Folates for medical use
阅读说明：本技术 医用叶酸盐 (Folates for medical use ) 是由 M·乌尔曼 G·维斯勒 A·博登穆勒 M·米勒 于 2018-08-29 设计创作，主要内容包括：本发明涉及无定形叶酸盐。该盐由叶酸阴离子和有机阳离子组成。叶酸阴离子选自5-甲酰基-(6S)-四氢叶酸、10-甲酰基-(6R)-四氢叶酸、5-甲基-(6S)-四氢叶酸、(6S)-四氢叶酸、5,10-亚甲基-(6R)-四氢叶酸及它们的氧化衍生物JK12A和Mefox,阳离子为选自精氨酸、胆碱、乙酰胆碱、1,1-二甲基-双胍、苯乙基-双胍、甜菜碱甲酯和二甲基氨基乙醇的有机化合物。阳离子是具有互补药理活性的有机化合物。(The present invention relates to amorphous folate. The salt consists of a folate anion and an organic cation. The folate anion is selected from the group consisting of 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5-methyl- (6S) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid and their oxidized derivatives JK12A and Mefox, and the cation is an organic compound selected from the group consisting of arginine, choline, acetylcholine, 1-dimethyl-biguanide, phenethyl-biguanide, betaine methyl ester and dimethylaminoethanol. The cation is an organic compound having complementary pharmacological activity.)
1. Amorphous folate consisting of a tetrahydrofolate anion and an organic cation, wherein the anion is selected from the group consisting of 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5-methyl- (6S) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid and its oxidized derivatives JK12A and Mefox, and the cation is an organic compound selected from the group consisting of: arginine, choline, acetylcholine, 1-dimethyl-biguanide, phenylethylbiguanide, betaine methyl ester and dimethylaminoethanol, the folate having a high solubility in a non-polar solvent having a relative polarity of 0.82 compared to water.
2. Amorphous folate according to claim 1, consisting of a tetrahydrofolate anion and an organic cation, wherein said anion is selected from the group consisting of 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5-methyl- (6S) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid and its oxidized derivatives JK12A and Mefox, and said cation is selected from the group consisting of di-arginine, di-choline, di-acetylcholine, di- (1, 1-dimethyl-biguanide), di-dimethylaminoethanol, di-betaine methyl ester and di- (phenethyl-biguanide).
3. Amorphous folate according to claim 1, consisting of a tetrahydrofolate anion and an organic cation, wherein said anion is selected from the group consisting of 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5-methyl- (6S) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid and its oxidized derivatives JK12A and Mefox, and said cation is selected from the group consisting of mono-arginine, mono-choline, mono-acetylcholine, mono- (1, 1-dimethyl-biguanide), mono-dimethylaminoethanol, mono-betaine methyl ester and mono- (phenethyl-biguanide).
4. The amorphous folate of claim 2, wherein said tetrahydrofolate anion is 5-formyl- (6S) -tetrahydrofolate, said organic cation is di-arginine, and said amorphous folate is at D2In O1The H-NMR shifts were:
5. the amorphous folate according to claim 2, wherein said tetrahydrofolate anion is 5-formyl- (6S) -tetrahydrofolate, said organic cation is di-choline, and said amorphous folate is at D2In O1The H-NMR shifts were:
。
6. The amorphous folate according to claim 2, wherein said tetrahydrofolate anion is 5-formyl- (6S) -tetrahydrofolate, said organic cation is bis- (1, 1-dimethyl-biguanide), and said amorphous folate is at D2In O1The H-NMR shifts were:
7. amorphous folate according to claim 2, characterized in thatWherein the tetrahydrofolate anion is 5-formyl- (6S) -tetrahydrofolate, the organic cation is bis- (phenethyl-biguanide), and the amorphous folate is at D2In O1The H-NMR shifts were:
。
8. The amorphous folate of claim 2, wherein said tetrahydrofolate anion is 5-methyl- (6S) -tetrahydrofolate, said organic cation is di-arginine, and said amorphous folate is at D2In O1The H-NMR shifts were:
。
9. The amorphous folate according to claim 2, wherein said tetrahydrofolate anion is 5-methyl- (6S) -tetrahydrofolate, said organic cation is di-choline, and said amorphous folate is at D2In O1The H-NMR shifts were:
10. the amorphous folate according to claim 2, wherein said tetrahydrofolate anion is 5-methyl- (6S) -tetrahydrofolate and said organic cation is bis- (1, 1-dimethyl-bisGuanidine), the amorphous folate is at D2In O1The H-NMR shifts were:
。
11. The amorphous folate according to claim 2, wherein said tetrahydrofolate anion is 5-methyl- (6S) -tetrahydrofolateThe organic cation is bis- (phenethyl-biguanide), and the amorphous folate is at D2In O1The H-NMR shifts were:
12. a pharmaceutical composition comprising at least one folate according to any one of claims 1 to 11 as the main active compound and at least one pharmaceutically acceptable excipient.
13. Use of at least one folate according to any one of claims 1 to 11, for the preparation of a medicament, food additive or nutritional supplement for the prevention and/or treatment of a defect or disease in which the administration of tetrahydrofolate leads to a positive stimulatory effect.
14. A method of preparing the amorphous folate of any one of claims 1 to 11, consisting of tetrahydrofolate anion and an organic cation, comprising the step of adding oxalic acid or a fluoride salt to an aqueous composition of an alkaline earth metal salt of folate.
Technical Field
The present invention relates to folate, to its preparation, and to compositions comprising folate.
Background
Depression and other mental health disorders such as dementia, autism, ADHD and alzheimer's disease, as well as chronic non-infectious diseases (NCD) such as type 2 diabetes, vascular disease and cancer, are an increasing burden on patients and medical systems, especially in view of the aging population. The causes of these different diseases are manifold; however, as a common risk factor, suboptimal folate status is found in the whole body or in specific tissues.
It is well known that the vitamins of the B-complex are involved in many metabolic processes of the human body, such as the conversion of carbohydrates to glucose, wherein glucose is metabolized to produce energy. These vitamins are more important in the breakdown of fats and proteins and play an important role in maintaining the tone of muscles in the inner wall of the digestive tract and promoting the health of the nervous system (e.g., eyes, skin, hair, liver).
Furthermore, it is well known that folic acid is mandatory in the production and maintenance of new cells. This is particularly important during periods of rapid cell division and growth, such as infancy and pregnancy. Folate is essential for replication of DNA. Therefore, folate deficiency can block DNA synthesis and cell division, most clinically affecting the bone marrow (i.e., the site of rapid cell turnover). Since synthesis of RNA and proteins is not hindered, large red blood cells (i.e., megaloblasts) are produced, resulting in giant cell anemia such as megaloblastic anemia, which is seen in celiac disease, nutritional anemia, pregnancy, infancy, or childhood. Therefore, folic acid is required in both adults (especially the elderly) and children to produce normal red blood cells and to prevent anemia. Folate also helps prevent DNA changes that may lead to cancer.
However, tetrahydrofolic acid and its derivatives are known to have a very high instability, in particular due to their sensitivity to oxidation. in particular, 5-formyltetrahydrofolic acid (Folinic acid), leucovorin (L eucovin) is of great importance as a pharmaceutical ingredient mainly in oncology, as a concomitant therapy for methotrexate and 5-fluorouracil treatment and in the treatment of folate-deficient anemia associated with pregnancy, antibiotic treatment etc. among folic acid and reduced folic acid, calcium salts can be said to be the most stable derivatives US 5817659 and US 6441168 disclose crystalline salts (preferably calcium salts) of 5-methyl- (6R, S) -, (6S) -or (6R) -tetrahydrofolic acid with at least one equivalent of crystal water per said acid 5-methyltetrahydrofolic acid is the only commercially available 5-methyltetrahydrofolic acid derivatives which can directly penetrate the blood/brain barrier without the need for further metabolism.
Various folates are known. Typically, these salts comprise folic acid and either inorganic cations (such as calcium and magnesium) or organic cations (such as glucosamine or galactosamine). These alkaline earth metal cations are so inert that they do not have any pharmacological effect on the human body per se. The rare solubility of this salt in aqueous solutions has been widely reported. WO2009/103334 describes a glucosamine salt of 5-methyl-tetrahydrofolate (5-MTHF-glucosamine) with good water solubility. Furthermore, the solubility of folate in non-polar solvents is also very limited. For example, 5-MTHF-glucosamine is described as a milky to light brown powder, very soluble in water (at 25 ℃), soluble in dilute acids or dilute bases, insoluble in organic solvents (EFSA journal 2013; 11 (10): 3358). Aqueous compositions with improved folic acid solubility and stability have been disclosed, for example in US9301922 and US 9642853. In addition, a number of folic acid compositions have been described which contain folic acid and other compounds such as vitamins, lysine, thiamine and other active ingredients. However, a stable folate having good solubility in water and non-polar solvents in combination with other active compounds would allow for a more versatile pharmaceutical composition.
Disclosure of Invention
It is an object of the present invention to provide folate, which binds other active compounds and exhibits good stability, high solubility in non-polar solvents and good water solubility.
This object is achieved by the folate as defined in claim 1 of the present invention. Further preferred embodiments are subject to the dependent claims.
The amorphous folate according to the invention consists of a tetrahydrofolate anion and an organic cation, wherein the anion is selected from the group consisting of 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5-methyl- (6S) -tetrahydrofolic acid, 5, 10-diformyl- (6S) -tetrahydrofolic acid, 5-methyl-10-formyl- (6S) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid and its oxidized derivatives JK12A and Mefox. Furthermore, the cation is an organic compound selected from the group consisting of: arginine, choline, acetylcholine, 1-dimethyl-biguanide, phenethylbiguanide, betaine methyl ester and dimethylaminoethanol. The folate has a high solubility in non-polar solvents, which have a relative polarity of 0.82 compared to water.
Solubility of a particular folate in a particular non-polar solvent of greater than 5 mass% (m/m) is considered high solubility in a non-polar solvent. The solubility was determined at 20 ℃.
The structure of Mefox is shown below:
the structure of JK12A is shown below:
both compounds Mefox and JK12A are oxidized derivatives of 5-methyl- (6S) -tetrahydrofolic acid.
In a preferred embodiment, the amorphous folate consists of tetrahydrofolate anion and an organic cation. The anion is selected from the group consisting of 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5-methyl- (6S) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid and its oxidized derivatives JK12A and Mefox. The cation is selected from the group consisting of di-arginine, di-choline, di-acetylcholine, di- (1, 1-dimethyl-biguanide), di- (phenethyl-biguanide), di-betaine methyl ester and di-dimethylaminoethanol.
In another embodiment, the amorphous folate consists of a tetrahydrofolate anion and an organic cation, wherein the anion is selected from the group consisting of 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5-methyl- (6S) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid and its oxidized derivatives JK12A and Mefox, and wherein the cation is selected from the group consisting of mono-arginine, mono-choline, mono-acetylcholine, mono- (1, 1-dimethyl-biguanide), mono- (phenethyl-biguanide), mono-betaine methyl ester and mono-dimethylaminoethanol.
In another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation mono-arginine, the amorphous tetrahydrofolate is in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous tetrahydrofolate is formed from the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate andorganic cationic mono- (1, 1-dimethyl-biguanide) composition, amorphous tetrahydrofolate in heavy water (D)2In O)1The H-NMR shifts were:
chemical Shift δ (1H) in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
8.51/7.8
Unimodal (s)/unimodal(s)
1H
7.54/7.50
Doublet (d)/doublet (d)
2H
6.62/6.57
Doublet (d)/doublet (d)
2H
4.73
Multiplet peak (m)
1H
4.27
Multiplet peak (m)
1H
3.42
Multiplet peak (m)
1H
3.28
Multiplet peak (m)
2H
3.16
Multiplet peak (m)
1H
2.92
Single peak(s)
6H
2.32
Multiplet peak (m)
2H
2.12
Multiplet peak (m)
1H
1.95
Multiplet peak (m)
1H
In another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation mono- (phenethyl-biguanide), the amorphous tetrahydrofolate being in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation mono-choline, in heavy water (D)2In O)1The H-NMR shifts were:
chemical Shift δ (1H) in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
8.50/7.80
Unimodal (s)/unimodal(s)
1H
7.54/7.50
Doublet (d)/doublet (d)
2H
6.62/6.57
Doublet (d)/doublet (d)
2H
4.73
Multiplet peak (m)
1H
4.27
Multiplet peak (m)
1H
3.93
Multiplet peak (m)
2H
3.43
Multiplet peak (m)
1H
3.39
Multiplet peak (m)
2H
3.27
Multiplet peak (m)
2H
3.16
Multiplet peak (m)
1H
3.07
Single peak(s)
9H
2.32
Multiplet peak (m)
2H
2.12
Multiplet peak (m)
1H
1.95
Multiplet peak (m)
1H
In another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-methyl- (6S) -tetrahydrofolate and the organic cation mono- (1, 1-dimethyl-biguanide), the amorphous tetrahydrofolate being in heavy water (D)2In O)1The H-NMR shifts were:
chemical Shift δ (1H) in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
7.59
Twin peaks (d)
2H
6.67
Twin peaks (d)
2H
4.25
Multiplet peak (m)
1H
3.57
Multiplet peak (m)
1H
3.38
Multiplet peak (m)
2H
3.20
Multiplet peak (m)
1H
3.13
Multiplet peak (m)
1H
2.95
Single peak(s)
6H
2.84
Multiplet peak (m)
1H
2.67
Single peak(s)
3H
2.26
Multiplet peak (m)
2H
2.10
Multiplet peak (m)
1H
1.96
Multiplet peak (m)
1H
In another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-methyl- (6S) -tetrahydrofolate and the organic cation mono- (phenethyl-biguanide), the amorphous tetrahydrofolate being in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-methyl- (6S) -tetrahydrofolate and the organic cation mono-choline in heavy water (D)2In O)1The H-NMR shifts were:
chemical Shift δ (1H) in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
7.56
Twin peaks (d)
2H
6.65
Twin peaks (d)
2H
4.20
Multiplet peak (m)
1H
3.95
Multiplet peak (m)
2H
3.43
Multiplet peak (m)
1H
3.40
Triplet (t)
2H
3.20
Twin peaks (d)
1H
3.08
Single peak(s)
9H
3.04
Multiplet peak (m)
1H
2.99
Multiplet peak (m)
1H
2.89
Multiplet peak (m)
1H
2.42
Single peak(s)
3H
2.20
Multiplet peak (m)
2H
2.04
Multiplet peak (m)
1H
1.91
Multiplet peak (m)
1H
In another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation mono-2-dimethylaminoethanol, in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-methyl- (6S) -tetrahydrofolate and the organic cation mono-arginine, the amorphous tetrahydrofolate in heavy water (D)2In O)1The H-NMR shifts were:
chemical Shift δ (1H) in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
7.59
Twin peaks (d)
2H
6.67
Twin peaks (d)
2H
4.25
Multiplet peak (m)
1H
3.69
Triplet (t)
1H
3.58
Multiplet peak (m)
1H
3.41
Multiplet peak (m)
2H
3.19
Multiplet peak (m)
5H
2.71
Single peak(s)
3H
2.27
Multiplet peak (m)
2H
2.10
Multiplet peak (m)
1H
1.96
Multiplet peak (m)
1H
1.83
Multiplet peak (m)
2H
1.60
Multiplet peak (m)
2H
In another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation mono-acetylcholine, in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous tetrahydrofolate is derived from tetrahydrofolateIon 5-methyl- (6S) -tetrahydrofolic acid and organic cation mono-acetylcholine, amorphous tetrahydrofolic acid salt in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous tetrahydrofolate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation di-arginine, the amorphous tetrahydrofolate is in heavy water (D)2In O)1The H-NMR shifts were:
chemical Shift δ (1H) in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
8.53/7.82
Unimodal (s)/unimodal(s)
1H
7.56/7.53
Doublet (d)/doublet (d)
2H
6.64/6.59
Doublet (d)/doublet (d)
2H
4.75
Multiplet peak (m)
1H
4.21
Multiplet peak (m)
1H
3.66
Triplet (t)
2H
3.44
Multiplet peak (m)
1H
3.30
Multiplet peak (m)
2H
3.17
Multiplet peak (m)
1H
3.11
Single peak(s)
4H
2.23
Multiplet peak (m)
2H
2.07
Multiplet peak (m)
1H
1.95
Multiplet peak (m)
1H
1.79
Multiplet peak (m)
4H
1.58
Multiplet peak (m)
4H
In another embodiment, the amorphous folate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation di-choline, and the amorphous tetrahydrofolate is in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous folate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation bis- (1, 1-dimethyl-biguanide), the amorphous tetrahydrofolate being in heavy water (D)2In O)1The H-NMR shifts were:
chemical shift δ(1H) Expressed in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
8.57/7.86
Unimodal (s)/unimodal(s)
1H
7.60/7.56
Doublet (d)/doublet (d)
2H
6.68/6.63
Doublet (d)/doublet (d)
2H
4.80
Multiplet peak (m)
1H
4.25
Multiplet peak (m)
1H
3.48
Multiplet peak (m)
1H
3.33
Multiplet peak (m)
2H
3.22
Multiplet peak (m)
1H
2.96
Single peak(s)
12H
2.25
Multiplet peak (m)
2H
2.10
Multiplet peak (m)
1H
1.96
Multiplet peak (m)
1H
In another embodiment, the amorphous folate consists of the tetrahydrofolate anion 5-formyl- (6S) -tetrahydrofolate and the organic cation bis- (phenethyl-biguanide), and is present in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous folate consists of the tetrahydrofolate anion 5-methyl- (6S) -tetrahydrofolate and the organic cation di-arginine, and the amorphous tetrahydrofolate is in heavy water (D)2In O)1The H-NMR shifts were:
in another embodiment, the amorphous folate consists of the tetrahydrofolate anion 5-methyl- (6S) -tetrahydrofolate and the organic cation bis- (1, 1-dimethyl-biguanide), the amorphous tetrahydrofolate being in heavy water (D)2In O)1The H-NMR shifts were:
chemical Shift δ (1H) in ppm
Multiple severity (Multiplicity)
Strength (Intensity)
7.59
Twin peaks (d)
2H
6.67
Twin peaks (d)
2H
4.25
Multiplet peak (m)
1H
3.44
Multiplet peak (m)
1H
3.24
Multiplet peak (m)
1H
3.10
Multiplet peak (m)
1H
3.01
Multiplet peak (m)
1H
2.95
Single peak(s)
12H
2.92
Multiplet peak (m)
1H
2.48
Single peak(s)
3H
2.25
Multiplet peak (m)
2H
2.10
Multiplet peak (m)
1H
1.97
Multiplet peak (m)
1H
In another embodiment, the amorphous folate consists of the tetrahydrofolate anion 5-methyl- (6S) -tetrahydrofolate and the organic cation bis- (phenethyl-biguanide), the amorphous tetrahydrofolate being in heavy water (D)2In O)1The H-NMR shifts were:
in another preferred embodiment, the pharmaceutical composition comprises at least one folate according to the invention as the main active compound. The composition further comprises at least one pharmaceutically acceptable excipient. The composition may comprise, for example, a buffer compound. Suitable and preferred buffer compounds are tromethamine and hydroxyethylpiperazine ethanesulfonic acid (HEPES). In addition, antioxidant compounds may be present in the composition. Preferred antioxidant compounds are thioglycerol, Dithiothreitol (DTT), and cysteine.
Furthermore, at least one folate according to the invention is used for the preparation of a medicament, food additive or nutritional supplement for the prevention and/or treatment of defects or diseases in which the administration of folate leads to a positive promoting effect. There are many disease states that are positively promoted by compositions comprising folate. Such diseases are, for example, pathophysiological, neurological and inflammatory diseases.
Additionally, a process for the preparation of the amorphous folate according to the invention is provided, comprising the step of adding oxalic acid or a fluoride salt to an aqueous composition of an alkaline earth metal salt of folic acid, said tetrahydrofolate consisting of folate anions and organic cations.
Detailed Description