CN109925279A - A kind of lipoic acid injection and preparation method thereof - Google Patents
A kind of lipoic acid injection and preparation method thereof Download PDFInfo
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- CN109925279A CN109925279A CN201711370758.4A CN201711370758A CN109925279A CN 109925279 A CN109925279 A CN 109925279A CN 201711370758 A CN201711370758 A CN 201711370758A CN 109925279 A CN109925279 A CN 109925279A
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- lipoic acid
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Abstract
The present invention provides a kind of highly-safe lipoic acid preparation, including lipoic acid, co-solvent component, which is characterized in that in the lipoic acid preparation, contain lipoic acid ethylenediamine disubstitution product, wherein lipoic acid ethylenediamine disubstitution product content is not higher than 0.2%.The lipoic acid preparation is highly-safe, and toxicity is low, can preferably play the clinical efficacy of lipoic acid.
Description
Technical field
The present invention relates to a kind of pharmaceutical preparations and preparation method thereof, and in particular to a kind of injection containing lipoic acid and its
Preparation method.
Background technique
Lipoic acid, its chemical name is (±) -5- [3- (1,2- dithiolane)]-valeric acid, molecular formula is
C8H14O2S2, molecular weight 206.33 is the natural products separated from pork liver for the first time by ReedShi, is pyruvic acid and three
The cofactors of the oxidative deamination reaction of α-ketoglutaric acid in carboxylic acid recycle.Lipoic acid has both fat-soluble and water-soluble characteristic,
It can be transported in body most tissues and play a role.
Lipoic acid is widely used in the neuropathy for the treatment of diabetes.Isolated test shows that this product can reduce nerve
The lipid oxidation phenomenon of tissue, may prevent the glycosylation of protein；And can inhibit aldose reductase, thus Portugal can be prevented
Grape sugar or galactolipin transform into sorbierite, so lipoic acid can prevent diabetes, control blood glucose and prevent from making because of hyperglycemia
At neuropathy.
Lipoic acid is at home and abroad widely applied at present, and earliest marketed drug lipoic acid is derived from Germany, in Europe, lipoic acid
Injection is the drug of the multiple Micro-circutation syndrome for the treatment of diabetes that is uniquely being identified and having listed；In China, the medicine in
The approval of import in 2000.In November, 2009, national new health insurance directory have included alpha-lipoic acid injection for the first time to adjust blood glucose
Medical insurance Class B drugs.
As lipoic acid is in clinical extensive use, the report of its adverse reaction is also gradually increased.In clinical application, sulphur
The adverse reaction of sad injection is relatively common, reaches 10% or so, shows as infusion site pain, phlebitis, arm acid more
Swollen, dizzy, headache, needle pricked pain etc..Therefore, how research reduces the adverse reaction rate of lipoic acid, reduces bad anti-
The severity answered is those skilled in the art's technical problem urgently to be resolved.
In addition, solubility is small in water for lipoic acid, and there is thermal instability may be thermal decomposited when the temperature is excessively high,
Higher impurity is generated, to influence its bioactivity and safety in utilization.Therefore, in the prior art, lipoic acid injection is long
Phase storage will appear related substance excessive problem, and clarity is unqualified, and pH value reduces, the poor influence drug effect of stability and drug peace
Quan Xing.
It would therefore be desirable to which a kind of can overcome the composition of lipoic acid thermal instability and relative ease can be real
Existing preparation method, improves the thermal stability of lipoic acid injection, and reduces the incidence of its adverse reaction.
Summary of the invention
For this purpose, applicant has conducted extensive research lipoic acid, and is found surprisingly that, and in production and storage process, sulphur
Lipoic acid ethylenediamine disubstitution product (molecule containing 1 structure of formula) this forefathers can be generated in sad preparation not it has been found that it is related
Substance, and this substance can not only reduce the drug effect of lipoic acid preparation, can also increase lipoic acid preparation to a certain extent in body
Interior toxicity.The lipoic acid ethylenediamine disubstitution product includes the various molecules containing such as 1 structure of formula.
Formula 1:
Therefore, it is necessary to control the amount in lipoic acid preparation in relation to substance to improve its drug effect and safety in utilization.
On the one hand, be based on object above, the present invention provides a kind of lipoic acid preparation, containing lipoic acid, preparation often use auxiliary material,
Lipoic acid ethylenediamine disubstitution product (molecule containing 1 structure of formula, such as its salt or hydrate), wherein lipoic acid ethylenediamine is double to be taken
0.001-0.2% (w/v) is not higher than for object content.Preferably, lipoic acid content and lipoic acid ethylenediamine disubstitution product content it
Than being not less than 25:1.
The preparation, which often uses auxiliary material, to be the one or more of cosolvent, isotonic regulator, buffer salt, antioxidant.
On the other hand, the present invention provides a kind of highly-safe lipoic acid injection, containing lipoic acid, co-solvent component,
It is characterized in that, also containing lipoic acid ethylenediamine disubstitution product in the lipoic acid injection, wherein lipoic acid ethylenediamine is double
Substituent is the molecule (such as its salt or hydrate) containing 1 structure of formula, the preferably molecule (C of formula 118H32O2N2S4), i.e. N, N'-
(ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide), content are not higher than 0.2% (w/v), preferably no greater than
0.01%, more preferably no higher than 0.002%, most preferably no more than 0.001%.
The ratio between content of lipoic acid and lipoic acid ethylenediamine disubstitution product is not less than 25:1, and the ratio between preferred content is not less than
150:1, the ratio between preferred content are not less than 1500:1.
In the lipoic acid injection, lipoic acid content (w/v) is not less than 1%, preferred content 2-10%, more preferable content
It is 2.5%.
In the lipoic acid injection, co-solvent component content (w/v) is not less than 0.1%, and preferred content is not less than
0.5%, preferred content is 0.5-0.8%.
The cosolvent is selected from one or more of combinations of ethylenediamine, tromethamine or meglumine.
Further, pharmaceutically acceptable isotonic regulator can also be contained in the lipoic acid injection, it is described etc.
Seep regulator be selected from sodium chloride, glucose, fructose, phosphate, citrate, mannitol, polyethylene glycol, propylene glycol one kind or
Several combinations.
Further, pharmaceutically acceptable buffer salt, the buffer salt can also be contained in the lipoic acid injection
One or more of combinations selected from such as citrate, phosphate, carbonate etc..
Further, pharmaceutically acceptable antioxidant, the antioxygen can also be contained in the lipoic acid injection
Agent is selected from one or more of combinations of such as cysteine, sodium hydrogensulfite, glycine, sodium sulfite.
The above-mentioned method for preparing lipoic acid injection, step include: (1) pretreatment: water for injection deaeration by nitrogen stripping, detection
Water oxygen level qualification (water oxygen level < 20ug/ml-50ug/ml)；Measure the water for injection for matching liquid total amount 3/4-4/5；(2)
Weigh: weigh lipoic acid by recipe quantity 102%-105%, by recipe quantity weigh cosolvent, lipoic acid ethylenediamine disubstitution product and
Sufficient active charcoal；(3) dilute to match method: cosolvent is added in prescription ratio, stirs evenly；(4) add raw material: continuing to stir (100r/
Min-500r/min it) is slowly added to recipe quantity lipoic acid and lipoic acid ethylenediamine disubstitution product under state, stirs evenly；(5) add
Enter auxiliary material: recipe quantity isotonic regulator, buffer salt, antioxidant being added as needed；(6) add active carbon: adding water to full dose,
Active carbon is added, stirs 20-40 minutes；(7) decarburization: through 4 μm of stud coarse filtration；(8) preliminary survey: measurement pH value, solution ph should be
8.0-8.3；(9) refined filtration: through 0.45 μm of filter membrane core by-pass filtration；(10) end-filtration: through 0.22 μm of filter membrane core second level degerming
Filter to encapsulating step, prepare start to filling beginning when be limited in 3-4.5 hours；(11) encapsulating: detection intermediate products, content
97.0-103.0%, pH value 8.0-8.3 are qualified products, and the intermediate products medical fluid that will test qualification is filling, and encapsulating starts to filling
Sealed knot beam when be limited in 12 hours；(12) nitrogen charging, sealing, sterilizing: 121 DEG C of moist heat sterilization 15-30min.Using above-mentioned system
The generation of injection lipoic acid ethylenediamine disubstitution product during storage can be effectively reduced in Preparation Method.
On the other hand, the present invention also provides a kind of alpha-lipoic acid liposome injection, including lipoic acid, phosphatide, octadecylamine,
Nonionic surfactant, citrate, mercaptoethanol, which is characterized in that in the alpha-lipoic acid liposome injection, contain sulphur
Sad ethylenediamine disubstitution product, wherein in mass, lipoic acid ethylenediamine disubstitution product content is not higher than 0.2% (w/v), excellent
Choosing is not higher than 0.01%, more preferably no higher than 0.001%.The ratio between lipoic acid and the content of lipoic acid ethylenediamine disubstitution product be not low
In 25:1, the ratio between preferred content is not less than 150:1, and the ratio between preferred content is not less than 1500:1.
In mass, 120 parts of the lipoic acid, 0-120 parts of phosphatidase 3,50-80 parts of octadecylamine, nonionic surfactant
It is 25-40 parts, 1-5 parts of citrate, mercaptoethanol is 3-12 parts, is dissolved in 1000 parts of waters for injection.
Preferably, the phosphatide is synthetic phospholipid, it is furthermore preferred that the synthetic phospholipid is Distearate Phosphatidylcholine.
Preferably, the nonionic surfactant is PLURONICS F87.
Preferably, the citrate is sodium citrate.
The above-mentioned method for preparing alpha-lipoic acid liposome injection, step include: that (1) is pungent by the lipoic acid of recipe quantity, sulphur
Sour ethylenediamine disubstitution product, octadecylamine, phosphatide and nonionic surfactant are dissolved in organic solvent, are uniformly mixed, decompression removes
Organic solvent is removed, lipid membrane is obtained；(2) under nitrogen protection, the citrate buffer solution of pH=5-7, vibration are added into bottle
It shakes, stirs 20-50 minutes, revolving speed 400-700r/min, make the complete aquation of immobilized artificial membrane, with the homogeneous emulsification 5-10 of tissue mashing machine
Minute, liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in revolving speed 9000-12000r/min；(3) citric acid is added
Sodium and mercaptoethanol, stirring, constant volume are filling to get lipoic acid liposome injection with 0.45 μm of filtering with microporous membrane.Using
The prescription of liposome and above-mentioned method for preparing lipidosome, can highly effective reduction preparation lipoic acid second two in storage
The generation of amine disubstitution product.
On the other hand, the present invention also provides a kind of preparation method of lipoic acid, step includes: (1) sodium sulfide solution
Configuration；(2) synthesis of lipoic acid ethyl ester；(3) lipoic acid second ester hydrolysis；(4) acidification of sodium thioctate；(5) lipoic acid is thick
The purifying of product.
Specifically, step (1) is, the vulcanized sodium of 20 parts by weight is dissolved in 80-120 parts by weight purified water, at 35 DEG C -45
DEG C dissolution, be configured to sodium sulfide solution；Step (2) is that 6,8-dichloro-octanoic acid second is added in the aqueous solvent of 100-200 parts by weight
20-80 parts of ester, 5-10 parts of sulphur, 2-5 parts of tetrabutylammonium bromide, are warming up to 85-86 DEG C, and the vulcanized sodium that step (1) is obtained is molten
Liquid is slowly added into this aqueous solution.After sodium sulfide solution is added, the reaction was continued 3-5 hours；After reaction, 60- is cooled to
It 65 DEG C, static about 60 minutes, is layered to get lipoic acid ethyl ester is arrived；Step (3) is the addition lipoic acid ethyl ester in aqueous solvent, then
20-25 parts of sodium hydroxides and 1-3 parts of tetrabutylammonium bromide are added, 200-400r/min stirring is warming up to 60 DEG C -80 DEG C, reaction
After, it filters while hot；Step (4) is that the filtrate that step (3) obtains is cooled to 5 DEG C -10 DEG C, then uses salt acid for adjusting pH
Lipoic acid crude product is obtained to 1-2 filtering；Step (5) is that the lipoic acid crude product that step (4) obtains is added to mixed solvent
In (hexamethylene: ethyl acetate=5-10:1 (w:w)), under 100-500r/min stirring, 40 DEG C -45 DEG C are heated to, heat preservation
30-50 minutes, it is completely dissolved lipoic acid crude product, 36 DEG C~40 DEG C of purified water is then added, is kept stirring 10-15 minutes,
After organic layer is heated to 40 DEG C -50 DEG C, silica gel is added in static 20-30 minutes removing water layer, after stirring 2h, is filtered.So
After be cooled to 16 DEG C -20 DEG C, stirring and crystallizing 1 hour, filter and obtain the wet fine work of lipoic acid, then at a temperature of 35 DEG C -40 DEG C
It is 12-24 hours dry.
On the other hand, the present invention also provides a kind of N, (1, the 2- dithiolane -3- valeryls of N'- (ethane -1,2- diyl) two
Amine) preparation method, in the preparation process in accordance with the present invention, each reaction is usually in atent solvent, at -20 DEG C to reflux temperature
It is carried out under (preferably 0-80 DEG C).Reaction time is usually 0.02 hour to 240 hours, and preferably 0.5 hour to 60 hours.
In a preferred example, route preparation can be synthesized as follows in formula (1) compound.
Synthetic route:
Compound 1a and 1b reacts 0.02-240 hours under coupling conditions, obtains compound (1).
Preferably, which carries out in atent solvent, it is furthermore preferred that the atent solvent is that inertia aprotic, polar is molten
Agent.Reaction temperature is between -20-80 DEG C.
Preferably, coupling agent N, N '-carbonyl dimidazoles (CDI), 1- ethyl -3- (3- dimethylamino) carbodiimide, 1-
Hydroxybenzotriazole (EDC/HOBT), hexafluorophosphoric acid (- 1 base of 7- azepine benzotriazole)-N, N, N ', N '-tetramethylurea, 1- hydroxyl
One or more of combinations of azepine benzotriazole.
Polar aprotic solvent includes but is not limited to: methylene chloride, chloroform, and 1,2 ,-dichloroethanes, ethyl acetate,
Tetrahydrofuran, ether, methyl tertiary butyl ether(MTBE), 1,6- dioxane, dimethylformamide, dimethyl sulfoxide, acetonitrile or combinations thereof.
Preferably, reaction condition is 0-40 DEG C, is reacted 0.5 hour to 60 hours.
N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) can be used as the inspection of lipoic acid preparation impurity
The standard items of survey.
On the other hand, it was found by the inventors of the present invention that two (1,2- dithiolane -3- penta of N, N'- (ethane -1,2- diyl)
Amide) can be used as lubricant applied to high polymer preparation in, reach improve lubricity, reduce friction, reduce Interface Adhesion
The purpose of performance；Application of dispersant be can simultaneously serve as in the preparation of pigment or filler.
The high polymer is preferably polyethylene (PE), polypropylene (PP), polystyrene (PS), acrylonitrile-styrene resin
(AS), acrylonitrile-butadiene-styrene (ABS) plastics (ABS), polyamide (PA), poly terephthalic acid class (PET), polycarbonate
(PC)。
On the other hand, the present invention also provides N in a kind of lipoic acid injection, N'- (ethane -1,2- diyl) two (1,2-
Dithiolane -3- pentanamide) detection method.This method N suitable for lipoic acid injection, N'- (ethane -1,2- diyl) two
The detection of (1,2- dithiolane -3- pentanamide), and convenient for operation, separating degree is high, favorable reproducibility.
N in the lipoic acid injection, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) are used
Liquid chromatography is detected.
Stationary phase using octadecylsilane chemically bonded silica as filler, preferably selection Kromasil 100-5C18 (4.6 ×
250mm, 5 μm) chromatographic column；Mobile phase is mixed by mobile phase A liquid with Mobile phase B, mobile phase A are as follows: 0.002-0.008mol/
L phosphate solution, preferably 0.005mol/L potassium dihydrogen phosphate, are 2-2.5 with phosphorus acid for adjusting pH value, it is furthermore preferred that pH
Value is adjusted to 2.3；Mobile phase B are as follows: acetonitrile-methanol (5-10:30-50), preferably acetonitrile-methanol (8:42)；Ladder is carried out by table 2
Degree elution；Flow velocity is 0.5-1.5ml/min, preferably 1.0ml/min；Column temperature is 20-40 DEG C, preferably 35 DEG C；Detection wavelength
For 200-250nm, preferably 215nm；When detection, with reference level N, two (1,2- dithiolane -3- of N'- (ethane -1,2- diyl)
Pentanamide) it is reference substance, the N in lipoic acid injection, two (1,2- bis- sulphur penta of N'- (ethane -1,2- diyl) are calculated with external standard method
Ring -3- pentanamide) content.
2 gradient elution of table flows phase concentration variation
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 60 | 40 |
| 15 | 60 | 40 |
| 30 | 30 | 70 |
| 110 | 30 | 70 |
| 111 | 60 | 40 |
| 120 | 60 | 40 |
Inventor is found surprisingly that, when the raising with lipoic acid ethylenediamine disubstitution product content, lipoic acid injection
Adverse reaction rate can be significantly increased, and specifically, the adverse reaction rate of user's nervous system significantly improves, specific table
It is now headache, dizziness etc..
On the other hand, the present invention also provides a kind of quality evaluating method of lipoic acid injection, the method is detection note
The content for penetrating lipoic acid ethylenediamine disubstitution product in liquid, when lipoic acid ethylenediamine disubstitution product content is more than in the injection
When 0.2%, rejected product is determined that it is.
Preferably, the lipoic acid ethylenediamine disubstitution product is N, two (1,2- bis- sulphur penta of N'- (ethane -1,2- diyl)
Ring -3- pentanamide).
On the other hand, the present invention also provides a kind of method for improving lipoic acid injection safety in utilization, the method is,
The content for detecting lipoic acid ethylenediamine disubstitution product in injection, when lipoic acid ethylenediamine disubstitution product content in the injection
When more than 0.2% (2g/L), rejected product is determined that it is, and abandon；When lipoic acid ethylenediamine disubstitution product in the injection
When content is no more than 0.2% (2g/L), if lipoic acid ethylenediamine disubstitution product content is x% (10x g/L), then the sulphur
Sad injection should use in (0.2-x)/0.00639 month after sensing.
Preferably, the lipoic acid ethylenediamine disubstitution product is N, two (1,2- bis- sulphur penta of N'- (ethane -1,2- diyl)
Ring -3- pentanamide).
Lipoic acid of the present invention can be (±)-lipoic acid, be also possible to (+)-lipoic acid.
Compared with prior art, the present invention has the advantage that
1. present invention finds related substances new in a kind of lipoic acid preparation, and adequately grind to its toxicity
Study carefully, while having speculated the relationship between this impurity and lipoic acid adverse reaction.
2. stability is good the present invention provides a kind of lipoic acid injection, highly-safe, toxicity is low, can be better
Play the clinical efficacy of lipoic acid.
3. the present invention provides in lipoic acid injection, to the detection method of lipoic acid ethylenediamine disubstitution product, separation
Degree is high, and specificity is good, can carry out accurate detection to the lipoic acid ethylenediamine disubstitution product in lipoic acid injection.
4. the lipoic acid of this method preparation, purity is higher, preparation the present invention provides a kind of preparation method of lipoic acid
The related content of material of preparation is small, improves the safety of preparation.
5. the present invention also provides a kind of preparation method of lipoic acid injection, the injection safety of this method preparation
Good, stability is high, and the related content of material of lipoic acid injection can be effectively reduced.
6. the present invention also provides a kind of method of lipoic acid injection quality evaluation, by being taken to lipoic acid ethylenediamine is double
For the detection of object content, the product that related content of material is higher than standard is determined as rejected product, to avoid the higher production of toxicity
Product are used by the patient, to improve the safety in utilization of lipoic acid injection.
7. the present invention also provides a kind of methods for improving lipoic acid injection safety in utilization, by lipoic acid second two
The detection of amine disubstitution product content, to judge that this batch of product can continue the time saved, to reduce wind when patient medication
Danger.
Detailed description of the invention:
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with
It obtains other drawings based on these drawings.
Fig. 1 is N, the mass spectrogram of N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide)
Fig. 2 is the liquid chromatogram of lipoic acid preparation
Specific embodiment
Below by embodiment, the present invention is further illustrated.It should be understood that the embodiment of the present invention is only to use
In illustrating the present invention, rather than limiting the invention, under concept thereof of the invention, simple modifications of the invention are belonged to
In the scope of protection of present invention.
Embodiment 1
The preparation of N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide)
CDI (9.0g, 55.7mmol) is dissolved in methylene chloride (150mL), is added lipoic acid (11.5g, 55.7mmol),
Transfer funnel etc. is rinsed with q. s. methylene chloride.After appropriate stirring, dropwise addition ethylenediamine/dichloromethane solution (2.2g ethylenediamine,
36.6mmol；100mL methylene chloride), it is stirred overnight at room temperature.Reaction solution is after pickling, alkali cleaning, washing, organic layer anhydrous slufuric acid
Sodium is dry, concentration, obtains N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) 7.6g, yield 62.4%,
For light yellow solid.MS m/z (ESI): 459.1 (M+23), 435.1 (M-1).
Embodiment 2
The preparation of N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide)
Reaction is added in lipoic acid (2.5g, 12.1mmol), triethylamine (2.45g, 24.2mmol), tetrahydrofuran (20mL)
In bottle, open stirring, be heated to 30-40 DEG C, be added dropwise methylchloroformate/tetrahydrofuran solution (1.71g methylchloroformate,
18.1mmol；2mL tetrahydrofuran), ethylenediamine/tetrahydrofuran solution (0.89g ethylenediamine, 14.8mmol are added dropwise after appropriate reaction；
2mL tetrahydrofuran), it reacts 5 hours.100mL methylene chloride is added into system, is washed with 30mL saturated sodium bicarbonate aqueous solution
2 times, organic layer concentration obtains N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) 1.53g, yield
57.8%, it is faint yellow solid.The mass spectrogram of product is as shown in Figure 1.MS m/z (ESI): 459.1 (M+23), 435.1 (M-1).
Embodiment 3
Prepare lipoic acid: (1): the vulcanized sodium for weighing 20g is dissolved in 80g purified water, is dissolved at 35 DEG C, is configured to vulcanized sodium
Solution；(2): 6,8-dichloro-octanoic acid ethyl ester 20g, sulphur 5g, tetrabutylammonium bromide 2g are added in 100g water, is warming up to 85 DEG C,
The sodium sulfide solution that step (1) obtains is slowly added into this aqueous solution.After sodium sulfide solution is added, the reaction was continued 3 hours；
After reaction, 60 DEG C are cooled to, static about 60 minutes, is layered to get lipoic acid ethyl ester is arrived；(3): sulphur being added in aqueous solvent
Ethyl caprilate adds sodium hydroxide 20g and tetrabutylammonium bromide 1g, 200r/min stirring, is warming up to 60 DEG C, reaction terminates
Afterwards, it filters while hot；(4): the filtrate that step (3) obtains being cooled to 5 DEG C, then with salt acid for adjusting pH to 1, filtering is to obtain sulphur
Sad crude product；(5): by the lipoic acid crude product that step (4) obtains be added to mixed solvent (hexamethylene: ethyl acetate=5:1 (w:
W) in), under 100r/min stirring, 40 DEG C is heated to, 30 minutes is kept the temperature, is completely dissolved lipoic acid crude product, is then added
36 DEG C of purified water is kept stirring 10 minutes, static 20 minutes removing water layers, after organic layer is heated to 40 DEG C, is added
Silica gel after stirring 2h, filters.16 DEG C are subsequently cooled to, stirring and crystallizing 1 hour, filters and obtains the wet fine work of lipoic acid, then exist
It is 12 hours dry at a temperature of 35 DEG C.
Embodiment 4
Prepare lipoic acid: (1): the vulcanized sodium for weighing 20g is dissolved in 120g purified water, is dissolved at 45 DEG C, is configured to vulcanize
Sodium solution；(2): 6,8-dichloro-octanoic acid ethyl ester 80g, sulphur 10g, tetrabutylammonium bromide 5g being added in 200g water, is warming up to 86
DEG C, the sodium sulfide solution that step (1) obtains is slowly added into this aqueous solution.After sodium sulfide solution is added, it is 5 small that the reaction was continued
When；After reaction, 65 DEG C are cooled to, static about 60 minutes, is layered to get lipoic acid ethyl ester is arrived；(3): adding in aqueous solvent
Enter lipoic acid ethyl ester, add sodium hydroxide 25g and tetrabutylammonium bromide 3g, 400r/min stirring, is warming up to 80 DEG C, reaction knot
Shu Hou is filtered while hot；(4): the filtrate that step (3) obtains being cooled to 10 DEG C, is then filtered with salt acid for adjusting pH to 2 to obtain the final product
To lipoic acid crude product；(5): by the lipoic acid crude product that step (4) obtains be added to mixed solvent (hexamethylene: ethyl acetate=10:
1 (w:w)) in, under 500r/min stirring, 45 DEG C are heated to, 50 minutes is kept the temperature, is completely dissolved lipoic acid crude product, then
40 DEG C of purified water is added, is kept stirring 15 minutes, static 30 minutes removing water layers, after organic layer is heated to 50 DEG C,
Silica gel is added, after stirring 2h, filters.20 DEG C are subsequently cooled to, stirring and crystallizing 1 hour, filters and obtains the wet fine work of lipoic acid, so
It is 24 hours dry at a temperature of 40 DEG C afterwards.
Embodiment 5
Prepare lipoic acid: (1): the vulcanized sodium for weighing 20g is dissolved in 100g purified water, is dissolved at 40 DEG C, is configured to vulcanize
Sodium solution；(2): 6,8-dichloro-octanoic acid ethyl ester 50g, sulphur 8g, tetrabutylammonium bromide 3g being added in 150g water, is warming up to 85
DEG C, the sodium sulfide solution that step (1) obtains is slowly added into this aqueous solution.After sodium sulfide solution is added, it is 4 small that the reaction was continued
When；After reaction, 62 DEG C are cooled to, static about 60 minutes, is layered to get lipoic acid ethyl ester is arrived；(3): adding in aqueous solvent
Enter lipoic acid ethyl ester, add sodium hydroxide 22g and tetrabutylammonium bromide 2g, 300r/min stirring, is warming up to 70 DEG C, reaction knot
Shu Hou is filtered while hot；(4): the filtrate that step (3) obtains being cooled to 6 DEG C, is then filtered with salt acid for adjusting pH to 1.5 to obtain the final product
To lipoic acid crude product；(5): the lipoic acid crude product that step (4) obtains is added to mixed solvent (hexamethylene: ethyl acetate=6:1
(w:w)) in, 42 DEG C are heated under 300r/min stirring, 40 minutes is kept the temperature, is completely dissolved lipoic acid crude product, then plus
The purified water for entering 37 DEG C is kept stirring 12 minutes, and removing water layer adds after organic layer is heated to 45 DEG C within static 25 minutes
Enter silica gel, after stirring 2h, filters.18 DEG C are subsequently cooled to, stirring and crystallizing 1 hour, filters and obtains the wet fine work of lipoic acid, then
It is 16 hours dry at a temperature of 37 DEG C.
Embodiment 6
Lipoic acid injection is prepared in the present inventive method.
(1) commercially available lipoic acid 20g, two (1,2- dithiolane -3- of ethylenediamine 5g, N, N'- (ethane -1,2- diyl) are weighed
Pentanamide) 2g；(2) pre-process: water for injection deaeration by nitrogen stripping, detection water oxygen level are qualified (water oxygen level < 20ug/ml)；
Measure the water for injection for matching liquid total amount 3/4, that is, 750ml；(3) it weighs: lipoic acid is weighed by recipe quantity 102%, by prescription
Amount weighs ethylenediamine, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) and sufficient active charcoal；(4) dilute
With method: ethylenediamine is added in prescription ratio, stirs evenly；(5) add raw material: slowly adding in the case where continuing stirring (100r/min) state
Enter recipe quantity lipoic acid, stirs evenly；(6) add active carbon: adding water to 1000ml, active carbon is added, stir 20 minutes；(7)
Decarburization: through 4 μm of stud coarse filtration；(8) preliminary survey: measurement pH value, solution ph should be 8.0-8.3；(9) refined filtration: through 0.45 μm of filter membrane
Core by-pass filtration；(10) end-filtration: through 0.22 μm of filter membrane core second level aseptic filtration to encapsulating step, preparation starts to filling to open
Beginning when be limited in 3 hours；(11) encapsulating: detection intermediate products, content 97.0-103.0%, pH value 8.0-8.3 are qualified produce
Product, will test that qualified intermediate products medical fluid is filling, encapsulating start to terminate to encapsulating when be limited in 12 hours；(12) nitrogen charging,
Sealing, sterilizing: 121 DEG C of moist heat sterilization 15min.Up to lipoic acid injection.
Embodiment 7
Lipoic acid injection is prepared in the present inventive method.
(1) commercially available lipoic acid 50g, two (1,2- dithiolane -3- of ethylenediamine 8g, N, N'- (ethane -1,2- diyl) are weighed
Pentanamide) 0.1g；(2) it pre-processes: water for injection deaeration by nitrogen stripping, the qualified (water oxygen level < 50ug/ of detection water oxygen level
ml)；Measure the water for injection for matching liquid total amount 4/5, that is, 800ml；(3) it weighs: weighing lipoic acid by recipe quantity 105%, press
Recipe quantity weighs ethylenediamine, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) and sufficient active charcoal；
(4) dilute to match method: ethylenediamine is added in prescription ratio, stirs evenly；(5) add raw material: continuing to stir under (500r/min) state
It is slowly added to recipe quantity lipoic acid, is stirred evenly；(6) add active carbon: adding water to 1000ml, active carbon is added, stir 40 points
Clock；(7) decarburization: through 4 μm of stud coarse filtration；(8) preliminary survey: measurement pH value, solution ph should be 8.0-8.3；(9) refined filtration: through 0.45 μ
M filter membrane core by-pass filtration；(10) end-filtration: through 0.22 μm of filter membrane core second level aseptic filtration to encapsulating step, preparation start to
Filling beginning when be limited in 4.5 hours；(11) encapsulating: detection intermediate products, content 97.0-103.0%, pH value 8.0-8.3
For qualified products, will test that qualified intermediate products medical fluid is filling, encapsulating start to terminate to encapsulating when be limited in 12 hours；
(12) nitrogen charging, sealing, sterilizing: 121 DEG C of moist heat sterilization 30min.Up to lipoic acid injection.
Embodiment 8
Lipoic acid injection is prepared in the present inventive method.
(1) commercially available lipoic acid 25g, two (1,2- dithiolane -3- of ethylenediamine 6g, N, N'- (ethane -1,2- diyl) are weighed
Pentanamide) 0.01g；(2) it pre-processes: water for injection deaeration by nitrogen stripping, the qualified (water oxygen level < 20ug/ of detection water oxygen level
ml)；Measure the water for injection for matching liquid total amount 3/4, that is, 750ml；(3) it weighs: weighing lipoic acid by recipe quantity 102%, press
Recipe quantity weighs ethylenediamine, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) and sufficient active charcoal；
(4) dilute to match method: ethylenediamine is added in prescription ratio, stirs evenly；(5) add raw material: continuing to stir under (300r/min) state
It is slowly added to recipe quantity lipoic acid, is stirred evenly；(6) add active carbon: adding water to 1000ml, active carbon is added, stir 30 points
Clock；(7) decarburization: through 4 μm of stud coarse filtration；(8) preliminary survey: measurement pH value, solution ph should be 8.0-8.3；(9) refined filtration: through 0.45 μ
M filter membrane core by-pass filtration；(10) end-filtration: through 0.22 μm of filter membrane core second level aseptic filtration to encapsulating step, preparation start to
Filling beginning when be limited in 4 hours；(11) encapsulating: detection intermediate products, content 97.0-103.0%, pH value 8.0-8.3 are
Qualified products, will test that qualified intermediate products medical fluid is filling, encapsulating start to terminate to encapsulating when be limited in 12 hours；(12)
Nitrogen charging, sealing, sterilizing: 121 DEG C of moist heat sterilization 30min.Up to lipoic acid injection.
Embodiment 9
Lipoic acid injection is prepared in the present inventive method.
(1) the lipoic acid 25g prepared in embodiment 3, ethylenediamine 6g, N, N'- (ethane -1,2- diyl) two (1,2- are weighed
Dithiolane -3- pentanamide) 0.01g；(2) pre-process: water for injection deaeration by nitrogen stripping, detection water oxygen level is qualified (to be contained in water
Oxygen amount < 20ug/ml)；Measure the water for injection for matching liquid total amount 3/4, that is, 750ml；(3) it weighs: being weighed by recipe quantity 102%
Lipoic acid weighs ethylenediamine, N by recipe quantity, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) and enough
Active carbon；(4) dilute to match method: ethylenediamine is added in prescription ratio, stirs evenly；(5) add raw material: continuing to stir (300r/min)
It is slowly added to recipe quantity lipoic acid under state, stirs evenly；(6) add active carbon: adding water to 1000ml, active carbon is added, stirs
It mixes 30 minutes；(7) decarburization: through 4 μm of stud coarse filtration；(8) preliminary survey: measurement pH value, solution ph should be 8.0-8.3；(9) refined filtration:
Through 0.45 μm of filter membrane core by-pass filtration；(10) it end-filtration: through 0.22 μm of filter membrane core second level aseptic filtration to encapsulating step, prepares
Start to filling beginning when be limited in 4 hours；(11) encapsulating: detection intermediate products, content 97.0-103.0%, pH value 8.0-
8.3 be qualified products, will test that qualified intermediate products medical fluid is filling, encapsulating start to terminate to encapsulating when be limited to 12 hours
It is interior；(12) nitrogen charging, sealing, sterilizing: 121 DEG C of moist heat sterilization 30min.Up to lipoic acid injection.
Embodiment 10
Lipoic acid injection is prepared in the present inventive method.
(1) the lipoic acid 25g prepared in embodiment 3, ethylenediamine 6g are weighed；(2) it pre-processes: water for injection deaeration by nitrogen stripping,
It is qualified (water oxygen level < 20ug/ml) to detect water oxygen level；Measure the water for injection for matching liquid total amount 3/4, that is, 750ml；
(3) it weighs: weighing lipoic acid by recipe quantity 102%, weigh ethylenediamine and sufficient active charcoal by recipe quantity；(4) dilute to match method: by place
Ethylenediamine is added in square ratio, stirs evenly；(5) add raw material: being slowly added to recipe quantity in the case where continuing stirring (300r/min) state
Lipoic acid stirs evenly；(6) add active carbon: adding water to 1000ml, active carbon is added, stir 30 minutes；(7) decarburization: through 4 μ
M stud coarse filtration；(8) preliminary survey: measurement pH value, solution ph should be 8.0-8.3；(9) refined filtration: through 0.45 μm of filter membrane core level-one mistake
Filter；(10) end-filtration: through 0.22 μm of filter membrane core second level aseptic filtration to encapsulating step, preparation starts the time limit to filling beginning
For in 4 hours；(11) encapsulating: detection intermediate products, content 97.0-103.0%, pH value 8.0-8.3 are qualified products, be will test
Qualified intermediate products medical fluid is filling, encapsulating start to terminate to encapsulating when be limited in 12 hours；(12) nitrogen charging, sealing, sterilizing:
121 DEG C of moist heat sterilization 30min.Up to lipoic acid injection.
Embodiment 11
Alpha-lipoic acid liposome injection is prepared in the present inventive method.
Weigh commercially available lipoic acid 120g, Distearate Phosphatidylcholine 30g, octadecylamine 50g, PLURONICS F87 25g, Chinese holly
Rafter acid sodium 1g, mercaptoethanol 3g, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) 2g.
(1) lipoic acid of recipe quantity, octadecylamine, Distearate Phosphatidylcholine, PLURONICS F87, N, N'- (ethane -1,
2- diyl) two (1,2- dithiolane -3- pentanamides) be dissolved in 1000ml volume ratio be 3: 1 ethyl alcohol and acetone mixed solvent,
It is uniformly mixed, in organic solvent is removed under reduced pressure on rotary film evaporator, obtains lipid membrane；
(2) under nitrogen protection, the sodium citrate buffer solution that 800mlpH is 5 is added into bottle, shaking stirs 20 points
Clock, revolving speed 400r/min make the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 5 minutes, revolving speed 9000r/
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in min；
(3) sodium citrate and mercaptoethanol is added, stirring is settled to 1000ml, with 0.45 μm of filtering with microporous membrane, fills
Dress is to get alpha-lipoic acid liposome injection.
Embodiment 12
Alpha-lipoic acid liposome injection is prepared in the present inventive method.
Weigh commercially available lipoic acid 120g, Distearate Phosphatidylcholine 120g, octadecylamine 80g, PLURONICS F87 40g, Chinese holly
Rafter acid sodium 5g, mercaptoethanol 12g, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) 0.1g.
(1) lipoic acid of recipe quantity, octadecylamine, Distearate Phosphatidylcholine, PLURONICS F87, N, N'- (ethane -1,
2- diyl) two (1,2- dithiolane -3- pentanamides) be dissolved in 1000ml volume ratio be 5: 1 ethyl alcohol and acetone mixed solvent,
It is uniformly mixed, in organic solvent is removed under reduced pressure on rotary film evaporator, obtains lipid membrane；
(2) under nitrogen protection, the sodium citrate buffer solution that 800mlpH is 7 is added into bottle, shaking stirs 50 points
Clock, revolving speed 700r/min make the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 10 minutes, revolving speed 12000r/
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in min；
(3) sodium citrate and mercaptoethanol is added, stirring is settled to 1000ml, with 0.45 μm of filtering with microporous membrane, fills
Dress is to get alpha-lipoic acid liposome injection.
Embodiment 13
Alpha-lipoic acid liposome injection is prepared in the present inventive method.
Weigh commercially available lipoic acid 120g, Distearate Phosphatidylcholine 50g, octadecylamine 60g, PLURONICS F87 30g, Chinese holly
Rafter acid sodium 3g, mercaptoethanol 6g, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) 0.01g.
(1) lipoic acid of recipe quantity, octadecylamine, Distearate Phosphatidylcholine, PLURONICS F87, N, N'- (ethane -1,
2- diyl) two (1,2- dithiolane -3- pentanamides) be dissolved in 1000ml volume ratio be 4: 1 ethyl alcohol and acetone mixed solvent,
It is uniformly mixed, in organic solvent is removed under reduced pressure on rotary film evaporator, obtains lipid membrane；
(2) under nitrogen protection, the sodium citrate buffer solution that 800mlpH is 6.6, shaking, stirring 40 are added into bottle
Minute, revolving speed 500r/min makes the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 8 minutes, revolving speed
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in 10000r/min；
(3) sodium citrate and mercaptoethanol is added, stirring is settled to 1000ml, with 0.45 μm of filtering with microporous membrane, fills
Dress is to get alpha-lipoic acid liposome injection.
Embodiment 14
Alpha-lipoic acid liposome injection is prepared in the present inventive method.
Weigh the lipoic acid 120g, Distearate Phosphatidylcholine 50g, octadecylamine 60g of the preparation of embodiment 3, poloxamer
18830g, sodium citrate 3g, mercaptoethanol 6g, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide)
0.01g。
(1) lipoic acid of recipe quantity, octadecylamine, Distearate Phosphatidylcholine, PLURONICS F87, N, N'- (ethane -1,
2- diyl) two (1,2- dithiolane -3- pentanamides) be dissolved in 1000ml volume ratio be 4: 1 ethyl alcohol and acetone mixed solvent,
It is uniformly mixed, in organic solvent is removed under reduced pressure on rotary film evaporator, obtains lipid membrane；
(2) under nitrogen protection, the sodium citrate buffer solution that 800mlpH is 6.6, shaking, stirring 40 are added into bottle
Minute, revolving speed 500r/min makes the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 8 minutes, revolving speed
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in 10000r/min；
(3) sodium citrate and mercaptoethanol is added, stirring is settled to 1000ml, with 0.45 μm of filtering with microporous membrane, fills
Dress is to get alpha-lipoic acid liposome injection.
Embodiment 15
Alpha-lipoic acid liposome injection is prepared in the present inventive method.
Weigh the lipoic acid 120g, Distearate Phosphatidylcholine 50g, octadecylamine 60g of the preparation of embodiment 3, poloxamer
18830g, sodium citrate 3g, mercaptoethanol 6g.
(1) lipoic acid of recipe quantity, octadecylamine, Distearate Phosphatidylcholine, PLURONICS F87 are dissolved in 1000ml volume
Than the mixed solvent of ethyl alcohol and acetone for 4: 1, it is uniformly mixed, in organic solvent is removed under reduced pressure on rotary film evaporator, obtains
To lipid membrane；
(2) under nitrogen protection, the sodium citrate buffer solution that 800mlpH is 6.6, shaking, stirring 40 are added into bottle
Minute, revolving speed 500r/min makes the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 8 minutes, revolving speed
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in 10000r/min；
(3) sodium citrate and mercaptoethanol is added, stirring is settled to 1000ml, with 0.45 μm of filtering with microporous membrane, fills
Dress is to get alpha-lipoic acid liposome injection.
Comparative example 1
Lipoic acid injection is prepared according to the method for CN103655469A, the specific steps are as follows:
Lipoic acid 15g, ethylenediamine 3.87g are weighed, water for injection adds to 600ml.
Preparation process: water for injection deaeration by nitrogen stripping, detection water oxygen level are qualified (water oxygen level < 50ug/ml).It measures
Water for injection with liquid total amount 3/4.Ethylenediamine is added in prescription ratio, stirs evenly.It is slowly added in the case where continuing stirring
Recipe quantity lipoic acid, stirs evenly.It adds water to full dose, active carbon is added, stir 30 minutes.Through 3 μm of stud filters (5 of Φ
Core) coarse filtration decarburization.Preliminary survey pH value is 8.15.Filter membrane core by-pass filtration, 0.22 μm of polyether sulfone folding are folded through 0.45 μm of polyether sulfone again
Folded filter membrane core level-one aseptic filtration.The aseptic filtration of filter membrane core second level is folded to encapsulating using 0.22 μm of polyether sulfone, and 121 DEG C damp and hot
Sterilize 15min to get.
Comparative example 2
Lipoic acid injection is prepared according to the method for CN102525930A, the specific steps are as follows:
Weigh lipoic acid 240g, Distearate Phosphatidylcholine 300g, octadecylamine 200g, PLURONICS F87 150g, chlorination
Sodium 18g, mercaptoethanol 10g.
(1) 240g lipoic acid, 200g octadecylamine, 300g Distearate Phosphatidylcholine and 150g PLURONICS F87 is molten
In the mixed solvent of ethyl alcohol and acetone that 1500ml volume ratio is 4: 1, it is uniformly mixed, in being removed under reduced pressure on rotary film evaporator
Organic solvent obtains lipid membrane；
(2) under nitrogen protection, the citrate buffer solution that 1500mlpH is 6.6, shaking, stirring 30 are added into bottle
Minute, revolving speed 500r/min makes the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 12 minutes, revolving speed
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in 8000r/min；
(3) 18g sodium chloride and 10g mercaptoethanol is added, stirring is settled to 2000ml, with 0.45 μm of filtering with microporous membrane,
It is filling to get 100 alpha-lipoic acid liposome injections.
Comparative example 3
Lipoic acid injection is prepared according to the method for CN103655469A, the specific steps are as follows:
Weigh lipoic acid 15g, (1, the 2- dithiolane -3- valeryl of ethylenediamine 3.87g, N, N'- (ethane -1,2- diyl) two
Amine) 4g, water for injection adds to 600ml.
Preparation process: water for injection deaeration by nitrogen stripping, detection water oxygen level are qualified (water oxygen level < 50ug/ml).It measures
Water for injection with liquid total amount 3/4.Ethylenediamine is added in prescription ratio, stirs evenly.It is slowly added in the case where continuing stirring
Recipe quantity lipoic acid and N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide), stir evenly.Add water to
Active carbon is added in full dose, stirs 30 minutes.Through 3 μm of stud filters of Φ (5 core) coarse filtration decarburization.Preliminary survey pH value is 8.15.It passes through again
0.45 μm of polyether sulfone folds filter membrane core by-pass filtration, 0.22 μm of polyether sulfone folds the level-one aseptic filtration of filter membrane core.Using 0.22 μ
M polyether sulfone folds the aseptic filtration of filter membrane core second level to encapsulating, 121 DEG C of moist heat sterilization 15min to get.
Comparative example 4
Lipoic acid injection is prepared according to the method for CN102525930A, the specific steps are as follows:
Weigh lipoic acid 240g, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) 8g, distearyl
Sour phosphatidyl choline 300g, octadecylamine 200g, PLURONICS F87 150g, sodium chloride 18g, mercaptoethanol 10g.
(1) by 240g lipoic acid, 20g N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide),
200g octadecylamine, 300g Distearate Phosphatidylcholine and 150g PLURONICS F87 are dissolved in the ethyl alcohol that 1500ml volume ratio is 4: 1
With the mixed solvent of acetone, it is uniformly mixed, in organic solvent is removed under reduced pressure on rotary film evaporator, obtains lipid membrane；
(2) under nitrogen protection, the citrate buffer solution that 1500mlpH is 6.6, shaking, stirring 30 are added into bottle
Minute, revolving speed 500r/min makes the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 12 minutes, revolving speed
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in 8000r/min；
(3) 18g sodium chloride and 10g mercaptoethanol is added, stirring is settled to 2000ml, with 0.45 μm of filtering with microporous membrane,
It is filling to get 100 alpha-lipoic acid liposome injections.
Comparative example 5
Lipoic acid injection is prepared using the method for the present invention.
(1) the lipoic acid 25g prepared in embodiment 3, ethylenediamine 6g, N, N'- (ethane -1,2- diyl) two (1,2- are weighed
Dithiolane -3- pentanamide) 4g；(2) it pre-processes: water for injection deaeration by nitrogen stripping, the qualified (water oxygen level of detection water oxygen level
<20ug/ml)；Measure the water for injection for matching liquid total amount 3/4, that is, 750ml；(3) it weighs: it is pungent to weigh sulphur by recipe quantity 102%
Acid weighs ethylenediamine, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) and sufficient active by recipe quantity
Charcoal；(4) dilute to match method: ethylenediamine is added in prescription ratio, stirs evenly；(5) add raw material: continuing to stir (300r/min) state
Under be slowly added to recipe quantity lipoic acid, stir evenly；(6) add active carbon: adding water to 1000ml, active carbon, stirring 30 is added
Minute；(7) decarburization: through 4 μm of stud coarse filtration；(8) preliminary survey: measurement pH value, solution ph should be 8.0-8.3；(9) refined filtration: warp
0.45 μm of filter membrane core by-pass filtration；(10) end-filtration: through 0.22 μm of filter membrane core second level aseptic filtration to encapsulating step, preparation is opened
Begin to filling beginning when be limited in 4 hours；(11) encapsulating: detection intermediate products, content 97.0-103.0%, pH value 8.0-
8.3 be qualified products, will test that qualified intermediate products medical fluid is filling, encapsulating start to terminate to encapsulating when be limited to 12 hours
It is interior；(12) nitrogen charging, sealing, sterilizing: 121 DEG C of moist heat sterilization 30min.Up to lipoic acid injection.
Comparative example 6
Alpha-lipoic acid liposome injection is prepared using the method for the present invention.
Weigh the lipoic acid 120g, Distearate Phosphatidylcholine 50g, octadecylamine 60g of the preparation of embodiment 3, poloxamer
18830g, sodium citrate 3g, mercaptoethanol 6g, N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) 4g.
(1) lipoic acid of recipe quantity, octadecylamine, Distearate Phosphatidylcholine, PLURONICS F87, N, N'- (ethane -1,
2- diyl) two (1,2- dithiolane -3- pentanamides) be dissolved in 1000ml volume ratio be 4: 1 ethyl alcohol and acetone mixed solvent,
It is uniformly mixed, in organic solvent is removed under reduced pressure on rotary film evaporator, obtains lipid membrane；
(2) under nitrogen protection, the sodium citrate buffer solution that 800mlpH is 6.6, shaking, stirring 40 are added into bottle
Minute, revolving speed 500r/min makes the complete aquation of immobilized artificial membrane, with the homogeneous emulsification of tissue mashing machine's high speed 8 minutes, revolving speed
Liposome turbid liquor is made with 0.45 μm of filtering with microporous membrane in 10000r/min；
(3) sodium citrate and mercaptoethanol is added, stirring is settled to 1000ml, with 0.45 μm of filtering with microporous membrane, fills
Dress is to get alpha-lipoic acid liposome injection.
Embodiment 16
Liquid chromatographic detection is carried out to the lipoic acid preparation of embodiment 8, embodiment 13, and carries out system suitability test.
Method particularly includes: Kromasil 100-5C18 (4.6 × 250mm, 5 μm) chromatographic column is selected, mobile phase is by mobile phase
A liquid is mixed with Mobile phase B, mobile phase A are as follows: 0.002mol/L potassium dihydrogen phosphate is 2 with phosphorus acid for adjusting pH value；Stream
Dynamic phase B are as follows: acetonitrile-methanol (5:50)；Gradient elution is carried out by table 3；Flow velocity is 0.5ml/min；Column temperature is 20 DEG C；Detection wavelength
For 200nm.
The results are shown in Table 4 for system suitability test.
3 gradient elution of table flows phase concentration variation
The system suitability of 4 this method of table
Embodiment 17
Liquid chromatographic detection is carried out to the lipoic acid preparation of embodiment 8, embodiment 13, and carries out system suitability test.
Method particularly includes: Kromasil 100-5C18 (4.6 × 250mm, 5 μm) chromatographic column is selected, mobile phase is by mobile phase
A liquid is mixed with Mobile phase B, mobile phase A are as follows: 0.008mol/L potassium dihydrogen phosphate is 2.5 with phosphorus acid for adjusting pH value；
Mobile phase B are as follows: acetonitrile-methanol (10:30)；Gradient elution is carried out by table 5；Flow velocity is 1.5ml/min；Column temperature is 40 DEG C；Detection
Wavelength is 250nm.
The results are shown in Table 6 for system suitability test.
5 gradient elution of table flows phase concentration variation
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 60 | 40 |
| 15 | 60 | 40 |
| 30 | 30 | 70 |
| 110 | 30 | 70 |
| 111 | 60 | 40 |
| 120 | 60 | 40 |
The system suitability of 6 this method of table
Embodiment 18
Liquid chromatographic detection is carried out to the lipoic acid preparation of embodiment 8, embodiment 13, and carries out system suitability test.
Method particularly includes: Kromasil 100-5C18 (4.6 × 250mm, 5 μm) chromatographic column is selected, mobile phase is by mobile phase
A liquid is mixed with Mobile phase B, mobile phase A are as follows: 0.005mol/L potassium dihydrogen phosphate is 2.3 with phosphorus acid for adjusting pH value；
Mobile phase B are as follows: acetonitrile-methanol (8:42)；Gradient elution is carried out by table 7；Flow velocity is 1.0ml/min；Column temperature is 35 DEG C；Detect wave
A length of 215nm.
Chromatogram to the lipoic acid preparation of embodiment 8 is as shown in Fig. 2, the peak of retention time 40.681 is N, N'- (second
Alkane -1,2- diyl) two (1,2- dithiolane -3- pentanamides), the peak of retention time 31.044 is lipoic acid.
The results are shown in Table 8 for system suitability test.According to 8 data of table it is found that the method for the present embodiment is preferably to examine
Survey method.
7 gradient elution of table flows phase concentration variation
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 60 | 40 |
| 15 | 60 | 40 |
| 30 | 30 | 70 |
| 110 | 30 | 70 |
| 111 | 60 | 40 |
| 120 | 60 | 40 |
The system suitability of 8 this method of table
Embodiment 19
Using the method for embodiment 18 to comparative example 1-6, lipoic acid and N in the preparation of embodiment 6-15, N'- (ethane-
1,2- diyl) contents of two (1,2- dithiolane -3- pentanamides) detected.The results are shown in Table 9.According to embodiment 8, in fact
The testing result for applying example 13 can prepare N, N'- (ethane -1,2- diyl) two (1,2- bis- using preparation method of the invention
Thiophane -3- pentanamide) content is less than 0.02g/L, the lipoic acid injection of even less than 0.01g/L, that is, N, N'- (second
Alkane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) contents are less than 0.002% (w/v), even less than 0.001% (w/
V), this is unable to reach in the prior art.
9 comparative example 1-6 of table, embodiment 6-15 preparation in lipoic acid and N, N'- (ethane -1,2- diyl) two (1,2-
Dithiolane -3- pentanamide) content
Embodiment 20
Long term toxicity test is carried out to lipoic acid preparation prepared by comparative example 1-6 and embodiment 6-15.(the agent of people and rat
6.25) measuring conversion ratio is
Healthy SD rat is taken, weight (125 ± 10) g, every group 50, half male and half female, it is daily that experimental group presses amount of formulation
The dosage of 0.5ml/kg is injected intravenously administration, and blank control group presses same amount of normal saline drug administration by injection.Observation one week before dispensing, respectively
Situations such as group animal activity, feed, excrement, is without exception, then starts to be administered.
It is administered once, continuous 60 days, weighs weekly primary on time daily, adjust dosage according to changes of weight.Carry out appearance
The overviews such as sign, behavioral activity, fecal character, appetite, changes of weight.
After to medication, plucks eyeball and blood is taken to carry out blood picture, blood bio-chemistry checking, five coring, liver, spleen, lung, kidney internal organs claim
Weight, calculates the organ weight/power ratio of every 100g weight.
Experimental result is as shown in table 10.
The lipoic acid preparation of table 10 comparative example 1-6 and embodiment 6-15 preparation carries out long term toxicity test result
Occurs the not normal rat of nervous system according to the experimental result of table 10, in comparative example 3-6, it is known that in lipoic acid
In preparation, the adverse reaction of rat occurs for (1, the 2- dithiolane -3- pentanamide) content of N, N'- (ethane -1,2- diyl) two
Rate has a certain impact, and the N of high-content, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) cause bad
The rising of reactivity.
Embodiment 21
The finished product that comparative example 1-6 and embodiment 6-15 is obtained, is placed in 20 DEG C and is kept in dark place 36 months.
Lipoic acid and N in above-mentioned lipoic acid preparation, two (1,2- dithiolane-of N'- (ethane -1,2- diyl) are detected respectively
3- pentanamide) content.
Detection method is as described in Example 18.
Testing result is as shown in table 11.
After 1120 DEG C of table are kept in dark place 36 months, comparative example 1-6, embodiment 6-15 preparation in lipoic acid and N, N'- (second
Alkane -1,2- diyl) two (1,2- dithiolane -3- pentanamides) content
As shown in Table 11, lipoic acid preparation (the embodiment 6- prepared using heretofore described preparation method
15), compared to the preparation method with comparative example 1, comparative example 2, two (1,2- bis- sulphur penta of the N in storage, N'- (ethane -1,2- diyl)
Ring -3- pentanamide) yield it is smaller (p < 0.01), the content of lipoic acid is influenced also smaller (p < 0.01).
The lipoic acid preparation prepared using conventional method is generating N, two (1,2- bis- sulphur penta of N'- (ethane -1,2- diyl)
Ring -3- pentanamide) while, the content of lipoic acid also necessarily reduces, and affects the curative effect of drug.
Wherein, the lipoic acid prepared using the method for the present invention (embodiment 3), after being prepared into corresponding preparations (embodiment 9,10,
Embodiment 14,15), the yield phase of the N in storage, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide)
For the preparation (embodiment 6-8, embodiment 11-13) of commercially available lipoic acid preparation, (p < 0.01) is substantially reduced.
Above-mentioned number is it was demonstrated that either using lipoic acid preparation method provided in the present invention or lipoic acid preparation
Preparation method can effectively reduce N during lipoic acid preparation storage, two (1,2- bis- sulphur of N'- (ethane -1,2- diyl)
Penta ring -3- pentanamide) yield, to guarantee the stabilization of lipoic acid drug content.
Embodiment 22
The finished product that comparative example 1-6 and embodiment 6-15 is obtained, is placed in 20 DEG C and is kept in dark place 36 months.To above-mentioned preparation by real
The method for applying example 20 carries out long term toxicity test.Experimental result is as shown in table 12.
After 1220 DEG C of table are kept in dark place 36 months, the lipoic acid preparation of comparative example 1-6 and embodiment 6-15 preparation carries out long-term
Toxicity test result
According to the experimental result of table 12 it is found that in lipoic acid preparation, two (1,2- bis- sulphur of N, N'- (ethane -1,2- diyl)
Penta ring -3- pentanamide) content have a significant impact to the adverse reaction rate of rat, the N of high-content, N'- (ethane -1,
2- diyl) two (1,2- dithiolane -3- pentanamides) cause adverse reaction rate to significantly rise.
By the experimental result of table 12 it is found that either comparative example 1-6 or embodiment 6,11, work as N, N'- (ethane -1,2-
Diyl) when being more than 2g/L (0.2%), quantity meeting occurs for rat bad reaction for the contents of two (1,2- dithiolane -3- pentanamides)
Be significantly increased (p < 0.01), and content is higher, and adverse reaction rate is higher；According to embodiment 7-10,12-13, work as N,
The content of N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) is more than that 0.1g/L (0.01%) is less than 2g/L
(0.2%) when, minimal amount of adverse reaction can occur for rat, considerably less than the embodiment (p < 0.01) that content is more than 2g/L；And
In embodiment 14,15, the content of N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) are less than 0.1g/L
(0.01%), then there is not adverse reaction.Further demonstrate N, N'- (ethane -1,2- diyl) in control lipoic acid preparation
The importance of two (1,2- dithiolane -3- pentanamide) contents, and demonstrate the stabilization of lipoic acid preparation prepared by the present invention
Property and safety.
Moreover, by 11 data of table it is found that under conditions of being kept in dark place for 20 DEG C, in lipoic acid preparation, N, N'- (ethane -1,
2- diyl) incrementss of two (1,2- dithiolane -3- pentanamides) are about 2.3g/L every 36 months, that is, 0.0639g/
The L/ month, that is, 0.00639%/moon, if (1, the 2- dithiolane -3- valeryl of a certain moment N, N'- (ethane -1,2- diyl) two
Amine) content be x%, then (1, the 2- dithiolane -3- valeryl of the N in certain lipoic acid preparations, N'- (ethane -1,2- diyl) two
Amine) content (0.2-x)/0.00639 month can reach 0.2% after sensing, significantly improve lipoic acid adverse reaction rate, therefore
It the use of lipoic acid preparation is after sensing safer usage mode in (0.2-x)/0.00639 month.
It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, may be used also
With several improvements and modifications are made to the present invention, these improvements and modifications also fall within the scope of protection of the claims of the present invention.
Claims (12)
1. a kind of lipoic acid preparation often uses auxiliary material, lipoic acid ethylenediamine disubstitution product (containing 1 structure of formula containing lipoic acid, preparation
Molecule), it is characterised in that: lipoic acid ethylenediamine disubstitution product content be not higher than 0.2% (w/v),
2. a kind of highly-safe lipoic acid injection, contains lipoic acid, co-solvent component, it is characterised in that: the lipoic acid
In injection, also contain lipoic acid ethylenediamine disubstitution product, wherein lipoic acid ethylenediamine disubstitution product is to contain 1 structure of formula
Molecule, content are not higher than 0.2% (w/v).
3. injection according to claim 2, it is characterised in that: lipoic acid ethylenediamine disubstitution product is the molecule of formula 1, i.e.,
N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide).
4. injection according to claim 2 or 3, it is characterised in that: lipoic acid ethylenediamine disubstitution product content is not higher than
0.01%.
5. injection according to claim 4, it is characterised in that: lipoic acid ethylenediamine disubstitution product content is not higher than
0.002%.
6. injection according to claim 2 or 3, it is characterised in that: lipoic acid and lipoic acid ethylenediamine disubstitution product
The ratio between content is not less than 25:1.
7. injection according to claim 6, it is characterised in that: the content of lipoic acid and lipoic acid ethylenediamine disubstitution product
The ratio between be not less than 150:1.
8. injection according to claim 2 or 3, it is characterised in that: in the lipoic acid injection, lipoic acid content
(w/v) it is not less than 1%, co-solvent component content (w/v) is not less than 0.1%.
9. the detection method of lipoic acid ethylenediamine disubstitution product in a kind of lipoic acid injection, the method is using liquid chromatogram
Method is detected, and stationary phase using octadecylsilane chemically bonded silica as filler, mixed by mobile phase A liquid with Mobile phase B by mobile phase
It closes, mobile phase A are as follows: 0.002-0.008mol/L phosphate solution is 2-2.5 with phosphorus acid for adjusting pH value；Mobile phase B are as follows:
Acetonitrile-methanol (5-10:30-50)；Gradient elution is carried out by table 1；Flow velocity is 0.5-1.5ml/min；Column temperature is 20-40 DEG C；Inspection
Survey wavelength is 200-250nm,
Table 1
10. a kind of quality evaluating method of lipoic acid injection, the method are, detect lipoic acid ethylenediamine pair in injection and take
Determine that it is unqualified when lipoic acid ethylenediamine disubstitution product content is more than 0.2% in the injection for the content of object
Product.
11. a kind of method for improving lipoic acid injection safety in utilization, the method are to detect lipoic acid second two in injection
The content of amine disubstitution product determines that it is not when lipoic acid ethylenediamine disubstitution product content is more than 0.2% in the injection
Qualified product, and abandon；When lipoic acid ethylenediamine disubstitution product content is no more than 0.2% in the injection, if lipoic acid
Ethylenediamine disubstitution product content is x%, then the lipoic acid injection should make in (0.2-x)/0.00639 month after sensing
With.
12.N, N'- (ethane -1,2- diyl) two (1,2- dithiolane -3- pentanamide) are as lipoic acid preparation defects inspecting
The application of standard items.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103655469A (en) * | 2013-12-19 | 2014-03-26 | 门毅 | Prescription and preparation technology of lipoic acid injection combination |
| CN105616343A (en) * | 2014-11-04 | 2016-06-01 | 蓬莱诺康药业有限公司 | Lipoic acid injection liquid and preparation method thereof |
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| CN103655469A (en) * | 2013-12-19 | 2014-03-26 | 门毅 | Prescription and preparation technology of lipoic acid injection combination |
| CN105616343A (en) * | 2014-11-04 | 2016-06-01 | 蓬莱诺康药业有限公司 | Lipoic acid injection liquid and preparation method thereof |
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