WO2016034145A1 - (6s)-5-甲基四氢叶酸或其盐组合物及其制备和应用 - Google Patents

(6s)-5-甲基四氢叶酸或其盐组合物及其制备和应用 Download PDF

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WO2016034145A1
WO2016034145A1 PCT/CN2015/088925 CN2015088925W WO2016034145A1 WO 2016034145 A1 WO2016034145 A1 WO 2016034145A1 CN 2015088925 W CN2015088925 W CN 2015088925W WO 2016034145 A1 WO2016034145 A1 WO 2016034145A1
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salt
methyltetrahydrofolate
vitamin
isovitamin
acid
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PCT/CN2015/088925
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English (en)
French (fr)
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成永之
成志
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连云港金康和信药业有限公司
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Priority to NZ730790A priority Critical patent/NZ730790A/en
Priority to EP15838962.7A priority patent/EP3190112A4/en
Priority to AU2015311370A priority patent/AU2015311370B2/en
Priority to JP2017512929A priority patent/JP6960853B2/ja
Priority to US15/509,014 priority patent/US10398697B2/en
Priority to KR1020177009202A priority patent/KR20170047394A/ko
Publication of WO2016034145A1 publication Critical patent/WO2016034145A1/zh

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Definitions

  • the invention relates to the field of pharmaceutical preparations, in particular to a composition and preparation method of (6S)-5-methyltetrahydrofolate or a salt thereof, and a solid of high purity (6S)-5-methyltetrahydrofolate or a salt thereof Solution composition and preparation method thereof, and application of (6S)-5-methyltetrahydrofolic acid and a salt thereof.
  • Applications include the use of a composition or solid solution of the above (6S)-5-methyltetrahydrofolate or a salt thereof; the addition of a pharmaceutically acceptable reducing substance as a stabilizer, which provides (6S)-5- Methyltetrahydrofolate products have excellent safety properties.
  • 5-methyltetrahydrofolate and its salts have been increasingly studied for physiological activities in humans. It is used as a supplement to human folic acid for the prevention and/or treatment of neurological diseases; combined with 5-fluorouracil or methotrexate for tumor therapy; and for vascular and cardiovascular diseases for pathophysiology; Major depression, severe senile dementia treatment and adjuvant therapy, etc., show good application prospects.
  • 5-methyltetrahydrofolate or its salts are very unstable and highly susceptible to degradation, especially to oxygen and moisture. Therefore, a composition of 5-methyltetrahydrofolate or a salt thereof, for example, a pharmaceutical preparation generally has a problem of poor stability, easy discoloration, deepening of color, a marked decrease in content, and an increase in degradation products.
  • the primary oxidation product is characterized as 4-hydroxy-5-methyltetrahydrofolate. It is inferred from the structure that this product has no toxic side effects.
  • the inventors developed an efficient method by first preparing a high-purity 5-methyltetrahydrofolate primary oxidation product and conducting a series of studies (PCT/CN2013/073959), and found that: unexpectedly, 5-A
  • the primary oxidation product of tetrahydrofolate or its salts is JK12A, and the structural formula is as follows:
  • JK12A refers to the initial oxidation product of 5-methyltetrahydrofolate.
  • the relative retention time of the main peak and the main peak is about 0.36 depending on the specific conditions, and the related literature is called 5-methyl- Dihydrofolate or 4-hydroxy-5-methyl-tetrahydrofolate.
  • 5-methyltetrahydropteric acid is another common impurity in the 5-methyltetrahydrofolate product, similar to the formation of JK12A, which is oxidized to become JK1303 of the following structure:
  • JK1303 has severe renal toxicity. JK1303 was administered to mice in a single dose of 1000 mg/kg. After 14 days, the renal tubules of the mice were severely necrotic, and a large number of vacuoles were found in the renal tubules. The toxicity of the primary oxidation product of 5-methyltetrahydropteroate also changed very significantly.
  • Patent CN102612358 discloses a 5-methyltetrahydrofolate stabilized granule which is increased in stability by embedding crystalline 5-methyltetrahydrofolate with wax.
  • Patent CN102813656 discloses a 5-methyltetrahydrofolate stable composition comprising one or more amino groups free of thiol groups to increase stability.
  • Patent US Pat. No. 6,441,168 discloses the significant enhancement of stability by preparing 5-methyltetrahydrofolate to form a crystalline form.
  • the crystal form has a relatively large particle size and slow dissolution, and it is often required to be milled first in the preparation process, which also tends to increase the risk of oxidation of 5-methyltetrahydrofolate.
  • Patent CN2012086794 discloses a new crystalline form of (6S)-5-methyltetrahydrofolate, which has good chemical stability on the one hand and a significant increase in solubility properties, so that no milling is required during the formulation process. However, the content of impurities such as JK12A is still high, and the amount of these impurities gradually increases with time.
  • microencapsulated 5-methyltetrahydrofolate is prepared by using materials such as cellulose and shellac to improve the stability of the active ingredient-5-methyltetrahydrofolate in the preparation product.
  • An object of the present invention is to provide a composition containing high purity (6S)-5-methyltetrahydrofolic acid or a salt thereof and a process for producing the same, based on the prior art.
  • Another object of the present invention is to provide a solid solution containing (6S)-5-methyltetrahydrofolate or a salt thereof and a stabilizer and a process for producing the same.
  • a third object of the present invention is to provide a pharmaceutical composition containing high purity (6S)-5-methyltetrahydrofolate or a salt thereof or containing the above solid solution.
  • a fourth object of the present invention is to provide a high-purity (6S)-5-methyltetrahydrofolate or a salt thereof or a solid solution containing (6S)-5-methyltetrahydrofolic acid or a salt thereof and a stabilizer in medicine use.
  • the object of the invention can be achieved by the following measures:
  • composition containing (6S)-5-methyltetrahydrofolate or a salt thereof wherein the content of (6S)-5-methyltetrahydrofolic acid or a salt thereof is not less than 98.0%, wherein the relevant impurity JK12A The content of not more than 0.1%, 5-methyltetrahydropteric acid should not be detected;
  • the main substance in the composition containing (6S)-5-methyltetrahydrofolic acid or a salt thereof of the present invention is (6S)-5-methyltetrahydrofolate or a salt thereof.
  • the content of (6S)-5-methyltetrahydrofolic acid or a salt thereof in the composition is the mass content or HPLC content of (6S)-5-methyltetrahydrofolic acid or a salt thereof, and is generally a mass content.
  • the content of (6S)-5-methyltetrahydrofolic acid or a salt thereof in the composition is not less than 99.0%, wherein the content of the related impurity JK12A is not more than 0.1%, 5-methyltetrahydropterylic acid Do not check out.
  • the content of (6S)-5-methyltetrahydrofolic acid or a salt thereof in the composition is not less than 99.5%, wherein the content of the related impurity JK12A is not more than 0.1%, 5-methyltetrahydro butterfly Acid should not be detected.
  • the content of (6S)-5-methyltetrahydrofolic acid or a salt thereof in the composition is not less than 99.8%, wherein the content of the relevant impurity JK12A is not more than 0.05%, 5-methyltetrahydro butterfly Acid should not be detected.
  • the (6S)-5-methyltetrahydrofolate in the present invention is selected from the group consisting of a potassium salt, a sodium salt, a calcium salt, a magnesium salt, a barium salt, a zinc salt, a D-glucosamine salt, and a D-galactosamine salt.
  • an arginine salt preferably a calcium salt, a D-glucosamine salt, a D-galactosamine salt or an arginine salt.
  • Method A is: directly reacting crude (6S)-5-methyltetrahydrofolate with an inverting reducing agent in water at a pH of 6-8, a temperature of 50-90 ° C, or performing the reaction under ultrasonic conditions. , after the reaction, the (6S)-5-methyltetrahydrofolate composition is obtained;
  • Method B is: reacting crude (6S)-5-methyltetrahydrofolate, an inverting reducing agent and a corresponding salt in water at a pH of 6-8, a temperature of 50-90 ° C, or performing the reaction under ultrasonic conditions. , after the reaction, the (6S)-5-methyltetrahydrofolate composition is obtained;
  • the inverter reducing agent in Method A or Method B is selected from the group consisting of vitamin C, vitamin C ester, vitamin C salt, isovitamin C, isovitamin C ester, isovitamin C salt, mercaptoethanol, cysteine, and decyl sulfonate.
  • the inverting reducing agent in Method A or Method B is selected from one or more of vitamin C, vitamin C ester, vitamin C salt, isovitamin C, isovitamin C ester or isovitamin C salt.
  • Kind preferably one or more of vitamin C, vitamin C sodium salt, vitamin C potassium salt, vitamin C calcium salt, vitamin C phosphate magnesium salt, vitamin C phosphate sodium salt or isovitamin C; most preferably vitamin C Or isovitamin C; the corresponding salt is a chloride salt, preferably calcium chloride, magnesium chloride, zinc chloride.
  • the reaction temperature is 60-80 ° C
  • the pH is 7-7.5, preferably the pH is adjusted with sodium hydroxide
  • the weight of the inverting reducing agent in water The concentration is not less than 2%, preferably not less than 20%, more preferably not less than 40%.
  • the sum of the content of the stabilizer and (6S)-5-methyltetrahydrofolate or a salt thereof in the chromatogram is greater than 99.5%; wherein the stabilizer is selected from the group consisting of vitamin C, vitamin C ester, vitamin C salt, and different One or more of vitamin C, isovitamin C ester, isovitamin C salt, dithiothreitol, reduced glutathione or lipoic acid.
  • the stabilizer and the (6S)-5-methyltetrahydrofolate in a liquid chromatogram of a solid solution containing (6S)-5-methyltetrahydrofolate or a salt thereof and a stabilizer The sum of the contents of the salt thereof or the salt thereof is more than 99.8%, preferably more than 99.9%; the (6S)-5-methyltetrahydrofolic acid or a salt thereof is the high purity (6S)-5-methyl group according to claim 1. Hydrogen folic acid or a salt thereof.
  • the stabilizer is selected from one or more of vitamin C, vitamin C ester, vitamin C salt, isovitamin C, isovitamin C ester or isovitamin C salt, preferably vitamin C, vitamin C sodium salt, vitamin C potassium salt, vitamin C calcium salt, vitamin C phosphate magnesium salt, vitamin C phosphate sodium salt, isovitamin C, isovitamin C sodium salt, isovitamin C potassium salt, isovitamin C calcium salt, One or more of the isovitamin C phosphate magnesium salt and the isovitamin C phosphate sodium salt, more preferably vitamin C or isovitamin C.
  • the weight ratio of the stabilizer to (6S)-5-methyltetrahydrofolate is from 0.2 to 1000:1, preferably from 1 to 10:1.
  • a method for preparing a solid solution containing the above (6S)-5-methyltetrahydrofolate or a salt thereof and a stabilizer which comprises the method C or the method D, wherein
  • Method C is: crude (6S)-5-methyltetrahydrofolate or the high-purity (6S)-5-methyltetrahydrofolate according to claim 1 and a stabilizer in water at a pH of 3 to 5, temperature The reaction is carried out directly at 50 to 90 ° C, or under ultrasonic conditions, and the reaction is precipitated and the solid is separated;
  • Method D is: crude (6S)-5-methyltetrahydrofolate or the high purity (6S)-5-methyltetrahydrofolate according to claim 1 and a stabilizer and corresponding salt in water at pH 6 ⁇ 8.
  • the reaction is carried out at a temperature of 50 to 90 ° C, or the reaction is carried out under ultrasonic conditions, and the solid is precipitated and separated after the reaction;
  • the stabilizer is selected from the group consisting of vitamin C, vitamin C ester, vitamin C salt, isovitamin C, isovitamin C ester, isovitamin C salt, dithiothreitol, reduced glutathione or lipoic acid
  • the corresponding salt is a chloride salt, preferably calcium chloride, magnesium chloride, zinc chloride.
  • the reaction temperature is 60-80 ° C, the pH is 3 to 4, and the pH is adjusted with hydrochloric acid; or the reaction temperature of the method D is 60- At 80 ° C, the pH is 7 to 7.5, and the pH is adjusted with sodium hydroxide.
  • a pharmaceutical composition comprising:
  • a reducing agent selected from the group consisting of vitamin C, vitamin C ester, vitamin C salt, isovitamin C, isovitamin C ester, isovitamin C salt, mercaptoethanol, cysteine, decyl sulfonic acid, One or more of dithiothreitol, reduced glutathione or lipoic acid.
  • the pharmaceutical composition is a common tablet, a sustained release tablet, a controlled release tablet, an effervescent tablet, a granule, a general capsule, a sustained release capsule, a controlled release capsule, a dry suspension, orally.
  • the weight ratio of the reducing agent to (6S)-5-methyltetrahydrofolic acid or a salt thereof in the pharmaceutical composition is from 0.2 to 1000:1, preferably from 1 to 10:1.
  • the high-purity (6S)-5-methyltetrahydrofolic acid or a salt thereof of the present invention or the solid solution containing the (6S)-5-methyltetrahydrofolic acid or a salt thereof and a stabilizer thereof for preparing or preventing breast-feeding Use of an animal for a drug associated with a deficiency of 5-methyltetrahydrofolate or a salt thereof.
  • Reversal refers to the phenomenon that (6S)-5-methyltetrahydrofolate and its salt change back to JK12A impurity (6S)-5-methyltetrahydrofolate and its salt; JK1303 changes back to 5-methyltetrahydropteric acid .
  • Salting out A method of adding a certain concentration of a neutral salt to a solution of a solid solute to improve crystallization, often improving yield and quality.
  • Solid solution A mixture of one or more solids uniformly dispersed in another solid.
  • (6S)-5-methyltetrahydrofolate and its salts are introduced and produced during storage, storage, and impurities; including 4-aminobenzoylglutamic acid, JK12A, (6S)-Mefox , pyrazin triazine derivative, tetrahydrofolate, 7,8-dihydrofolate, folic acid, 5,10-methyltetrahydrofolate, 5-methyltetrahydropteric acid, N2-methylamino-tetrahydrofolate And their salt.
  • Purity refers to the ratio of the peak area of (6S)-5-methyltetrahydrofolate in liquid chromatography to the sum of the peak areas of (6S)-5-methyltetrahydrofolate and all related substances.
  • the present inventors have surprisingly found that most of the oxidative degradation impurities such as JK12A can be reversed to (6S)-5-methyltetrahydrofolate using a relatively high concentration of a reducing reducing agent, and JK1303 is reversed to 5-methyltetrahydrogen. Butyric acid. Subsequent salting out can precipitate high purity (6S)-5-methyltetrahydrofolate, resulting in enormous high purity at higher yields, which was not reported in the literature.
  • (6S)-5-methyltetrahydrofolate and its salt can be prepared with a purity of more than 98.0%, or even close to 100.0%.
  • the related impurities include JK12A, JK1303, and 5-methyltetrahydropteric acid. Reducing, even below, the detection limit of liquid chromatography, which is not possible with the prior art.
  • the invention also provides a preparation method of (6S)-5-methyltetrahydrofolic acid or a salt composition thereof: (6S)-5-methyltetrahydrofolic acid or a crude salt thereof is first dissolved in water or an aqueous solvent Treatment with an inverter reducing agent to invert most of the oxidative degradation impurities into (6S)-5-methyltetrahydrofolate, and then precipitate high purity (6S)-5-A by salting out or other methods. Tetrahydrofolate or a salt thereof.
  • the inverter reducing agent selected by the invention is as described above, and the inverting reducing agent for reversing the oxidized impurities has moderate reduction property, and the reducing property is too high to cause reduction degradation, and if it is too low, the effect is not obtained.
  • the inverting reducing agent has a weight concentration in the solvent of Process A, Process B, and the above-described composition of not less than 2%, preferably not less than 20%, more preferably not less than 40%.
  • the salting-out salting-out agent of the present invention may be a common salt in addition to the above-mentioned reducing agent, for example, by adding a neutral inorganic salt such as sodium chloride, potassium chloride or magnesium sulfate. In the present invention, it is preferred to increase the concentration of the reducing agent to achieve the salting out effect.
  • the present invention provides a solid solution consisting of (6S)-5-methyltetrahydrofolate or a salt thereof and a stabilizer, and a stabilizer in the liquid chromatogram and (6S)-5-methyltetra
  • the sum of the hydrogen folate levels is greater than 99.5%, preferably greater than 99.8%, more preferably greater than 99.9%.
  • the above content is calculated by the peak area in the liquid chromatogram.
  • the solid solution is a dispersion form excellent in uniformity and can effectively protect the active ingredient of the present invention.
  • the solid solution of the present invention maintains a high degree of homogeneity of (6S)-5-methyltetrahydrofolic acid and a stabilizer, thereby ensuring a stabilizing effect.
  • (6S)-5-methyltetrahydrofolate is a C-type crystal
  • the particles are uniform and fine
  • the particle size distribution range is narrow
  • a highly uniform solid solution can be formed with the stabilizer. This is difficult to achieve with the prior art.
  • the solid solution of the present invention is (6S)-5-methyltetrahydrofolate and a salt thereof and vitamin C, or isovitamin C, or lipoic acid, or reduced glutathione. , or a solid solution of dithiothreitol and their salts.
  • a solid solution prepared in accordance with the present invention exhibits only two typical absorption peaks in its HPLC profile and exhibits a high degree of purity.
  • the stabilizer in the solid solution and (6S)-5-methyltetrahydrofolate are uniformly dispersed, which can better exert the stabilizing effect.
  • a stabilizer may be additionally added, and other additives may be added as needed.
  • the invention also provides a preparation method of another solid solution: firstly dissolving (6S)-5-methyltetrahydrofolic acid or a salt thereof in water or an aqueous solvent, adding a stabilizer to reverse the impurities, and then stabilizing agent and 6S)-5-methyltetrahydrofolate and its salts are precipitated together, filtered and dried. Further, (6S)-5-methyltetrahydrofolic acid and a stabilizer are dispersed in water, dissolved by stirring with sodium hydroxide, and then the corresponding salt is added, crystallized, filtered, and dried.
  • the present invention still further provides a safe use of (6S)-5-methyltetrahydrofolate or a salt thereof, comprising: a. using a combination of (6S)-5-methyltetrahydrofolate or a salt thereof as described above Or a solid solution as described above; b. adding a pharmaceutically acceptable reducing substance as a stabilizer.
  • a safe use of a preferred (6S)-5-methyltetrahydrofolate or a salt thereof wherein the stabilizer is selected from the group consisting of vitamin C and its esters and salts, isovitamin C and its esters and salts, disulfide Threitol, reduced glutathione, lipoic acid, preferably vitamin C and its esters and salts, isovitamin C and its esters and salts, more preferably sodium, potassium and calcium salts of vitamin C and isovitamin C, Vitamin C phosphate magnesium salt, vitamin C phosphate sodium salt, most preferably vitamin C and isovitamin C.
  • the stabilizer is selected from the group consisting of vitamin C and its esters and salts, isovitamin C and its esters and salts, disulfide Threitol, reduced glutathione, lipoic acid, preferably vitamin C and its esters and salts, isovitamin C and its esters and salts, more preferably sodium, potassium and calcium salts of vitamin C and isovita
  • the combination of the stabilizer and (6S)-5-methyltetrahydrofolate is directly mixed as a solid or dissolved first.
  • the solution is then precipitated together or prepared as a eutectic or solid solution, preferably as a solid solution.
  • the weight ratio of the stabilizer to (6S)-5-methyltetrahydrofolate is 0.2 to 1000:1; Preferably it is 1 to 10:1.
  • a preferred embodiment is to use vitamin C and its salt as both a reducing agent, a salting-out agent and a stabilizer as described above.
  • vitamin C and its salts are both a reducing agent for reversing the oxidation impurities and a salting-out agent in the purification process, and a stabilizer for (6S)-5-methyltetrahydrofolate.
  • Vitamin C and its salts remaining in the preparation process do not need to be specifically removed. In the subsequent process, the required amount of vitamin C may be added as needed.
  • the use of reduced glutathione, lipoic acid and its single isomer, dithiothreitol, isovitamin C, etc. can achieve similar effects.
  • these materials are also suitable as reducing agents, salting-out agents and stabilizers.
  • (6S)-5-methyltetrahydrofolate or a salt thereof as described above is applied to a compound preparation of (6S)-5-methyltetrahydrofolate or a combination of other active ingredients.
  • (6S)-5-methyltetrahydrofolate or a salt thereof as described above for the preparation of a medicament for treating or preventing a disease associated with a deficiency of 5-methyltetrahydrofolate or a salt thereof in a mammal.
  • the present invention also provides the use of (6S)-5-methyltetrahydrofolate or a salt thereof for the safe use of a medicament for the treatment or prevention of a disease associated with the deficiency of 5-methyltetrahydrofolate or a salt thereof in a mammal.
  • Diseases associated with the deficiency of 5-methyltetrahydrofolate or a salt thereof include, for example, subacute encephalitis associated with dementia and vacuolar myopathy, cerebral acid deficiency, pathophysiological vascular and cardiovascular diseases, for example Premature occlusive arterial disease, severe vascular disease in infancy and childhood, progressive stenosis, intermittent claudication, renal vascular hypertension, ischemic cerebrovascular disease, premature retinal artery and retinal vein occlusion, Cerebral occlusive arterial disease, occlusive peripheral arterial disease, premature death due to thromboembolic disease and/or ischemic heart disease; autoimmune diseases such as psoriasis, celiac disease, arthritis and inflammatory conditions; due to folic acid A erythroblastic anemia caused by salt deficiency, intestinal malabsorption, an antidote for folic acid antagonists (such as methotrexate, pyrimethamine or trimethoprim); used to
  • the selection principle of the reducing agent, the salting-out agent and the stabilizer according to the present invention is firstly chemically safe and does not cause degradation of the product; secondly, it is physiologically relatively safe, and the safety in the food and medicine fields is preferably fully verified. Substance. It can be understood that the amount of the reducing agent, the salting-out agent and the stabilizer are required to exceed a minimum amount, otherwise the result may not be sufficient to reverse the impurities, the salting-out effect is poor or/and the stabilizing effect is not good. It will also be appreciated that, in general, there is no strict upper limit to the amount of reducing agent or stabilizer.
  • the impurities of the (6S)-5-methyltetrahydrofolic acid and the salt thereof according to the present invention are JK12A and 5-methyltetrahydropteric acid, and their structures are respectively
  • 5-methyltetrahydropteroate can be converted into JK1303 with severe renal toxicity in a high yield after oxidation, and thus requires strict control.
  • the invention has the beneficial effects of high purity (6S)-5-methyltetrahydrofolic acid or a salt thereof and a preparation method thereof, and a solid solution of high purity (6S)-5-methyltetrahydrofolic acid or a salt thereof And its preparation, and the safe use of (6S)-5-methyltetrahydrofolic acid and its salts; in these embodiments, (6S)-5-A through the ingenious application of pharmaceutically acceptable reducing agents
  • the harmful chemicals that may be produced in the tetrahydrofolate or its salt product are reversed back to the product itself, thereby achieving the purpose of safely using (6S)-5-methyltetrahydrofolate or its salt product.
  • Figure 1 is an enlarged partial anatomical view showing the presence of a large number of vacuoles after severe necrosis of the renal tubules in 14 days after administration of JK1303.
  • HPLC detection conditions of the present invention are:
  • Mobile phase A 7.80 g of NaH 2 PO 4 ⁇ 2H 2 O was weighed and dissolved in 1000 ml of water, and the pH was adjusted to 6.5 with a 32% NaOH solution.
  • Mobile phase B 5.07 g of NaH 2 PO 4 ⁇ 2H 2 O was weighed, dissolved in 650 ml of water, added to 350 ml of methanol, and the pH was adjusted to 8.0 with a 32% NaOH solution.
  • sample solution Take appropriate amount of sample, and distill it with low temperature water at a temperature of 2-8 degrees (distilled with sodium sulfite under nitrogen protection and collected and stored and allowed to cool to 2-8 ° C). It was dissolved under nitrogen to prepare a solution having a concentration of about 0.5 mg/ml, which is now available.
  • ultrasonic waves are used as an auxiliary means, which can significantly improve the reversal and crystallization speed.
  • test data in the examples only involve 5-methyltetrahydrofolate and its salts and related substances.
  • JK12A 0.1 g of JK12A (HPLC: 95.91%) was taken, 10 g of water was added, and the pH was adjusted to 7.2 with 10% sodium hydroxide with stirring, and the solid was completely dissolved. Then, 0.8 g of dithiothreitol was slowly added, and after stirring for 1 hour, 97.53% of 5-methyltetrahydrofolate and 0.72% of JK12A remained in the reaction mixture.
  • JK12A 0.1 g of JK12A (HPLC: 95.91%) was taken, 10 g of water was added, and the pH was adjusted to 7.2 with 10% sodium hydroxide with stirring, and the solid was completely dissolved. Then, 0.5 g of cysteine was slowly added, and after stirring for 1 hour, a sample was detected to detect 96.64% of 5-methyltetrahydrofolate in the reaction solution, and the residue of JK12A was 0.85%.
  • JK12A HPLC: 74.04%
  • 0.2 g of mercaptoethanol was added, and it was added to 5 ml of water, and the pH was adjusted to 7.5 with 30% sodium hydroxide with stirring, and dissolved. After stirring for 1 hour, the reaction solution was sampled to detect 60.92% of 5-methyltetrahydrofolate and 0.59% of JK12A.
  • JK12A 0.1 g of JK12A (HPLC: 74.04%), 0.2 g of sodium decylsulfonate was added, and it was added to 5 ml of water, and the pH was adjusted to 7.5 with 30% sodium hydroxide with stirring, and dissolved. After stirring for 1 hour, a sample was taken to detect 72.85% of 5-methyltetrahydrofolate in the reaction solution, and the residue of JK12A was 0.61%.
  • the base was adjusted to maintain pH 7.0, stirred at 60 ° C for 3 hours, and the reaction solution was sampled and detected (5-methyltetrahydrofolate: 96.49%; JK12A: 0%; 6(S)-Mefox: 0%;)
  • JK1303 0.1 g of JK1303 (HPLC: 96.7%) was taken, 10 g of water was added, and the pH was adjusted to 7.0 with 10% sodium hydroxide under stirring, and the solid was completely dissolved. Then, 1 gram of vitamin C sodium was slowly added, and after stirring for 1 hour, a sample of 96.6% of 5-methyltetrahydropteroic acid was detected in the reaction solution, and the residue of JK1303 was 0.
  • the content is the mass content, the same below.
  • 1S-5-methyltetrahydrofolate 1g, 1g of vitamin C, 0.5g of NaCl, was added to 10ml of water, neutralized to 7.0 by a 30% aqueous solution of sodium hydroxide, and dissolved. After stirring for 1 h, 1 g of a 40% aqueous solution of calcium chloride was added, and ultrasonication was carried out at 70 ° C. After 1 h, a solid was precipitated, filtered, washed, and the filter cake was dried in vacuo to give 0.52 g of a solid composition of 6S-5-methyltetrahydrofolate.
  • 6S-5-methyltetrahydrofolate 0.5 g, 10 mg of vitamin C, 0.1 g of sodium chloride, added to 10 ml of water, neutralized to 7.5 with 30% aqueous sodium hydroxide solution, dissolved, and transparent. Under nitrogen protection, stirring for 1 h, adding 1 g of 20% aqueous calcium chloride solution, and transferring to 70 ° C for ultrasonic crystallization, after 1.0 h, white particles were precipitated. Filtration, washing, and filter cake were dried in vacuo to give 0.38 g of 6S-5-methyltetrahydrofolate as a solid.
  • 1S-5-methyltetrahydrofolate 1g 3g of isovitamin C, added to 20ml of water, neutralized to 7.0 with 30% aqueous sodium hydroxide solution, dissolved, and transparent. After stirring for 1 h, 1 g of a 40% aqueous solution of calcium chloride was added, and ultrasonication was carried out at 60 ° C. After 1 h, a solid was precipitated, filtered, washed, and the filter cake was dried in vacuo to give 0.80 g of a 6S-5-methyltetrahydrofolate calcium salt composition.
  • 6R 1 g of S-5-methyltetrahydrofolate, 10 g of vitamin C, added to 15 ml of water, neutralized to 7.0 with 30% aqueous sodium hydroxide solution, dissolved, and transparent. 1 g of a 40% aqueous solution of calcium chloride was added, ultrasonically crystallized at 70 ° C, filtered, and then washed with water, and dried under vacuum to give 0.54 g of a 6R, S-5-methyltetrahydrofolate calcium salt composition.
  • 6S-5-methyltetrahydrofolate 0.5 g, 1.0 g of lipoic acid, was added to 10 ml of water, neutralized to 7.5 with 30% aqueous sodium hydroxide solution, and dissolved. After stirring for 1 h, add 1 g of 20% aqueous calcium chloride solution, transfer to 70 ° C for ultrasonic crystallization, 1 h to obtain a solid solution, filter, dry, liquid phase detection (using ELSD detector) 6S-5-methyltetrahydrofolate Calcium salt: 27.69%, lipoic acid 72.26%.
  • 6S-5-methyltetrahydrofolate 0.5g, 1.5g reduced glutathione, added to 10ml water, neutralized to 7.5 with 30% sodium hydroxide solution, dissolved, stirred for 1h, added 1g 20% calcium chloride The aqueous solution was transferred to 70 ° C for ultrasonic crystallization. After 1.0 h, white particles were precipitated, filtered, dried, and liquid phase tested (using ELSD detector). 6S-5-methyltetrahydrofolate calcium salt: 37.62%, reduced gluten Glycopeptide 62.33%.
  • 6S-5-methyltetrahydrofolate 0.5g, 3.0g vitamin C, added to 10ml water, neutralized to 7.5 with 30% sodium hydroxide aqueous solution, dissolved, stirred for 1h, add appropriate 6N hydrochloric acid aqueous solution, adjust pH to 4.0, 50 ° C stirring and crystallization, 1.0 h after the precipitation of white particles, filtration, drying, liquid phase detection of 6S-5-methyltetrahydrofolate: 26.80%, vitamin C: 73.17%.

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Abstract

一种(6S)-5-甲基四氢叶酸或其盐组合物及其制备和应用，其中(6S)-5-甲基四氢叶酸或其盐的含量不小于98.0％，相关杂质JK12A的含量不大于0.1％，5-甲基四氢蝶酸不得检出。

Description

(6S)-5-甲基四氢叶酸或其盐组合物及其制备和应用 技术领域
本发明为药物制剂领域，具体涉及一种(6S)-5-甲基四氢叶酸或其盐的组合物和制备方法，高纯度(6S)-5-甲基四氢叶酸或其盐的固体溶液组合物及其制备方法，以及(6S)-5-甲基四氢叶酸及其盐的应用。其应用包括使用上述(6S)-5-甲基四氢叶酸或其盐的组合物或者固体溶液；添加药学上可以接受的还原性物质作为稳定剂，该安全用法提供了(6S)-5-甲基四氢叶酸制品优良的安全性能。
背景技术
近年来，5-甲基四氢叶酸及其盐对于人体生理活性的研究越来越多。人们将其作为人体叶酸的补充剂用于预防和/或治疗神经学上的疾病；联合5-氟尿嘧啶或者甲氨蝶呤应用于肿瘤治疗；以及应用于病理生理学的血管和心血管疾病；应用于重度抑郁，重度老年性痴呆的治疗和辅助治疗等等，展现了良好的应用前景。
然而，5-甲基四氢叶酸或其盐非常不稳定，极易降解，特别是对氧和水分高度敏感。因此5-甲基四氢叶酸或其盐的组合物例如药物制剂普遍存在稳定性较差，容易变色，色泽加深、含量明显下降，降解产物增加的问题。
关于5-甲基四氢叶酸或者其活性单一异构体(6S)-5-甲基四氢叶酸的氧化降解，已有很多论文涉及。不幸的是，根据本发明人的一项研究，之前很多关于5-甲基四氢叶酸初级氧化产物的论述是片面甚至是错误的。这些结论误导人们降低了对5-甲基四氢叶酸质量的要求。按照Ratanasthien K等人在Serum folates in man中(Ratanasthien K.,Blair J.A.,Leeming R.J.,Cooke W.T.and Melikian V.(1977)..J.Clin.Path.30:438-448)；Blair J.A等人在Autoxidation of 5-Methyl-5,6,7,8-tetrahydrofolic acid(Blair J.A.,Pearson A.J.and Robb A.J.(1975)..J.C.S.Perkin II,p.18-21.)中；欧洲食品安全局在Opinion of the Scientific Panel on Food Additives,Flavourings,Processing Aids and Materials in Contact with Food on a request from the Commission related to Calcium L-Methylfolate Question(The EFSA Journal (2004)135,1-20)的论述，5-甲基四氢叶酸在氧化剂作用下首先氧化为5-甲基二氢叶酸。
Figure PCTCN2015088925-appb-000001
在其它文献中，初级氧化产物被定性为4-羟基-5-甲基四氢叶酸。人们根据结构推断这个产物是没有毒副作用的。
发明人开发出高效的方法，首先制备出了高纯度5-甲基四氢叶酸初级氧化产物，并进行了一系列研究(PCT/CN2013/073959)，结果发现：出乎意料的，5-甲基四氢叶酸或其盐类的初级氧化产物为JK12A，结构式如下：
Figure PCTCN2015088925-appb-000002
JK12A是指5-甲基四氢叶酸的初氧化产物，本发明的检测条件下，依具体条件的差异，其和主峰的相对保留时间在0.36左右，相关文献中被称为5-甲基-二氢叶酸或者4-羟基-5-甲基-四氢叶酸。
这是一个全新结构的产物.这个发现推翻了之前的，包含上述文献中的结论。一般说来,结构上的较大变化，将会引起生理活性的较大变化。随后的研究发现，JK12A对T淋巴细胞增殖具有显著的抑制作用。一般认为，这将引起人类免疫力的降低。
Figure PCTCN2015088925-appb-000003
Figure PCTCN2015088925-appb-000004
对JK12A的一项急性毒性研究发现，在2000毫克每公斤的剂量下，受试小白鼠在1分钟内均当场死亡，但解剖发现肝脏肾脏没有显著变化，可能该化合物有其他毒性靶标，且急性毒性显著。
发明人的研究发现，5-甲基四氢蝶酸是5-甲基四氢叶酸产物中的另外一个常见杂质，类似于JK12A的形成，该杂质经氧化后变成了如下结构的JK1303：
Figure PCTCN2015088925-appb-000005
随后的研究发现JK1303有严重的肾脏毒性。对小鼠按照1000毫克/公斤剂量进行单次给药给予JK1303，14天后小鼠肾小管严重坏死，解剖发现其肾小管出现大量空泡。5-甲基四氢蝶酸的初级氧化产物的毒性同样发生了非常显著的变化。
以上的实验表明，在包含5-甲基四氢叶酸及其盐的产品中，由5-甲基四氢叶酸及其盐氧化降解产生的杂质JK12A，5-甲基蝶酸的氧化产物JK1303对人体都是显著有害的，不能够也不应该被忽视。因此开发5-甲基四氢叶酸或其盐的安全可靠的应用方案，非常必要。
专利CN102612358公布了一种包含5-甲基四氢叶酸盐稳定颗粒剂，通过将结晶5-甲基四氢叶酸盐用蜡包埋以增加其稳定性。
专利CN102813656公布了一种包含5-甲基四氢叶酸盐稳定组合物，包含一种或者多种不含巯基的氨基酸以增加稳定性。
专利US6441168公布了通过将5-甲基四氢叶酸钙制备成某一晶型来显著增强其稳定性。但是该晶型颗粒度比较大，溶解缓慢，在制剂生产过程中往往需要先碾磨，这往往也会增大5-甲基四氢叶酸被氧化的风险。
专利CN2012086794公布了一种(6S)-5-甲基四氢叶酸钙的新晶型，该晶型一方面化学稳定性良好，同时溶解性能也有显著提高，从而在制剂过程中不需要碾磨。然而，JK12A等杂质含量仍然较高，且随着时间推移这些杂质的量也逐渐增加。
还有文献报道，用纤维素、虫胶等材料制备微胶囊包埋5-甲基四氢叶酸，以提高制剂产品中活性成分-5-甲基四氢叶酸的稳定性。
然而，所有这些措施仍然不能把JK12A控制到足够安全的水平。经实验检测，目前市售的5-甲基四氢叶酸原料药级产品中，结晶形态的5-甲基四氢叶酸总杂质一般在1-4％，JK12A达到0.3-3.0％；市售无定形产品中，总杂质甚至高达10％以上，严重威胁到人们的用药安全，这些事实表明，当前所有这些原料药产品中包含的JK12A在内的杂质没有被正确对待。高度的化学不稳定性，也使得应用中难以精确控制剂量。比如，在0.451毫克这个常见剂量下，不稳定使得实际剂量很容易出现较大偏差。
发明内容
本发明的目的是在现在技术的基础上，提供一种含有高纯度(6S)-5-甲基四氢叶酸或其盐的组合物及其制备方法。
本发明的另一目的是提供一种含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液及其制备方法。
本发明第三个目的是提供一种含有高纯度(6S)-5-甲基四氢叶酸或其盐或者含有上述固体溶液的药物组合物。
本发明第四个目的是提供高纯度(6S)-5-甲基四氢叶酸或其盐或者含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液在医药方面的用途。
本发明的目的可以通过以下措施达到：
一种含有(6S)-5-甲基四氢叶酸或其盐的组合物，该组合物中(6S)-5-甲基四氢叶酸或其盐的含量不小于98.0％，其中相关杂质JK12A的含量不大于0.1％,5-甲基四氢蝶酸不得检出；
Figure PCTCN2015088925-appb-000006
本发明的含有(6S)-5-甲基四氢叶酸或其盐的组合物中的主要物质为(6S)-5-甲基四氢叶酸或其盐。该组合物中(6S)-5-甲基四氢叶酸或其盐的含量为(6S)-5-甲基四氢叶酸或其盐的质量含量或HPLC含量，一般为质量含量。
在一种方案中，该组合物中(6S)-5-甲基四氢叶酸或其盐的含量不小于99.0％，其中相关杂质JK12A的含量不大于0.1％,5-甲基四氢蝶酸不得检出。
在另一种方案中，该组合物中(6S)-5-甲基四氢叶酸或其盐的含量不小于99.5％，其中相关杂质JK12A的含量不大于0.1％,5-甲基四氢蝶酸不得检出。
在另一种方案中，该组合物中(6S)-5-甲基四氢叶酸或其盐的含量不小于99.8％，其中相关杂质JK12A的含量不大于0.05％,5-甲基四氢蝶酸不得检出。
本发明中的(6S)-5-甲基四氢叶酸盐选自钾盐、钠盐、钙盐、镁盐、钡盐、锌盐、D-葡糖胺盐、D-半乳糖胺盐或精氨酸盐，优选钙盐、D-葡糖胺盐、D-半乳糖胺盐或精氨酸盐。
一种上述(6S)-5-甲基四氢叶酸或其盐的组合物的制备方法，其包括方法A或方法B；其中，
方法A为：将(6S)-5-甲基四氢叶酸盐粗品与逆变还原剂在水中于pH值6～8、温度50～90℃下直接进行反应，或者在超声条件下进行反应，反应后即得到(6S)-5-甲基四氢叶酸盐组合物；
方法B为：将(6S)-5-甲基四氢叶酸粗品、逆变还原剂和相应盐在水中于pH值6～8、温度50～90℃下进行反应，或者在超声条件下进行反应，反应后即得到(6S)-5-甲基四氢叶酸盐组合物；
方法A或方法B中的逆变还原剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯、异维生素C盐、巯基乙醇、半胱氨酸、巯乙基磺酸、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种。
在一种优选方案中，方法A或方法B中的逆变还原剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯或异维生素C盐中的一种或几种；优选维生素C、维生素C钠盐、维生素C钾盐、维生素C钙盐、维生素C磷酸酯镁盐、维生素C磷酸酯钠盐或异维生素C中的一种或几种；最优选维生素C或异维生素C；所述相应盐为氯化盐，优选氯化钙、氯化镁、氯化锌。
在一种优选方案中，所述方法A或B的反应过程中，反应温度为60～80℃，pH值为7～7.5，优选用氢氧化钠调节pH值；逆变还原剂在水中的重量浓度不小于2％，优选不小于20％，更优选不小于40％。
一种含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液，其主要含有(6S)-5-甲基四氢叶酸或其盐以及稳定剂，该固体溶液的液相色谱图谱中稳定剂和(6S)-5-甲基四氢叶酸盐或其盐的含量之和大于99.5％；其中所述稳定剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯、异维生素C盐、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种。
在一种优选方案中，含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液的液相色谱图谱中稳定剂和(6S)-5-甲基四氢叶酸盐或其盐的含量之和大于99.8％,优选大于99.9％；所述(6S)-5-甲基四氢叶酸或其盐为权利要求1所述的高纯度(6S)-5-甲基四氢叶酸或其盐。
在一种优选方案中，所述稳定剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯或异维生素C盐中的一种或几种，优选维生素C、维生素C钠盐、维生素C钾盐、维生素C钙盐、维生素C磷酸酯镁盐、维生素C磷酸酯钠盐、异维生素C、异维生素C钠盐、异维生素C钾盐、异维生素C钙盐、异维生素C磷酸酯镁盐、异维生素C磷酸酯钠盐中的一种或几种，更优选维生素C或异维生素C。稳定剂和(6S)-5-甲基四氢叶酸盐的重量比为0.2～1000：1，优选1～10：1。
一种上述含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液的制备方法，其包括方法C或方法D，其中，
方法C为：将(6S)-5-甲基四氢叶酸粗品或权利要求1所述的高纯度(6S)-5-甲基四氢叶酸以及稳定剂在水中于pH值3～5、温度50～90℃下直接进行反应，或者在超声条件下进行反应，反应后析出并分离固体；
方法D为：将(6S)-5-甲基四氢叶酸粗品或权利要求1所述的高纯度(6S)-5-甲基四氢叶酸以及稳定剂和相应盐在水中于pH值6～8、温度50～90℃下进行反应，或者在超声条件下进行反应，反应后析出并分离固体；
其中所述稳定剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯、异维生素C盐、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种；所述相应盐为氯化盐，优选氯化钙、氯化镁、氯化锌。
在一种优选方案中，所述方法C的反应过程中，反应温度为60～80℃，pH值为3～4，用盐酸调节pH值；或方法D的反应过程中，反应温度为60～80℃，pH值为7～7.5，用氢氧化钠调节pH值。
一种药物组合物，其包括：
a)上述的含有(6S)-5-甲基四氢叶酸或其盐的组合物，，或者上述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液；和
b)还原剂，所述还原剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯、异维生素C盐、巯基乙醇、半胱氨酸、巯乙基磺酸、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种。
在一种方案中，该药物组合物为普通片剂、缓释片剂、控释片剂、泡腾片剂、颗粒剂、普通胶囊、缓释胶囊、控释胶囊、干混悬剂、口服溶液、口服混悬液、注射剂或冻干粉针剂。
在一种方案中，该药物组合物中的还原剂和(6S)-5-甲基四氢叶酸或其盐的重量比为0.2～1000：1，优选1～10：1。
本发明的高纯度(6S)-5-甲基四氢叶酸或其盐或所述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液在制备治疗或预防哺乳动物与5-甲基四氢叶酸或其盐缺乏有关的疾病的药物中的用途。
名词解释：
若无特别说明，本发明申请文本中所使用的以下名词具有以下含义。
逆转：指(6S)-5-甲基四氢叶酸及其盐中JK12A杂质变回(6S)-5-甲基四氢叶酸及其盐；JK1303变回5-甲基四氢蝶酸的现象。
盐析：固体溶质的溶液中添加一定浓度的中性盐从而改善结晶的方法，经常能够提高收率和质量。
固体溶液：一种或多种固体均匀分散在另一种固体中形成的混合物。
相关物质：(6S)-5-甲基四氢叶酸及其盐在生产，使用，储存过程中带入和产生的杂质；包括4-氨基苯甲酰谷氨酸，JK12A，(6S)-Mefox，吡嗪并三嗪衍生物，四氢叶酸，7,8-二氢叶酸，叶酸，5，10-甲基四氢叶酸，5-甲基四氢蝶酸，N2-甲氨基-四氢叶酸和它们的 盐。
纯度：指在液相色谱中(6S)-5-甲基四氢叶酸峰面积在(6S)-5-甲基四氢叶酸和所有相关物质峰面积总和中所占的比例。
含量：若无特别说明，本文所述含量一般指的是物质的质量含量。
在以前的研究报道中，人们尝试在(6S)-5-甲基四氢叶酸溶液中加入一些除氧剂比如巯基乙醇，抗坏血酸，亚硫酸钠等去除氧气，以阻止持续的氧化降解。他们的结论是，生产中应用这些物质是不可行的，也没有必要。这些研究中，使用低浓度还原剂就可以达到除氧的目的。但这样的浓度无法实现本专利所述氧化杂质的充分逆转。金康方案中使用高浓度还原剂改变了这一状况。
本发明人惊奇的发现，使用较高浓度的还原性适中的还原剂能够实现大部分氧化降解杂质比如JK12A逆转为(6S)-5-甲基四氢叶酸，JK1303逆转为5-甲基四氢蝶酸。之后的盐析作用可以析出高纯度的(6S)-5-甲基四氢叶酸盐，从而在较高收率下得到了不可思议的高纯度，这是以往文献中没有报道过的。
Figure PCTCN2015088925-appb-000007
使用这个方法，可以制备出纯度超过98.0％，甚至接近100.0％纯度的(6S)-5-甲基四氢叶酸及其盐，相关杂质包括JK12A，JK1303，5-甲基四氢蝶酸均大大降低，甚至低于液相色谱的检测限度，这是现有技术无法实现的。
高纯度(6S)-5-甲基四氢叶酸及其盐成功制备的重要意义不仅在于制造技术的显著进步，关键在于纠正和避免了以往因研究缺失造成的对(6S)-5-甲基四氢叶酸及其盐质量要求方面的误导而导致的后果。随着(6S)‐5‐甲基四氢叶酸应用剂量的逐渐扩大，这个意义将更加明显。有一个数据可以说明问题:目前公开的急性毒性试验数据中,(6S)‐5‐甲基四氢叶钙大鼠LD50(半数致死 量)为2000毫克/公斤；采用了发明人的技术,规避有害杂质后的产品,对比试验中15000毫克/公斤剂量老鼠均无异常反应,所以只能得到MTD(最大耐受量)大于15克/公斤的结论。新技术对产品安全性的贡献是不言而喻的。
本发明还提供了一种(6S)-5-甲基四氢叶酸或其盐组合物的制备方法：(6S)-5-甲基四氢叶酸或其盐粗品先溶于水或者含水溶剂中，用逆变还原剂进行处理，使大部分氧化降解杂质逆变为(6S)-5-甲基四氢叶酸盐，然后通过盐析作用或其他方法析出高纯度(6S)-5-甲基四氢叶酸或其盐。
本发明选用的逆变还原剂如上所述，该用于氧化杂质逆转的逆变还原剂要有适中的还原性，还原性过高造成还原降解,过低则达不到效果。
在一种实施方式中，逆变还原剂在方法A、方法B以及上述的组合物制备中，溶剂中的重量浓度不小于2％，优选不小于20％，更优选不小于40％。
上述中使用的盐析这一词汇和生物化学中通常的含义稍有不同，但是效果类似：可以提高结晶收率；强化了结晶母液对杂质的富集效果从而使产品纯度达到一个不可思议的高度。
本发明所述起盐析作用的盐析剂除上述的还原剂,还可以可为常用盐类，比如通过添加氯化钠，氯化钾，硫酸镁等中性无机盐。本发明中优选增大还原剂浓度来达到盐析效果。
另一方面，本发明所提供的由(6S)-5-甲基四氢叶酸或其盐和稳定剂组成的固体溶液，其液相色谱图谱中稳定剂和(6S)-5-甲基四氢叶酸盐含量之和大于99.5％,优选大于99.8％,更优选大于99.9％。上述的含量通过液相色谱图谱中的峰面积计算。
固体溶液作为一种均一性优良的分散形式，能够有效保护本发明的活性成分。
本发明中的固体溶液，使(6S)-5-甲基四氢叶酸与稳定剂保持了高度均一性，从而确保稳定效果。典型实施例中，(6S)-5-甲基四氢叶酸钙为C型结晶，颗粒均匀细小，粒度分布范围狭窄，能够和稳定剂形成高度均匀的固体溶液。这是现有技术难以实现的。
在一种优选的实施方式中，本发明所述固体溶液为(6S)-5-甲基四氢叶酸及其盐与维生素C，或异维生素C，或硫辛酸，或还原型谷胱甘肽，或二硫苏糖醇和它们的盐的固体溶液。
典型情况下，按照本发明制备的固体溶液，在其HPLC图谱中仅有两个典型吸收峰，表现出高度的纯净。
固体溶液中稳定剂和(6S)-5-甲基四氢叶酸均匀分散，可以更好的发挥稳定效果。
本发明中的固体溶液产品中，可以额外添加稳定剂，它还可以根据需要添加其他助剂。
本发明还提供了另一种固体溶液的制备方法：将(6S)-5-甲基四氢叶酸或其盐先溶于水或者含水溶剂中，加入稳定剂使杂质逆转，然后稳定剂和(6S)-5-甲基四氢叶酸及其盐一起析出，过滤，烘干。进一步的，(6S)-5-甲基四氢叶酸和稳定剂分散于水中，滴加氢氧化钠搅拌溶解，然后加入相应的盐，析晶，过滤，烘干。
本发明还进一步提供了(6S)-5-甲基四氢叶酸或其盐的安全用法，包括：a.使用以上所述的含(6S)-5-甲基四氢叶酸或其盐的组合物或以上所述固体溶液；b.添加药学上可以接受的还原性物质作为稳定剂。
在一种优选的(6S)-5-甲基四氢叶酸或其盐的安全用法，其中，所述稳定剂选自维生素C及其酯和盐、异维生素C及其酯和盐，二硫苏糖醇、还原型谷胱甘肽、硫辛酸，优选维生素C及其酯和盐、异维生素C及其酯和盐，更优选维生素C和异维生素C的钠盐、钾盐、钙盐，维生素C磷酸酯镁盐，维生素C磷酸酯钠盐，最优选维生素C和异维生素C。
在一种优选的(6S)-5-甲基四氢叶酸或其盐的安全用法中，稳定剂与(6S)-5-甲基四氢叶酸的组合方式为固体直接混合，或者先溶于溶液中然后一起析出，或者制备成共晶或者固体溶液，优选制备成固体溶液。
上述(6S)-5-甲基四氢叶酸或其盐的安全用法中，稳定剂的使用中断了(6S)-5-甲基四氢叶酸及其盐继续氧化的进程。这个方案可以有效控制JK12A及其他有害降解产物对人体的负面影响，精确控制应用剂量。我们把该安全用法也称为金康方案(METHOD JK)。金康方案中，两个部分单独使用也可以改善(6S)-5-甲基四氢叶酸制品的安全特性。
在一种优选的(6S)-5-甲基四氢叶酸或其盐的安全用法，所述稳定剂和(6S)-5-甲基四氢叶酸的重量比例为0.2～1000：1；更优选1～10：1。
本发明所述的安全用法中，一种优选的实施方案是：使用维生素C及其盐同时作为上文所述还原剂，盐析剂，稳定剂。换言之，维生素C及其盐既是实现氧化杂质逆转的还原剂，又是提纯过程中的盐析剂，同时又是(6S)-5-甲基四氢叶酸的稳定剂。制备过程残留的维生素C及其盐，不需要专门去除。在其后的过程中，根据需要添加需要量的维生素C即可。类似的,使用还原型谷胱甘肽,硫辛酸及其单一异构体,二硫苏糖醇，异维生素C等都可以达到类似效果。按照逻辑,这些物质单独用作还原剂,盐析剂和稳定剂也是合适的。
以上所述的(6S)-5-甲基四氢叶酸或其盐的安全用法，应用于(6S)-5-甲基四氢叶酸单方制剂或者和其他活性成分组成的复方制剂。优选普通片剂、缓释片剂、控释片剂、泡腾 片剂、颗粒剂、普通胶囊、缓释胶囊、控释胶囊、干混悬剂、口服溶液、口服混悬液、注射剂、冻干粉针剂等。更优选普通片剂、普通胶囊、口服溶液、注射剂、冻干粉针剂。
以上所述的(6S)-5-甲基四氢叶酸或其盐的安全用法,在制备治疗或预防哺乳动物与5-甲基四氢叶酸或其盐缺乏有关的疾病的药物中的用途。
本发明还提供了(6S)-5-甲基四氢叶酸或其盐的安全用法在制备治疗或预防哺乳动物与5-甲基四氢叶酸或其盐缺乏有关的疾病的药物中的用途。与5-甲基四氢叶酸或其盐缺乏有关的疾病包括，例如，与痴呆和空泡性的脊髓病相关的亚急性脑炎,脑叶酸缺乏症,病理生理学的血管和心血管疾病，例如过早蔽塞性动脉疾病、在婴儿期和儿童时期中严重的血管疾病、进行性动脉狭窄、间歇性跛行、肾血管高血压、缺血性脑血管疾病、过早视网膜动脉和视网膜静脉闭塞、脑闭塞性动脉疾病、闭塞性外周动脉疾病、由于血栓栓塞疾病和/或缺血性心脏病导致的过早死亡；自身免疫疾病，例如，牛皮癣、乳糜泻、关节炎和炎症病况；由于叶酸盐缺乏导致的巨成红细胞性贫血、肠吸收障碍，用作叶酸拮抗剂(如甲氨蝶呤、乙胺嘧啶或甲氧苄啶等)的解毒剂；用于预防甲氨蝶呤过量或大剂量治疗后所引起的严重毒性作用，用于降低女性流产和/或生产具有神经管缺陷、唇裂缺陷和/或腭裂缺陷的胎儿的风险，用于保持和/或正常高半胱氨酸水平和/或代谢；DNA和RNA的合成和/或功能和/或变化的改变和细胞合成的改变；抑郁症。
本发明所述的还原剂、盐析剂、稳定剂的选用原则首先是化学上的安全，不会使产品发生降解；其次是生理上相对安全，优先选择在食品和医药领域安全性得到充分验证的物质。可以理解，还原剂，盐析剂，稳定剂的用量都要求超出一个最低用量值，否则可能造成不能使杂质充分逆转、盐析效果不佳或/和稳定效果不佳的后果。还可以理解，一般地，还原剂,稳定剂的用量没有严格的上限。
本发明所述的(6S)-5-甲基四氢叶酸及其盐的杂质有JK12A，5-甲基四氢蝶酸，它们的结构分别为
Figure PCTCN2015088925-appb-000008
Figure PCTCN2015088925-appb-000009
其中的5-甲基四氢蝶酸氧化后可以高收率的转化为有严重肾脏毒性的JK1303，因而也需要严格控制。
发明的有益效果是：本发明提供的高纯度(6S)-5-甲基四氢叶酸或其盐和其制备方法，高纯度(6S)-5-甲基四氢叶酸或其盐的固体溶液及其制备方法，以及(6S)-5-甲基四氢叶酸及其盐的安全用法；在这些实施方案中，都通过对药学可接受还原剂的巧妙应用，将(6S)-5-甲基四氢叶酸或其盐产品中可能产生的有害化学品逆转回产品本身，从而达到安全使用(6S)-5-甲基四氢叶酸或其盐产品的目的。
附图说明
图1为JK1303给药14天后小鼠肾小管严重坏死后出现大量空泡的解剖局部放大图。
具体实施方式
为了更好的理解本发明的技术方案，下面结合本发明的具体实施例对本发明的技术方案作进一步说明，但其不限制本发明。
本发明的HPLC检测条件是：
色谱条件：
色谱柱 C18-250*4.6mm-5um
检测波长 UV 280nm
流速 1.0ml/min
进样量 20ul
柱温 30℃
流动相配制：
流动相A：称取7.80g NaH2PO4·2H2O，溶于1000ml水中，用32％NaOH溶液调节pH至6.5。
流动相B：称取5.07g NaH2PO4·2H2O，溶于650ml水中，加入350ml甲醇，用32％NaOH溶液调节pH至8.0。
梯度程序:
T(min) A％ B％
0 100 0
14 45 55
17 0 100
22 0 100
31 100 0
样品溶液制备：取样品适量，在2-8度温度下以低温水(氮气保护下加入亚硫酸钠蒸馏并收集和保存并放冷至2-8℃)。在氮气下溶解制成浓度约0.5mg/ml的溶液，现用现制。
本发明的某些实施例中使用了超声波作为辅助手段,可以明显提高逆转和结晶速度.
为方便说明主要问题，实施例中检测数据只涉及5-甲基四氢叶酸及其盐及其相关物质。
实施例1 JK12A制备
在氮气保护下取10克6S-5-甲基四氢叶酸于反应瓶中，加入80克水，搅拌下用10％氢氧化钠调溶液pH至7.3，固体完全溶清，加入5克活性炭，敞口搅拌过夜反应。HPLC检测原料反应完毕，过滤，滤液用50％的醋酸调pH至4.0，析出晶体，过滤，滤饼分别用乙醇、丙酮洗涤，真空干燥得到6.0克黄色固体JK12A，化学纯度99.42％。
实施例2 5-甲基四氢蝶酸的制备
将6S-5-甲基四氢叶酸61克缓慢加入25％氢氧化钠的水溶液600ml中，升温至回流温度，过夜反应，原料反应完毕后，降温至室温，过滤除杂，滤液用浓硫酸调节PH9-10， 析出杂质，降温至10-15℃，过滤，滤液继续用浓硫酸调节PH 2，析出固体，60℃搅拌半小时，趁热过滤，水洗，干燥，得到44g白色固体，纯度96％。
实施例3 JK1303的制备
取5克6S-5-甲基四氢蝶酸投入三口瓶中，加入50g水，充分搅拌，用10％氢氧化钠溶液调PH至9.0，完全溶清，加3g活性炭，氧气球密闭，搅拌过夜，过滤，滤饼用水洗涤，滤液用50％的醋酸水溶液调PH至4，搅拌析晶30min.过滤，滤饼用水洗两次，30ml丙酮打浆两次，30℃真空干燥，得3.04g产物。HPLC纯度94.3％
实施例4. JK12A的逆转
取0.1克JK12A(HPLC:95.91％)，加入10克水，搅拌下用10％氢氧化钠调pH至7.2，固体全部溶解。然后缓慢加入0.8克二硫苏糖醇，搅拌1h后，取样检测反应液中5-甲基四氢叶酸97.53％，JK12A残留0.72％。
实施例5. JK12A的逆转
取0.1克JK12A(HPLC:95.91％)，加入10克水，搅拌下用10％氢氧化钠调pH至7.2，固体全部溶解。然后缓慢加入0.5克半胱氨酸，搅拌1h后，取样检测反应液中5-甲基四氢叶酸96.64％，JK12A残留0.85％。
实施例6. JK12A的逆转
理学 取0.1克JK12A(HPLC:74.04％)，0.2g巯基乙醇，加入到5ml水中，搅拌下用30％氢氧化钠调pH至7.5，溶解。继续搅拌1h后，取样检测反应液中5-甲基四氢叶酸60.92％，JK12A残留0.59％。
实施例7. JK12A的逆转
取0.1克JK12A(HPLC:74.04％)，0.2g巯乙基磺酸钠，加入到5ml水中，搅拌下用30％氢氧化钠调pH至7.5，溶解。继续搅拌1h后，取样检测反应液中5-甲基四氢叶酸72.85％，JK12A残留0.61％。
实施例8 6(S)-Mefox和JK12A混合物的制备和转化
在氮气保护下取1克6S-5-甲基四氢叶酸于反应瓶中，加入8克水，搅拌，用10％氢氧化钠调溶液pH至7.3，固体完全溶清，加入3克活性炭，敞口搅拌反应10小时，HPLC检测反应液(5-甲基四氢叶酸:12.20％；JK12A:83.51％,6(S)-Mefox:0.57％)；过滤，滤液中加维生素C 10克，液碱调节保持pH7.0，60℃搅拌3小时，取样检测反应液(5-甲基四氢叶酸:96.49％；JK12A:0％；6(S)-Mefox:0％；)
实施例9 JK1303逆转
取0.1克JK1303(HPLC:96.7％)，加入10克水，搅拌下用10％氢氧化钠调pH至7.0，固体全部溶解。然后缓慢加入1克维生素C钠，搅拌1h后，取样检测反应液中5-甲基四氢蝶酸96.6％，JK1303残留为0。
实施例10 6S-5-甲基四氢叶酸钙盐组合物
50ml水，50g维生素C，氢氧化钠中和到7.0，溶解，透明。分批添加6S-5-甲基四氢叶酸钙盐粗品1克。65℃超声2h，过滤，洗涤，真空干燥，得到6S-5-甲基四氢叶酸钙盐组合物0.72g。
相关物质 6S-5-甲基四氢叶酸钙盐粗品/含量 最终产品/含量
JK12A 0.49％ —
5-甲基四氢叶酸 99.26％ 99.9％
5-甲基四氢蝶酸 0.14％ —
其他杂质 0.11％ 0.01
含量为质量含量，下同。
实施例11 6S-5-甲基四氢叶酸钙盐组合物
6S-5-甲基四氢叶酸15克，150g维生素C，加入到225ml水中，30％的氢氧化钠水溶液中和到7.0，溶解。搅拌1h后加15g 40％的氯化钙水溶液，70℃超声析晶，3h后析出白色颗粒，过滤，然后水洗，滤饼真空干燥，得10.1g 6S-5-甲基四氢叶酸钙盐固体组合物。
相关物质 6S-5-甲基四氢叶酸粗品/含量 最终产品/含量
JK12A 0.22％ 0.03％
6(S)-Mefox 0.02％ 0.01％
5-甲基四氢叶酸 99.40％ 99.95％
5-甲基四氢蝶酸 0.03％ —
其他杂质 0.33％ 0.01％
实施例12 6S-5-甲基四氢叶酸钙盐组合物
6S-5-甲基四氢叶酸1g，1g维生素C，0.5gNaCl，加入到10ml水中，30％的氢氧化钠水溶液中和到7.0，溶解。搅拌1h后加1g40％的氯化钙水溶液，70℃超声，1h后析出固体，过滤，洗涤，滤饼真空干燥，得0.52g 6S-5-甲基四氢叶酸钙盐固体组合物。
相关物质 6S-5-甲基四氢叶酸粗品/含量 最终产品/含量
JK12A 0.22％ 0.07％
6(S)-Mefox 0.02％ 0.01％
5-甲基四氢叶酸 99.40％ 99.75％
5-甲基四氢蝶酸 0.03％ —
其他杂质 0.33％ 0.15％
实施例13 6S-5-甲基四氢叶酸钙盐组合物
6S-5-甲基四氢叶酸0.5克，10mg维生素C，0.1g氯化钠，加入到10ml水中，30％的氢氧化钠水溶液中和到7.5，溶解，透明。氮气保护下，搅拌1h，再加1g 20％的氯化钙水溶液，转入70℃超声析晶，1.0h后析出白色颗粒。过滤，洗涤，滤饼真空干燥，得0.38g 6S-5-甲基四氢叶酸钙盐固体组合物。
相关物质 6S-5-甲基四氢叶酸粗品/含量 最终产品/含量
JK12A 0.22％ 0.09％
6(S)-Mefox 0.02％  
5-甲基四氢叶酸 99.40％ 99.63％
5-甲基四氢蝶酸 0.03％ —
其他杂质 0.33％ 0.27％
实施例14 6S-5-甲基四氢叶酸钙盐组合物
6S-5-甲基四氢叶酸1g，3g异维生素C，加入到20ml水中，30％的氢氧化钠水溶液中和到7.0，溶解，透明。搅拌1h后加1g40％的氯化钙水溶液，60℃超声，1h后析出固体，过滤，洗涤，滤饼真空干燥，得0.80g 6S-5-甲基四氢叶酸钙盐组合物。
相关物质 6S-5-甲基四氢叶酸粗品/含量 最终产品/含量
JK12A 0.22％ -
6(S)-Mefox 0.02％ 0.05％
5-甲基四氢叶酸 99.40％ 99.75％
5-甲基四氢蝶酸 0.03％ -
其他杂质 0.33％ 0.18％
实施例15 6S-5-甲基四氢叶酸组合物
取0.5g实施例11的(6S)-5-甲基四氢叶酸钙盐组合物，加热至40℃，用盐酸溶液调节至pH4.0，维持体系的pH不变，搅拌1h析晶，冷却至室温，过滤，洗涤。25℃真空烘干，得到0.23g(6S)-5-甲基四氢叶酸组合物，HPLC检测((6S)-5-甲基四氢叶酸：99.80％， JK12A0％,6(S)-Mefox：0.07％，5-甲基四氢蝶酸0％)。
相关物质 6S-5-甲基四氢叶酸钙组合物/含量 最终产品/含量
JK12A 0.03％ -
6(S)-Mefox 0.01％ -
5-甲基四氢叶酸 99.95％ 99.80％
5-甲基四氢蝶酸 — -
其他杂质 0.01％ 0.20％
实施例16 6R,S 5-甲基四氢叶酸钙盐组合物
6R,S-5-甲基四氢叶酸1克，10g维生素C，加入到15ml水中，30％的氢氧化钠水溶液中和到7.0，溶解，透明。加1g 40％的氯化钙水溶液，70℃超声析晶，过滤，然后水洗，真空干燥，得到0.54g 6R,S-5-甲基四氢叶酸钙盐组合物。
相关物质 6R，S-5-甲基四氢叶酸粗品/含量 最终产品/含量
JK12A 0.21％ —
6(S)-Mefox — —
5-甲基四氢叶酸 99.15％ 99.59％
5-甲基四氢蝶酸 0.28％ 0.08％
其他杂质 0.36％ 0.33％
实施例17 6R,S-5-甲基四氢叶酸钠盐组合物
6R,S-5-甲基四氢叶酸1克，5g维生素C，加入到20ml水中，30％的氢氧化钠水溶液中和到7.0，溶解，透明，60℃下隔氧反应2小时。然后将上述反应液滴加至200ml乙醇中析出钠盐，过滤，洗涤，真空干燥，得到0.52g 6R,S-5-甲基四氢叶酸钠盐组合物。
相关物质 6R，S-5-甲基四氢叶酸粗品/含量 最终产品/含量
JK12A 0.21％ —
6(S)-Mefox — —
5-甲基四氢叶酸 99.15％ 99.64％
5-甲基四氢蝶酸 0.28％ 0.12％
其他杂质 0.36％ 0.34％
实施例18 6S-5-甲基四氢叶酸精氨酸盐组合物
5g维生素C，加入5ml水，氢氧化钠中和到7.0，溶解，透明。加6S-5-甲基四氢叶 酸精氨酸盐粗品0.5g，溶清，60℃超声搅拌2h，将上述反应液滴加至50ml乙醇中，搅拌析出固体，过滤，检测固体，得到6S-5-甲基四氢叶酸精氨酸盐组合物0.26g。
相关物质 5-甲基四氢叶酸精氨酸盐粗品/含量 最终产品/含量
JK12A 0.76％ —
6(S)-Mefox 1.30％ 0.27
5-甲基四氢叶酸 97.69％ 99.70％
5-甲基四氢蝶酸 0.05％ —
其他杂质 0.20％ 0.03％
实施例19 6S-5-甲基四氢叶酸D-葡萄糖胺盐组合物
5g维生素C，加入10ml水，氢氧化钠中和到7.0，溶解，透明。加6S-5-甲基四氢叶酸D-葡萄糖胺盐粗品2.0g，溶清，60℃超声搅拌2h，缓慢滴加乙醇，析出固体，过滤，乙醇重结晶，得到6S-5-甲基四氢叶酸D-葡萄糖胺盐组合物。
Figure PCTCN2015088925-appb-000010
实施例20 6S-5-甲基四氢叶酸钙盐和维生素C的固体溶液
6S-5-甲基四氢叶酸1克，2.0g维生素C，加入到20ml水中，30％的氢氧化钠水溶液中和到7.5，溶解。搅拌1h后加1g 40％的氯化钙水溶液，转入70℃超声析晶，1.0h后析出白色颗粒，过滤，烘干，液相检测6S-5-甲基四氢叶酸钙盐：94.89％，维生素C：5.11％。
实施例21 6S-5-甲基四氢叶酸钙盐和异维生素C的固体溶液
6S-5-甲基四氢叶酸0.1克，0.3g异维生素C，加入到1.5ml水中，30％的氢氧化钠水溶液中和到7.5，溶解。搅拌1h后加1g40％的氯化钙水溶液，转入90℃搅拌析晶，1.0h后析出白色颗粒，过滤，烘干，液相检测6S-5-甲基四氢叶酸钙盐：29.29％，异维生素C：70.66％。
实施例22 6S-5-甲基四氢叶酸钙盐和硫辛酸的固体溶液
6S-5-甲基四氢叶酸0.5克，1.0g硫辛酸，加入到10ml水中，30％的氢氧化钠水溶液中和到7.5，溶解。搅拌1h后加1g 20％的氯化钙水溶液，转入70℃超声析晶，1h后得到固体溶液，过滤，烘干，液相检测(使用ELSD检测器)6S-5-甲基四氢叶酸钙盐：27.69％，硫辛酸72.26％。
实施例23 5-甲基四氢叶酸钙和还原型谷胱甘肽的固体溶液
6S-5-甲基四氢叶酸0.5克，1.5g还原型谷胱甘肽，加入到10ml水中，30％的氢氧化钠水溶液中和到7.5，溶解，搅拌1h，加1g20％的氯化钙水溶液，转入70℃超声析晶，1.0h后析出白色颗粒，过滤，烘干，液相检测(使用ELSD检测器)6S-5-甲基四氢叶酸钙盐：37.62％，还原型谷胱甘肽62.33％。
实施例24 (6S)-5-甲基四氢叶酸和维生素C的固体溶液
6S-5-甲基四氢叶酸0.5克，3.0g维生素C，加入到10ml水中，30％的氢氧化钠水溶液中和到7.5，溶解，搅拌1h，滴加适当6N的盐酸水溶液，调节pH至4.0，50℃搅拌析晶，1.0h后析出白色颗粒，过滤，烘干，液相检测6S-5-甲基四氢叶酸：26.80％，维生素C：73.17％。
实施例25 (6S)-5-甲基四氢叶酸锌和维生素C的固体溶液
将6S-5-甲基四氢叶酸0.5克，1.5g维生素C，加入到10ml水中，20％的氢氧化钠水溶液中和，至pH7.5时，反应液溶清。加入0.8g50％的氯化锌水溶液，结晶2小时，氮气保护下过滤，真空干燥.液相检测含(6S)-5-甲基四氢叶酸锌88.7％,维生素C：11.0％。
实施例26(6S)-5-甲基四氢叶酸钙和维生素C磷酸酯镁的固体溶液
将6S-5-甲基四氢叶酸0.5克，2.0g维生素C磷酸酯镁，加入到10ml水中，20％的氢氧化钠水溶液中和到7.5，溶清。加入0.6g50％的氯化钙水溶液，转入70℃超声中反应，1.5h后析出白色颗粒，氮气保护下过滤，烘干.液相检测含(6S)-5-甲基四氢叶酸钙62.4％,维生素C：37.4％。
实施例27 金康方案产品6S-5-甲基四氢叶酸钙盐与维生素C的组合物稳定性
将1g实施例11的6S-5-甲基四氢叶酸钙盐组合物与不同重量比例的维生素C，充分研磨混合,放置于25℃，湿度40％的恒温箱中，定期检测6S-5-甲基四氢叶酸钙和JK12A。
Figure PCTCN2015088925-appb-000011

Claims (14)

  1. 一种含有(6S)-5-甲基四氢叶酸或其盐的组合物，其特征在于该组合物中(6S)-5-甲基四氢叶酸或其盐的含量不小于98.0％，其中相关杂质JK12A的含量不大于0.1％,5-甲基四氢蝶酸不得检出；
    Figure PCTCN2015088925-appb-100001
  2. 根据权利要求1所述的含有(6S)-5-甲基四氢叶酸或其盐的组合物，其特征在于(6S)-5-甲基四氢叶酸或其盐的含量不小于99.0％，其中相关杂质JK12A的含量不大于0.1％,5-甲基四氢蝶酸不得检出；优选(6S)-5-甲基四氢叶酸或其盐的含量不小于99.5％，其中相关杂质JK12A的含量不大于0.1％,5-甲基四氢蝶酸不得检出；更优选(6S)-5-甲基四氢叶酸或其盐的含量不小于99.8％，其中相关杂质JK12A的含量不大于0.05％,5-甲基四氢蝶酸不得检出。
  3. 根据权利要求1所述的含有(6S)-5-甲基四氢叶酸或其盐的组合物，其特征在于所述(6S)-5-甲基四氢叶酸盐选自钾盐、钠盐、钙盐、镁盐、钡盐、锌盐、D-葡糖胺盐、D-半乳糖胺盐或精氨酸盐，优选钙盐、D-葡糖胺盐、D-半乳糖胺盐或精氨酸盐。
  4. 一种权利要求1或2所述的高纯度(6S)-5-甲基四氢叶酸或其盐的组合物的制备方法，其特征在于其包括方法A或方法B；其中，
    方法A为：将(6S)-5-甲基四氢叶酸盐粗品与逆变还原剂在水中于pH值6～8、温度50～90℃下直接进行反应，或者在超声条件下进行反应，反应后即得到(6S)-5-甲基四氢叶酸盐组合物；
    方法B为：将(6S)-5-甲基四氢叶酸粗品、逆变还原剂和相应盐在水中于pH值6～8、温度50～90℃下进行反应，或者在超声条件下进行反应，反应后即得到(6S)-5-甲基四氢叶酸盐组合物；
    所述逆变还原剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C 酯、异维生素C盐、巯基乙醇、半胱氨酸、巯乙基磺酸、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种。
  5. 根据权利要求4所述的高纯度(6S)-5-甲基四氢叶酸或其盐的组合物的制备方法，其特征在于所述逆变还原剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯或异维生素C盐中的一种或几种；优选维生素C、维生素C钠盐、维生素C钾盐、维生素C钙盐、维生素C磷酸酯镁盐、维生素C磷酸酯钠盐或异维生素C中的一种或几种；最优选维生素C或异维生素C；所述相应盐为氯化盐，优选氯化钙、氯化镁、氯化锌。
  6. 根据权利要求4所述的高纯度(6S)-5-甲基四氢叶酸或其盐的组合物的制备方法，其特征在于所述方法A或B的反应过程中，反应温度为60～80℃，pH值为7～7.5，优选用氢氧化钠调节pH值；逆变还原剂在水中的重量浓度不小于2％，优选不小于20％，更优选不小于40％。
  7. 一种含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液，其特征在于其主要含有(6S)-5-甲基四氢叶酸或其盐以及稳定剂，该固体溶液的液相色谱图谱中稳定剂和(6S)-5-甲基四氢叶酸盐或其盐的含量之和大于99.5％；其中所述稳定剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯、异维生素C盐、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种。
  8. 根据权利要求7所述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液，其特征在于该固体溶液的液相色谱图谱(6S)-5-甲基四氢叶酸盐或其盐的含量之和大于99.8％,优选大于99.9％；所述(6S)-5-甲基四氢叶酸或其盐为权利要求1所述的高纯度(6S)-5-甲基四氢叶酸或其盐。
  9. 根据权利要求7所述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液，其特征在于所述稳定剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯或异维生素C盐中的一种或几种，优选维生素C、维生素C钠盐、维生素C钾盐、维生素C钙盐、维生素C磷酸酯镁盐、维生素C磷酸酯钠盐、异维生素C、异维生素C钠盐、异维生素C钾盐、异维生素C钙盐、异维生素C磷酸酯镁盐、异维生素C磷酸酯钠盐中的一种或几种，更优选维生素C或异维生素C；稳定剂和(6S)-5-甲基四氢叶酸盐的重量比为0.2～1000：1，优选1～10：1。
  10. 权利要求7所述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液的制备方法，其特征在于其包括方法C或方法D，其中，
    方法C为：将(6S)-5-甲基四氢叶酸粗品或权利要求1所述的高纯度(6S)-5-甲基四氢叶酸以及稳定剂在水中于pH值3～5、温度50～90℃下直接进行反应，或者在超声条件下进行反应，反应后析出并分离固体；
    方法D为：将(6S)-5-甲基四氢叶酸粗品或权利要求1所述的高纯度(6S)-5-甲基四氢叶酸以及稳定剂和相应盐在水中于pH值6～8、温度50～90℃下进行反应，或者在超声条件下进行反应，反应后析出并分离固体；
    其中所述稳定剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯、异维生素C盐、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种；所述相应盐为氯化盐，优选氯化钙、氯化镁、氯化锌。
  11. 权利要求10所述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液的制备方法，其特征在于所述方法C的反应过程中，反应温度为60～80℃，pH值为3～4，用盐酸调节pH值；方法D的反应过程中，反应温度为60～80℃，pH值为7～7.5，用氢氧化钠调节pH值。
  12. 一种药物组合物，其特征在于其包括：
    a)权利要求1所述的含有(6S)-5-甲基四氢叶酸或其盐的组合物，或者权利要求6所述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液；和
    b)还原剂，所述还原剂选自维生素C、维生素C酯、维生素C盐、异维生素C、异维生素C酯、异维生素C盐、巯基乙醇、半胱氨酸、巯乙基磺酸、二硫苏糖醇、还原型谷胱甘肽或硫辛酸中的一种或几种。
  13. 根据权利要求12所述的药物组合物，其特征在于该组合物为普通片剂、缓释片剂、控释片剂、泡腾片剂、颗粒剂、普通胶囊、缓释胶囊、控释胶囊、干混悬剂、口服溶液、口服混悬液、注射剂或冻干粉针剂；其中还原剂和(6S)-5-甲基四氢叶酸或其盐的重量比为0.2～1000：1，优选1～10：1。
  14. 权利要求1所述的高纯度(6S)-5-甲基四氢叶酸或其盐或权利要求6所述的含有(6S)-5-甲基四氢叶酸或其盐和稳定剂的固体溶液在制备治疗或预防哺乳动物与5-甲基四氢叶酸或其盐缺乏有关的疾病的药物中的用途。
PCT/CN2015/088925 2014-09-04 2015-09-04 (6s)-5-甲基四氢叶酸或其盐组合物及其制备和应用 WO2016034145A1 (zh)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3512507A4 (en) * 2016-10-20 2019-11-13 Grunbiotics Pty Ltd ORAL MEDICINAL FORMULATIONS
CN115656372A (zh) * 2022-10-27 2023-01-31 北京斯利安药业有限公司 一种s-四氢叶酸异构体手性分析方法

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109864978B (zh) * 2017-12-04 2022-07-12 深圳奥萨制药有限公司 一种5-甲基四氢叶酸的缓释制剂及其制备方法
CN109939077B (zh) * 2017-12-04 2022-07-22 深圳奥萨制药有限公司 一种含有5-甲基四氢叶酸的控释制剂
CN108926544B (zh) * 2018-08-19 2021-08-06 张奉明 一种四代叶酸片及其制备方法
CN109164182B (zh) * 2018-09-19 2021-06-11 无锡紫杉药业有限公司 一种对l-四氢叶酸对甲苯磺酸盐（6s）光学纯度的分析检测方法
CN110237046B (zh) * 2019-03-29 2021-07-30 福格森（武汉）生物科技股份有限公司 一种l-5-甲基四氢叶酸微囊的制备方法
EP4360635A1 (en) * 2021-06-24 2024-05-01 Lianyungang Jinkang Hexin Pharmaceutical Co. Ltd. Application of folic acid derivatives in preparation of drugs for treating contact lens discomfort and xerophthalmia
CN114957257A (zh) * 2022-03-21 2022-08-30 河北冀衡药业股份有限公司 一种5-甲基四氢叶酸钙的制备方法
CN115611899A (zh) * 2022-10-25 2023-01-17 无锡紫杉药业股份有限公司 一种l-5-甲基四氢蝶酸的制备方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009103334A1 (en) * 2008-02-20 2009-08-27 Gnosis S.P.A. Folates, compositions and uses thereof
CN102702200A (zh) * 2012-04-25 2012-10-03 连云港金康和信药业有限公司 (6rs)-5-甲基四氢叶酸钙盐晶型及其制备方法
CN102775408A (zh) * 2012-01-20 2012-11-14 连云港金康医药科技有限公司 稳定的无定型5-甲基四氢叶酸盐及其制备方法
WO2013107236A1 (zh) * 2012-01-20 2013-07-25 连云港金康和信药业有限公司 （6s）-5-甲基四氢叶酸盐晶型及其制备方法
CN103450202A (zh) * 2012-06-04 2013-12-18 南京莱因医药科技有限公司 L-5-甲基四氢叶酸与有机碱加成盐的方法
CN103694239A (zh) * 2012-09-27 2014-04-02 上海科胜药物研发有限公司 一种5-甲基-（6s）-四氢叶酸晶型a及其制备方法

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5124452A (en) * 1978-07-10 1992-06-23 Bioresearch S.P.A. Process for producing d,1-5-methyltetrahydrofolic acid and its salts
CH693905A5 (de) 1999-04-15 2004-04-15 Eprova Ag Stabile kristalline Salze von 5-Methyltetrahydrofolsäure.
UY32836A (es) 2009-08-12 2011-03-31 Bayer Schering Pharma Ag Partículas estabilizadas que comprenden 5-metil-(6s)-tetrahidrofolato
EP2837632B1 (en) * 2012-04-13 2016-05-18 Lianyungang Jinkang Hexin Pharmaceutical Co. Ltd. Derivatives of triazabicyclo[3.2.1]octane useful for the treatment of proliferative diseases
CN103664945B (zh) * 2012-09-07 2016-01-20 南京莱因医药科技有限公司 L-5-甲基四氢叶酸氨基酸盐的制备方法
CN102813656B (zh) 2012-09-13 2014-05-07 广东岭南制药有限公司 5-甲基四氢叶酸或其盐的稳定药物组合物
EP2781214A1 (en) * 2013-03-22 2014-09-24 Chemo Research, S.L. Formulation of amorphous calcium L-5-methyltetrahydrofolate (L-5-MTHF-Ca)
CN107812195B (zh) * 2014-09-04 2021-04-20 连云港金康和信药业有限公司 (6s)‐5‐甲基‐四氢叶酸钙盐的稳定药物组合物

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009103334A1 (en) * 2008-02-20 2009-08-27 Gnosis S.P.A. Folates, compositions and uses thereof
CN102775408A (zh) * 2012-01-20 2012-11-14 连云港金康医药科技有限公司 稳定的无定型5-甲基四氢叶酸盐及其制备方法
WO2013107236A1 (zh) * 2012-01-20 2013-07-25 连云港金康和信药业有限公司 （6s）-5-甲基四氢叶酸盐晶型及其制备方法
CN102702200A (zh) * 2012-04-25 2012-10-03 连云港金康和信药业有限公司 (6rs)-5-甲基四氢叶酸钙盐晶型及其制备方法
CN103450202A (zh) * 2012-06-04 2013-12-18 南京莱因医药科技有限公司 L-5-甲基四氢叶酸与有机碱加成盐的方法
CN103694239A (zh) * 2012-09-27 2014-04-02 上海科胜药物研发有限公司 一种5-甲基-（6s）-四氢叶酸晶型a及其制备方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP3190112A4 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3512507A4 (en) * 2016-10-20 2019-11-13 Grunbiotics Pty Ltd ORAL MEDICINAL FORMULATIONS
CN115656372A (zh) * 2022-10-27 2023-01-31 北京斯利安药业有限公司 一种s-四氢叶酸异构体手性分析方法

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