HK1118729B - Pharmazeutische zusammensetzung enthaltend gestagene und/oder estrogene und 5-methyl-(6s)-tetrahydrofolat - Google Patents

Description

Pharmaceutical composition containing progestogen and/or estrogen and 5-methyl- (6S) -tetrahydrofolic acid

The invention relates to pharmaceutical compositions containing a progestogen, an estrogen and 5-methyl- (6S) -tetrahydrofolic acid, which are useful as oral contraceptives and also prevent conditions and malformations caused by folic acid deficiency, while at the same time not masking vitamin B₁₂Symptoms of deficiency.

Pharmaceutical companies active in the field of birth control have been striving to improve existing contraceptives. This includes not only increasing contraceptive reliability through the development of new substances, but also improving ease of use. In contrast, new methods of combining contraception with disease prevention are also being sought.

Some diseases are thought to be associated with folate deficiency. Thus, for example, administration of folates in the form of folic acid can minimize the risk of cardiovascular disorders and certain malignant disorders, such as breast or colon cancer.

Fetal development defects are particularly serious consequences of folate deficiency in women of reproductive age. Thus, women with low folate levels have an increased risk of developing infants with congenital malformations such as neural tube defects, ventricular valve defects, and genitourinary defects, as compared to women with sufficiently high folate levels.

Neural tube defects are the most common congenital malformations of the central nervous system. They are caused by incomplete neural tube closure at about the third to fourth weeks of embryonic development. Neural tube defects include spina bifida (in some cases with meningeal or spinal meningeal bulging), brain bulging, and anencephaly, characterized by partial or total loss of brain area. Children with a brain-free deformity are virtually non-viable.

A spina bifida is characterized by incomplete closure of the vertebral arch. The consequences of this depend on the nature of the injury, being life-long disabilities in the form of various sensory and motor deficits-thus, for example, two thirds of children and adults rely on wheelchairs due to muscle paralysis. Treatment requires coverage of the defect, shunt drainage of cerebrospinal fluid, and lengthy orthopedic and neurological rehabilitation. Average medical charge is 500000 per child

It is assumed that approximately 250000 neonates in the world have neural tube defects. Neonatal defects are approximately 1-2 per 1000 neonates in germany and the united states. In germany, about 500 live infants suffer from neural tube defects every year, and another 500 cases terminate pregnancy based on prenatal ultrasound diagnosis.

A sufficiently high folate level at conception and in the early stages of pregnancy is critical to avoid neural tube defects. It is generally believed that a folate level of at least 906nmol/l in the red blood cells is desirable to reduce the frequency of neural tube defects.

Ingestion of folic acid at the correct time before and after conception is known to reduce neural tube defects by 50-70%. Folate fortified foods practiced in the united states have significantly reduced the incidence of neural tube defects, by over 50% in canada and chile.

However, voluntary addition of folic acid to food and ingestion of folic acid products, as for example in germany, do not allow all women of childbearing age to reach a sufficient level. First, many women are unaware of the risk of neural tube defects and the likelihood of ingesting folic acid to minimize the corresponding risk. Thus, in many countries, far less than 10% of them take folic acid product before and after conception time. Second, despite the increasing convenience of modern contraceptive methods, many pregnancies are unplanned pregnancies-estimated to reach 50% in the united states (inst. of Medicine 1998, NEJM2004), so that the ready intake of folic acid products prior to conception is also excluded from the outset. In addition, for example, in the United states, about 5-8% of users do not reliably take oral contraceptives.

Thus, the purpose of patent US6,190,693(Kafrissen et al) is to prevent certain conditions which can be treated with folic acid in the users of oral contraceptives. Kafrissen accomplishes this by adding folic acid to an oral contraceptive. He discloses a method for administering folic acid using a pharmaceutical composition comprising a conventional substance having contraceptive activity and folic acid.

However, the introduction of folic acid into oral contraceptives presents a serious health risk in itself, as folic acid may mask vitamin B₁₂Lack early, but still treatable symptoms such as megaloblastic anemia. This is because of the vitamin B₁₂The blood symptoms caused by the deficiency can be well treated by the additional administration of folic acid, making it difficult to detect vitamin B₁₂Lack, or no detectable vitamin B at all₁₂Deficiency, thus results in no diagnosis of vitamin B₁₂And (4) lack. Neuropsychiatric symptoms such as paresthesia and ataxia are then also untreated and may instead deteriorate irreversibly.

The object of patent application WO03/070255(Coelingh Bennink) is therefore to avoid the masking of vitamin B in the users of folic acid-containing oral contraceptives₁₂Lack of health risks due to symptoms. Coelingh Bennink is prepared by adding vitamin B to an oral contraceptive₁₂To achieve this object. He discloses a kit for an oral hormonal contraceptive comprising an estrogen and/or a progestin, tetrahydrofolic acid and essential vitamin B₁₂。

Another problem associated with the administration of 5-methyl- (6S) -tetrahydrofolate-free folic acid and tetrahydrofolate products is the genetic polymorphism of methylenetetrahydrofolate reductase (MTHFR C677T), which is heterozygous in about 55% of the Caucasian population and homozygous in about 10-15%. This polymorphism results in a decrease in the activity of methylenetetrahydrofolate reductase and therefore affected women are unable to fully metabolize supplied folate and tetrahydrofolate into 5-methyl- (6S) -tetrahydrofolate which is active in vivo. This polymorphism is a recognized risk factor for conditions caused by folate deficiency, particularly for neural tube defects.

Another problem that causes difficulties is that folic acid is not a naturally occurring substance in food. To be biologically active, it must first be metabolically converted to 7, 8-dihydrofolate and (6S) -tetrahydrofolate by dihydrofolate reductase. The metabolic capacity to convert previtamin folate into its active reduced form, in particular the first activation step, is limited and also subject to great variability. Since dihydrofolate reductase does not function in the metabolism of L-methyl folate (metafolin), it is expected that drugs that inhibit dihydrofolate reductase, such as methotrexate, should not interact with dihydrofolate reductase.

Also to provide an adequate supply of folic acid to women suffering from a methylene tetrahydrofolate reductase deficiency, EP0898965 (muller et al) proposes to use 5-methyl- (6S) -tetrahydrofolic acid or a suitable pharmaceutically acceptable salt as a dietary supplement or as a pharmaceutical ingredient. Furthermore, EP1044975a1 discloses stable crystalline salts of 5-methyl- (6S) -tetrahydrofolic acid and a process for their preparation.

Most pregnancies are known to occur shortly after withdrawal of contraceptives (Farrow et al, HumanReproduction vol.17, No. 10, pp.2754-2761, 2002). If the administration is irregular and unreliable, conception may occur even during the administration period. In addition, it is known that even after one person stops taking additional folic acid, it may benefit for about 90 additional days (ACRHD): The public health issues, including The safety and functional clinical experience, associated with The combining of The folic acid and The organic contextual inter-a single combination product, 12/15/2003; meeting discipline, problem 4). However, a prerequisite is that, in addition to a normal diet, a sufficiently large amount of folic acid has been taken a sufficiently long time before. This so-called tissue depot effect can be seen by an increase in folate levels in the red blood cells.

In addition, low folate/high homocysteine levels are also known to be associated with multiple spontaneous abortions (Merlen et al, Obstet. et. Gynecol.2000, 95: pp.519-524).

The object of the present invention is to produce an oral contraceptive which, although it prevents diseases caused by folic acid deficiency, at the same time does not mask vitamin B₁₂Symptoms of deficiency. It is a further object of the present invention to disclose a dosing regimen which ensures that the user of the pharmaceutical composition of the present invention is reliably protected from disorders or malformations caused by folate deficiency, in particular neural tube defects, for a period of time after termination. Both objectives also apply in the case of homozygous and heterozygous polymorphisms of the enzyme methylenetetrahydrofolate reductase in the user, which negatively influence the utilization of folate and thus its biological activity in the prevention of neural tube defects.

The present invention achieves this object by a pharmaceutical composition containing one or more progestogens and/or estrogens and 5-methyl- (6S) -tetrahydrofolic acid, together with pharmaceutically acceptable excipients and carriers.

The present invention is based on the insight that by administering 5-methyl- (6S) -tetrahydrofolate alone, disorders caused by folate deficiency are treated and prevented without even masking vitamin B₁₂Symptoms of deficiency are possible, which is surprising with respect to WO 03/070255. Thus, the administration of vitamin B is no longer required₁₂To avoid the health risks described in WO 03/070255. Despite the administration of 5-methyl- (6S) -tetrahydrofolic acid, physicians are still able to diagnose and optionally treat vitamin B₁₂And (4) lack.

In vitamin B₁₂In the absence of vitamin B, it is of course possible to administer vitamin B additionally₁₂. In addition, it is also possible to optionally add other vitamins such as vitamin B₆Or vitamin B₂. The present invention is further based on the insight that the use of 5-methyl- (6S) -tetrahydrofolate alone in a contraceptive, unlike administration of folic acid or other tetrahydrofolic acid, enables the body to be spared even in the case of homozygous or heterozygous polymorphism of methylenetetrahydrofolate reductaseThe limited, full use of the folate component and thus the full use of its biological activity in preventing congenital malformations caused by folate deficiency is surprising in relation to WO 03/070255.

5-methyl- (6S) -tetrahydrofolate is metabolically synthesized from 5, 10-methylene- (6R) -tetrahydrofolate (see FIG. 1). This biochemical reaction is catalyzed by methylenetetrahydrofolate reductase (MTHFR), a number of genetic mutants of which are known, some of which exhibit limited biological activity (MTHFR C677T polymorphism). In the next step 5-methyl- (6S) -tetrahydrofolate is converted into tetrahydrofolate catalysed by Methionine Synthase (MS). This transfers the 5-methyl group of 5-methyl- (6S) -tetrahydrofolate to the amino acid homocysteine (Hcy) which converts it to the amino acid methionine (Met). This vitamin B₁₂The dependent response is also known as homocysteine methylation in homocysteine metabolism. 5-methyl- (6S) -tetrahydrofolate plays a particular role in the reduced folate group, since 5-methyl- (6S) -tetrahydrofolate can only be converted into tetrahydrofolate by the homocysteine methylation reaction. Tetrahydrofolic acid is the actual carrier molecule for a one-carbon unit in multiple oxidation states. In metabolism, 5-methyl- (6S) -tetrahydrofolate can only be synthesized from 5, 10-methylene- (6R) -tetrahydrofolate and can only be further metabolized by conversion to tetrahydrofolate. The first enzymatic reaction (MTHFR) is irreversible under physiological conditions and the second enzymatic reaction (MS) is vitamin B₁₂Dependent, which means that vitamin B, if present₁₂In the absence, 5-methyl- (6S) -tetrahydrofolate accumulates and cannot be further metabolized. This phenomenon is also known as methyl trap. Only 5-methyl- (6S) -tetrahydrofolic acid has this special property, whereas none of the other oxidized and reduced folic acids like folic acid, 7, 8-dihydrofolic acid, (6S) -tetrahydrofolic acid, 5-formyl- (6S) -tetrahydrofolic acid, 10-formyl- (6R) -tetrahydrofolic acid, 5, 10-methenyl- (6R) -tetrahydrofolic acid, 5, 10-methylene- (6R) -tetrahydrofolic acid, 5-formimino- (6S) -tetrahydrofolic acid has this property. 5-methyl- (6S) -tetrahydrofolic acid is non-masked vitamin B₁₂The only naturally occurring folic acid that is lacking. This is true for 5-methyl- (6S) -tetrahydrofolate and oral administrationThe combined use of contraceptives is very important and is one aspect of the present invention.

The progestagens that may be used in the pharmaceutical composition of the present invention are the following: levonorgestrel, norgestimate, nordehydrohydroxyprogesterone, dydrogesterone, drospirenone, 3-beta-hydroxydesogestrel, 3-ketodesogestrel (═ etonogestrel), 17-deacetylnorgestimate, 19-norprogesterone, acetoxypregnenolone, allylestrenol, Amgestone, chlormadestone, cyproterone, demegestone, desogestrel, dihydroprogesterone (dihydrogesterone), demegesterone, ethisterone, norgestrel, fluogesterone acetate, Gastrinone, gestodene, hydroxytryptone, medroxyprogesterone, hydroxyprogesterone, lynestrenol (@ ethisterone), mecestone, medroxyprogesterone, norelgestrome, nomegestrol (═ dehydrogesterone), isogestrel, norgestrel (including d-norgestrel and dl-norethindrone), pregnenone, norgestrel, Norethindrone, progesterone, quingestrol, (17 α) -17-hydroxy-11-methylene-19-norpregna-4, 15-dien-20-yn-3-one ((17alpha) -17-hydroxy-11-methyl-19-norpregna-4, 15-dien-20-yn-3-one), tibolone, trimegestone, acetophen-algestrol, nesterone (Nestorone), prometrone, 17-hydroxyprogesterone esters, 19-nor-17 hydroxyprogesterone, 17 α -ethynyltestosterone, 17 α -ethynyl-19-nortestosterone, d-17 β -acetoxy-13 β -ethyl-17 α -ethynyl-4-en-3-one oxime or compounds disclosed in WO00/66570, in particular tanaproget. Levonorgestrel, norgestimate, nordehydrohydroxyprogesterone, drospirenone, dydrogesterone are preferred. Drospirenone is particularly preferred.

Suitable estrogens are ethinyl estradiol, mestranol, quinestrol (quinestranol), estradiol, estrone, estrane, estriol, estetrol and conjugated equine estrogens. In this respect, ethinyl estradiol, estradiol and mestranol are preferred, with ethinyl estradiol being particularly preferred.

The respective progestogen and/or estrogen employed in the present invention is in an amount corresponding to that generally known in contraceptives.

For example, generally for progestogens, the following are mentioned:

drospirenone 0.5-5mg

Levonorgestrel 30-250 mug

180 mu g of norgestimate

0.5-1mg of norethindrone acetate

Cyproterone acetate 1-2mg

Desogestrel 20-150 mug

Dienogest 2-3mg

60-75 mug of gestodene

Tibolone 2.5mg

Drospirenone is preferably administered according to the invention in an amount of, for example, 0.5 to 5mg, particularly preferably 3mg, per day.

For example, the amounts of estrogen used according to the invention are as follows:

ethinyl estradiol 10-50 mu g

Estradiol 1-4mg

Mestranol 50 mug

Ethinyl estradiol is preferably administered according to the invention in an amount of, for example, 10 to 50 μ g, particularly preferably 10 to 30 μ g, very particularly preferably 20 to 30 μ g, per day.

Reference to 5-methyl- (6S) -tetrahydrofolic acid in the form according to the invention refers to the free acid form and the pharmaceutically acceptable salt form, as well as to modified forms of 5-methyl- (6S) -tetrahydrofolic acid (N- [4- [ [ (2-amino-1, 4, 5, 6, 7, 8-hexahydro-4-oxo-5-methyl- (6S) -pteridinyl) methyl ] amino ] benzoyl ] -L-glutamic acid).

Pharmaceutically acceptable salts mean pharmacologically and pharmaceutically acceptable salts. Such pharmacologically and pharmaceutically acceptable salts may be alkali metal or alkaline earth metal salts, preferably sodium, potassium, magnesium or calcium salts. Calcium salts are particularly preferred.

Particularly preferred 5-methyl- (6S) -tetrahydrofolic acid (L-methylfolic acid), for example the calcium salt, according to the invention is used in an amount of 0.1 to 10mg, preferably 0.4 to 1mg, particularly preferably 451. mu.g (corresponding to 400. mu.g folic acid or 416. mu.g 5-methyl- (6S) -tetrahydrofolic acid).

The crystalline modification disclosed in EP1044975 is preferred as a modification of 5-methyl- (6S) -tetrahydrofolic acid.

Optionally vitamin B may be present₆Or vitamin B₂. However, corresponding additions are not necessary for the implementation of the invention. Vitamin B₆In the usual dosage, 1mg to 5mg, preferably 1mg to 3mg per day can be used. Vitamin B₂1mg to 5mg, preferably 1mg to 2mg per day can be used in the usual dosage, and 2 to 5mg per day can be used in the high dosage.

The progestogen and/or estrogen herein is a substance that has contraceptive efficacy. 5-methyl- (6S) -tetrahydrofolic acid is added as vitamin to prevent disorders and malformations caused by folic acid deficiency without masking the possible vitamin B₁₂Symptoms of deficiency. Furthermore, women who have limited folate metabolism due to their reduced MTHFR enzyme activity (MTHFR C677T polymorphism) and reduced folate levels also benefit from 5-methyl- (6S) -tetrahydrofolate.

In a preferred variant of the invention drospirenone is administered in an amount of from 0.5 to 5mg, preferably 3mg, daily, ethinylestradiol in an amount of from 10 to 50 μ g, preferably from 10 to 30 μ g, particularly preferably from 20 to 30 μ g. In this preferred variant of the invention, the calcium salt of 5-methyl- (6S) -tetrahydrofolic acid is present in an amount of 0.1 to 10mg, preferably 0.4 to 1mg, particularly preferably 451. mu.g (corresponding to 400. mu.g folic acid).

The formulation of pharmaceutical products based on the new pharmaceutical compositions is carried out by methods known per se: the active ingredients are processed with carrier substances, fillers, disintegration-influencing substances, binders, wetting agents, lubricants, absorbents, diluents, masking flavors (masking flavors), colorants and the like used in pharmaceutical technology and converted into the desired administration form, which also includes sustained-release forms.

In the medicament of the invention, the estrogen and the progestogen and 5-methyl- (6S) -tetrahydrofolate can be present in a combined dosage unit. However, it is also possible to formulate the estrogen and the progestin as one and the 5-methyl- (6S) -tetrahydrofolate as the other in separate dosage units.

Vitamin B₁₂And 5-methyl- (6S) -tetrahydrofolic acid are unstable in the presence of atmospheric oxygen and moisture. Attempts were made to combine ethinyl estradiol with vitamin B₁₂When formulated together, the two materials were found to be incompatible with each other. The measurement of the incompatibility between the target formulation ingredients is carried out by thermal analysis methods (DSC, differential scanning calorimetry). This can be used to identify incompatibilities by low melting enthalpies and melting temperatures. These are caused by, for example, a decrease in the proportion of crystalline material and an increase in impurities. In the assay, the excipients or active ingredients were investigated in each case with vitamin B₁₂And compatibility was tested under the influence of various gases and temperatures. In said study, vitamin B₁₂Showing a strong interaction with ethinyl estradiol. The results of the incompatibility measurements can be seen in table 1.

Table 1: summary of compatibility studies

Substance(s)	Compatibility	Compatibility types	Note
				Drospirenone	+	Is mostly good	O2-sensitivity
Ethinyl estradiol	---	Strong interaction	O2Very sensitive
				Ethinyl estradiol beta-cyclodextrin compound	+	Is mostly better below 60 DEG C	O2-sensitive, moisture-sensitive
Lactose	+	Is mostly better below 60 DEG C	O2-sensitive, moisture-sensitive
				Corn starch	++	Well below 60 deg.C	O2-sensitive, moisture-sensitive
Modified corn starch	++	Well below 60 deg.C	O2-sensitive, moisture-sensitive
				Polyvinylpyrrolidone	+	Is mostly better below 60 DEG C	O2-sensitive, moisture-sensitive
Magnesium stearate	++/-	Inert, preferably below 60 ℃	O2-sensitive, moisture-sensitive
				Hydroxypropyl methylcellulose	++	Well below 60 deg.C	O2-sensitive, moisture-sensitive
Hydroxypropyl cellulose	++	Well below 60 deg.C	O2-sensitive, moisture-sensitive
				Maltodextrin	+/--	Inert, preferably below 60 ℃	O2-sensitive, moisture-sensitive
Polyethylene glycol 6000	-	Interaction with moisture	O2-sensitive, moisture-sensitive
				Coating mixture	+	Is mostly better below 60 DEG C	O2-sensitive, moisture-sensitive

Symbol:

compatibility expected to be good below the stated temperature

+ compatible below said temperature

+ +/-inert compatible, compatible with possible good below said temperature

+/- -inert compatible, below which temperature it seems compatible

- (- - - -) (Strong) interaction, incompatibility

Polyvinylpyrrolidone (PVP) is particularly suitable because of its wetting properties for hormone preparations (Moneghini et al, Int J Pharm 175, 1998, 177-. But the 5-methyl- (6S) -tetrahydrofolate formulation with PVP increased the degradation rate of 5-methyl- (6S) -tetrahydrofolate (compare tables 2 and 3; method 3).

Thus, another object of the present application achieved by the present invention is to enable the preparation of 5-methyl- (6S) -tetrahydrofolic acid and optionally vitamin B₁₂Stable ethinylestradiol formulations in the presence are possible.

It has been found that ethinylestradiol can surprisingly be prevented from reacting with vitamin B by using ethinylestradiol in the formulation as a beta-cyclodextrin complex (ethinylestradiol being a beta-cyclodextrin inclusion compound; for the preparation, compare WO02/49675)₁₂Incompatibility between them.

The corresponding formulations of the invention are described in example 1 (comparative compositions A, B and D).

They furthermore contain a mixture of corn starch and modified corn starch. The starch consists of amylose and amylopectin. Both are polysaccharides based on alpha-glucose units. However, instead of corn starch, it is also possible to use, for example, rice starch, potato starch or wheat starch in pharmaceutical preparations. Starch is used in swollen, suspended or dissolved form as a binding liquid or as a solid. It may be unmodified or partially modified. The preferred corn starch for use in the present invention has an empirical formula of (C)₆H₁₀O₅)_nWherein n is 300-. The molecular weight is 50000-160000.

Only part of the starch used in the pharmaceutical preparation acts as pure filler. Another part of which serves as an adhesive. According to the invention, 1 to 5%, preferably 1.8 to 3%, of the tablet weight should be added as a binder in the form of corn starch. In addition to corn starch, it is also possible to use starch or starch compounds such as maltodextrin or cellulose derivatives, for example carboxymethylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose or methylcellulose, as binders. The present invention preferably uses low-substituted cellulose derivatives. The viscosity in a 2% aqueous solution is 1-20 mPas. Derivatives having a viscosity of from 2 to 20mPas are preferred according to the invention, derivatives having a viscosity of from 3 to 6mPas being particularly preferred.

The portion of the corn starch used in the preferred formulations of the invention may be replaced by low-substituted Hydroxypropylcellulose (HPC) in a concentration of 0.5-5% (w/w), preferably 1-3% (w/w), particularly preferably 2% (w/w). In this case, the hydroxypropylcellulose has low substitution when the hydroxyls of the hydroxypropylcellulose are esterified or etherified at not less than 5% and not more than 16%.

Table 2 shows the% content of 5-methyl- (6S) -tetrahydrofolate in each tablet used immediately after preparation as a function of binder at a given 100% content. The indicated 5-methyl- (6S) -tetrahydrofolate content was measured in the content uniformity assay (CUT). The formulation studied was prepared by the following (method 2): mixing the ingredients, granulating with corn starch as binder, absorbing 5-methyl- (6S) -tetrahydrofolic acid after the granulation process is completed, mixing again, and tabletting. In contrast, polyvinylpyrrolidone was added to the formulation of method 3 as a binder instead of corn starch. The 5-methyl- (6S) -tetrahydrofolate content of the formulation prepared by method 3 is low.

Table 2: l-methyl Folic acid content immediately after preparation as a function of binder

	L-methyl Folic acid content absorption, PVP (method 3)	L-methyl Folic acid content absorption, corn starch (method 2)
			Average	90.5％	96.1％

Table 3 shows the content of 5-methyl- (6S) -tetrahydrofolic acid as a function of the adhesive at use after one month of storage at a given temperature and humidity. Table 2 demonstrates that the tendency of 5-methyl- (6S) -tetrahydrofolic acid formulated with PVP to be less stable is demonstrated, particularly when stored at a temperature of 40 ℃ and a relative humidity (rH) of 75%.

Table 3: l-methyl Folic acid content after storage as a function of binder

	25 ℃/60% rH absorption PVP (method 3)	25 deg.C/60% rH absorption corn starch (method 2)	40 ℃/75% rH absorption PVP (method 3)	40 deg.C/75% rH absorption corn starch (method 2)
					Open vial	89.5％	92.1％	37.7％	67.7％

Oral formulations are typically prepared by granulation, tableting and coating. However, since 5-methyl- (6S) -tetrahydrofolic acid is sensitive to oxygen and moisture, degradation occurs even during granulation. However, further degradation of 5-methyl- (6S) -tetrahydrofolic acid during storage is particularly noticeable. In a formulation where all the components of the drug including 5-methyl- (6S) -tetrahydrofolic acid were first mixed and then granulated only as usual, the 5-methyl- (6S) -tetrahydrofolic acid remaining after 1 month storage in a closed vial at 40 ℃ and 75% relative humidity was only 60% of the initial use (compare Table 5). Losses in the granulation can be reduced by absorbing 5-methyl- (6S) -tetrahydrofolic acid only after the granulation process is completed. Thus, the dry mixture in the preparation stabilizes 5-methyl- (6S) -tetrahydrofolic acid. However, in addition, surprisingly, this also has a further stabilizing effect on the shelf life. The 5-methyl- (6S) -tetrahydrofolate content of the prepared formulations was above 90% after storage for the same time under the same conditions (compare Table 5).

Table 4 shows the% content of 5-methyl- (6S) -tetrahydrofolate in each tablet used immediately after preparation as a function of the preparation method. Method 1 differs from method 2 in the time of addition of 5-methyl- (6S) -tetrahydrofolate in the preparation of the tablets studied. In method 1, 5-methyl- (6S) -tetrahydrofolic acid is present in the mixture even during granulation, whereas in method 2, 5-methyl- (6S) -tetrahydrofolic acid is absorbed only after granulation. The 5-methyl- (6S) -tetrahydrofolate content of the preparation prepared by method 1 is significantly lower.

Table 4: function of the content of L-methyl Folic acid immediately after preparation on the preparation method

	L-MethylFolic acid content granulation (method 1)	L-methyl Folic acid content absorption (method 2)
			Average	88.5％	96.1％
Coefficient of distribution	6.1	2.5

Table 5 shows the content of 5-methyl- (6S) -tetrahydrofolic acid as a function of the preparation method, after one month of storage at a given temperature and humidity. Table 4 demonstrates that the tendency of the formulation to be less stable when 5-methyl- (6S) -tetrahydrofolate is added prior to granulation is demonstrated, particularly when stored at 40 ℃ and 75% relative humidity (rH).

Table 5: function of the content of stored L-methyl Folic acid to the preparation method

	25 ℃/60% rH granulation (method 1)	25 ℃/60% rH absorption (method 2)	40 ℃/75% rH granulation (method 1)	40 ℃/75% rH absorption (method 2)
					Open vial	63.2％	92.1％	43.4％	67.7％
Closed vial	74.5％	92.5％	58.9％	90.1％

It is known that the release occurs more slowly with dry mixtures than with granulation. However, it was surprisingly found that the release was not delayed, but actually accelerated, by the dry mixture of 5-methyl- (6S) -tetrahydrofolic acid. To this end, these tablets were studied in an in vitro dissolution test conducted in a 0.03% aqueous solution of ascorbic acid at 50rpm and 37 ℃ using a USP paddle apparatus. Table 6 shows the results of the in vitro dissolution test.

Table 6: % dissolution

Time [ min ]]	5-methyl- (6S) -tetrahydrofolate method 1 dissolution [% ]]	5-methyl- (6S) -tetrahydrofolic acid method 2 dissolution [% ]]
			0	0	0
10	59.2	81.4
			15	66.8	89.3
30	73.1	91.3
			45	76.7	91.1
60	75.8	91.2

Regular intake of the pharmaceutical composition of the invention containing a particularly preferred dose of 451 μ g of 5-methyl- (6S) -tetrahydrofolate calcium salt per day leads to an increase in folate concentrations in the serum and in the erythrocytes until a steady state is reached. The corresponding kinetics of folic acid invasion of erythrocytes are described by a half-life of 6-10 weeks. Based on this half-life, it is expected that a steady state red blood cell folate level of about 97% can be reached after about 5 half-lives (corresponding to about 30 to 50 weeks). If daily intake of the pharmaceutical composition of the present invention is continued, the red blood cell folate level remains within the steady state concentration range. After cessation of ingestion of the pharmaceutical composition of the present invention, the red blood cell folate levels will also slowly decline over a half-life of about 6 to 10 weeks. Thus, even without continued ingestion of the pharmaceutical composition of the present invention, the red blood cell folate levels remain above the limit 906nmol/l normally considered sufficient to prevent neural tube defects for weeks. Thus, the product of the invention ensures a reduced risk of disorders and congenital malformations caused by folate deficiency, even after the long-term intake of the drug of the invention ("contraceptive") has ceased.

The invention also relates to 5-methyl- (6S) -tetrahydrofolic acid, one or more estrogens and/or progestogens and optionally vitamin B₆And/or vitamin B₂And the use of pharmaceutically acceptable excipients and carriers for the manufacture of a medicament for reducing the risk of a condition caused by folate deficiency and a congenital malformation caused by folate deficiency after prolonged continuous ingestion of the medicament for at least 8 weeks prior to termination.

In addition, the invention relates to a kit comprising at least 20 daily dosage units containing the medicament according to the invention and at least one pharmaceutical composition comprising 5-methyl- (6S) -tetrahydrofolic acid and optionally vitamin B₁₂Vitamin B₆And/or vitamin B₂Wherein the number of all dosage units present in the kit is at least 28 and the dosage units are arranged such that dosage units containing the medicament of the invention should be taken first and dosage units containing neither estrogen nor progestin should be taken second. In the case of the first-mentioned at least 20 daily dosage units containing the medicament of the invention, it is also possible to formulate 5-methyl- (6S) -tetrahydrofolic acid separately and arrange it spatially as a further dosage unitAnd (c) means for making it clear from the arrangement that the two dosage units are taken in combination.

The different kits of the invention which are further developed are reflected in claims 18 to 22, 38, 39 and 40.

In particular, the medicament of the invention may also be administered in a so-called prolonged regimen according to claims 43 to 50. This means that for more than 28 consecutive days, the extended use period is completed by administering 1 to 7 days of dosage units containing 5-methyl- (6S) -tetrahydrofolate alone or 1 to 7 days of placebo (dosage unit active) or 1 to 7 days of empty days of contraceptive (no dosage units administered).

The following examples explain the subject matter of the invention in more detail, without intending to limit it.

Example 1:

the composition of the tablet of the present invention (80mg) can be seen in Table 7.

Table 7: composition of the tablets of the invention

Composition (I)	Measurement of
							Composition of	A	B	C	D
Drospirenone ethinylestradiol*L-methyl Folic acid vitamin B12Lactose monohydrate corn starch**Modified corn starch magnesium stearate		3mg0.03mg0.451mg--to80mg16.40mg2mg***9.60mg0.80mg	3mg0.02mg0.451mg--to80mg16.40mg2mg***9.60mg0.80mg	----0.451mg--to80mg16.40mg2mg***9.60mg0.80mg	3mg0.03mg0.451mg0.1mgto80mg16.40mg2mg***9.60mg0.80mg

^*: optionally ethinylestradiol-beta-cyclodextrin complex; the stated amounts refer in this case to uncomplexed ethinylestradiol. If ethinyl estradiol-beta-cyclodextrin complex is used, 10 times the amount should be used. This is because the content of ethinylestradiol in the β -cyclodextrin complex is about 9.5 to 12.5% (compare WO 02/49675).

^**: marking^**The corn starch fraction of (a) may be replaced with other binders such as 1.6mg of low substituted hydroxypropyl cellulose.

^***: corn starch as a binder^**The amount of (B) may also be, for example, 1.8 mg.

The oral preparation is prepared by the following steps: mixing the above components, granulating with corn starch as binder part, absorbing 5-methyl- (6S) -tetrahydrofolic acid calcium salt after the granulation process is completed, mixing, tabletting, and coating.

Example 2:

blood was taken every 8 weeks from 80 healthy young fertile women and the red blood cell folate levels were determined using validated microbiological, immunological or instrumental (e.g. HPLC, LC-MS/MS) methods or a suitable combination of these methods.

8 weeks after the first blood sample (screening phase),

451 μ g of 5-methyl- (6S) -tetrahydrofolate calcium salt was administered daily over a period of 40 weeks, or:

drospirenone 3mg, ethinylestradiol 30 μ g and calcium 5-methyl- (6S) -tetrahydrofolate 451 μ g (tablets of composition A in example 1) were administered simultaneously daily for the first 21 days of each cycle. 451. mu.g of 5-methyl- (6S) -tetrahydrofolate calcium salt (composition C) was administered for 7 days following this period. A further 21 days (second period) of 3mg drospirenone, 30. mu.g ethinylestradiol and 451. mu.g 5-methyl- (6S) -tetrahydrofolate calcium salt (composition A), a further 7 days of only 451. mu.g 5-methyl- (6S) -tetrahydrofolate calcium salt (composition C) and so on (medication phase).

After 48 weeks no further 5-methyl- (6S) -tetrahydrofolate was administered. Alternatively, the administration of drospirenone and ethinylestradiol may be discontinued for another 40 weeks or the same.

The final blood sample was collected after 88 weeks. Due to the long-term nature of the study, the withdrawal rate can reach 50%.

Example 3:

blood was taken every 8 weeks from 80 healthy young fertile women and the red blood cell folate levels were determined using validated microbiological, immunological or instrumental (e.g. HPLC, LC-MS/MS) methods or a suitable combination of these methods.

At 8 weeks after the first blood sample, drospirenone 3mg, ethinylestradiol 20 μ g and calcium 5-methyl- (6S) -tetrahydrofolate 451 μ g (composition B) were in each case administered simultaneously on the first 24 days of each cycle over a period of 40 weeks. The administration of 451. mu.g of 5-methyl- (6S) -tetrahydrofolate calcium salt (composition C) was continued for 7 days at the immediately following stage. A further 21 days (second period) of 3mg drospirenone and 20. mu.g ethinylestradiol and 451. mu.g 5-methyl- (6S) -tetrahydrofolate calcium salt (composition B) were administered, a further 7 days of only 451. mu.g 5-methyl- (6S) -tetrahydrofolate calcium salt (composition C) were administered, and so on.

After 48 weeks, the administration of 5-methyl- (6S) -tetrahydrofolate was discontinued, but rather the administration of drospirenone and ethinylestradiol was continued for another 40 weeks or was interrupted as such.

The final blood sample was collected after 88 weeks. Due to the long-term nature of the study, the withdrawal rate can reach 50%.

The initial erythrocyte folate level in the subject is about 500 to 700nmol/l, which varies with eating habits, but in each case is less than 906 nmol/l. This value increases on the following day when the dietary habits remain the same, reaching a value of about 906nmol/l only after 6 to 8 weeks, i.e. after the second week. After a sustained administration for at least 30 weeks (corresponding to the lower half-life limit of 5), the red blood cell folate levels reached about 1200 to 1600nmol/l (steady state), although the diet remained the same. After termination of the administration of 5-methyl- (6S) -tetrahydrofolate, the level of erythrocyte folate decreases continuously. It is expected that starting from an average steady-state concentration of 1400nmol/l and maintaining the same eating habits, the red blood cell folate levels drop to below 906nmol/l in the eleventh to thirteenth weeks after cessation of the pharmaceutical composition of the invention, and therefore the concentration in the lowest red blood cells is generally sufficient to prevent neural tube defects in the eleventh to thirteenth weeks after cessation of the pharmaceutical composition of the invention.

Example 4: long term folate study

Blood was drawn every 2 weeks from 180 healthy young fertile women, half of which received an fortified folate diet, and the red blood cell folate levels were determined using validated microbiological, immunological or instrumental (e.g. HPLC, LC-MS/MS) methods or a suitable combination of these methods.

At 8 weeks after the first blood draw, the first group of 90 women received over a 24-week period:

drospirenone 3mg, ethinylestradiol 30 μ g and calcium 5-methyl- (6S) -tetrahydrofolate 451 μ g were administered simultaneously daily for the first 21 days of each cycle. 451. mu.g of 5-methyl- (6S) -tetrahydrofolate, calcium salt, was administered for 7 days immediately thereafter. A further 21 days (second cycle) of 3mg drospirenone, 30. mu.g ethinylestradiol and 451. mu.g 5-methyl- (6S) -tetrahydrofolate calcium salt, a further 7 days of only 451. mu.g 5-methyl- (6S) -tetrahydrofolate calcium salt and so on (drug treatment phase).

Group 90 women received as a control group administration of 3mg drospirenone, 30 μ g ethinylestradiol and 400 μ g folic acid according to the same dosing schedule.

In both cases the last blood was collected after 24 weeks. Followed by an observation period of 20 weeks during which the contraceptive product Yasmin is administeredFor 20 weeks, i.e. 3mg drospirenone and 30 μ g ethinylestradiol were administered simultaneously daily on the first 21 days of each cycle, and immediately thereafter no active substance was administered for 7 days (placebo or no dose). The withdrawal rate can reach 30 percent.

The starting erythrocyte folate level in the subject is less than 906 nmol/l. This value increases if the eating habits remain the same on the following days, in most women after 6 to 8 weeks a value of about 906nmol/l is reached upon administration of the pharmaceutical composition according to the invention. After 24 weeks of continuous dosing and maintaining the same dietary habits, red blood cell folate levels were achieved in both groups, indicating equivalence between the two treatment groups (bioequivalence criterion 80-125%). After termination of the administration of 5-methyl- (6S) -tetrahydrofolate, the level of erythrocyte folate decreases continuously. The time at which the red blood cell folate levels dropped below the accepted 906nmol/l threshold, which is generally considered sufficient to avoid neural tube defects, was confirmed.

Most women in the first group still showed sufficient levels of red cell folate 3 months after termination of intake.

Claims (46)

1. A medicament, comprising;

-5-methyl- (6S) -tetrahydrofolic acid

-one or more estrogens selected from: ethinylestradiol, mestranol and estradiol, and/or

At least one progestin selected from the group consisting of: levonorgestrel, norgestimate, nordehydrohydroxyprogesterone, dydrogesterone, drospirenone, cyproterone, desogestrel, dienogest, gestodene, and tibolone,

-optionally vitamin B₆And/or vitamin B₂

-and a pharmaceutically acceptable excipient/carrier,

absence of vitamin B₁₂。

2. The medicament of claim 1, comprising a crystalline calcium salt of 5-methyl- (6S) -tetrahydrofolic acid.

3. The medicament of claim 1, comprising 5-methyl- (6S) -tetrahydrofolic acid, drospirenone, and ethinylestradiol.

4. A medicament according to claim 3, comprising a daily dose of 0.1 to 10mg of 5-methyl- (6S) -tetrahydrofolic acid.

5. A medicament according to claim 3, comprising a daily dose of 0.4 to 1mg of 5-methyl- (6S) -tetrahydrofolic acid.

6. A medicament according to claim 3, comprising a daily dose of 451 μ g of 5-methyl- (6S) -tetrahydrofolate calcium salt.

7. A medicament according to claim 3, comprising drospirenone in a daily dose from 0.5 to 5 mg.

8. A medicament according to claim 3, comprising drospirenone in a daily dose of 3 mg.

9. A medicament according to claim 3, comprising ethinylestradiol in a daily dose of 10 to 50 μ g.

10. A medicament according to claim 3, comprising ethinylestradiol in a daily dose of 10 to 30 μ g.

11. A medicament according to claim 3, comprising ethinylestradiol at a daily dose of 20 μ g.

12. A medicament according to claim 3, comprising ethinylestradiol at a daily dose of 30 μ g.

13. The medicament of claim 3, comprising

A daily dose of 451. mu.g of 5-methyl- (6S) -tetrahydrofolate calcium,

drospirenone in a daily dose of 3mg, and

ethinyl estradiol at a daily dose of 20 μ g.

14. The medicament of claim 3, comprising

A daily dose of 451. mu.g of 5-methyl- (6S) -tetrahydrofolate calcium,

drospirenone in a daily dose of 3mg, and

ethinylestradiol in a daily dose of 30 μ g.

15. A kit comprising

-at least 20 daily dosage units containing a medicament according to any one of claims 1 to 14, and

-at least one compound containing 5-methyl- (6S) -tetrahydrofolic acid and optionally vitamin B₆And/or vitamin B₂The daily dosage unit of (a) is,

-wherein the number of all dosage units present in the kit is at least 28, and

-the arrangement of the dosage units is such that dosage units containing the medicament of any one of the preceding claims should be taken first and then dosage units containing only 5-methyl- (6S) -tetrahydrofolate.

16. The kit of claim 15, comprising

-20-30 daily dosage units containing a medicament according to any one of claims 1-14, and

1-10 daily dosage units containing 5-methyl- (6S) -tetrahydrofolate.

17. The kit of claim 15, comprising

-21-26 daily dosage units containing a medicament according to any one of claims 1-14, and

2-7 daily dosage units containing 5-methyl- (6S) -tetrahydrofolate,

-wherein the number of all dosage units present in the kit is 28.

18. The kit of claim 15, comprising

-21 daily dosage units containing a medicament according to any one of claims 1 to 14, and

7 daily dosage units containing 5-methyl- (6S) -tetrahydrofolate.

19. The kit of claim 15, comprising

-24 daily dosage units containing a medicament according to any one of claims 1 to 14, and

4 daily dosage units containing 5-methyl- (6S) -tetrahydrofolate.

20. A kit according to claim 15, comprising 451 μ g of 5-methyl- (6S) -tetrahydrofolate calcium salt per daily dosage unit.

21. The use of the following combination for the preparation of a medicament,

-5-methyl- (6S) -tetrahydrofolic acid,

-one or more estrogens selected from: ethinyl estradiol, mestranol, and a diol, and/or

At least one progestin selected from the group consisting of: levonorgestrel, norgestimate, nordehydrohydroxyprogesterone, dydrogesterone, drospirenone, cyproterone, desogestrel, dienogest, gestodene, and tibolone,

-optionally vitamin B₆And/or vitamin B₂，

-and a pharmaceutically acceptable excipient/carrier,

absence of vitamin B₁₂，

The medicament is for use in reducing the risk of conditions caused by folate deficiency and congenital malformations caused by folate deficiency after prolonged continuous intake of the medicament for at least 6-10 weeks prior to termination.

22. The use of claim 21 for reducing the risk of neural tube defects.

23. The use of claim 21 for reducing cardiac defects.

24. The use of claim 23, wherein the cardiac defect is a ventricular valve defect.

25. The use according to claim 21 for reducing the risk of urinary tract malformations.

26. The use according to claim 21 for reducing the risk of cleft lip, maxillofacial and palatine cleft.

27. The use of claim 21 for reducing the risk of spontaneous abortion.

28. The use of claim 21 for reducing malignant disorders.

29. The use of claim 21, wherein the malignant disorder is breast cancer or colon cancer.

30. The use of claim 21 for reducing the risk of a cardiovascular disorder.

31. Use according to one of claims 22 to 30, with 5-methyl- (6S) -tetrahydrofolic acid, drospirenone and ethinylestradiol.

32. Use according to one of claims 22 to 30, with 451 μ g of 5-methyl- (6S) -tetrahydrofolate calcium salt, 3mg of drospirenone and 20 μ g of ethinylestradiol.

33. Use according to one of claims 22 to 30, with 451 μ g of 5-methyl- (6S) -tetrahydrofolate calcium salt, 3mg of drospirenone and 30 μ g of ethinylestradiol.

34. The kit of claim 15, comprising

-21 daily dosage units containing the medicament of claim 13, and

7 daily dosage units containing 5-methyl- (6S) -tetrahydrofolate.

35. The kit of claim 15, comprising

-24 daily dosage units containing the medicament of claim 13, and

4 daily dosage units containing 5-methyl- (6S) -tetrahydrofolate.

36. The kit of claim 15, comprising

-21 daily dosage units containing the medicament of claim 14, and

7 daily dosage units containing 5-methyl- (6S) -tetrahydrofolate.

37. A process for the formulation of a medicament according to any one of claims 1 to 14, characterised in that 5-methyl- (6S) -tetrahydrofolate is absorbed only after granulation.

38. The process according to claim 37, characterized in that low-substituted hydroxypropylcellulose is used as binder.

39. A kit according to claim 15 comprising more than 28 daily dosage units, wherein at least 28 daily dosage units contain a medicament according to any of claims 1-14, wherein at least one daily dosage unit contains 5-methyl- (6S) -tetrahydrofolate, and wherein the dosage units are arranged such that dosage units containing a medicament according to any of claims 1-14 should be taken first, and dosage units containing only 5-methyl- (6S) -tetrahydrofolate should be taken second.

40. The kit of claim 39, wherein the number of dosage units containing a medicament according to any one of claims 1 to 14 is 28 plus 21, 22, 23, 24, 25, 26 or 27 or an integer multiple of 28 plus 21, 22, 23, 24, 25, 26 or 27 and wherein the number of daily dosage units containing only 5-methyl- (6S) -tetrahydrofolate is 7, 6, 5, 4, 3, 2 or 1.

41. The kit of claim 40, wherein the multiple is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

42. A kit comprising more than 28 daily dosage units, wherein at least 28 daily dosage units contain a medicament according to any of claims 1-14, wherein at least one daily dosage unit is a placebo or a blank contraceptive day, and wherein the dosage units are arranged such that a dosage unit containing a medicament according to any of claims 1-14 should be taken first.

43. The kit of claim 42, wherein the number of dosage units containing the medicament of any one of claims 1 to 14 is 28 plus 21, 22, 23, 24, 25, 26 or 27 or an integral multiple of 28 plus 21, 22, 23, 24, 25, 26 or 27, and wherein the number of placebo or blank contraceptive days is 7, 6, 5, 4, 3, 2 or 1.

44. The kit of claim 43, wherein the multiple is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

45. The kit of any one of claims 39 to 44, wherein the drug-containing dosage unit comprises a drug of claim 13.

46. The kit of any one of claims 39 to 44, wherein the drug-containing dosage unit comprises a drug of claim 14.