WO2013131270A1 - Scutellarin hydrate and preparation method and use thereof - Google Patents

Description

[Name of invention established by ISA according to Rule 37.2] scutellarin hydrate, preparation method and use thereof Technical Field

 The present invention relates to the field of medical technology, and in particular to a cardio-cerebral vascular therapeutic drug, scutellarin derivative, a preparation method thereof and use thereof.

Background Art

The scutellarin is present in the Chinese herbal medicine scorpion flower. From the pharmacological point of view, scutellarin can improve microcirculation, increase cerebral blood flow, improve blood-brain barrier permeability, improve blood rheology, reduce whole blood viscosity, and resist Platelet aggregation is significant, promotes fibrinolytic activity, reduces fibrinogen, prevents thrombolysis; prevents calcium overload and extracellular excitatory amino acid accumulation in nerve cells, inhibits protein kinase C displacement activation or overactivation, reduces brain edema, and delays Ischemic brain cell death, increased trophic myocardial blood flow, improved myocardial microcirculation, and a certain role in lowering blood lipids. Currently used clinically for the treatment of cerebral thrombosis, cerebral hemorrhage and its sequelae; coronary heart disease, angina pectoris, arrhythmia; hyperviscosity; refractory neck syndrome, vertebral basilar artery insufficiency and other ischemia and microcirculation disorders The disease has important application value in the treatment of cardiovascular and cerebrovascular diseases. At present, the published literature only reports scutellarin (C ₂₁ H ₁₈ O _{12 ,} CAS No. : 27740-01-8, molecular weight: 462.37). So far, there is no publicly available literature report at home and abroad. Crystalline hydrate C ₂₁ H ₁₈ O ₁₂ ·nH ₂ O , n=1.75 ～ 2.0] and its preparation method and use.

Technical Problem

The invention relates to a scutellarin derivative, a preparation method thereof and use thereof, and further relates to a scutellarin crystal hydrate [ C ₂₁ H ₁₈ O ₁₂ ·nH ₂ O , n=1.75 ～ 2.0 , n It can be 1.75, 1.8, 2.0 or a number between them] and its preparation method and use.

Technical Solution

 The preparation of scutellarin crystal hydrate includes the following methods:

Method A In a reaction vessel, 2 to 20 times (by volume) of water, or a C ₁ -C ₆ low molecular alcohol, a C ₂ -C ₆ low molecular nitrile, C ₃ -C one or more lower ketones _8, C ₂ -C ₈ esters of low molecular weight in the extraction (including mention cold, heat extraction, or ultrasonic extraction, microwave extraction or reflux extraction, or the like) once to several times a solvent (1~6 times), combine the extracts, filter, concentrate the filtrate, dilute the concentrate with water, adjust the pH to 6.5-9 with alkali (the base used includes sodium carbonate, sodium hydrogencarbonate, sodium hydroxide, hydrogen phosphate) a commonly chemically defined pharmaceutically acceptable Lewis base or an alkaline solution thereof, such as disodium, sodium phosphate, arginine, lysine, a low molecular amine such as C ₁ -C ₁₂ , filtered, passed through a silica gel column Or macroporous adsorption resin, or polyamide column chromatography or glucose gel resin, water or C ₁ -C ₆ low molecular alcohol, C ₃ -C ₈ lower ketone, C ₂ -C ₆ low molecular nitrile, etc. One or more of them are eluted, and the filtrate is adjusted to pH 1-5 with acid (the acid used includes hydrochloric acid, sulfuric acid, phosphoric acid, carbonic acid, phosphorus). Lewis acid or an acidic solution of sodium dihydrogen chemically defined, lactic acid, and other low molecular weight of C ₁ -C _8), stirred, placed to precipitate sufficiently precipitate, the resulting precipitate was filtered, washed with water and a C ₁ -C ₆ low molecular weight alcohols, C ₂ -C ₈ esters of low molecular weight, C ₃ -C ₈ lower ketones, C ₂ -C ₆ of a low molecular weight nitrile, C ₂ -C ₆ ether, a lower, C ₁ -C ₆ lower halo of One or more of a hydrocarbon, a C ₁ -C ₈ low molecular acid or the like is a crystallization solvent, which is recrystallized one or several times, filtered, washed with water or a C ₃ -C ₈ lower ketone, C ₂ -C ₆ One or several washings of the low molecular nitrile, and drying the obtained solid to obtain scutellarin crystal hydrate;

 Or method B

In the reaction vessel, the scutellaria whole plant is extracted with one or more of 2 to 20 times (volume ratio) of water, an alkaline solution or a C ₁ -C ₆ low molecular alcohol, etc. (including Cold extraction, heat extraction, desorption or ultrasonic extraction, or microwave extraction or reflux extraction, etc.) once to several times (1 to 6 times), filtering, combining the filtrate, concentrating the filtrate, diluting the concentrate with water, adjusting the pH with alkali Between 6.5 and 9 (the base used includes sodium carbonate, sodium hydrogencarbonate, sodium hydroxide, disodium hydrogen phosphate, sodium phosphate, potassium carbonate, sodium citrate, arginine, lysine, C ₁ -C ₁₂ a low-molecular amine such as a generally chemically defined pharmaceutically acceptable Lewis base or an alkaline solution thereof, filtered, filtrated through a silica gel column and a macroporous adsorption resin, or a polyamide column chromatography or a glucose gel resin Elution with one or more of water or C ₁ -C ₆ low molecular alcohol, C ₃ -C ₈ lower ketone, C ₂ -C ₆ low molecular nitrile, etc., and the filtrate is adjusted to pH 1 with acid. 1-5 (acid used include hydrochloric acid, sulfuric acid, phosphoric acid, carbonic acid, sodium dihydrogen phosphate, lactic acid, C ₁ -C ₈ lower the Lewis acid or an acidic solution chemically defined protic acid, etc.), is placed, to precipitate sufficiently precipitate, the resulting precipitate was filtered, washed with water and a lower alcohol C ₁ -C ₆ is, C ₂ -C ₈ esters of low molecular weight, C _3- C ₈ lower ketone, C ₂ -C ₆ low molecular nitrile, C ₂ -C ₆ lower ether, C ₁ -C ₆ lower halogenated hydrocarbon, C ₁ -C ₈ low molecular acid, etc. One or more are crystallization solvents, recrystallized one or several times, filtered, washed with water, and the obtained solid is dried to obtain scutellarin crystal hydrate;

 In the extraction process according to the present invention, other preparation procedures may be carried out after the enamel medicinal material powder is sufficiently immersed in an extraction solvent as is conventional. The filtering method in the preparation process of the present invention may be one of using a filter cloth, a funnel, a fused glass filter, a sand core filter rod, a plate and frame filter press, a microporous filter membrane, a ceramic membrane or an ultrafiltration or Several filtration methods are performed once or several times (1~6 times). In preparation Scutellaria In the method or process for removing impurities by ultrafiltration, the ultrafilter may be selected from a flat plate type, a coil type, a tube type, a hollow fiber type or a round box type, and preferably a roll type and a hollow fiber type ultrafilter. The ultrafiltration membrane is used to remove impurities with a molecular weight of 4,000 to 60,000, and an ultrafiltration membrane having a molecular weight of 6,000 to 30,000 is preferable, and an ultrafiltration membrane having a relative molecular mass of 6,000 to 20,000 is more preferable.

In the preparation process of the two preparation methods of the scutellarin crystal hydrate of the present invention, the extract is adjusted to have a pH of 6.8 to 9 with a base (the base used includes sodium carbonate, sodium hydrogencarbonate, sodium hydroxide, hydrogen phosphate). a common chemically defined pharmaceutically acceptable Lewis base or an alkaline solution thereof, such as disodium, sodium phosphate, potassium carbonate, sodium citrate, a low molecular amine of C ₁ -C ₁₂ , etc., filtered, and the filtrate is subjected to silica gel chromatography. Column and macroporous adsorption resin, or polyamide column chromatography or glucose gel resin, water or C ₁ -C ₆ low molecular alcohol, C ₃ -C ₈ lower ketone, C ₂ -C ₈ low molecular ester, One or more of the C ₂ -C ₆ low molecular weight nitriles are eluted, and the filtrate is adjusted to pH 1-5 with an acid (the acid used includes hydrochloric acid, sulfuric acid, phosphoric acid, carbonic acid, sodium dihydrogen phosphate, lactic acid, C ₁ - a chemically defined Lewis acid such as a low molecular acid such as C ₈ or an acidic solution thereof, placed, and the precipitate is subjected to a process of analysis, which can be diluted with water after the precipitation of the acid precipitate in the preparation process The above process is carried out 1 to 3 times for purification purposes, and then heavy Crystallization process.

 The macroporous adsorption resin according to the present invention may include a macroporous adsorption resin of the following types: D-101 type, DA-201 type, MD- 05271, GDX-105, CAD-40, XAD-4, SIP, D-, X-5, AB-8, LD-605, LD-601, DM- Type 130, R-A type, CHA-101 type, WLD type (mixed type), H-107 type, NKA-9 type, etc.

The different steps of Process A and Method B of the hydrates of the present invention, including those in the extraction and refining processes, can also be used crosswise for the preparation of the compounds of the present invention.

In the present invention, a lower alcohol or a lower alcohol is defined by the number of carbon atoms is C ₁ -C ₆ (i.e.: alcohols of 1 to 6 carbon atoms), such as methanol, ethanol, isopropanol, butanol and the like; C ₂ - The carbon number of the lower nitrile of C ₆ is defined as C ₂ -C ₆ , such as acetonitrile, propionitrile or the like; the number of carbon atoms of the lower ether or low molecular ether is defined as C ₂ -C ₈ , such as diethyl ether, diisopropyl ether, butyl Ether, etc.; the lower halogenated hydrocarbon has a carbon number defined as C ₁ -C ₆ (ie, 1 to 6 carbon atoms), including dichloromethane, dichloroethane, chloroform, etc.; the lower alkyl ester has a carbon number defined as C ₂ - C ₈ (ie 2-8 carbon atoms), including butyl acetate, ethyl acetate, ethyl formate, etc.; the number of carbon atoms of a low molecular linear or branched alkane or cycloalkane is defined as C ₅ - C ₁₀ (ie 5-10 carbon atoms), including pentane, n-hexane, cyclohexane, petroleum ether, etc.; the number of carbon atoms of the low molecular aromatic hydrocarbon is defined as C ₆ -C ₁₂ , including benzene, toluene, etc.; C ₃ - The low molecular ketone of C ₈ is defined as a ketone of 3-8 carbon atoms, including acetone, methyl ethyl ketone, pentanone, ketone, isohexanone, etc.; the carbon number of the low molecular acid of C ₁ -C ₈ is defined as 1 8 carbon sources Organic acids, including formic acid, acetic acid, propionic acid, lactic acid, citric acid, maleic acid, tartaric acid and the like; low molecular weight amine is defined as C ₁ -C ₁₂ organic amine 1 to 12 carbon atoms include dimethylamine, Diethylamine, triethylamine, cyclohexylamine, dicyclohexylamine, tert-butylamine, N-methylmorpholine, pyridine, picoline, chiral or racemic basic amino acids, such as L-arginine, DL - lysine, etc.; for any type of labeling method for the number of carbon atoms described as 'low molecular' compounds, as long as it appears once in the text of this application, any other unlabeled description of the same type of compound as 'low molecule' The number of carbon atoms is consistent with the number already indicated herein.

a crystallization or recrystallization solvent of scutellarin hydrate, preferably water with methanol, ethanol, isopropanol, n-butanol, acetone, methyl ethyl ketone, isohexanone, methyl acetate, ethyl acetate, acetic acid, dichloromethane, One or more of chloroform, diethyl ether, diisopropyl ether, tetrahydrofuran, petroleum ether, benzene, or a certain proportion of water in an organic solvent.

 The product of the present invention can be dried at different temperatures (e.g., 20-100 ° C), drying time (1). Hours to several days), or with other desiccants (including silica gel, phosphorus pentoxide, anhydrous calcium chloride, anhydrous sodium sulfate, etc.) under ambient conditions, or using atmospheric or decompression methods for the final The product is dried. Its drying temperature is preferably 3 0 to 70 °C.

 The moisture measurement of the present invention uses the Karl Fischer method, and 10% is available. The imidazole anhydrous methanol is used as a solvent, and the melting point of the scutellarin hydrate of the present invention is measured by a melting point apparatus, and the melting point analyzer is not corrected.

 The scutellarin of the present invention may have different crystal forms, for example, scutellarin prepared from different solvent systems for crystallization or recrystallization systems. The X-ray powder diffraction pattern of 1.75, 2 hydrates can vary. The solution of the crystalline hydrate in the present invention or the crystalline hydrate in the example is in the same HPLC The chromatographic retention time of its main peak is consistent under conditions.

 In one aspect, the invention provides a different crystalline hydrate of scutellarin.

 In another aspect, the invention provides a process for the preparation of different crystalline hydrates.

In another aspect, the invention provides a pharmaceutical composition comprising any one or more of scutellarin hydrate prepared by the method of the invention, and one or more pharmaceutically acceptable excipients.

The invention further provides a process for the preparation of a pharmaceutical formulation comprising the combination of any one or more of scutellarin hydrate prepared by the method of the invention and at least one or a pharmaceutically acceptable excipient.

 The invention further provides a crystalline hydrate of scutellarin, which is prepared for treatment and is suitable for preparation Human and mammalian coronary heart disease, angina pectoris, myocardial infarction, arrhythmia, ischemic brain disease, cerebral thrombosis, cerebral hemorrhage and its sequelae, vascular dementia, Peripheral circulatory disorders, including various arterial occlusive diseases, vasculitis, microcirculatory disorders caused by diabetes, hypertension, hyperlipidemia, Hyperviscosity; application of refractory cervical syndrome, vertebrobasilar insufficiency and other diseases of ischemia and microcirculatory disorders for the treatment or prevention of diseases.

The use of the scutellarin hydrate of the present invention: the scutellarin hydrate of the present invention is used for preparing a lyophilized powder injection preparation for injection, a powder injection preparation, a large infusion preparation, a small water injection, a solid preparation and the like.

The scutellarin hydrate of the invention is used for preparing solid preparation tablets (including ordinary tablets, controlled sustained release tablets and capsules, buccal tablets, dispersible tablets, orally disintegrating tablets, etc.), capsules (including ordinary capsules, and slowing down) Release capsules, etc.), granules, suspensions, pills, etc., which may contain pharmaceutically acceptable fillers such as starch, modified starch, lactose, microcrystalline cellulose, cyclodextrin, sorbitol, mannitol, Xylitol, calcium phosphate, calcium hydrogen phosphate, amino acid, polyethylene glycol 4000 - 20000 , poloxamer, gelatin, stearic acid, sodium stearate, etc.; pharmaceutically acceptable disintegrating agents, such as starch, modified starch, microcrystalline cellulose, sodium carboxymethyl starch, crosslinked polyvinylpyrrolidone, Low-substituted hydroxypropylcellulose, surfactant, pharmaceutically acceptable wetting agent and binder, such as gelatinized starch, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyethylene Pyrrolidone, alginic acid and salts thereof; pharmaceutically acceptable lubricants and glidants such as stearic acid, magnesium stearate, polyethylene glycol 4000 - 8000, talc, micronized silica gel, magnesium lauryl sulfate, etc.; pharmaceutically acceptable sweeteners and flavors such as sucrose, aspartame, cyclamate, sodium saccharin, sucralose, edible flavors A pharmaceutically acceptable suspending stabilizer, such as Xanthan gum, guar gum, gelatin, gum arabic, methyl cellulose, ethyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose, tragacanth, sodium alginate Etc; other pharmaceutically acceptable stabilizers, such as beta- Cyclodextrin, etc.

 The crystalline hydrate of the present invention is different from The deliquescent of the anhydrate prevents the air from being blocked during the treatment, and the crystallization hydrate has good slidability, thereby improving the operability of the preparation; and preparing the solid preparation It has good dissolution properties, making it easy to be absorbed into the blood circulation, improving bioavailability, and facilitating its rapid function.

 Scutellarin hydrate injection, the preparation method is:

 The preparation method of the lyophilized powder injection preparation is as follows: taking scutellarin hydrate , may be added with a pharmaceutically acceptable co-solvent, lyophilized support or co-agent, stabilizer, water for injection, stirring to dissolve, if necessary, the pH can be adjusted to 4.5 to 7.5 with a pharmaceutically acceptable acid base, plus activity carbon 0.005 ~ 1% (W / V) stirring for 15 ~ 45min, filtration, hydration, sterile filtration, press 5 ~ 400mg / The bottle (in terms of the main drug) is dispensed, freeze-dried, and tamponed to obtain a finished product.

 Scutellarin hydrate hydrate small-volume injection and preparation process thereof: scutellarin hydrate Adding water for injection and a pharmaceutically acceptable additive, for example, a pharmaceutically acceptable cosolvent, a pH adjuster, a pharmaceutically acceptable antioxidant, an inert gas, filtration, sterilization, and a sterile small volume injection , its pH is at Between 4.5 and 7.5.

The pharmaceutically acceptable lyophilized support or co-agent may contain lactose, glucose, mannitol, sorbitol, xylitol, dextran, amino acids or salts thereof (including glycine, taurine, arginine, etc.), phosphoric acid One or more of sodium dihydrogenate, disodium hydrogen phosphate, sodium deoxycholate, and the like.

 Its pharmaceutically acceptable pH The regulator may be a pharmaceutically acceptable inorganic or organic acid, an inorganic base or an organic base, or a Lewis acid or a base in a broad sense, and may contain one or more of them, and may be hydrochloric acid, phosphoric acid, propionic acid, acetic acid or the like. Acetate, such as sodium acetate, lactic acid and lactic acid pharmaceutically acceptable salt, citric acid pharmaceutically acceptable salt, sodium carbonate, sodium hydrogencarbonate, potassium hydrogencarbonate, sodium hydroxide, potassium hydroxide, phosphate, tartaric acid and medicinal use thereof Salt, borax, boric acid, succinic acid, caproic acid, adipic acid, fumaric acid, maleic acid, trishydroxyaminomethane, diethanolamine, ethanolamine, isopropanolamine, diisopropanolamine, 2-Amino-2-(hydroxymethyl) 1 ,3-propanediolamine, 1,2-hexanediamine, N- Methylglucamine, diisopropylamine and salts thereof, polyhydroxycarboxylic acids and pharmaceutically acceptable salts such as glucuronic acid, gluconic acid, lactobionic acid, malic acid, threonic acid, glucoheptonic acid, amino acids and amino acid salts, etc. One or several of them.

The pharmaceutically acceptable antioxidants and stabilizers thereof may be sulfurous acid, sulfite, bisulfite, pyrosulfite, dithionite, thiosulfate, organic sulfur compound thiourea, glutathione Glycopeptide, dimercaptopropanol, thioglycolic acid and salt, thiolactic acid and salt, thiodipropionic acid and salt, phenolic compounds such as gallic acid and salt, caffeic acid and its salts, ferulic acid and its salts, Di-tert-butyl-p-phenol, 2,5-dihydroxybenzoic acid, 2,5-dihydroxybenzoate, amino acid and salts thereof; ascorbic acid and ascorbate, isoascorbic acid and isoascorbate, nicotinamide, tartaric acid, nitric acid Salt, phosphate, pharmaceutically acceptable salt, citrate, EDTA and EDTA salts, such as EDTA calcium sodium, EDTA tetrasodium, N-bis(2-hydroxyethyl)glycine, α-cyclodextrin, β-cyclodextrin Refined, γ-cyclodextrin, glucosyl-cyclodextrin (G ₁ -CYD ), maltosyl-β-cyclodextrin, hydroxypropyl β-cyclodextrin, 2-hydroxypropyl β-cyclodextrin (2-HP-β-CYD), 3-hydroxypropyl β-cyclodextrin (3-HP-β-CYD), sulfobutylether-β-cyclodextrin One or several (SBE-β-CD), such as _{(SBE 7 -β-CD),} SBE 4 -β-CD and the like.

The pharmaceutically acceptable co-solvent may be sodium carbonate, potassium carbonate, sodium hydrogencarbonate, disodium hydrogen phosphate, sodium phosphate, dipotassium hydrogen phosphate, sodium lactate, sodium citrate, N-methyl glucosamine, N-B. Glucosamine, nicotinamide, ethylenediamine, diethylamine, ethanolamine (including diethanolamine, triethanolamine, etc.), trishydroxyaminomethane, L or D or DL type amino acids (including L or D or DL type arginine, Pharmaceutically acceptable organic amines such as lysine, citrulline, histidine, etc., Tween 20-80, poloxamer (including poloxamer 124, 188, 237, 338, 407, etc.), Polyethylene glycol 200-2000, ethanol, ethylene glycol, propylene glycol, glycerol, glucosyl-cyclodextrin (G ₁ -CYD ), maltosyl-β-cyclodextrin, 2-hydroxypropyl β- One or more of cyclodextrin, 3-hydroxypropyl β-cyclodextrin, sulfobutylether-β-cyclodextrin, and water.

 The lyophilized support or adjuvant may be glucose, fructose, xylitol, sorbitol, mannitol, invert sugar, maltose, dextran, N- Methylglucamine, N-ethylglucosamine, nicotinamide, ethylenediamine, diethylamine, ethanolamine (including diethanolamine, triethanolamine, etc.), disodium hydrogen phosphate, sodium phosphate, dipotassium hydrogen phosphate, sodium lactate, Sodium citrate, L or D Or DL type amino acids (including L or D or DL type arginine, lysine, citrulline, histidine, etc.), Tween 20-80, poloxamer (including poloxamer 124, 188) , 237, 338, 407, etc.), polyethylene glycol 200-2000, ethanol, ethylene glycol, glycerol, 2-hydroxypropyl β-cyclodextrin, 3-hydroxypropyl β-cyclodextrin ( One or more of 3-HP-β-CYD), sulfobutylether-β-cyclodextrin, and the like.

The pharmaceutically acceptable isotonicity adjusting agent may be one or more of glucose, fructose, xylitol, sorbitol, mannitol, invert sugar, maltose, dextran, sodium chloride, potassium chloride, sodium lactate, and the like. .

 De-heating and sterilization methods can be added to the amount of liquid From 0.005 to 3% of activated carbon to heat source, microporous membrane sterilization and hot pressing sterilization, ultrafiltration can also be used to remove bacteria and heat sources. In the ultrafiltration method, the ultrafilter may be a flat plate type, a coil type, a tube type, a hollow fiber type or a round box type, etc., preferably a roll type and a hollow fiber type ultrafilter, and the molecular weight of the interception is 50,000 to 300,000. After removing most of the heat-generating substances and bacteria, the filter removes the remaining heat source by using an ultrafiltration membrane with a molecular weight of 4,000 to 30,000, preferably an ultrafiltration membrane having a molecular weight of 6000 to 30,000.

The scutellarin crystal hydrate of the invention is suitable for preparing coronary heart disease, angina pectoris, myocardial infarction, arrhythmia, ischemic brain disease, cerebral thrombosis, cerebral embolism, cerebral hemorrhage and sequelae thereof, and vascularity in human and mammals dementia Peripheral circulatory disorders such as various arterial occlusive diseases, vasculitis, microcirculatory disorders caused by diabetes, refractory neck syndrome, vertebral basilar artery insufficiency and other ischemia and diseases with microcirculatory disorders, high viscosity Blood The use of drugs for the treatment or prevention of diseases.

 Dosage Usage: Under normal circumstances, in adults, intramuscular injection of scutellarin crystal hydrate: 5-200 mg / time, 1-3 times a day. Intravenous injection: 5-400 mg/time, diluted with 25% glucose injection. Intravenous infusion: 5-400mg, with 5% glucose injection or 0.9% sodium chloride injection 250 ~ 500ml Dilute, 1-2 times a day. Use more than half of the children. Dosage for parenteral administration: 10 to 70 kg body weight of human or animal, generally 10 to 1000 mg scutellarin crystal hydrate / day, minute 1-3 times of administration; children are halved or more.

Advantageous Effects

 The scutellarin containing crystal water obtained by the present invention, surprisingly, scutellarin containing crystal water The hygroscopicity is much lower than that of scutellarin, which does not contain crystal water, and scutellarin, which contains crystal water. The hydrate is more stable than the water without crystal water, is easy to store and transport, and has good water solubility at room temperature, and is easy to be made into a water-soluble preparation. Further, the deliquescent of the anhydrate causes the air to be prevented from blocking or the like during the treatment, and the hydrate has a good slidability, thereby improving the operability of the preparation.

Crystalline solids have higher chemical stability and physical stability than amorphous forms and low crystallinity forms, and they may also exhibit improved hygroscopicity, bulk properties, and or flow properties.

The discovery of useful pharmaceutically useful compounds for new crystalline hydrates provides a new opportunity to enhance the action characteristics of pharmaceutical products, which expands the formulation of pharmaceutical formulations by which the formulation scientist designs, for example, a drug having a target release profile or other desired properties. A library of materials, the field requires scutellarin hydrate.

 Surprisingly, characteristic, thermal analysis of the hydrate of the invention The (TG-DTA) map has a corresponding endothermic peak under the weightless platform, and the thermal analysis map shows the crystalline hydrate of scutellarin, such as 1.75, 2.0 hydrate, and the like.

The scutellarin hydrate of the present invention is a yellow or light yellow powder and can be stably stored. Refer to the Chinese Pharmacopoeia for the wettability test: take about 1g of scutellarin anhydrate and the hydrate of the present invention, placed in a dry constant weight watch glass, accurately weighed, 25 ° C, relative humidity of 75%, respectively The samples were sampled at 0h and 48h, and the percentage of wet weight gain was calculated. The results showed that the anhydrate-free wettability was much higher than that of the hydrate of the present invention, and the crystal hydrate of the scutellarin of the present invention was better. Stable storage, under the conditions of RH75% and 40 °C, the crystallization of scutellarin and the anhydrate of scutellarin were sealed in a vial and accelerated stability test. Chromatographic conditions: Chromatography Column: Kromasil C _l8 (200 mm × 4.6 mm, 5 μ m), mobile phase: methanol-0.1% phosphoric acid (40:60), detection wavelength 335 nm, flow rate 1 ml/min, and the magnitude of the increase in related substances was measured. The results are shown in Tables 1 to 2.

Table 1. Results of the wettigation test Sampling time (48 hours) % weight gain compared to 0 hours Scutellarin 2.0 hydrate 0.32% Scutellarin 1.75 hydrate 0.67% Scutellaria 6.89%

Table 2. Results of accelerated stability test Sampling time (June) Multiples of related substances compared to 0 months Scutellarin 2.0 hydrate 0.35 Scutellarin 1.75 hydrate 0.38 Scutellaria 1.61

 It is explained that the scutellarin crystal hydrate of the present invention has better storage stability.

Description of Drawings

 Figure 1 is a thermal analysis of scutellarin 1.75 hydrate.

 Figure 2 is a thermal analysis of scutellarin 2.0 hydrate.

Best Mode

Thermal analysis test conditions: Setaram Setsys 16, sample volume of about 3-10mg, heating rate: 10K / min, N ₂ flow rate: 50ml / min, temperature: room temperature ~ 400 °C.

 Surprisingly, characteristically, about 50 of the thermal analysis (TG-DTA or TG-DSC) map of the hydrate of the present invention ~ 165 °C or so weight loss platform has a corresponding endothermic peak.

Example 1 Preparation of scutellarin 1.75 Crystalline hydrate In the extraction tank, 2 kg of dried sorghum whole grass powder of about 40 mesh was refluxed three times with 200 ml of methanol, 12 L of ethanol and 2 L of water as a mixed solvent, and filtered. The mixture is coarsely filtered, and the extracts are combined, filtered through a 0.45-1 μm microporous membrane, and filtered through an ultrafiltration membrane with a molecular weight of 10,000 to 30,000. The filtrate is concentrated to almost no alcohol, and then adjusted with sodium bicarbonate solution. The pH of the diluent is about 7.2, filtered, and the filtrate is passed through D101 type macroporous adsorption resin, washed with water, and the eluate is collected, filtered, and the filtrate is adjusted to pH=2~3 with dilute phosphoric acid, and allowed to stand for analysis, and filtered. The precipitate was washed with water to be nearly neutral, washed with ethanol and chloroform, and suction filtered. The obtained solid was recrystallized twice with a solvent of methanol (2 ml, acetone 150 ml, chloroform 5 ml, water 10 ml), filtered, washed with water, and dried at about 45 ° C vacuum drying. 24 hours, yellow powder 1.1g, melting point: 193 °C gray (ELECTROTHERMAL MELTING POINT APPARATUS, uncorrected); UV: take 5 μg / ml of methanol solution, measured by spectrophotometry, found at wavelength 284nm and 335nm Maximum absorption at HPLC; HPLC: The main peak has the same retention time as the main peak of the commercially available scutellarin injection; ESI-MS: m/z: 461; infrared spectrum: ν ^KBr _max cm ^-1 3380 (width), 2920 1721, 1661, 1608, 1557, 1498, 1467, 1362, 1249, 1215, 1184, 1084, 1043, 923, 846, 740, 619, the moisture content is 6.61% by Karl Fischer method, thermal analysis: platform weight loss is about 6.37%, This is in the range of error with the sample containing 1.75 crystal water (theoretical value 6.39%) (see Figure 1), theoretical value of elemental analysis: C 51.07%, H4.39%; measured value: C 51.19%, H4. 46%.

Example 2 Preparation of scutellarin 2.0 hydrate In the extraction tank, 4 kg of 40 kg of scutellaria whole plant dry powder was refluxed 3 times with 32 L of ethanol and 4 L of water as a mixed solvent, and the mixture was filtered, and the filtrate was combined. Concentrated to almost no alcohol, diluted with water, and then adjust the pH of the diluted solution to 7.5 with sodium carbonate solution, filter with 0.2-1μm ceramic membrane, the filtrate is adsorbed with D101 macroporous resin, washed with water, the eluent is collected, filtered, and the filtrate is filtered. Adjust pH=2~3 with acetic acid, let stand, add to the precipitate for analysis, filter, precipitate with water to cause near neutrality, then wash with ethanol, chloroform, suction filtration, the obtained solid is diluted with water, and sodium carbonate solution is added under stirring. The solution is dissolved, and the pH of the diluent is adjusted to about 7.5, and the ultrafiltration membrane (polysulfone hollow fiber membrane module, fiber pore diameter of about 0.11 m1, inner diameter of about 1.1 mm) with molecular weight cut off of 8000 to 20000 is used for filtration. Acetic acid adjusted pH=2～3, allowed to stand, to be precipitated and analyzed, filtered, precipitated with water to cause near neutrality, washed with ethanol and chloroform, suction filtration, the obtained solid was methanol 2ml, ether 10ml, acetone 300ml, positive Butanol 5ml, water 30ml The solvent was recrystallized twice, filtered, washed with water, and dried under vacuum at 45 ° C for 6 hours to obtain a yellow powder of 2.87 g. Melting point: 192 ° C gray (uncorrected), UV spectrum: 5 μg/ml of methanol solution According to the spectrophotometric method, it is found that there is maximum absorption at wavelengths of 284 nm and 335 nm, and minimum absorption at 310 nm; infrared spectrum: ν ^KBr _max cm ^-1 3513 , 3376 (width), 2920 , 1721 , 1661 , 1608 , 1574 , 1498, 1467, 1442, 1361, 1249, 1225, 1183, 1150, 1083, 1043, 902, 845, 740, 619, 589, ESI-MS: m/z: 461; Karlscher's method for determining moisture 7.74%, heat Analysis: The weight loss of the platform is about 7.26%, which is within the error range of the sample containing 2 crystal water (theoretical value 7.23%) (see Figure 2). The theoretical value of elemental analysis: C 50.61%, H4.45%; Value: C 50.76%, H4.57%.

Example 3 Preparation of scutellarin 2 hydrate In the extraction tank, 2 kg of scutellaria whole plant dry powder of about 40 mesh was fully wetted with 80 L of ethanol and 3 L, and then ultrasonically extracted with ethanol 16 L and water 3 L as a mixed solvent. Next, the extracts were combined, filtered through a filter cloth, and filtered through a 0.45 μm microporous membrane, using an ultrafiltration membrane (polysulfone hollow fiber membrane module with a molecular pore size of about 0.001 to 30,000). About 1.1mm) Filtration, the filtrate is concentrated under reduced pressure to almost no alcohol, diluted with water, and then the pH of the diluted solution is adjusted to about 7.5 with sodium carbonate solution, filtered, D101 macroporous resin adsorption, water washing, collecting the eluent, Filtration, the filtrate is adjusted to pH=2~3 with acetic acid, and it is allowed to stand for analysis. After filtration, the precipitate is washed with water to cause near neutrality, washed with ethanol and chloroform, and suction filtered. The obtained solid is diluted with water and stirred with phosphoric acid. The sodium solution is dissolved, and the pH of the diluent is adjusted to about 7.5. The membrane is filtered with an ultrafiltration membrane with a molecular weight of 8,000 to 20,000. The filtrate is adjusted to pH=2 to 3 with acetic acid, and allowed to stand for analysis. The precipitate is washed with water to cause near neutrality. The mixture was washed with ethanol and chloroform, and suction filtered. The obtained solid was recrystallized twice with methanol (1 ml, isopropyl alcohol 5 ml, acetone 120 ml, acetic acid 5 ml, and water 10 ml), filtered, washed with water, and dried at 50 ° C for 4 hours under vacuum. Yellow powder 1.21g; Melting point: 189 °C gray (uncorrected), UV spectrum: 10 μg/ml methanol solution, maximum absorption at 284nm and 335nm, minimum absorption at 310nm; HPLC: main peak and Commercially available scutellarin injection main peak retention time is the same; infrared spectrum: ν ^KBr _max cm ^-1 3513, 3376 (width), 2920, 1721, 1661, 1608, 1574, 1498, 1467, 1442, 1361, 1249, 1225 , 1183, 1150, 1083, 1043, 902, 845, 740, 619, 589, ESI-MS: m/z: 461; Karlscher's method for determining moisture is 7.68%, thermal analysis for TG-DSC: platform weight loss is about 7.06%, This is in the range of error with the sample containing 2 crystal water (theoretical value 7.23%); theoretical value of elemental analysis: C 50.61%, H4.45%; measured value: C 50.55%, H4.61%.

 Example 4 Preparation of scutellarin 1.75 hydrate In the extraction tank, a 40-mesh lamp was planted. 3kg of dry powder was fully wetted with 60% ethanol 4L, then ultrasonically extracted with saturated sodium bicarbonate solution 180ml, ethanol 18L, water 4L as a mixed solvent, and the extract was combined, filtered through a filter cloth, and filtered through a 0.45 μm ceramic membrane. Filter with an ultrafiltration membrane with a molecular weight of 10,000 to 30,000, and adjust the pH of the dilution to 7.5 with sodium carbonate solution or acetic acid. Silica gel column, washed with water, the resulting liquid is passed through D101 macroporous adsorption resin, Washed, collected eluate, filtered, the filtrate was adjusted with acetic acid pH = 2 ~ 3, allowed to stand, to be precipitated and analyzed, filtered, precipitated with water to cause near neutral, washed with ethanol, chloroform, suction filtration, the resulting solid Dilute with water, add sodium carbonate solution to dissolve it under stirring, and adjust the pH of the diluent to about 7.5. Use an ultrafiltration membrane (polysulfone hollow fiber membrane module) with a molecular weight of 8,000 to 20,000. , The fiber pore size is about 0.Ol1μm, the inner diameter is about 1.1mm). The filtrate is adjusted with acetic acid to adjust the pH=2~3, and it is allowed to stand. After the precipitation is analyzed, it is filtered, and the precipitate is washed with water to cause near neutrality, and then ethanol and isohexanone are used again. Washed, suction filtered, used for solids 1 ml of methanol, 5 ml of isopropanol, 160 ml of acetone, 5 ml of ethyl acetate, 3 ml of acetonitrile, 10 ml of water were crystallized as a solvent, filtered, washed with water, and dried under vacuum at 50 ° C. Hour, yellow powder 1.56g; melting point: 194 °C grayed out (uncorrected), UV spectrum: 10 μg/ml methanol solution, wavelength 284nm and 335nm There is maximum absorption, minimum absorption at 310nm; HPLC: main chromatographic peak and commercial scutellarin injection main peak retention time is the same; ESI-MS: m / z: 461; The Karl Fischer method measures moisture of 6.79%, and the thermal analysis of TG-DSC: platform weight loss is about 6.32%, which is within the error range of the sample containing 1.75 crystal water (theoretical value 6.39%); Elemental analysis Theoretical value: C 51.07%, H4.39%; Found: C 50.95%, H4.51%. A small amount of sample was dried at 140 ° C in the presence of phosphorus pentoxide for 3 h to obtain an anhydride.

 Example 5 Preparation of scutellarin 2 hydrate freeze-dried powder needle scutellarin dihydrate 5g (calculated as anhydrous), sodium carbonate 0.6g, mannitol 5g, sodium dihydrogen phosphate 0.2g, sodium ascorbate 0.2g, EDTA calcium sodium 0.02g, plus 20-50 ° C water for injection about 250ml, stir to dissolve, adjust the pH with 5.5 ~ 7.1 with phosphoric acid and disodium hydrogen phosphate solution, add activated carbon 0.01 ~ 0.5% (W / V Stir for 15-30min, filter, make up water for injection to 300ml, filter with 0.22 micron microporous membrane or filter with ultrafiltration membrane with molecular weight of 6000~20000. 25, 50mg / bottle or 100mg / bottle (according to scutellaria sulphate anhydrate) divided, vacuum freeze-dried, tampon, to obtain lyophilized products.

 Example 6 Preparation of scutellarin 2 hydrate freeze-dried powder needle scutellarin hydrate 5g sodium bicarbonate 1g Add mannitol 30g, sodium ascorbate 1g, EDTA disodium 0.02g, add about 850ml of 20-50 °C water for injection, stir to dissolve, adjust pH with citric acid and sodium citrate solution For about 7.0, add activated carbon 0.3% (W / V) for 20min, filter, make water for injection to 1000ml, add activated carbon 0.01 ~ 0.5% (W / V Stir for 15-30min, filter, filter with 0.22 micron microporous membrane, press 5, 10, 20 or 50mg/ The bottle (according to the scutellaria sulphate anhydrate) is packaged, vacuum freeze-dried, tamponed, and lyophilized.

 Example 7 Preparation of scutellarin 2 hydrate freeze-dried powder Needle scutellarin hydrate 5g (as anhydrous), L- Arginine 1.8g, sorbitol 15g, sodium dihydrogen phosphate 0.5g, sodium tartrate 0.2g, cysteine hydrochloride 0.1g, EDTA calcium sodium 0.02g, plus 30-50 ° C water for injection about 400ml, stir to dissolve, adjust the pH with tartaric acid and sodium tartrate solution to 5.6 ~ 6.5, add activated carbon 0.5% (W / V) and stir for 30min , filter, make up water to 500ml, filter with 0.22 micron microporous membrane or filter with ultrafiltration membrane with molecular weight of 6000 ~ 20000, press 20, 50mg / Bottles or 100mg/bottle (as anhydrous) are packaged, vacuum freeze-dried, and tamponed to obtain a lyophilized product.

 Example 8 Preparation of scutellarin 2 hydrate freeze-dried powder needle scutellarin 2 hydrate 5g (According to anhydrous matter), lysine 1.47g, mannitol 10g, sodium dihydrogen phosphate 0.5g, sodium cysteine hydrochloride 0.1g, EDTA disodium 0.05g, plus 30-50 ° C water for injection about 420ml, stir to dissolve, adjust the pH with citric acid and sodium citrate solution to 5.6 ~ 6.2, add 0.5% (W / V) activated carbon stirring 30min, filter, make up water to 500ml, filter with 0.22 micron microporous membrane or filter with ultrafiltration membrane with molecular weight of 6000 ~ 20000, press 5, 10 20, 50mg/bottle or 100mg/bottle (according to scutellaria sulphate anhydrate), vacuum freeze-drying, tamping, lyophilized product.

 Example 9 Preparation of scutellarin 2 hydrate injection Injection scutellarin hydrate (according to dry product) 10g , disodium hydrogen phosphate 4g, sodium citrate 2g, sodium hydrogen sulfite 1g, propylene glycol 10ml, EDTA disodium 0.1g Under the strong light, add water for injection, pass nitrogen, stir to dissolve, adjust the pH of citric acid and sodium citrate solution to 5.6 ~ 6.3, add activated carbon 0.1% (W / V) and stir 15 ~ 45min, filter, hydrate to about 2000ml, filter with 0.22 micron microporous membrane or use the molecular weight of interception 6000 ~ 20000 The ultrafiltration membrane is filtered, the solution is saturated with nitrogen, and the finished product is sterilized according to the main drug 10, 20, 25, 40 mg, 50 mg or 100 mg/min.

 Example 10 Preparation of scutellarin 1.75 hydrate Injection preparation scutellarin hydrate (according to dry product) 5g , Tween-80 20ml, cysteine hydrochloride 1g, propylene glycol 20ml, EDTA disodium 0.1g Under the strong light, add water for injection, pass nitrogen, stir to dissolve, adjust the pH of citric acid and sodium citrate solution to 5.6 ~ 6.3, add activated carbon 0.1% (W / V) and stir for 45min. , filter, hydrate to about 2000ml, filter with 0.22 micron microporous membrane or filter with ultrafiltration membrane with molecular weight of 6000 ~ 20000, the solution is saturated with nitrogen, according to the main drug 2.5, 5, 10mg / branch, sterilized finished product.

 Example 11 Preparation of scutellarin 2 hydrate injection Injection scutellarin 2 hydrate (by dry product) 10g , arginine 4g, sodium citrate 2g, cysteine hydrochloride 1g, propylene glycol 20ml, EDTA disodium 0.2g Under the strong light, add water for injection, pass nitrogen, stir to dissolve, adjust the pH of citric acid and sodium citrate solution to 5.6 ~ 6.3, add activated carbon 0.1% (W / V) and stir for 35min, Filter, hydrate to about 5000ml, filter with 0.22 micron microporous membrane, the solution is saturated with nitrogen, according to the main drug 10, 20, 25, 40mg or 50mg / Packed and sterilized to obtain finished products.

 Example 12 Preparation of scutellarin 2 hydrate injection Glucose 250g Add water for injection, stir to dissolve completely, add 0.5% activated carbon, heat for 10-30 minutes, filter and decarbonize through sand filter; scutellarin 2 hydrate (according to dry product) 5g Mix well with the above filtrate, add 3 g of sodium dihydrogen phosphate, 1 g of sodium tartrate, 20 ml of glycerol, 1 g of sulfobutyl-β-cyclodextrin, disodium EDTA 0.05 g After the dissolution is complete, add water for injection to 5000ml, adjust the pH value to 4.5-6.0 with 1-3M hydrochloric acid or lactic acid or citric acid and sodium citrate solution, and add 0.05. % of activated carbon, heated and stirred for about 10-30 minutes, filtered and decarbonized, and then filtered once or twice through a 0.22 um microporous membrane, the solution is saturated with high-purity nitrogen, and the solution is tested according to the main drug 25 50mg or 100mg potting, sterilization, packaging is available.

 Example 13 scutellarin 1.75 hydrate Preparation of sodium chloride infusion: scutellarin 51.7 hydrate (as anhydrous) 25g, sodium chloride 425g, sodium citrate 5g, EDTA disodium 1g Add water for injection, the solution is saturated with high-purity nitrogen, stir to completely dissolve, adjust the pH value with 5.0~6.5 with citric acid solution and sodium citrate solution, add water for injection to 50L, add liquid volume 0.05% activated carbon, stirred for about 10-30 minutes, filtered for decarbonization, and then subjected to 0.22um membrane fine filtration or with a molecular weight of 6,000 to 20,000. Ultrafiltration membrane filtration, semi-finished test, after its content, pH value and clarity are qualified, under high purity nitrogen, filled in 50ml, 100ml or 250ml In bottles or plastic bags, sterilize and package.

 Example 14 Preparation of scutellarin 2 hydrate Sodium chloride infusion: scutellarin 2 hydrate (as anhydrous) 20g, sodium chloride 425g, sodium tartrate 5g, EDTA disodium 0.5g Add water for injection, the solution is saturated with high-purity nitrogen, stir to completely dissolve, adjust the pH value with 5.0-6.5 with tartaric acid solution and sodium tartrate solution, add water for injection to 50L, add liquid volume 0.05% activated carbon, stirred for about 10-30 minutes, filtered and decarburized, and then finely filtered through a 0.22um microporous membrane for 2 times. After semi-finished test, its content and pH are determined. After passing the value and clarity, it can be filled in a 50ml, 100ml or 250ml bottle or plastic bag under high purity nitrogen, sterilized and packaged.

 Example 15 scutellarin hydrate tablet (50 mg/tablet)

 Prescription: scutellarin 2 hydrate 50g

 Microcrystalline cellulose 135g

 Lactose 10g

 Sodium Carboxymethyl Starch 5g

 --cyclodextrin 5g

 5 % PVP 30 (50% aqueous ethanol solution)

 Magnesium stearate 2g

 Use scutellarin 2 hydrate, microcrystalline cellulose, lactose, β-cyclodextrin, sodium carboxymethyl starch over 100 mesh sieve, with 5 % PVP 30's 50% ethanol solution is made of soft binder. It is sieved through 18 - 24 mesh, dried, and sieved through 14 - 20 mesh. Add magnesium stearate and mix.

 Example 16 Preparation of scutellarin hydrate capsule (50 mg/capsule)

 Prescription: scutellarin 2 hydrate 50g

 Microcrystalline cellulose 65g

 Lactose 10g

 Sodium Carboxymethyl Starch 5g

 --cyclodextrin 5g

 5 % PVP 30 (50% aqueous ethanol solution)

 Magnesium stearate 2g

 盏 盏 乙 2 2 水 hydrate, microcrystalline cellulose, lactose, β-cyclodextrin, sodium carboxymethyl starch over 100 mesh sieve, with 5 % PVP 30's 50% aqueous solution of ethanol is made of soft material made of adhesive. It is sieved through 18 - 24 mesh, dried, and sieved through 14 - 20 mesh. Mix magnesium stearate and fill the capsules.

 Example 17 Preparation of scutellarin hydrate dispersible tablet (50 mg/tablet)

 Prescription: scutellarin 2 hydrate 50g

 Microcrystalline cellulose 135g

 Lactose 10g

 Polyethylene glycol 6000 3g

 Low substituted hydroxypropyl cellulose 10g

 Sodium Carboxymethyl Starch 5g

 --cyclodextrin 5g

 Aspartan 1g

 5 % PVP 30 (50% aqueous ethanol solution)

 Magnesium stearate 2g

 It is scutellarin 2 hydrate, microcrystalline cellulose, lactose, polyethylene glycol 6000, low-substituted hydroxypropyl cellulose, β-cyclodextrin, Sodium carboxymethyl starch and aspartame were passed through a 100 mesh sieve, and a soft material of 5 % PVP 30 in 50% ethanol was used as a binder. The granules were sieved through 18 - 24 mesh, dried, and passed 14 - After 20 mesh sieves, add magnesium stearate that has passed through a 100 mesh sieve and mix.

 Example 18 Preparation of scutellarin hydrate capsule (50 mg/capsule)

 Prescription: scutellarin 1.75 hydrate 50g

 Microcrystalline cellulose 65g

 Corn starch 10g

 Low substituted hydroxypropyl cellulose 5g

 --cyclodextrin 5g

 5 % PVP 30 (50% aqueous ethanol solution)

 Micronized silica gel 2g

 Using scutellarin hydrate, microcrystalline cellulose, corn starch, β-cyclodextrin, low-substituted hydroxypropyl cellulose through a 100 mesh sieve, 5% of PVP 30's 50% ethanol solution is made of soft binder. It is sieved through 18 - 24 mesh, dried, and sieved through 14 - 20 mesh. Mix the micro-silica gel and fill the capsules.

 Example 19 Preparation of scutellarin 1.75 hydrate tablet (50 mg/tablet)

 Prescription: scutellarin 1.75 hydrate 50g

 Mannitol 120g

 Lactose 25g

 Sodium Carboxymethyl Starch 5g

 --cyclodextrin 5g

 5 % PVP 30 (50% aqueous ethanol solution)

 Magnesium stearate 2g

 Use scutellarin hydrate, mannitol, lactose, β-cyclodextrin, sodium carboxymethyl starch over 100 mesh sieve, with 5% PVP 30% of 50% ethanol aqueous solution is made of soft material for binder. It is sieved through 18-24 mesh, dried, and sieved through 14-20 mesh. After adding magnesium stearate, it is mixed and compressed.

 Example 20 Preparation of scutellarin hydrate granules (100 mg/pack)

 Prescription: scutellarin hydrate hydrate 100g (based on anhydrous matter)

 Mannitol 185g

 Lactose 710g

 Methyl cellulose 3g

 Xanthan gum 5g

 Solid food flavor 1g

 5 % hydroxypropyl methylcellulose

 Use scutellarin hydrate, mannitol, lactose, methylcellulose, xanthan gum, food flavor over 100 mesh sieve, use 5 % of hydroxypropyl methylcellulose in 50% ethanol solution is made of soft material made of binder, granulated by 18 - 24 mesh, dried at 60 ° C or below, after 14 - 20 mesh sieve, after centrifugation Packaging.

 Example 21 Preparation of scutellarin hydrate suspension (25 mg/pack)

 Prescription: scutellarin hydrate 25g (based on anhydrous matter)

 Mannitol 165g

 Lactose 780g

 Xanthan gum 8g

 Solid food flavor 1g

 5 % hydroxypropyl methylcellulose

 The scutellarin hydrate, mannitol, lactose, and food flavor are passed through a 100 mesh sieve, using 50% of 5% hydroxypropyl methylcellulose. Appropriate amount of ethanol aqueous solution is soft material made of adhesive. It is sieved through 18 - 24 mesh, dried below 60 °C, and sieved through 14 - 20 mesh. The xanthan gum passed through a 100 mesh sieve was mixed with the above granules and packaged.

 Example 22 Preparation of scutellarin hydrate pellets

 Prescription: scutellarin 2 hydrate 25g (based on anhydrous matter)

 Polyethylene glycol 6000 325g

 Polyethylene glycol 4000 30g

 Stearic acid 20g

 The polyethylene glycol 6000, polyethylene glycol 4000, and stearic acid are heated to melt, and the saponin is passed through a 100 mesh sieve. The hydrate is added thereto, stirred and mixed, and dropped into vegetable oil or dimethicone under heat preservation, and then cooled to a pellet, and the dropping pills are collected, and dried.

 Example 23

 The pharmacological test is as follows:

 48 Wistar male rats weighing 250-300 g were randomly divided into 4 groups: sham operation group, ischemia reperfusion group And scutellarin 1.75 hydrate group, scutellarin 2 hydrate group, 12 in each group. The drug group of the present invention: using scutellarin hydrate The saline was administered intraperitoneally at a dose of 75 mg/kg twice daily for 7 days, and the last dose was 60 minutes after cerebral ischemia surgery; sham operation and ischemia reperfusion The groups were given an equal volume of physiological saline.

 Rats were anesthetized with 10% chloral hydrate (3.5 ml/kg) intraperitoneally and fixed in supine position. Zea was used in the literature. Longa line 拴 improved method modeling [Journal of Shanghai Jiaotong University (Medical Science) 2007, 27 (10): 1218-1222]; the sham operation group was inserted only 10mm, and the rest of the steps were the same as the model group. After the animal was awakened, the surgical side Homer sign and the contralateral dyskinesia appeared. After successful modeling, the drug group of the present invention was administered twice a day, the dose was the same as before, the sham operation group and ischemia The reperfusion group was given an equal volume of physiological saline. The line was withdrawn 2 hours after model establishment, and the rats were sacrificed 24 hours after reperfusion, and the brain was quickly weighed. 10% brain tissue homogenate prepared with physiological saline at low temperature, low temperature centrifugation, 3000 After centrifugation for 10 min at r/min, the precipitate was discarded, and the supernatant was taken. The SOD and MDA were determined according to the specifications of the commercially available superoxide dismutase SOD, malondialdehyde MDA, nitric oxide NO, and nitric oxide synthase NOS kit. , NO, NOS content or activity, the results are shown in Table 3.

 Table 3. Effects of the drug of the present invention on NO, NOS, MDA and SOD in brain tissue of rats with cerebral ischemia-reperfusion

 NO (μmol/L)  NOS (U/ml)  MDA (nmol/ml)  SOD (U/ml)  mock surgical group  17 ±4.26  16 ±4.64  1.47 ±0.53  218 ±24.07  Ischemia reperfusion group  26 ± 6.92  26 ±6.07  2.91 ±0.66  134 ± 19.25  Scutellarin 1.75 hydrate group  20 ±6.32  20 ±5.51  2.07 ±0.78  181 ±23.61  Scutellarin 2 hydrate group  21 ±7.54  19 ±5.74  2.02 ±0.71  179 ±19.36

 The results showed that ischemia reperfusion was compared with the sham operation group. The SOD activity in the brain tissue of the model group was significantly decreased, and the MDA, NO content and NOS activity were significantly increased (P<0.01). Compared with the model group, the activity of SOD in the brain tissue of the drug of the present invention was significantly increased, the activity of NOS was decreased, and the contents of MDA and NO were significantly decreased (P<0.01).

With the deepening of the understanding of the pathological mechanism of ischemic brain injury, it has been found that oxygen free radicals, excitatory amino acids, nitric oxide (NO), monoamine transmitters, Ca ²⁺ overload and other factors are involved in the damage process. . In ischemic brain injury, a large amount of oxygen free radicals are produced during ischemia. SOD is the main enzymatic defense system against oxygen free radicals in the cell. It removes superoxide anion radicals by disproportionation and protects the organism from free radical attack. It has been confirmed that SOD can significantly alleviate the damage caused by cerebral ischemia, not only to give anti-cerebral ischemia injury to exogenous SOD, but also to antagonize or promote cerebral ischemia by knocking out or transferring SOD gene into transgenic animals. Damage. It can be seen that the level of SOD directly reflects the body's ability to scavenge oxygen free radicals.

A variety of different oxygen free radicals attack the unsaturated fatty acids in the biofilm, triggering lipid peroxidation, forming a series of lipid free radicals and their degradation products MDA. MDA acts as a metabolite of lipid peroxidation of oxygen free radicals and biofilm unsaturated fatty acids, and its content changes indirectly to reflect changes in oxygen free radical content in tissues.

Studies have shown that NO plays an important role in the pathogenesis of cerebral ischemic injury and myocardial ischemic injury. NO content increases significantly in the first few minutes after ischemia, then slowly decreases, and NO increases again during reperfusion. The mechanism may be: neuronal damage after cerebral ischemia, enhanced cell membrane depolarization process, increased excitatory amino acid production such as presynaptic glutamate, increased extracellular glutamate concentration, and activation of N-methyl -D-aspartate (NMDA) receptor, NMDA receptor activation increases post-synaptic Ca ²⁺ influx, activates nitric oxide synthase NOS, promotes NO production; second, after cerebral ischemia, A large consumption of ATP leads to a decrease in energy metabolism disorder and cAMP-dependent protein kinase activity, and an increase in NOS activity due to dephosphorylation of NOS, thereby promoting NO production.

Nitric oxide synthase is the rate-limiting enzyme NOS for nitric oxide synthesis. In the early stage of cerebral ischemia, NOS-mediated NO promotes blood supply in the ischemic area by dilating blood vessels, which has a short-term protective effect, but with the large amount of NO, NOS in the ischemic area mediates neurotoxic effects. It will soon be dominant, which will aggravate cerebral ischemic injury. In the advanced stage, cerebral ischemic brain tissue damage, inflammatory response stimulate macrophages, neuroglia, neurons and so on can produce inducible NOS in large quantities, and inducible NOS can be slow. The NO is produced in a long-lasting manner, and excessive NO is produced and released, which aggravates neuronal damage. Therefore, reducing the production of inducible NOS can significantly reduce the release of NO, thereby reducing the cytotoxic effect of NO and protecting brain cells.

The results of this study showed that the levels of NO and NOS in brain tissue increased significantly after cerebral ischemia-reperfusion injury in rats, indicating that NO and NOS are involved in the pathogenesis of cerebral ischemia-reperfusion injury. The contents of NO and NOS decreased significantly. It is indicated that the drug of the present invention exerts a protective effect on cerebral ischemia-reperfusion injury or cerebral infarction by regulating NOS activity and NO content.

Cerebral ischemia and reperfusion injury can lead to lipid peroxidation of brain cell membrane, produce excessive oxygen free radicals, and damage brain cell membrane. In cerebral ischemia, due to insufficient supply of tissue oxygen and energy metabolism, ATP production is reduced, ion pump failure, Na ⁺ -K ⁺ -ATPase activity is reduced, a large amount of Na ⁺ influx, K ⁺ outflow, Cl ^- and H ₂ O passively enters the cell, causing acute osmotic swelling of nerve cells to die. Na ⁺ influx and K ⁺ outflow lead to depolarization of cell membrane potential, voltage-dependent Ca ²⁺ channel opening, and massive Ca ²⁺ influx; and due to the release of K+, protein kinase C and transmitter, receptors The dependent Ca ²⁺ channel is open and there is a large influx of Ca ²⁺ . Intracellular calcium overload can lead to increased oxygen free radical production, increased arachidonic acid metabolism, increased excitatory amino acid transmitter release, etc. Excitatory amino acids can cause intracellular Ca ²⁺ overload to cause neuronal apoptosis, including oxygen free radicals. Increase is an important factor in ischemia-reperfusion injury. In addition, neuronal damage caused by increased free radicals caused by cerebral ischemia, increased calcium influx, increased nitric oxide caused by cerebral ischemia, and induced NOS are also important factors in vascular dementia. The anti-oxidation effect of cerebral ischemia, anti-free radical action, and reduction of nitric oxide production caused by ischemia provide a basis for the treatment of vascular dementia.

 Therefore, the medicament of the invention can significantly reduce the content of lipid peroxide MDA and NO in the brain tissue of the ischemic area, and improve The content of SOD in rat brain tissue increases the ability to scavenge free radicals and reduces the activity of NOS, which can alleviate the damage of brain tissue caused by free radical reaction. This shows that The drug of the invention inhibits the formation of free radicals and the like, protects the cell membrane structure, helps to reduce cerebral vascular permeability and improve blood rheology index. Improve brain microcirculation, play a protective role on the brain against the damage of oxygen free radicals on brain tissue after cerebral ischemia and reperfusion. Can be used for treatment or prevention Ischemic encephalopathy, cerebral infarction, cerebral thrombosis and its sequelae, vascular dementia, improving the brain microcirculation, and can be used to prepare drugs for the treatment and prevention of the corresponding diseases.

It is to be understood that many variations of the details are possible from the point of view of the present invention, which is not intended to limit the scope and spirit of the invention, and the invention is not limited to the embodiments described above.

Mode for Invention

Industrial Applicability

Sequence List Text

Claims (1)

1 . A cardio-cerebral vascular therapeutic drug, characterized in that the cardio-cerebral vascular therapeutic drug is a scutellarin derivative, namely scutellarin hydrate, and the molecular formula thereof is C ₂₁ H ₁₈ O ₁₂ ·nH ₂ O , n A number between 1.75 and 2.0. The cardio-cerebral vascular therapeutic agent according to claim 1, wherein the scutellarin hydrate is scutellarin 2.0 hydrate . The cardio-cerebral vascular therapeutic agent according to claim 1, wherein the scutellarin hydrate is scutellarin 1.75 hydrate . A method for preparing scutellarin hydrate according to claim 1, wherein the steps are as follows: Method A In the reaction vessel, the scutellaria flower is used in a volume ratio of 2 to 20 times the volume of water, or a C ₁ -C ₆ low molecular alcohol, a C ₂ -C ₆ low molecular nitrile, or a C3-C8 low grade. One or more of the ketone, C ₂ -C ₈ low molecular ester and the like are extracted once to several times as a solvent, and the extract is filtered and combined, the filtrate is concentrated, the concentrate is diluted with water, and the pH is adjusted to 6.5 with alkali. Between 9, filtration, silica gel column, or macroporous adsorption resin, or polyamide column chromatography or glucose gel resin, water or C ₁ -C ₆ low molecular alcohol, C ₃ -C ₈ lower ketone One or more of C ₂ -C ₆ low molecular weight nitriles are eluted, the filtrate is adjusted to pH 1-5 with acid, stirred, placed, and the precipitate is charged and analyzed, and the resulting precipitate is filtered, and water and C ₁ -C a low molecular alcohol of ₆ , a C ₂ -C ₈ low molecular ester, a C ₃ -C ₈ lower ketone, a C ₂ -C ₆ low molecular nitrile, a C ₂ -C ₆ lower ether, a C ₁ -C ₆ One or more of a lower halogenated hydrocarbon or a C ₁ -C ₈ low molecular acid or the like is a crystallization solvent, subjected to recrystallization once or several times, filtered, washed with water or a C ₁ -C ₆ low molecular alcohol, or C ₃ - C ₈ low level Washing one or more of a ketone, a C ₂ -C ₆ low molecular nitrile, etc., and drying the obtained solid to obtain a scutellarin crystal hydrate; Or method B In the reaction vessel, the scutellaria whole plant is extracted once or several times with one or more of water, an alkaline solution or a low molecular weight alcohol of C ₁ -C ₆ in a volume ratio of 2 to 20 times. Next, filter, combine the filtrate, concentrate the filtrate, dilute the concentrate with water, adjust the pH to 6.5-9 with acid or alkali, filter, filter the silica gel column and macroporous adsorption resin, or polyamide column chromatography or Glucose gel resin, eluted with water or _one or more of C ₁ -C ₆ low molecular alcohol, C ₃ -C ₈ lower ketone, C ₂ -C ₆ low molecular nitrile, etc. Adjusting the pH to 1 to 5, placing, precipitating the precipitate, filtering the resulting precipitate, using water or C ₁ -C ₆ low molecular alcohol, C ₂ -C ₈ low molecular ester, C ₃ -C ₈ lower ketone, One or more of C ₂ -C ₆ low molecular nitrile, C ₂ -C ₆ lower ether, C ₁ -C ₆ lower halogenated hydrocarbon, or C ₁ -C ₈ low molecular acid, etc. The solvent is subjected to recrystallization once or several times, filtered, washed with water, and the obtained solid is dried to obtain scutellarin crystal hydrate. 5. The use of scutellarin hydrate according to claim 1, characterized in that: for preparing cerulein hydrate containing The pharmaceutical composition for preparing a pharmaceutical composition comprising scutellarin hydrate comprises: a preparation for lyophilized powder injection for injection, or a sterile powder preparation, or a large infusion preparation, or a small water needle An injection or a solid preparation, and the solid preparation includes a tablet, a capsule, a granule, a suspension, and a dropping pellet. 6. The use of scutellarin hydrate according to claim 1, characterized in that it is suitable for preparation Human and mammalian coronary heart disease, angina pectoris, myocardial infarction, arrhythmia, ischemic brain disease, cerebral thrombosis, cerebral hemorrhage and its sequelae, vascular dementia, Peripheral circulatory disorders, including various arterial occlusive diseases, vasculitis, microcirculatory disorders caused by diabetes, hypertension, hyperlipidemia, Hyperviscosity; application of refractory cervical syndrome, vertebrobasilar insufficiency and other diseases of ischemia and microcirculatory disorders for the treatment or prevention of diseases. 7. The method for preparing scutellarin hydrate according to claim 4 , characterized in that the filtration method in the preparation process of the present invention comprises using a filter cloth, a funnel, a fused glass filter, a sand core filter rod, a plate and frame filter press, a microporous membrane, a ceramic membrane or a super One or several filtration methods such as filtration are performed one or several times. The method for preparing scutellarin hydrate according to claim 4, wherein in the preparation of the two methods of claim 4, the pH of the extract is adjusted to 6.8 to 9 by alkali. In between, filtration, filtrate through silica gel column and macroporous adsorption resin, or polyamide column chromatography or glucose gel resin, water or C ₁ -C ₆ low molecular alcohol, C ₃ -C ₈ lower ketone, C One or more of the low molecular weight ester of ₂ -C _{8 and} the low molecular weight nitrile of C ₂ -C ₆ are eluted, and the filtrate is adjusted to pH 1-5 with an acid, and the precipitate is charged to analyze the process of the process. In the preparation process, the precipitate obtained by filtering the acid precipitate may be diluted with water and then the above process is repeated for 1 to 3 times. The method for preparing scutellarin hydrate according to claim 4, wherein the recrystallization solvent is selected from the group consisting of Water, and one of the following solvents: methanol, ethanol, isopropanol, acetone, methyl ethyl ketone, isohexanone, acetonitrile, tetrahydrofuran, methyl acetate, ethyl acetate, acetic acid, dichloromethane, chloroform, diethyl ether, petroleum ether, benzene Kind or several.

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