CN110642909A

CN110642909A - Troxerutin compound

Info

Publication number: CN110642909A
Application number: CN201910991486.2A
Authority: CN
Inventors: 王明
Original assignee: Shaanxi Fotboll Pharmaceutical Co Ltd; Yining Elxing Intellectual Property Service Co Ltd; Hainan Dons Pharmaceutical Technology Co Ltd
Current assignee: Shaanxi Fotboll Pharmaceutical Co Ltd; Yining Elxing Intellectual Property Service Co Ltd; Hainan Dons Pharmaceutical Technology Co Ltd
Priority date: 2019-10-18
Filing date: 2019-10-18
Publication date: 2020-01-03

Abstract

The invention discloses a troxerutin compound prepared by an advanced online process control technology. The troxerutin compound is determined by X-ray powder diffraction, and an X-ray powder diffraction pattern expressed by a 2 theta diffraction angle shows characteristic diffraction peaks at the positions of 8.15 +/-0.2 degrees, 9.24 +/-0.2 degrees, 12.31 +/-0.2 degrees, 17.10 +/-0.2 degrees, 17.92 +/-0.2 degrees, 18.73 +/-0.2 degrees and 23.45 +/-0.2 degrees of 2 theta. The compound has the characteristics of high purity, low impurity content, good fluidity and good stability.

Description

Troxerutin compound

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a troxerutin compound prepared by an advanced online process control technology.

Background

The chemical name of Troxerutin (Troxerutin) is: 7, 3 ', 4' -troxerutin, molecular formula: c₃₃H₄₂O₁₉Molecular weight: 742.69, CAS number: 7085-55-4, the structural formula is as follows:

the product is a hydroxy ethyl rutin mixture containing troxerutin (7, 3 ', 4' -trihydroxyethyl rutin). The troxerutin content is not less than 88.0% (calculated on anhydrous substance) for injection. The product is yellow or yellowish green powder; no odor; has hygroscopicity, is soluble in water, slightly soluble in methanol, slightly soluble in ethanol, and insoluble in chloroform; the absorption maxima were determined spectrophotometrically at 254nm and 347nm, and the absorption minima at 283 nm.

Troxerutin, namely trioxyethylrutin, troxerutin, vitamin P4, and hydroxyethyl rutin, is a semisynthetic flavone compound prepared by hydroxyethylating rutin, and has effects of inhibiting erythrocyte and platelet aggregation, preventing thrombosis, increasing oxygen content in blood, improving microcirculation, and promoting angiogenesis to promote collateral circulation. It has protective effect on endothelial cells, can resist vascular injury caused by 5-hydroxytryptamine and bradykinin, increase resistance of capillary vessels, reduce permeability of capillary vessels, has the effect of preventing edema caused by rise of vascular permeability, and has remarkable protective effect on acute ischemic brain injury; it also has radioprotective, antiinflammatory, antiallergic, and antiulcer effects. The Chinese medicine composition is clinically applicable to hemiplegia, aphasia, pre-myocardial infarction syndrome, arteriosclerosis, central retinitis, thrombophlebitis, varicose veins, edema caused by increase of vascular permeability and the like caused by cerebral thrombosis and cerebral embolism.

The prior troxerutin dosage forms comprise granules, capsules, oral liquid, tablets, injection, freeze-dried powder injection and the like, wherein the oral preparation has low bioavailability, and the injection has higher bioavailability compared with the oral preparation, however, the stability of the injection and the freeze-dried powder injection prepared at present is still poor, the content of troxerutin is obviously reduced after long-time standing, and other hydroxyethyl rutin derivatives as impurities are obviously increased due to degradation, so that the quality requirements are not met, therefore, the effective period of the injection in the prior art is about one year and half under the shading condition, because the production and circulation links of the medicine need a certain time, the actual usable time is very short, the effective use period is easily exceeded, and the medicine cannot be used, there is therefore a need to improve the overall stability of the formulation to facilitate clinical use and to facilitate production and storage.

At present, troxerutin products in the market generally contain 80.0 percent (for oral administration) and 88.0 percent (for injection) of troxerutin as main components. The troxerutin for injection contains 12.0% of impurities, and the existing impurities bring great hidden troubles to clinical medication, wherein the reported adverse reactions caused by troxerutin injection are more than allergic reactions. Therefore, the purity of the troxerutin raw material medicine is improved, and the method has important practical significance.

Because the flavone structure of rutin has four active hydroxyl groups, under the alkaline condition, the four hydroxyl groups and ethylene oxide can generate Williamson etherification nucleophilic substitution reaction, although the four hydroxyl groups have different activities, the difference is small, and single troxerutin is difficult to obtain in the reaction.

The preparation method of high-content troxerutin reported in the literature generally comprises the following steps: (1) in the patent with the application number of 200610065718.4, water is used as a solvent, sodium hydroxide is used as a catalyst, ethylene oxide is used as an etherification reagent under normal pressure, the pH value of a reaction liquid end point is regulated and controlled to be 9.5-10.3 by resin in the reaction process, and the content of the obtained product is more than 85.0 percent; (2) in a patent with the application number of 201210443466.X, under high pressure, methanol is used as a solvent, sodium hydroxide is used as a catalyst, and ethylene oxide is used as an etherification reagent, so that the content of the obtained product is 88.0-93.0%; (3) in the patent with application number 201310028281.7, water or water-alcohol mixture is used as solvent, sodium hydroxide or potassium hydroxide is used as catalyst, epoxy ethane is used as etherification reagent, the reaction is carried out in two stages, the first stage generates the product mainly containing dihydroxyethyl rutin and trihydroxyethyl rutin, and the second stage adds borax for protecting C₅-OH, the proportion of the tetrahydroxyethyl rutin is reduced, and the content of the obtained troxerutin reaches more than 90.0 percent; (4) jiangzhengxiang and the like take troxerutin with the content of 81% as a raw material, purify the troxerutin by adopting a preferential crystallization method, dissolve a troxerutin crude product into a mixed solvent of methanol and isopropanol with the volume ratio of 27:1 at the temperature higher than 65 ℃, and then cool to 20-25 ℃, so that the content of the obtained troxerutin reaches more than 95%; (5) in the patent with application number 200310104132.0, methanol is used as a solvent, pyridine is used as a catalyst, the obtained crude troxerutin is refined by using an organic solvent (methanol and ethanol), and the content of the finally obtained product reaches 90.0 percent; (6) li Jun et al use C18 column, water-acetonitrile (volume ratio is 83:17) as mobile phase, separate and purify troxerutin raw materials by preparative HPLC to obtain troxerutin reference substance, in the patent with purity up to 99.0% (7) publication No. CN104177461A, rutin is first prepared into 7-monohydroxyethylrutin, then the 7-monohydroxyethylrutin is hydroxyethylated, the obtained crude troxerutin is refined to finally obtain troxerutin with content of 98.0%, troxerutin with content higher than 98.0% is obtained by the method, and cost is 2000-3000 yuan/kg.

In the existing reports, the method for increasing the content of troxerutin to more than 98.0% only needs chromatography, is complicated to operate, has high cost, and is not suitable for industrial popularization and application.

In order to solve the problems, it is important to prepare a troxerutin compound and a preparation thereof, wherein the troxerutin compound has good purity, low impurity content and stable properties.

The invention mainly aims at the problems, the applicant sufficiently considers the factors such as temperature, pH value, reaction time, additives and the like in the refining and refining processes of the initial raw materials to obtain the troxerutin compound which has high purity, low impurity content, higher safety and stability, the production process is simple, and the used raw materials, reagents and the like are all products which are cheap and easy to obtain, so that the troxerutin compound is suitable for industrial mass production.

Disclosure of Invention

In order to overcome the defects of low purity and high impurity content of troxerutin prepared by the prior art, the invention provides a troxerutin compound and a preparation method thereof, wherein the troxerutin compound is obtained by performing detailed parameter control on each step by utilizing an advanced online process control technology, and an X-ray powder diffraction pattern expressed by a 2 theta diffraction angle shows characteristic diffraction peaks at 8.15 +/-0.2 degrees, 9.24 +/-0.2 degrees, 12.31 +/-0.2 degrees, 17.10 +/-0.2 degrees, 17.92 +/-0.2 degrees, 18.73 +/-0.2 degrees and 23.45 +/-0.2 degrees. The method obviously improves the control level of the crystal form, solvent residue and impurities of the troxerutin, the crystal form is unique, and the content of the solvent residue and the impurities is far lower than that of the existing product.

The preparation prepared by the compound provided by the invention is simple in preparation process, does not need any excipient, and has better stability and small side effect.

The inventors of the present invention have intensively studied and found that a troxerutin compound having a high purity and a low impurity content is preferably obtained by a preparation method comprising the steps of:

(1) mixing and reacting dimethylformamide, anhydrous potassium carbonate and rutin, heating, dropwise adding 2-bromoethanol, keeping the temperature for reaction until the rutin disappears, cooling to room temperature, dropwise adding ethyl acetate, filtering, and drying to obtain a yellow solid troxerutin crude product;

(2) adding the troxerutin crude product into a mixed solution of dimethylformamide and acetone, heating, stirring for dissolving, filtering, cooling to room temperature, crystallizing overnight, filtering, pulping wet product with acetone, filtering, and drying to obtain a primary refined troxerutin product;

(3) and adding the refined troxerutin product into the mixed solution of isopropanol and purified water, heating, stirring and dissolving, cooling to room temperature, keeping the temperature, stirring, filtering, washing with anhydrous methanol, and drying to obtain the refined troxerutin product.

As a preferred embodiment of the invention, in the step (1), the heating temperature is 80-85 ℃, and the drying temperature is 50-60 ℃;

in a preferred embodiment of the invention, in the step (2), the mixture is heated to 65-70 ℃, stirred and dissolved, and the drying temperature is 50-60 ℃;

as a preferred embodiment of the invention, in the step (3), the mixture is heated to 80 ℃, stirred and dissolved, cooled to room temperature, kept warm and stirred for 6 hours, and dried at 50-60 ℃.

The invention is characterized in that the advanced on-line process control technology is utilized to carry out detailed parameter control on each step, and the specific parameters are as follows:

the specific process in the step (1) of the invention is as follows: mixing and reacting dimethylformamide, anhydrous potassium carbonate and rutin, heating to 80-85 ℃, dropwise adding 2-bromoethanol, keeping the temperature for reaction until the rutin disappears, cooling to room temperature, dropwise adding ethyl acetate, filtering, and drying at 50-60 ℃ to obtain a yellow solid troxerutin crude product; wherein the adding proportion of each material is dimethylformamide: anhydrous potassium carbonate: rutin: 2-bromoethanol ═ 4.1:0.9:1: 0.62.

The specific process in the step (2) of the invention is as follows: adding the troxerutin crude product into a mixed solution of dimethylformamide and acetone, heating to 65-70 ℃, stirring for dissolving, filtering, cooling to room temperature, crystallizing overnight, filtering, pulping a wet product with acetone for 2 hours, filtering, and drying at 50-60 ℃ to obtain a troxerutin primary refined product; wherein the adding proportion of each material is troxerutin crude product: dimethylformamide: acetone is 1:3: 5.

The specific process in the step (3) of the invention is as follows: adding the refined product of the troxerutin into a mixed solution of isopropanol and purified water, heating to 80 ℃, stirring for dissolving, cooling to room temperature, keeping the temperature, stirring for 6 hours, filtering, washing for 3 times by using anhydrous methanol, and drying at 50-60 ℃ to obtain a refined product of the troxerutin; wherein the adding proportion of each material is a refined product of troxerutin for the first time: isopropyl alcohol: purified water 1:5: 1.1.

The invention controls the on-line process parameters of the troxerutin preparation process, and the final product quality is achieved only if the operation of each step is detailed. The crystal form is unique, the solvent residue and impurities are lower than those of the existing product, the fluidity is better, the split charging of the preparation is convenient, and the stability is better.

The invention also provides a troxerutin injection preparation which is prepared by preparing and subpackaging the prepared compound, wherein the specification is 2ml:100 mg.

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

fig. 1 is an X-ray powder diffraction pattern of the troxerutin compound obtained in example 1.

FIG. 2 is a diagram of selective electron diffraction analysis of the troxerutin compound obtained in example 1.

Detailed Description

The following examples are further illustrative of the present invention and are in no way intended to limit the scope of the invention. The present invention is further illustrated in detail below with reference to examples, but it should be understood by those skilled in the art that the present invention is not limited to these examples and the preparation method used. Also, equivalent substitutions, combinations, improvements or modifications of the invention may be made by those skilled in the art based on the description of the invention, but these are included in the scope of the invention.

EXAMPLE 1 preparation of troxerutin Compounds

(1) Mixing 2500g of dimethylformamide, 550g of anhydrous potassium carbonate and 610g of rutin for reaction, heating to 80-85 ℃, dropwise adding 381g of 2-bromoethanol, keeping the temperature for reaction until the rutin disappears, cooling to room temperature, dropwise adding 7500g of ethyl acetate, filtering, and drying at 50-60 ℃ to obtain 684g of yellow solid troxerutin crude product, wherein the yield is 92.1%.

(2) 684g of the troxerutin crude product is added into mixed liquid of 2052g of dimethylformamide and 3420g of acetone, the mixed liquid is heated to 65-70 ℃, stirred and dissolved, filtered, cooled to room temperature, crystallized overnight, filtered, the wet product is pulped with 2280g of acetone for 2 hours, filtered and dried at 50-60 ℃ to obtain 647.7g of a troxerutin primary refined product, and the yield is 94.7%.

(3) Adding 647.7g of the primary refined troxerutin product into a mixed solution of 3239g of isopropanol and 712.5g of purified water, heating to 80 ℃, stirring and dissolving, cooling to room temperature, keeping the temperature and stirring for 6 hours, filtering, washing for 3 times by using anhydrous methanol, and drying at 50-60 ℃ to obtain 606.2g of the secondary refined troxerutin product, wherein the yield is 93.6%; the content is 99.2%.

And (3) crystal form determination results: example 1 the X-ray powder diffraction pattern of troxerutin shows characteristic diffraction peaks at 2 theta diffraction angles of 8.15 ± 0.2 °, 9.24 ± 0.2 °, 12.31 ± 0.2 °, 17.10 ± 0.2 °, 17.92 ± 0.2 °, 18.73 ± 0.2 °, 23.45 ± 0.2 °. Refer specifically to FIG. 1 of the specification.

Example 2 preparation of troxerutin Compounds

(1) Mixing and reacting 3280g of dimethylformamide, 720g of anhydrous potassium carbonate and 800g of rutin, heating to 80-85 ℃, dropwise adding 496g of 2-bromoethanol, keeping the temperature for reaction until the rutin disappears, cooling to room temperature, dropwise adding 10000g of ethyl acetate, filtering, and drying at 50-60 ℃ to obtain 908.7g of crude yellow solid troxerutin product with the yield of 93.3%.

(2) Adding 908.7g of troxerutin crude product into a mixed solution of 2726g of dimethylformamide and 4546g of acetone, heating to 65-70 ℃, stirring for dissolving, filtering, cooling to room temperature, crystallizing overnight, filtering, pulping wet products with 3031g of acetone for 2 hours, filtering, and drying at 50-60 ℃ to obtain 865.1g of troxerutin primary refined product, wherein the yield is 95.2%.

(3) Adding 865.1g of the primary refined troxerutin product into a mixed solution of 4326g of isopropanol and 951.6g of purified water, heating to 80 ℃, stirring for dissolving, cooling to room temperature, keeping the temperature and stirring for 6 hours, filtering, washing for 3 times by using anhydrous methanol, and drying at 50-60 ℃ to obtain 821g of a secondary refined troxerutin product with the yield of 94.9%; the content is 98.9%.

Example 3 preparation of troxerutin injection

Troxerutin injection is prepared by adopting the troxerutin compound raw material prepared in the example 1, and the specification is 2ml and 100 mg.

Prescription: (calculated according to 1000 batches)

The preparation process comprises the following steps:

1. preparing materials: 2/3 total amount of water for injection is added into the mixing tank, and troxerutin is dissolved in the water for injection. Dissolving edetate disodium in a small amount of water for injection, and adding into the above solution; adding medicinal carbon (2g/2000ml), heating, stirring for dissolving, adding injectable water to full dose, stirring thoroughly until the medicinal liquid is uniform, testing, filtering, and bottling.

2. Filling, filling nitrogen, sealing and checking the filling quantity according to requirements.

3. Sterilizing and detecting leakage.

Comparative example 1 preparation of troxerutin Compound

Troxerutin is prepared according to the existing preparation method route (I) mentioned in the background technology.

Adding 2.01Kg of rutin, 2.0Kg of water and 1.64g of sodium hydroxide into a reaction kettle at 75 ℃, and introducing nitrogen to drive off oxygen; and introducing ethylene oxide, monitoring the pH value of the solution, reducing the reaction temperature to 60 ℃ when the pH value of the reaction solution is 9.1, adding 200g of resin, and continuing to react until the pH value of the reaction solution is 10.0, and stopping adding the ethylene oxide. Adding hydrochloric acid aqueous solution with the ratio of 1: 1 to adjust the pH value of the solution to be about 4.5, carrying out hot filtration, concentration and vacuum drying to obtain 2.41Kg of troxerutin, wherein the content of 3 ', 4', 7-troxerutin is 89.3% by HPLC detection.

Comparative example 2 preparation of troxerutin injection

Troxerutin injection is prepared by adopting the troxerutin compound raw material prepared in the comparative example 1, wherein the specification is 2ml and 100 mg.

Prescription: (calculated according to 1000 batches)

The preparation process comprises the following steps:

3. Sterilizing and detecting leakage.

Comparative example 3 preparation of troxerutin Compound

According to the preparation method of the patent CN106892950A, but the preparation parameters are different.

(1) Adding 2.5Kg (63mol) of sodium hydroxide into 350L of distilled water, stirring for dissolving, adding 20.0Kg (33mol) of crude rutin, heating to 40 deg.C, stirring for dissolving, reacting for 3 hours, adding 800g of activated carbon, and vacuum filtering while hot to obtain clear filtrate. The filtrate was adjusted to pH 3.0 with hydrochloric acid. Slowly cooling to room temperature, controlling the cooling time to be 1h, standing for 1h at room temperature, performing suction filtration, leaching a filter cake with 50L of water, wherein the yield is 82.7%, and the purity is 90.1%.

(2) Adding 2.1Kg of rutin obtained in the step (1) and 112g of natural polymer sodium alginate into 8.5Kg of anhydrous methanol, then introducing 850g of ethylene oxide, sealing the autoclave, starting heating and stirring, heating to 50 ℃, keeping the temperature for reaction for 3h, rapidly cooling when the proportion of the troxerutin reaches 88% by HPLC (high performance liquid chromatography), stopping the reaction, adjusting the pH value of the reaction solution to 3 by using hydrochloric acid, standing for crystallization for 8h, performing suction filtration and drying to obtain 1.0Kg of crystals, wherein the yield is 47.6%.

(3) To 0.9Kg of the crystals obtained in (2), 18L of anhydrous methanol was added, and the mixture was heated to complete dissolution at reflux temperature: refluxing at 60 deg.C: 30 minutes, adding 30g of activated carbon for injection, carrying out suction filtration while the solution is hot, and cooling the filtrate to 28 ℃ for crystal growth for 7 hours. Performing suction filtration and drying to obtain 0.66Kg of troxerutin, wherein the yield is 73.3 percent, the total yield is 31.4 percent, and the content is 72.5 percent.

Comparative example 4 preparation of troxerutin injection

Troxerutin injection is prepared by adopting the troxerutin compound raw material prepared in the comparative example 3, and the specification is 2ml:100 mg.

Prescription: (calculated according to 1000 batches)

The preparation process comprises the following steps:

3. Sterilizing and detecting leakage.

Test example 1 purity measurement

The inventors carried out purity tests on inventive example 1 and comparative examples 1 and 3.

And (3) purity detection result:

examples	Purity (%)
		Example 1	99.2
Comparative example 1	89.3
		Comparative example 3	72.5

And (4) conclusion: the purity of the troxerutin compound prepared by the invention is obviously higher than that of troxerutin prepared by a comparative example, which indicates that the impurity content is far lower than that of similar products.

Test example 2 stability test

The inventors conducted an accelerated stability investigation test on troxerutin injection prepared in example 3 of the present invention and comparative examples 2 and 4. The examination conditions were 40 ℃ 2 ℃ and 75% + -5% relative humidity. Standing for 6 months, and sampling at the end of 0, 1, 2, 3, and 6 months. The investigation indexes are characters, clarity, solution color, pH value, related substances, visible foreign matters and content.

And (4) conclusion: from the results, through the 6-month accelerated test, each detection index of the sample prepared in the embodiment 3 of the invention is obviously superior to that of the products prepared in the comparative examples 2 and 4, and the invention fully shows that the troxerutin compound preparation prepared by the online process control technology has better stability and better quality than similar products.

Test example 3 solvent residue

The present inventors examined the solvent residue in inventive example 1 and comparative examples 1 and 3.

Examples	Ethylene oxide	Methanol
			Example 1	1ppm	Not detected out
Comparative example 1	4ppm	Not detected out
			Comparative example 3	6ppm	Not detected out

And (4) conclusion: the solvent residue in the troxerutin compound prepared in example 1 of the present invention is lower than that in comparative examples 1 and 3.

Test example 4 selected area electron diffraction analysis

By carrying out selective electron diffraction analysis on the compound in the embodiment 1 of the invention, the low-magnification morphology gives out a regular crystal shape, diffraction point spots are regularly arranged, and the compound is judged to be a single crystal, which shows that the product has a unique crystal form.

Refer specifically to figure 2 of the specification.

Claims

1. A troxerutin compound of the structure:

the characteristic diffraction peak is shown at the 2 theta of 8.15 +/-0.2 degrees, 9.24 +/-0.2 degrees, 12.31 +/-0.2 degrees, 17.10 +/-0.2 degrees, 17.92 +/-0.2 degrees, 18.73 +/-0.2 degrees, 23.45 +/-0.2 degrees in an X-ray powder diffraction pattern expressed by the 2 theta diffraction angle.

2. The process for the preparation of a troxerutin compound according to claim 1, which comprises the steps of:

3. The method according to claim 2, wherein the steps are subjected to detailed parameter control by an on-line control technique, which comprises the following steps:

in the step (1), the heating temperature is 80-85 ℃, and the drying temperature is 50-60 ℃;

heating to 65-70 ℃, stirring and dissolving in the step (2), and drying at 50-60 ℃;

and (4) heating to 80 ℃, stirring for dissolving, cooling to room temperature, keeping the temperature and stirring for 6 hours, and drying at 50-60 ℃.

4. A troxerutin injection preparation characterized by containing the troxerutin compound described in claim 1 in a size of 2ml to 100 mg.

5. A troxerutin injection preparation characterized by containing a troxerutin compound prepared by the method described in any one of claims 2 to 3, in a specification of 2ml to 100 mg.