solvent extraction, the described organic solvent choosing for extracting
From dichloromethane, ethyl acetate or normal hexane；Then remove organic solvent, gained solid organic solvent carries out purification and obtains final product,
The described organic solvent for purification is acetonitrile, methanol, ethanol, benzyl alcohol or its aqueous solution.
8. method according to claim 7, is characterised by, methods described further includes：Containing formula I or (II) chemical combination
After the eluent of thing removes acetonitrile, aqueous phase dichloromethane extraction and then dichloromethane is evaporated, products therefrom aqueous acetonitrile
Liquid dissolves, and putting at room temperature until separating out solid, filtering, obtaining final product, described acetonitrile solution is 90% acetonitrile solution.
9. method according to claim 1 is it is characterised in that described step b) high temperature fail temperature is for 100～180
℃.
10. method according to claim 1 it is characterised in that in described step b) standing time be 60～180min.
11. methods according to claim 1 are it is characterised in that methods described selectively includes thunder U.S.A is replaced further
Formula (I) in amine raw material and/or formula (II) compound carry out HPLC or LS-MS detection.
12. methods according to claim 11 are it is characterised in that described HPLC and/or LC-MS chromatographic condition is：
Chromatographic column：Hypersil GOLD 5μm 250×4.6mm
Eluant A:0.1% formic acid solution；Eluant B:Acetonitrile
Flow velocity:0.8ml/min；
Sampling volume：20μL；
Detector：288nm(UV)/MS.
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| CN102924410A (en) * | 2012-10-29 | 2013-02-13 | 华润赛科药业有限责任公司 | Preparation method and intermediate of ramelteon |
| CN103664849A (en) * | 2012-08-31 | 2014-03-26 | 上海阳帆医药科技有限公司 | Method for preparing 2-(1,6,7,8-tetrahydro-2H-indeno[5,4-b]furan-8-idenethamine |
| CN103880794A (en) * | 2012-12-21 | 2014-06-25 | 上海阳帆医药科技有限公司 | Preparation method for key intermediate of ramelteon |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103664849A (en) * | 2012-08-31 | 2014-03-26 | 上海阳帆医药科技有限公司 | Method for preparing 2-(1,6,7,8-tetrahydro-2H-indeno[5,4-b]furan-8-idenethamine |
| CN102924410A (en) * | 2012-10-29 | 2013-02-13 | 华润赛科药业有限责任公司 | Preparation method and intermediate of ramelteon |
| CN103880794A (en) * | 2012-12-21 | 2014-06-25 | 上海阳帆医药科技有限公司 | Preparation method for key intermediate of ramelteon |
Non-Patent Citations (1)
| Title |
|---|
| Metabolism of ramelteon in human liver microsomes and correlation with the effect of fluvoxamine on ramelteon pharmacokinetics;Obach, R. Scott et al.;《Drug Metabolism and Disposition》;20101231;第38卷(第8期);1381-1391 * |
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