CN108452010A - A kind of production technology, pharmaceutical preparation, content determination and the clinical application of anti-ischemic angiocardiopathy and cerebrovascular disease medicine pellet phenol - Google Patents

Abstract

The invention discloses the production technologies of full water-soluble pellet phenol：Stir extraction 3 times (10,5,5min), merging extracting solution, refined filtration soon in 25 DEG C/80~85 DEG C water with 5 times of amounts, filtrate is adjusted to pH3.0 4.0 with dilute hydrochloric acid, stands 2~4h, refined filtration, filtrate caustic lye of soda tune pH is 5.0 6.0, No. 2 macroreticular resins of separation are crossed, water, purified water, injection water, low-concentration ethanol wash successively, Diluted Alcohol elution, eluent tune pH is 2.0 2.5, No. 3 macroreticular resins of chromatography are crossed, 50% ethanol elution after washing is spray-dried to obtain the white extremely faint yellow red phenol of class；Disclose injection pellet phenol, serial novel form；Content is calculated by external standard method with 6 reference substances, the sum of remaining peak area makees object of reference with tanshin polyphenolic acid B and calculates content, and sum of the two is red phenol content；Also disclose the effect of 15mg/kg pellet phenol prevention and treatment cerebral arterial thrombosis curative effect is substantially better than 20mg/kg Nimodipines.

Description

A kind of production technology, pharmaceutical preparation, the content of anti-ischemic angiocardiopathy and cerebrovascular disease medicine pellet phenol Measuring method and clinical application

Technical field

The present invention relates to pharmaceutical technology fields, and in particular to a kind of preparation of anti-cerebral ischemia Reperfusion injury vulnerary pellet phenol, medicine Compositions and its content assaying method and clinical application.

Background technology

In recent years, Cerebral Haemorrhage Invasion Rate increases year by year, is one of current three big fatal diseases, has high incidence, height The characteristics of illness rate, high mortality, high disability rate, high relapse rate.Cerebral apoplexy is divided into ischemic cerebral apoplexy and hemorrhagic apoplexy, Ischemic Stroke incidence is far above hemorrhagic apoplexy, accounts for 70% of cerebral apoplexy or more, ischemic tissue of brain is extensive under certain condition , there is more serious brain disorder and structural damage in multiple blood flow, referred to as cerebral ischemia re-pouring injured (Ischemia-Reperfusion Injury, IRI).The main path for the treatment of cerebral arterial thrombosis is that the dredging cerebrovascular restores Blood supply, and mitigate ischemical reperfusion injury and have become indispensable pith in central aspects.

Radix Salviae Miltiorrhizae is labiate Radix Salviae Miltiorrhizae (Salviae miltiorrhiza Bunge) dry root and rhizome.Bitter, property It is slightly cold, converge to heart and liver channels.It is inducing meastruation to relieve menalgia with promoting blood circulation, the effect of relieving restlessness that clears away heart-fire, cool blood to disappear carbuncle.For chest impediment and cardialgia, gastral cavity Abdomen hypochondriac pain, the accumulation of lump in the abdomen lump in the abdomen, hot numbness pain, dysphoria and insomnia, irregular menstruation, dysmenorrhoea Amenorrhea, sore swell and ache curative.There are prevention and/or treatment Cerebral ischemia, inhibits the effects that Cataractogenesis at myocardial ischemia.

Inventor has obtained a kind of " preparation of Salvia root P.E, pharmaceutical preparation and content assaying method " patent (patent Number：ZL200510037786.5), which has continued to use " boiling " in " water boiling and precipitation with ethanol " classical technique, has got rid of " alcohol It is heavy ".Therefore, the aspects such as the matter of this product, amount and production technology have very big difference, patent extraction process to save with red sage formulation, Purity is good, and the amount of obtaining is high, at low cost, and the dispatch of drug evaluation center requires to rename, and requires to formulate the bound of red phenol content, This product title is checked and approved through State Food and Drug Administration and is set to " red phenol ", is distinguished with existing red sage formulation with showing.But Long-time boiling in technique, not only consumes mass energy, but also also causes principal component degradation.In this regard, inventor is not through for many years Slack research and innovation, achieve breakthrough：Room temperature/80~85 DEG C ultra-short Time stirs extraction soon, for the first time 10min, and the 2nd, 3 Secondary each extraction 5min, principal component degradation reduce 10~15%, and yield improves 10% or more, optimize extraction water consumption, extraction effect Rate is increased to 2.4 times；Resin model is optimized, purity is made to improve 20% or more；Change refined filtration mode, filter efficiency improves 40 Times；It is that dilute refined solution room temperature crosses column enrichment to change decompression thermal concentration, further saves the energy, and extracting and developing purifying concentrates full mistake Journey does not heat and/or seldom, according to incompletely statistics, produces Danshen root injection per year by the whole nation and calculates, can save standard coal 58.368 Ten thousand tons, 63.037 hundred million yuan of RMB is saved, reduces 152.92416 ten thousand tons of carbon dioxide of discharge, 4962.816 tons of sulfur dioxide, nitrogen 4319.232 tons of oxide greatly reduces pollution of the herbal pharmaceutical industry to environment, and yield is nearly 3 times of former patent, realizes Red phenol extraction technique greenization.This patent is the continuation and supplement of preceding patent.New patent compared with former patent, be shown in by main feature Table 1.

The main distinction of 1 new patent of table and former patent

The production technology of red phenol has produced amplification verification as a trial, and each critical technical parameter is reproducible, and feasibility is high, new special Sharp technique makes substantial progress in energy-saving and emission-reduction, product purity, the aspects such as production cycle, cost, quality standard, existing for Chinese medicine Generationization has stepped solid step forward.

Invention content

The present invention has found out in Radix Salviae Miltiorrhizae the generation and conversion of each ingredient of red phenol and its principal component, different geographical, similarly Domain difference cultivation condition, different collecting seasons, different drying modes or the Radix Salviae Miltiorrhizae of different concocting method productions, each ingredient of red phenol Ratio and content be different, even differ widely.Main active tanshin polyphenolic acid B and its relevant major part in Radix Salviae Miltiorrhizae Ingredient be all and/or only " termination of Radix Salviae Miltiorrhizae photosynthesis " i.e. acquire after, " small molecule substrates " certain in Radix Salviae Miltiorrhizae are in " certain " macromolecular " tanshin polyphenolic acid B is generated under the action of enzyme " or tanshin polyphenolic acid B degrades generate danshensu, protocatechualdehyde, caffeic acid, purple again Oxalic acid, salviandic acid A etc..Its synthesis condition and method have or using the solarization of sun light intensity or heating, dryings, or reinforce processing, or adjust Whole extraction process changes cold carry and is carried for Hot, target component can be made to reach requirement.In all methods adjust Extracting temperature be it is most effective, Most convenient, most economical method.This all operation should all be appropriate, cannot be excessive, excessive that Related Component can be made to degrade and reduce.Cause This, before the red phenol of production carrying out trial test is necessary, that is, it is appropriate to acquire representative sample, is ground into fine powder, weighs Danshen powder 20g, adds water/80~85 DEG C hot water 100g, and room temperature/low temperature quickly stirs extraction 10min, filtering, take filter residue 1 plus water/ 80~85 DEG C of hot water 100g, room temperature/low temperature quickly stir extraction 5min, filtering, filter residue 2 plus water/80~85 DEG C hot water 80g, room Temperature/low temperature quickly stirs extraction 5min, and filtering merges filtrate three times, places 3~4 hours, makes to be cooled to room temperature, secondary filter, essence Filtrate is 3.5~4.0 with 18% hydrochloric acid tune pH, is placed 6~15 hours, secondary filter, and 20 μ l injection HPLC chromatograms of subsequent filtrate are taken Instrument records chromatogram, and 3.0% should be not less than by calculating red phenol content in Radix Salviae Miltiorrhizae, and compared with compareing HPLC Fingerprinting of Danshen Crude Drugs, phase It should be not less than 90.0% like degree, meet enterprise's internal control quality standard of Radix Salviae Miltiorrhizae.

If danshen powder has clear improvement in 80~85 DEG C of thermal agitation extraction results, but still does not meet inner controlling standard of enterprise, Hot temperature raising degree then can or be properly increased, or heat is appropriately extended and carries the time, makes up to requirement.Even if enterprise's internal control matter is still not achieved Amount standard such as continues with and also can get qualified red phenol, but yield can be substantially reduced, and considers from economics, then the Radix Salviae Miltiorrhizae is not It is suitable for preparing the raw material of red phenol, or even is unable to hyoscine.

Can it be to prepare the important step of red phenol that qualified extracting solution be obtained, and further work is exactly to purify, concentrate and do It is dry.

The primary above-mentioned red sage root extract of column purification excessively is crossed No. 2 macroporous resin columns and (blade diameter length ratio=325: 1200), is used successively 5BV purified waters, 3BV3% ethyl alcohol, 3BV10% ethyl alcohol washing, then use 3~5BV20% ethanol elutions, flow velocity be 0.3~0.5BV/ H collects the eluent containing red phenol, is 2.00~2.50 with 18% hydrochloric acid tune pH, when necessary refined filtration, and column enrichment is crossed for secondary；

The secondary above-mentioned pH of column enrichment that crosses is that 2.00~2.50 refined solutions cross No. 3 resin columns (blade diameter length ratio=325: 300) enrichment (concentration), 5BV injections washing, drain water in column, washed with 0.8BV30% alcohol, after draining, then with 2BV50% alcohol elute, collection Eluent is slightly concentrated, keeps a concentration of 150~200mg/ml of red phenol, secondary filter to be spray dried to complete；

It is spray-dried above-mentioned concentrating and purifying liquid to dry using spray of negative pressure, inlet air temperature is 105~110 DEG C, leaving air temp It is 75 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate, obtains yellowish~yellow powder pellet phenol, scale Weight, calculated yield.

The invention also discloses one group of Radix Salviae Miltiorrhizae full water-soluble ingredients " red phenol ", rather than a few content in Radix Salviae Miltiorrhizae is very Few ingredient.

The invention also discloses ultra-short Time room temperature/low temperature stirring extraction, efficiently separate purifying, not heated resin column richness Collection reduces heat-sensitive drug degradation, or makes it possible the sterling of preparation temperature-sensitive, makes red phenol technique greenization to greatest extent.

The invention also discloses how from certain different quality Radix Salviae Miltiorrhizae can prepare HPLC finger-prints and the Radix Salviae Miltiorrhizae that compares The new method of red phenol of the HPLC fingerprint similarities more than 90.0%.

Pellet phenol preparation method favorable reproducibility disclosed by the invention, the product of trial production are used《Chromatographic fingerprints of Chinese materia medica is similar Degree judges software》It calculates, the finger-print of red phenol and the fingerprint similarity of Radix Salviae Miltiorrhizae are more than 90.0%, rather than several marks Property ingredient retention time or relative retention time it is close.

The invention also discloses using the method that no less than 6~8 reference substances measure red phenol content, 6~8 component contents The sum of no less than 95%.

The invention also discloses the new pharmaceutical compositions of red phenol and its new preparation process and new technologies.

The invention discloses the new pharmaceutical compositions of " red phenol " in prevention and treatment ischemic cardiovascular disease, cerebrovascular disease The clinical application of sick and stroke at convalescence etc..

Description of the drawings

Fig. 1, fresh salvia miltiorrhiza room temperature quickly stir extracting solution HPLC collection of illustrative plates (with compare collection of illustrative plates ratio, hence it is evident that it is dissimilar, be not included in Calculate similarity).

Fig. 2, fresh salvia miltiorrhiza add 80~85 DEG C of hot water quickly to stir the HPLC collection of illustrative plates of extracting solution.

Fig. 3, Radix Salviae Miltiorrhizae boil the HPLC collection of illustrative plates of quickly stirring extracting solution.

Fig. 4, " red phenol " the HPLC collection of illustrative plates produced as a trial, work done in the manner of a certain author are abstract of description attached drawing.

The HPLC collection of illustrative plates of Fig. 5, " the red phenol dripping pill " produced as a trial.

The HPLC collection of illustrative plates of Fig. 6, " the injection pellet phenol " produced as a trial.

Fig. 7, Radix Salviae Miltiorrhizae aqueous extract, 80~85 DEG C of hot water quickly stir extracting solution, Radix Salviae Miltiorrhizae boils quickly stirring extracting solution, pellet The HPLC collection of illustrative plates similarity analysis report of phenol, red phenol dripping pill, injection pellet phenol.

Fig. 8, Salvia miltiorrhiza Bge water leach cooking liquid, 80~85 DEG C of hot water quickly stir extracting solution, red phenol, injection pellet phenol, red phenol dripping pill HPLC collection of illustrative plates similarity calculation results.

Note：Standard control HPLC collection of illustrative plates is synthesized by 9 collection of illustrative plates, can not be printed, cannot be provided.

Specific implementation mode

Further describe the present invention by the following examples, it should be understood that these embodiments are only used for illustration Purpose is never limited in protection scope of the present invention.

The preparation of the red phenol of embodiment 1

Trial test：Qingdao production Radix Salviae Miltiorrhizae (spring excavates, heated sultry drying and processing mistake, places, and crushes) powder 20g is taken, is added Water 100g, room temperature quickly stir 10min, filtering, and filter residue 1 plus water 100g, room temperature quickly stir 5min, filter, filter residue 2 plus water 80g, room temperature quickly stir 5min, and filtering merges filtrate, secondary filter three times, take 20 μ l injection HPLC chromatogram instrument of subsequent filtrate, note Chromatogram is recorded, it is 3.35% that external standard method, which calculates red phenol content, and compared with compareing HPLC Fingerprinting of Danshen Crude Drugs, similarity is 98.6%.Meet the internal control quality standard of Radix Salviae Miltiorrhizae.

It takes above-mentioned trial test with the danshen powder 50kg criticized, adds water 250kg, quickly stir 10min, centrifugal rejection filter, filter Slag 1 plus water 250kg, 5min, centrifugal rejection filter, filter residue 2 plus water 200kg are quickly stirred, 5min is quickly stirred, centrifugal rejection filter merges Filtrate three times places 4h, makes to be cooled to room temperature, secondary filter, 18% hydrochloric acid tune pH of refined filtration liquid is 3.65, places 15h, accurate mistake Filter, filtrate are 5.61 with 10% caustic lye of soda tune pH, wait for once crossing column purification.

Primary column purification said extracted liquid of crossing crosses No. 2 macroporous resin columns (blade diameter length ratio=325: 1200), pure with 5BV successively Change water 3BV3% alcohol, the washing of 3BV10% alcohol, the elution of 2BV18% alcohol, 500 ± 100ml/min of flow velocity collects dilute alcohol eluent 160L is 2.2 with 18% salt acid for adjusting pH, waits for that secondary column of crossing is enriched with

The above-mentioned refined solution of the secondary enrichment of column excessively is crossed No. 3 resin columns and (blade diameter length ratio=325: 300) is enriched with, with 5BV waters for injection Washing, drain water in column, wash with 0.8BV30% alcohol, after draining, then with 1BV50% alcohol elution, collection 20L eluents, it is slightly dense Contracting, keeps a concentration of 188mg/ml of red phenol, secondary filter to be spray dried.

It is spray-dried above-mentioned refined solution spray of negative pressure drying, spray drying parameters design：Inlet air temperature 105 DEG C~110 DEG C, 70 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate of leaving air temp obtains yellow powder, weight 1.265kg, yield 2.53%.

The preparation of the red phenol of embodiment 2

Trial test：Qingdao production Radix Salviae Miltiorrhizae (autumn excavates, and dries, and crushes) powder 20g is taken, water 100g, room temperature is added quickly to stir 10min, filtering, filter residue 1 plus water 100g, room temperature quickly stir 5min, filter, and filter residue 2 plus water 80g, room temperature quickly stir 5min, Filtering merges filtrate, secondary filter three times, takes 20 μ l injection HPLC chromatogram instrument of subsequent filtrate, records chromatogram, external standard method calculates red Phenol content is 2.53%, and compared with compareing HPLC Fingerprinting of Danshen Crude Drugs, hence it is evident that dissimilar.The content of tanshin polyphenolic acid B is also apparent inclined It is low, do not meet the internal control quality standard of Radix Salviae Miltiorrhizae.

Another take adds 80~85 DEG C of hot water 100g with trial test with the Qingdao production danshen powder 20g criticized, quickly stirs 10min, mistake Filter, filter residue 1 plus 80~85 DEG C of hot water 100g, quickly stir 5min, filter, and filter residue 2 plus 80~85 DEG C of hot water 80g are quickly stirred 15min, filtering merge filtrate three times, let cool 3~4h, are cooled to room temperature, and secondary filter takes 20 μ l injection HPLC chromatograms of subsequent filtrate Instrument records chromatogram, and it is 3.49% to calculate red phenol content in Radix Salviae Miltiorrhizae with external standard method, and with standard control HPLC Fingerprinting of Danshen Crude Drugs Compare, similarity 99.83%, meets Radix Salviae Miltiorrhizae internal control quality standard.

It takes above-mentioned trial test with the danshen powder 50kg criticized, adds 80~85 DEG C of hot water 250kg, quickly stir 10min, Centrifugal rejection filter, filter residue 1 plus 80~85 DEG C of hot water 250kg, quickly stir 5min, centrifugal rejection filter, filter residue 2 plus 80~85 DEG C of hot water 200kg quickly stirs 5min, centrifugal rejection filter, and filtrate is extracted in merging three times, is placed 4h, is made to be cooled to room temperature, secondary filter, refined filtration 18% hydrochloric acid tune pH of liquid is 3.65, places 15h, secondary filter, and filtrate is 5.62 with 10% caustic lye of soda tune pH, is waited for primary Cross column purification.

Primary column purification said extracted liquid of crossing crosses No. 2 macroporous resin columns (blade diameter length ratio=325: 1200), pure with 5BV successively Change water, 3BV3% alcohol, the washing of 3BV10% alcohol, the elution of 3BV18% alcohol, 500 ± 100ml/min of flow velocity collects dilute alcohol eluent 160L is 2.2 with 18% salt acid for adjusting pH, waits for that secondary column of crossing is enriched with

The secondary above-mentioned refined solution of column enrichment of crossing crosses No. 3 resin columns (blade diameter length ratio=325: 300), with the washing of 5BV injections Wash, drain water in column, washed with 0.8BV30% alcohol, after draining, then with 1BV50% alcohol elute, collect 20L eluents, it is slightly dense Contracting, makes a concentration of 188mg/ml of red phenol, when necessary, secondary filter is to be spray dried.

It is spray-dried above-mentioned concentrating and purifying liquid to be dried with spray of negative pressure, spray drying parameters design：105 DEG C of inlet air temperature ~110 DEG C, 70 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate of leaving air temp obtains yellow powder, weight 1.275kg, yield 2.55%.

The preparation of the red phenol of embodiment 3

2 danshen powder 50.1kg of Example is extracted, adds 80~85 DEG C of hot water 250kg, quickly stirs 10min, centrifugal drying Filter, filter residue 1 plus 80~85 DEG C of hot water 250kg quickly stir 5min, centrifugal rejection filter, filter residue 2 plus 80~85 DEG C of hot water 200kg, soon Speed stirring 5min, centrifugal rejection filter merge filtrate three times, place 4h, make to be cooled to room temperature, secondary filter, 18% hydrochloric acid of refined filtration liquid It is 3.61 to adjust pH, places 15h, secondary filter, and filtrate is 5.63 with 10% caustic lye of soda tune pH, waits for once crossing column purification.

Primary column purification said extracted liquid of crossing crosses No. 2 macroporous resin columns (blade diameter length ratio=325: 1200), pure with 5BV successively Change water, 3BV3% alcohol, the washing of 3BV10% alcohol, the elution of 2BV18% alcohol, 500 ± 100ml/min of flow velocity collects dilute alcohol eluent 160L is 2.2 with 18% hydrochloric acid tune pH, waits for that secondary column of crossing is enriched with

The secondary above-mentioned refined solution of column enrichment of crossing crosses No. 3 resin columns (blade diameter length ratio=325: 300), with the washing of 5BV injections It washs, drains water in column, washed with 0.8BV30% alcohol, then after draining, eluted with 1BV50% alcohol, collect 20L eluents, it is slightly dense Contracting, makes a concentration of 191mg/ml of red phenol, refined filtration, to be spray dried when necessary.

It is spray-dried above-mentioned concentrating and purifying liquid to be dried with spray of negative pressure, spray drying parameters design：105 DEG C of inlet air temperature ~110 DEG C, 70 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate of leaving air temp obtains yellow powder, weight 1.287kg, yield 2.574%.

The preparation of the red phenol of embodiment 4

2 danshen powder 50.2kg of Example is extracted, adds 80~85 DEG C of hot water 250kg, quickly stirs 10min, centrifugal drying Filter, filter residue 1 plus 80~85 DEG C of hot water 250kg quickly stir 5min, centrifugal rejection filter, filter residue 2 plus 80~85 DEG C of hot water 200kg, soon Speed stirring 5min, centrifugal rejection filter merge filtrate three times, place 4h, make to be cooled to room temperature, secondary filter, 18% hydrochloric acid of refined filtration liquid It is 3.61 to adjust pH, is placed 11 hours, secondary filter, and filtrate is 5.7 with 10% caustic lye of soda tune pH, waits for once crossing column purification.

Primary column purification said extracted liquid of crossing crosses No. 2 macroporous resin columns (blade diameter length ratio=325: 1200), pure with 5BV successively Change water, 3BV3% alcohol, the washing of 3BV10% alcohol, the elution of 2BV18% alcohol, 500 ± 100ml/min of flow velocity collects dilute alcohol eluent 160L is 2.3 with 18% hydrochloric acid tune pH, waits for that secondary column of crossing is enriched with

The secondary above-mentioned refined solution of column enrichment of crossing crosses No. 3 resin columns (blade diameter length ratio=325: 300), with the washing of 5BV injections It washs, drains water in column, washed with 0.8BV30% alcohol, after draining, eluted with 1BV50% alcohol, collect 20L eluents, slightly concentrate, Make a concentration of 177mg/ml of red phenol, refined filtration, to be spray dried when necessary.

It is spray-dried above-mentioned refined solution to be dried with spray of negative pressure, spray drying parameters design：Inlet air temperature 105 DEG C~110 DEG C, 70 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate of leaving air temp obtains yellow powder, weight 1.259kg, yield 2.518%.

2~4 result of embodiment shows the relatively wilfully poor smaller of its purity and yield

The preparation of the red phenol of embodiment 5

Trial test：Radix Salviae Miltiorrhizae (is produced from Qingdao, and spring excavation is dried, crushed) powder 20g is taken, water 100g, room temperature is added quickly to stir 10min, filtering, filter residue 1 plus water 100g, room temperature quickly stir 5min, filter, and filter residue 2 plus water 80g, room temperature quickly stir 5min, Filtering merges filtrate three times, places 4h, and secondary filter takes 20 μ l injection HPLC chromatogram instrument of subsequent filtrate, chromatogram recorded, by outer It is 2.89% that mark method, which calculates red phenol content, and compared with standard control HPLC Fingerprinting of Danshen Crude Drugs, hence it is evident that dissimilar.Tanshin polyphenolic acid B Content it is also apparent relatively low, do not meet the internal control quality standard of Radix Salviae Miltiorrhizae.

Another take adds 80~85 DEG C of hot water 100g with trial test with the Qingdao production danshen powder 20g criticized, quickly stirs 10min, mistake Filter, filter residue 1 plus 80~85 DEG C of hot water 100g, quickly stir 5min, filter, and filter residue 2 plus 80~85 DEG C of hot water 80g are quickly stirred 5min, filtering merge filtrate three times, place 4h, make to be cooled to room temperature, and secondary filter takes 20 μ l injection HPLC chromatogram instrument of subsequent filtrate, Chromatogram is recorded, it is 3.68% to calculate red phenol content in Radix Salviae Miltiorrhizae with external standard method, and compared with compareing HPLC Fingerprinting of Danshen Crude Drugs, phase It is 99.88% like degree, meets Radix Salviae Miltiorrhizae internal control quality standard.

It takes and produces danshen powder 50kg with the Qingdao criticized with trial test, add 80~85 DEG C of hot water 250kg, quickly stir 10min, centrifugal rejection filter, filter residue 1 plus 80~85 DEG C of hot water 250kg, quickly stir 5min, centrifugal rejection filter, filter residue 2 plus 80~85 DEG C hot water 200kg quickly stirs 5min, and centrifugal rejection filter merges filtrate three times, places 4h, makes to be cooled to room temperature, secondary filter, essence Filtrate is 3.65 with 18% hydrochloric acid tune pH, places 15h, secondary filter, and filtrate is 5.62 with 10% caustic lye of soda tune pH, waits for one It is secondary to cross column purification.

Primary column purification said extracted liquid of crossing crosses No. 2 macroporous resin columns (blade diameter length ratio=325: 1200), pure with 5BV successively Change water, 3BV3% alcohol, 3BV10% alcohol, 2BV18% alcohol to wash, 500 ± 100ml/min of flow velocity collects dilute alcohol eluent 160L, uses 18% salt acid for adjusting pH is 2.2, waits for that secondary column of crossing is enriched with

The secondary above-mentioned refined solution of column enrichment of crossing crosses No. 3 resin columns (blade diameter length ratio=325: 300), with the washing of 5BV injections It washs, drains water in column, washed with 0.8BV30% alcohol, after draining, eluted with 1BV50% alcohol, collect 20L eluents, slightly concentrate, Keep a concentration of 188mg/ml of red phenol, secondary filter to be spray dried.

It is spray-dried above-mentioned refined solution to be dried with spray of negative pressure, spray drying parameters design：Inlet air temperature 105 DEG C~110 DEG C, 70 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate of leaving air temp obtains yellow powder, weight 1.275kg, yield 2.55%.

The preparation of the red phenol of embodiment 6

The danshen powder 50kg for extracting Example 5, adds 80~85 DEG C of hot water 250kg, quickly stirs 10min, centrifugal drying Filter；It takesFilter residue 1 plus 80~85 DEG C of hot water 250kg, quickly stir 5min, and centrifugal rejection filter takes filter residue 2 plus 80~85 DEG C of hot water 200kg stirs 5min, and centrifugal rejection filter merges filtrate three times, places 4h, makes to be cooled to room temperature, secondary filter, and refined filtration liquid is with 18% Hydrochloric acid tune pH is 3.65, is placed 16 hours (standing overnight), secondary filter, and filtrate is 5.6 with 10% caustic lye of soda tune pH, is waited for Primary column purification of crossing is used.

Primary column purification said extracted liquid of crossing crosses No. 2 macroporous resin columns (blade diameter length ratio=325: 1200), pure with 5BV successively Change water, 3BV3% ethyl alcohol, the washing of 3BV10% ethyl alcohol, with 4BV20% ethanol elutions, flow velocity is 0.3~0.5BV/h, collects elution Liquid 160L is 2.35 with 18% hydrochloric acid tune pH, obtains refined solution, and column enrichment is crossed for secondary.

It is secondary cross column enrichment take above-mentioned refined solution cross No. 3 resin columns (blade diameter length ratio=325: 300), 5BV purified waters/injection It is washed with water, drains, washed with 0.8BV30% alcohol, after draining, eluted with 2BV50% alcohol de-, collect eluent 20L, it is slightly dense Contracting, makes a concentration of 150~200mg/m of red phenoll, it lets cool to room temperature, when necessary secondary filter, it is to be spray dried；

Spray drying concentrates and purifies liquid and is dried with spray of negative pressure, and inlet air temperature is 105~110 DEG C, and leaving air temp is 70 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate obtain pale yellow powder, and weight 1.205kg yields are 2.41%

The preparation of the red phenol of embodiment 7

5 danshen powder 50kg of Example is extracted, 250kg is added, quickly stirs 10min, centrifugal rejection filter, filter residue 1 at room temperature Adding water 250kg, room temperature quickly stirs 5min, centrifugal rejection filter, filter residue 2 plus water 250kg, and room temperature quickly stirs 5min, centrifugal rejection filter, Merging filtrate three times, secondary filter, 18% hydrochloric acid tune pH of refined filtration liquid is 3.5~4.0, is placed 6~11 hours, secondary filter, Filtrate is 5.5~6.0 with 10% caustic lye of soda tune pH, waits for once crossing column purification.

Primary column purification said extracted liquid of crossing crosses No. 2 macroporous resin columns (blade diameter length ratio=325: 1200), pure with 5BV successively Change water, 3BV3% alcohol, 3BV10% alcohol, 2BV18% alcohol to wash, flow velocity 500 ± 100ml/ people collect dilute alcohol washing lotion about 160L, use 18% salt acid for adjusting pH is 2.00~2.50, waits for secondary column excessively.

The secondary above-mentioned refined solution of column enrichment of crossing crosses No. 3 resin columns (blade diameter length ratio=325: 300), the washing of 5BV injections drips Water in dry column is washed with 0.8BV30% alcohol, after draining, is washed with 1BV50% alcohol, and first 10L is discarded, and is received 20L eluents, is slightly concentrated Make a concentration of 175mg/ml of red phenol, lets cool to room temperature, it is to be spray dried.

It is spray-dried above-mentioned concentrating and purifying liquid to be dried with spray of negative pressure, spray drying parameters design：105 DEG C of inlet air temperature ~110 DEG C, 70 ± 5 DEG C, 250~300HZ of rotating speed of shower nozzle, 15~30rp/min of charging rate of leaving air temp obtains pale yellow powder, Weight 1.231kg yields are 2.462%.

5~7 result of embodiment shows that the relative deviation of its purity and yield is smaller；2~7 same places of production of display, difference are adopted Season is received, the product purity of acquisition and the relative deviation of yield are also smaller.

The preparation of 8 injection pellet phenol of embodiment

Prescription

Note：Red phenol used in following embodiment is that the red phenol of 2~7 gained of above example is sufficiently mixed uniformly, crosses 80 mesh sieve, warp Detection, red phenol content are 98.55%, and moisture 1.43% is pure accordingly to weigh red phenol and feed intake.

Technical process：

Red phenol and mannitol is taken to set in beaker.

Add freshly prepared water for injection to dissolve and is diluted to 1500ml.

Add 0.05% needle-use activated carbon, stir 10min, crosses 0.2 μ l filtering with microporous membrane.

It is sub-packed in 10ml cillin bottles, every bottle of 1.5ml, void plug rubber plug.

Precooling 3h at a temperature of setting -50 DEG C

The sample of precooling is set after being freeze-dried 16h in freeze drying box, is then gradually heating to 25 DEG C, continues drying 3h。

Rubber plug is compressed under vacuum, is taken out, Zha Gai, is labelled, is packed, is cased, storage.

The full review of sampling.

The preparation of the red phenol dripping pill of embodiment 9

Capsule core prescription

Coating fluid prescription

5000 ball of capsule core

Opadry 15.0g

Purify 150ml

Preparation process

Preprocessing raw material and auxiliary material：Red phenol is dried after sieving with 100 mesh sieve, and PEG6000, PEG4000 are finely ground respectively to be sieved with 100 mesh sieve, Europe Ba Daiyong purified waters stir 45 minutes or more, be made 10% suspension it is spare.

Melting：Red phenol, PEG6000, PEG4000 are weighed in prescription ratio, is uniformly mixed, is melted in 72 DEG C of ± 2 DEG C of water-baths, Electric stirring makes liquid be uniformly mixed.

Liquid shifts：Above-mentioned uniform liquid is transferred quickly in the feed liquid filling of 70 DEG C of preheatings.

Dripping：Regulation and control drop speed, makes liquid at the uniform velocity instill in 0 DEG C of 200cs methyl-silicone oils, collects ball.

Oil removing：Wipe oil removing.

Coating：Capsule core is taken, is set in coating pan, Opadry liquid is equably sprayed onto by pot rotating speed 15-20r/min with spray gun Capsule core surface, warm wind heating, makes capsule core dry (capsule core temperature is at 30~35 DEG C), dripping pill film coating weightening 1~3%,.

Packaging：The red phenol dripping pill that will be coated, is packed with high-density polyethylene bottle, and every bottle of 150 balls, every bottle is used in connection with Bright book is packed into capsule, after the requirement packaging worked out, storage.

Full inspection：Sampling is by the red phenol dripping pill quality standard full inspection drafted

The preparation of 10 compound pellet phenol dripping pill of embodiment

Capsule core prescription

It is coated prescription

Capsule core 10000

Opadry 32g

Preparation process

Preprocessing raw material and auxiliary material：It is sieved with 100 mesh sieve after Macrogol 6000 is finely ground；Mistake after arasaponin, natural borneol are finely ground 100 mesh sieve, as bulk pharmaceutical chemicals.Opadry is stirred 45 minutes or more with purified water, be made 10% suspension it is spare.

Melting：Red phenol, arasaponin, natural borneol, PEG6000 are weighed in prescription ratio to set in 75 DEG C of material tanks, in 72 DEG C of ± 2 DEG C of water-baths, melting electric stirring make liquid be uniformly mixed.

Pill：Pill dripping machine fills 12 hole drip trays, water dropper internal diameter 1.5mm, and 75 DEG C of drip tray temperature keeps the temperature 75 DEG C, drips away from 4cm, cold Lime set is 200cs dimethicones, 0 DEG C of condensate temperature, nozzle temperature 510 DEG C, normal dripping.

Oil removing：Above-mentioned dripping pill is taken to be wrapped in except oil removing in oilcloth.

Sieve ball：Undesirable set of dripping pill is screened out.

Coating：The dripping pill for taking appearance rounding ball re-qualified, sets in coating pan, pot rotating speed 5-15r/min, with spray gun by Europe bar Capsule core surface is equably sprayed onto for liquid, Hot-blast Heating makes capsule core drying (keep capsule core temperature 3035 DEG C).Dripping pill Coating weight gain 23%.

Packing, packaging, storage：By 70 every bottle, 10 bottles are filled per box, per 100 box of box installed, is labelled, storage.

Full inspection：Sampling is by the compound pellet phenol dripping pill quality standard full inspection drafted.

The preparation of the red phenol soft capsule of embodiment 11

Softgel shell prescription (weight ratio, softgel shell wall thickness 0.60.8)：

Content prescription：

Preparation process

Major parameter is set

Technical process

Preprocessing raw material and auxiliary material

It gets out relevant auxiliary materials by softgel shell prescription.Get out related supplementary material by content prescription, and will wherein solid it is former Auxiliary material crushes, and crosses 120 mesh sieve.

It is prepared by softgel shell

Swelling, takes gelatin to set in melten gel tank, and water and glycerine, room temperature is added to be swollen 2h.

Melten gel leads to 80 DEG C of hot water to melten gel tank interlayer, when melten gel tank content is up to 70 DEG C, maintains 30 minutes.

Fumaric acid and ethylparaben is added, stirs evenly

Water circulation type aspiration pump decompression degassing 15 minutes, to ensure the softgel shell bubble-free made.

Encapsulating machine is opened, the relevant parameter of softgel shell is adjusted, air compressor machine is opened, glue is made to enter encapsulating machine glue skin, And associated technical parameters are finely tuned, the thickness for making that rubber is made is 0.80 ± 0.05mm, and makes the rubber consistency of thickness of the right and left.

The relative position for adjusting two compacting idler wheels, to suppress capsule.

Content is prepared in prescription ratio, takes red phenol, sodium caprate, sodium pyrosulfite to mix well, the 1 of Tween 80 is added, The mixture of 2- propylene glycol, stirs evenly, and adds PEG400, after fully stirring evenly, is added to content storage tank, for use.

It suppresses soft capsule and starts sprinkler body switch, and adjust flow to make to meet sets requirement, that is, carry out preparing red phenol flexible glue Capsule.

Screening and oil removing reject that shape is not whole, and filling is discontented with etc. after undesirable set of soft capsule, are wrapped in except being removed in oilcloth Oil.

It dispenses and is dispensed, labelled with soft capsule of the packaging through screening and oil removing, packed, be put in storage.

Full inspection sampling carries out full inspection by the red phenol soft capsule quality standard drafted.

The red phenol of embodiment 12 delays the preparation of controlled release micro pill

The pellet of 1 normal dissolution rate of prescription

Technical process

Pretreatment takes red phenol to sieve with 100 mesh sieve.

Batch mixing pellet phenol (sieving with 100 mesh sieve), starch, pregelatinized starch, carboxyrnethyl starch sodium (interior plus 2.5g), microcrystalline cellulose It is uniformly mixed according to the principle of equal increments.

Soft or hard appropriate softwood is made with 70% ethanol wet in softwood processed.

Squeeze with it is round as a ball by softwood by being squeezed into item, using cut it is round as a ball, pellet.

It is dry to set pellet in vacuum drying chamber in 35 DEG C of vacuum drying.

Coating take pellet packet green film-coating soluble in the stomach to get.

Clinical research quality standard full inspection is pressed in full inspection sampling, and each Testing index should meet regulation.

Prescription 2 is sustained the pellet of 6h

Technical process

Preprocessing raw material and auxiliary material takes red phenol, lactose, sodium cyclamate, hydroxypropyl methylcellulose, carbomer to cross 80 mesh sieve respectively, gathers Tie up ketone K₃₀It is configured to 5% (w/v) solution with 75% ethyl alcohol.

Softwood processed weighs red phenol, lactose, hydroxypropyl methylcellulose, carbomer, and 30 POVIDONE K 30 BP/USP is added in mixing₃₀Softwood is made in liquid.

Squeeze with it is round as a ball by softwood by being squeezed into item, then after cutting, it is round as a ball.

Dry to set pellet in 35 DEG C of vacuum drying in vacuum drying chamber, removal fine powder is to get pellet.

Coating take pellet packet yellow acrylic resin IV film-coatings to get.

Clinical research quality standard full inspection is pressed in full inspection sampling, and each Testing index should meet regulation.

Prescription 3 is sustained the pellet of 12h

*：6% solution is made of 80% ethyl alcohol for ethyl cellulose, makees adhesive.80% ethanol consumption can fit on demand Work as adjustment.

Technical process

Preprocessing raw material and auxiliary material pellet phenol crosses 80 mesh sieve, and hydroxypropyl methylcellulose K100M CR (HPMC K100M CR) cross 80 mesh sieve； Ethyl cellulose (EC) impregnates the solution that 6% (w/v) is made in dissolving with 80% ethyl alcohol, spare.

Softwood processed weighs red phenol by prescription and hydroxypropyl methyl cellulose K100M is uniformly mixed, and 6% EC liquid is added, instead Multiple rubbing makes into uniform softwood

It squeezes round as a ball by squeezing spheronizator, is squeezed into item, cutting, round as a ball to get wet pellet.

Pellet drying sets wet pellet dry in drying box.

Coating take pellet packet red acrylic resin I film-coatings to get.

Clinical research quality standard full inspection is pressed in full inspection sampling, and each Testing index should meet regulation.

Red phenol delays controlled release micro pill capsules/tablets

Mixed ball weighs prescription 1,2,3 pellet 100g, 90g, 120g respectively, mixes well.

It is encapsulated that mixing pellet is taken to be packed into suitable softgel shell, per ball content weight 310mg, labelled amount 150mg.

Packing is dispensed with packaging, labelling, packaging, storage.

Clinical research quality standard full inspection is pressed in full inspection sampling, and indices should all meet regulation.

Content assaying method

Content assaying method the present invention relates to red phenol is used in conjunction with high-precision mass spectrum using UHPLC, to red phenol it is each at It point is positioned by molecular weight, selects the foundation of content and the relatively high ingredient of activity alternatively reference substance.With danshensu, former youngster 6 tea aldehyde, Rosmarinic acid, alkannic acid, tanshin polyphenolic acid B, salviandic acid A reference substances press external standard method and calculate each component content respectively, 6 The content of ingredient accounts for about the 93-97% of labelled amount.The sum of remaining small peak area is made, with reference to product, to calculate by external standard method with tanshin polyphenolic acid B Content accounts for about the 2-6% of labelled amount.The sum of two parts content is red phenol content, the content of red phenol be the 98% of labelled amount with On.

The clinical application of red phenol

The invention further relates to red phenol preparations in the new application for preparing anti-cerebral ischemia reperfusion injury：Pass through rat artery embolism Model (MCAO) establishes cerebral ischemia re-pouring animal model, breast in observed behavior, brain infarction area, brain water content, brain tissue Acidohydrogenase (LDH), creatine kinase (CK), superoxide dismutase (SOD) and malonaldehyde (MDA), IL-6, IL-1 β, TNF-α Level, rat cortical lesions situation after HE dyeing observation MCAO modelings and administration.As a result it proves to give rat pellet phenol ip 15mg/kg has the function of significant anti-cerebral ischemia reperfusion injury, and is substantially better than cerebral arterial thrombosis first-line treatment chemical drug The effect of Nimodipine ip 20mg/kg, can be used for preparing prevention and/or the treatment ischemic of anti-cerebral ischemia reperfusion injury The drug of cerebral apoplexy.

Claims (4)

1. a kind of preparation of anti-cerebral ischemia Reperfusion injury vulnerary pellet phenol is specifically related to one group of full water-soluble ingredient " red phenol " production Technique

(1) production technology of the red phenol of claim 1 is specifically related to the pretreatment and its crushing of Radix Salviae Miltiorrhizae, the more particularly to HPLC of medicinal material The pre- measurement of finger-print, pre- measured value are related to the selection of the specific embodiment of extraction operation.

(2) production technology of the red phenol of claim 1 is specifically related to take Danshen Root that the fast stirring of 5 times of water for measuring 25 DEG C/85 DEG C is added to carry It takes, first time extraction 10 minutes, respectively extracts 5 minutes for the second time, for the third time.The Extracting temperature of optimization is with the time with the HPLC of Radix Salviae Miltiorrhizae Finger-print prompt ingredient and its content status and select.

(3) Extracting temperature of the red phenol production technology of claim (2) with the time is selected according to the result of trial test, is carried when 25 DEG C Take peak number, peak intensity that liquid HPLC is composed has apparent difference compared with the HPLC spectrums of control Radix Salviae Miltiorrhizae, then separately danshen powder should be taken 80 DEG C~85 DEG C of extractions, then it may obtain satisfied result, it may be necessary to when properly increasing Extracting temperature, or extraction is appropriately extended Between.

(4) production technology of the red phenol of claim 1 places 3~4h, makes to be cooled to room temperature, accurate mistake more particularly to extracting solution is merged Filter, it is 2.5.~5.5 that refined filtration liquid is acidified pH with dilute hydrochloric acid, and the pH of optimization is 3.5.~4.0.Then acidifying solution is placed and is precipitated, Standing time is 4~16h, and optimization is 4~6h of placement, then for purifying after secondary filter.

(5) before the production technology of the red phenol of claim 1 was specifically related to resin column separating purification, refined filtration liquid should adjust pH and be 5.0~6.4, the pH that should adjust of optimization is 5.5~5.9..

(6) production technology of the red phenol of claim 1 is specifically related to pH be that 5.5~5.9 refined filtration liquid cross D101, D301, separation 2 Number, No. 3 macroreticular resins of chromatography isolated and purified, preferred resin be separation No. 2, analysis No. 3 macroreticular resins of chromatography, then according to Secondary water, purified water, injection water, 0.5%~25% medicinal alcohol gradient wash, the gradient wash medicinal alcohol of optimization are a concentration of 1%~12%.

(7) production technology of the red phenol of claim 1 is specifically related to be eluted with 12%~30% medicinal alcohol, the eluent of optimization For 14%~22% ethyl alcohol.

(8) be specifically related to will be dilute with D101, D301, No. 3 separation 2, chromatography macroreticular resins for the production technology of the red phenol of claim 1 Medicinal alcohol elutes liquid enrichment, and the resin of optimization is chromatography 3, detaches No. 2 macroreticular resins, excessively should be by dilute medicinal use before resin column It is 1.5~3.0 that ethanol eluate, which adjusts pH, and the pH of optimization is 2.0~2.5.

(9) production technology of the red phenol of claim 1 is each to determine more particularly to sampling each red phenol intermediates content of HPLC detections Stage work target.

(10) production technology of the red phenol of claim 1 adjusts it containing medicinal more particularly to satisfactory red phenol ethanol is merged After concentration of alcohol is 60% or so, secondary filter.

(11) production technology of the red phenol of claim 1 is controlled more particularly to taking 60% medicinal alcohol pellet phenol refined filtration liquid to be spray-dried Inlet air temperature is 80 DEG C~130 DEG C, and leaving air temp is 50~60, obtains off-white color or the red phenol of yellowish or yellow powder.

(12) " red phenol " made from claim 1 refer to the active ingredient that red phenol contains be in red rooted salvia all it is water-soluble at Point, rather than a few ingredient, it is promulgated according to State Food and Drug Administration《Chromatographic fingerprints of Chinese materia medica similarity Judge software》It calculates, for the finger-print of red phenol compared with compareing Radix Salviae Miltiorrhizae aqueous extract finger-print, similarity is more than 95%.

2. a kind of production technology of anti-cerebral ischemia Reperfusion injury vulnerary pellet phenol of claim 1 is combined more particularly to one group of pellet phenol agents It is prepared by object

(13) the red phenol agents composition of claim 2 contains the full water-soluble ingredient pellet phenol that claim 1 is produced from Radix Salviae Miltiorrhizae Pharmaceutically acceptable pharmaceutical carrier.

(14) claim 2 pharmaceutical composition contain can injection injection, such as injection pellet phenol, red phenol injection is red Phenol drip solution etc..

(15) claim 2 pharmaceutical composition contain can pro ore serial solid pharmaceutical preparation and novel form, such as red phenol piece, red phenol Capsule, red phenol granule, red phenol soft capsule, red phenol dripping pill, red phenol delay controlled release micro pill, and red phenol micro-capsule, red phenol micro emulsion and compound are red Each dosage form of phenol etc..

(16) claim 2 pharmaceutical composition contains the various preparations that can be for external application, such as red phenol paste, red phenol liniment, red phenol drop Eye agent, red phenol nasal drop, red phenol aerosol, red phenol gelling agent etc..

(17) claim 2 pharmaceutical composition also contains various other dosage forms of pharmaceutically acceptable and novel form etc..

(18 claim 2 pharmaceutical compositions also contain various pharmaceutically acceptable various specifications, such as 30mg, 60mg, 120mg, 240mg etc..

(19) pharmaceutical carrier of claim 2 pharmaceutical composition can be one or more solid, liquid and aerosol that can be compatible Agent solvent etc., dosage form depend on administering mode；Preparation process can also be varied.These are for a person skilled in the art It will be apparent that all within the scope of this patent is protected.

3. a kind of preparation of anti-cerebral ischemia Reperfusion injury vulnerary pellet phenol of claim 1 is measured more particularly to red phenol and its containing for preparation Determine method

(20) content of the red phenol of claim 3 is one group of multicomponent active component, and content assaying method includes with multiple right It compares according to product, is calculated by external standard method；The sum of part peak area other than reference substance makees object of reference with tanshin polyphenolic acid B, based on external standard method Content is calculated, the sum of two parts content is red phenol content.Reference substance number is more, and assay value is closer to actual value.

(21) major part is compareed with 6~8 reference substances in the content assaying method of the red phenol of claim 3, based on external standard method Content is calculated, total content is 92%~97%.

(22) the sum of peak area other than the part that reference substance compares in the content assaying method of the red phenol of claim 3 is with danshinolic acid B makees object of reference, and by the content that external standard method calculates, total content is 3%~6%.

(23) content of the red phenol of claim 3 should be not less than 98.5%.

4. the clinic that a kind of production technology of anti-cerebral ischemia Reperfusion injury vulnerary pellet phenol of claim 1 is specifically related to red phenol preparation is answered With

(24) the red phenol clinical application of claim 4 relate generally to ischemic Cardial or cerebral vascular diseases and stroke at convalescence obtain it is clinical Using.

(25) clinical application of the red phenol of claim 4 be specifically related to red phenol system arrange each preparation for Cardial or cerebral vascular diseases, gynaecology, Paediatrics, scald section, ENT dept. and diabetes, tumour, antibacterial, anti-inflammatory, anti-oxidant, enhancing is immune, improves the applications such as memory.

(26) clinical application of the red phenol of claim 4 is more particularly to promoting blood circulation and removing blood stasis, stasis-dispelling and pain-killing, promoting blood circulation, activating collaterals, relieving restlessness that clears away heart-fire, expansion Blood vessel, the prevention and treatment for increasing each clinically relevant disease of coronary artery and brain blood flow etc. " function and cure mainly ".