EP2841074A1

EP2841074A1 - Intrauterine application of 18-methyl-15ss,16ss-methylene-19-nor-20-spirox-4-en-3-one systems, intrauterine systems containing 18-methyl-15ss,16ss-methylene-19-nor-20-spirox-4-en-3-one, as well as the use thereof in contraception and gynaecological therapy

Info

Publication number: EP2841074A1
Application number: EP13717289.6A
Authority: EP
Inventors: Norbert Schmees; Lars RÖSE; Tuula VALO; Katja Prelle; Reinhard Nubbemeyer; Henriikka Korolainen; Harri Jukarainen
Original assignee: Bayer Pharma AG
Current assignee: Bayer Pharma AG
Priority date: 2012-04-23
Filing date: 2013-04-19
Publication date: 2015-03-04
Also published as: HK1205000A1; TN2014000444A1; AU2013254840A1; WO2013160213A1; IL235095A0; MX2014012849A; BR112014026193A2; MA37444A1; SG11201406582XA; CN104254333A; CO7111255A2; JP2015514791A; CR20140490A; DOP2014000241A; IN2014DN07839A; CL2014002836A1; CA2871003A1; PH12014502372A1; CU20140121A7; AR090799A1

Abstract

The present invention describes the intrauterine application of 18-methyl-15ß,16ß-methylene-19-nor-20-spirox-4-en-3-one of the general formula (1) in contraception and gynaecological therapy, as well as an intrauterine system containing compounds of formula (1).

Description

Intrauterine application of

18-methyl-15SS, 16ss-methylene-19-nor-20-Spirox-4-en-3-ones,

containing intrauterine systems

18-methyl-15β, 16β-methylene-19-nor-20-spirox-en-3-ones, and their use in contraception and gynecological therapy

The present invention relates to the in the claims

marked article, i. the intrauterine application of 18-ethyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-ones of the formula (I) and an intrauterine system containing 18-ethyl-15β, 16β-methylene 19-nor-20-spirox-4-ene-3-one of general formula I,

Formula (1) wherein R ⁶ and R ^{7 may be} a hydrogen atom or together may be an α-methylene group.

The invention thus relates to the intrauterine application of 18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-one (compound A) or 18-methyl-6a, 7a, 15β , 16β-dimethylene-19-nor-20-spirox-4-en-3-one (compound B)

(A) (B) , and an intrauterine system (IUS) containing one of said compounds.

Another object of the invention relates to the use of an IUS containing the substance (A) or (B) in contraception and in gynecological therapy.

Under gynecological therapy, e.g. the treatment of endometriosis, endometrial hyperplasia, inflammation of the endometrium (endometritis), uterine pain and dysmenorrhea, the use being excluded in the treatment of menorrhagia [also known as hypermenorrhea or heavy enstrual bleeding (HMB)] and other forms of uterine bleeding disorders ,

Another object of the invention relates to the use of 18-ethyl-15ß, 16ß-methylene-19-nor-20-spirox-4-en-3-one (compound A) in contraception and gynecological therapy.

The inventively employable progestins, 18-methyl-15ß, 16ß-methylene-19-nor-20-spirox-4-en-3-one (A) or 18-methyl-6a, 7a, 15ß, 16ß-dimethylene -19-nor-20-spirox-4-en-3-one (B), and their preparation, are described in WO 2008/000521, the first-mentioned compound (A) being disclosed there only as an intermediate.

Compound B and other substances described in WO 2008/000521 are used in pharmaceutical preparations for contraception and in therapy for the treatment of premenstrual disorders, such as headache, depressive moods, water retention and astodynia. In addition to oral and transdermal dosage forms, WO2008 / 000521 also discloses parenteral oily injection solutions. However, intrauterine use and the use of the compounds in an intrauterine system (IUS) are not described in WO 2008/000521.

Hormone-based contraceptive approaches, find due to the convenient

Application and high contraceptive safety among the users a wide acceptance. Among these, oral contraceptives (pill) are the most widely used contraceptive method in many countries. Nevertheless, hormone-containing, in particular estrogen-containing contraceptives in the public and the literature due to potential risks (such as a slightly increased risk of thrombosis,

Loss of libido, nausea and headache) are also critically discussed ¹ .

[0009] As a promising new contraceptive method, hormones are administered intrauterine, via a corresponding intrauterine system. Mention should be made first of all of ^Mirena® , a levonorgestrel-containing intrauterine system (IUS), which continuously releases the active substance over a period of up to five years. The product is used for contraception and in the treatment of increased menstrual bleeding (menorrhagia or hypermenorrhea). This product is described inter alia in EP 0652738 B1 and EP 0652737 B1.

The basis for the contraceptive effect of Mirena ^® is essentially the thickening of the cervical mucus and the local effect of Levonorgestrels, which leads to a strong antiproliferative effect on the endometrium. In addition, levonorgestrel alters the utero-tubal milieu and impairs sperm motility and function.

Although the contraceptive effect of Mirena ^{® is} significantly caused by the local action, can also be found in Mirena ^® due to the relatively high systemic stability of levonorgestrel (active ingredient in Mirena ^® )

Plasma drug levels of on average by 206 pg / ml ² . Although this value is below that of orally administered levonorgestrel-containing contraceptives, it is still so high that it inhibits ovulation in about 20% of users in the first year of application and the known systemic side effects such as acne, depressive moods, May result in chest pain or decreased libido ³ .

Even some first-time users complain about an irregular

Bleeding behavior in the initial phase, ie immediately after the onset of IUS (so-called "spotting") .This spotting may last up to a few months before either no or very little and rare bleeding occurs ⁴ .

[0013] Benign be Ovarialszysten as another common side effect of Mirena ^® described ^fifth

¹ Bitzer et al. Contraception 84 (201 1) 342-356

² see expert information Mirena March 201 1 - EN / 9

³ Lähteenmäki P. et al. Steroids 2000 65: 693-697

⁴ Suvisaari J, Lähteenmäki P. - Contraception 1996 Oct; 54 (4): 201-8.

⁵ Product Monograph - Mirena 8th edition August 2009; Finland: Schering AG and Leiras Oy In addition to the aforementioned hormonal (contraceptive) methods, there is a wide range of non-hormone based approaches and products, such as natural ones

Contraceptive methods (e.g., hormone level measurement or temperature method), mechanical methods (e.g., condom or diaphragm), or chemical methods (e.g., spermicides). Unfortunately, none of the alternative methods (apart from irreversible sterilization) has nearly the same contraceptive safety achieved with hormone-based methods.

The object of the present invention is therefore to provide a contraceptive method which, with comparable high contraceptive safety, as achieved by the known hormone-based contraceptive methods, has even better tolerability.

The faster achievement of a regular bleeding behavior, i.

less mocking, is another object of the invention ⁶

The object is achieved by the intrauterine application of compounds of the formula (I)

Formula (1) wherein R ⁶ and R ^{7 is} a hydrogen atom or together an α-methylene group, namely by the intrauterine application of 18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-ene-3 or 18-methyl-6a, 7a, 15β, 16β-dimethylene-19-nor-20-spirox-4-en-3-one.

Surprisingly, in intrauterine application of 18-methyl-15ß, 16ß-methyl-19-nor-20-spirox-4-en-3-one or 18-methyl-6a, 7a, 15ß, 16ß-bis-methylene 19-nor-20-spirox-4-en-3-one in the rat a differentiating effect between local (uterine) and systemic (peripheral tissue) effect, are shown. This effect could be achieved by comparing the local effects in the uterus

(Weight gain see Example 1, Figs 1/8 and 2/8) and by the systemic effect, such as e.g. the lowering of LH level in ovariectomized rats (Figures 3/8 and 4/8).

Also, the substances in comparison to LNG increased local potency, as in the gene expression experiment based on the strong induction

corresponding marker genes could be shown. Thus, the anti-estrogenic effect of progestagens on the uterus, etc. mediated via IGFBP-1. In Fig. 5/8 and 6/8 it is shown that the gene expression of IGFBP-1 is already induced by compound A at an approximately 7x lower release rate from the IUS than levonorgestrel.

The pronounced strong dissociation local vs. systemic and the high gestagenic efficacy of the substances is sufficient to induce a contraceptive effect solely due to the local effects. Systemically caused

Side effects, such as those that occur when other progestins are used, can thus be avoided or at least greatly reduced. Also, due to the possible higher local concentration of progestin, a faster adjusting and better bleeding control is to be expected.

As could be further demonstrated in comparative transactivation studies (see Example 2), the substances used according to the invention have an at least 10-fold lower androgenic activity compared to LNG. This property, compounded with the pronounced dissociation local vs. systemic, shows that no systemic androgenic effects (eg acne) are to be expected even with very high dose local uterine applications compared to levonorgestrel, even if systemic concentrations comparable to levonorgestrel in Mirena ^® applications should occur.

Due to the above properties of the substances A and B, the substances are excellent for intrauterine use in gynecological therapy but especially in contraception. An intrauterine

Administration by means of an intrauterine system is preferred.

As an intrauterine system, a polymer system can be used, as used for example in Mirena ^® .

⁶ Andersson et al. Contraception 1994, 49: 56-71 The preparation of an IUS is known to the person skilled in the art and is carried out as described, for example, in EP 0 652 738 B1.

Thus, the active ingredient (A) or (B) is first processed with a polymeric carrier material to a central rod (core). The active ingredient may be in any proportion with the polymeric carrier material, e.g. Polydimethylsiloxane (POMS), mixed.

The core thus produced, after molding, i. after this

Vulcanization is usually surrounded in a second step by a membrane based on a polymer that ensures a uniform dosage over a long period of time. The desired release rate ("release rate") can be controlled by selecting the polymer and the thickness of the membrane.

The polymer used for the membrane are in principle the same polymers as for the core (the central rod) in question. To mention here are e.g. Polydimethylsiloxane, which may optionally be fluorinated or other mixtures of polymers. The membrane thickness is preferably in the range of 0.5 mm.

The membrane is applied by a made of the desired polymer tube (membrane) is first brought to swell in a solvent and then pressed into the still swollen tube of drug-containing core. Preferably, the tube ends are then still closed with a plug, which preferably consists of the same material as the tube / membrane, in order to counteract "bleeding" of the active substance at the tube ends, which can lead to a "burst effect" in the application , Instead of the plugs, the tube can also be glued with silicone. Systems which can be used according to the invention are those which release a daily dose in the range from 1 to 500 μg of the respective active ingredient (A) or (B).

In this case, the release rate of active ingredient (A) may be chosen half as high as that of active ingredient (B) due to its higher efficacy.

For the active ingredient (A) thus results in a preferred dose range of 1- 200 g / day, more preferably 1-100 g / day, in particular from 2-50 g / day. Active ingredient (B) is preferably in the range between 2-500pg / day, more preferably from 2-200pg / day, in particular from 5-100 g / day, dosed.

Another object of the invention thus relates to an intrauterine system containing the active ingredient (A) or (B), as well as the application of the intrauterine system in the contraception.

The following examples serve to illustrate the invention.

The preparation of the inventively employable progestins, 18-methyl-15ß, 16ß-methylene-19-nor-20-spirox-4-en-3-one (compound A) or 18-methyl-6α, 7a, 15ß. 16β-dimethylene-19-nor-20-spirox-4-en-3-one (compound B) is carried out as described in WO 2008/000521 (Compound A: Example 14 f; Compound B: Example 2).

The method of preparation of the active substance loaded rods, which in the

The rat experiment described below was carried out analogously to the preparation of the drug reservoirs, as described, for example, for a human-suitable IUS (see, for example, EP 0 652 738 B1). As the polymer for the production of the rod, polysiloxanes and modified polysiloxane polymers can be used (for example, see EP 0652738 B1, WO 00/29464 and WO 00/00550).

In particular, a drug-loaded core was first prepared by using a mixture of polyethylene oxide block polydimethylsiloxane copolymer (PEO -b-PDMS), polydimethylsiloxane and 10 weight percent of the active ingredient (here the respective progestin A or B) using a Pt (O) -divinyltetramethyldisiloxane catalyst was vulcanized.

Instead of PEO-b-PDMS also polydimethylsiloxane (PDMS) can be used, was used as the catalyst for vulcanization bis (2,4-dichlorobenzoyl) peroxide.

For the preparation of the active substance-containing core, a vertical piston unit was used with a corresponding nozzle head. The nozzle head was dimensioned so that the active substance-containing core has an outer diameter of about 1 mm.

The active ingredient-containing core prepared in this way is subsequently treated with a membrane which consists of PDMS, polytrifluoropropylmethylsiloxanes (PTFPMS) or a PTFPMS / PDMS elastomeric mixture (75% of PTFPMS, 25% PDMS), jacketed. The membrane material had an inner diameter of ~ 1 mm and a

Outer diameter of -1.5 mm.

The sheathing was carried out by cutting the core and the membrane to the length of 10-15 mm, the membrane being slightly longer (about 1 mm at each end) than the core, so that the ends of the membrane after the insertion of the core can be closed with a small stopper. In order to allow the introduction of the core into the membrane, it was first swelled in a cyclohexane or acetone-hexane mixture. In the swollen membrane then the active substance-containing core was inserted. The ends of the tubes are either glued with silicone or closed with a small stopper made of PTFPMS. Example 1

The local uterine action of progestin versus systemic

Side effects (dissociation) have been studied in rat studies. The uterus of ovariectomized rats respond with decidualization and weight gain to the implantation of progestin-containing IUS (rods). The local progestin effects were also determined by gene expression changes.

Serum levels of luteinizing hormone (LH) serve to detect systemic effects of locally applied progestin. Basal serum LH levels of ovariectomized rats are elevated compared to levels of intact control animals. Unwanted systemic efficacy of uterine-administered progestin can be demonstrated by a decrease in LH levels.

Method:

Female ovariectomized rats were treated with estradiol (E2) for three days (0.2 g / day / animal, dosing was subcutaneous). On day 4, each animal was implanted with an IUS (rod) in the right uterine horn. The left uterine horn remained untreated as an internal comparison. The E2 applications were continued at a daily dose of 0.1 g / animal to increase the responsiveness of the uterus (preservation of progesterone). Receptor expression) to ensure progestins. On days 4, 10 and 17, blood was drawn for LH level determinations.

Carrying out the gene expression analyzes:

The uterine tissue was purified in 800 μL RLT lysis buffer (Qiagen, Hilden, Germany, # 79216) using the Precellys24 homogenizer (Peqlab, Erlangen,

Germany; 2.8mm ceramic beads; # 91-PCS-CK28, 2x 6000rpm) homogenized. 400μΙ of the homogenate obtained were used for the isolation of the total RNA. For this, the QIAsymphony RNA kit (Qiagen, # 931636) was used

QIAsymphony SP robot used for automated sample processing. The reverse transcription of 1 pg to 4pg total RNA was performed under

Use of the SuperScript III First Strand Synthesis System (Invitrogen, Carlsbad, USA; # 18080-051) according to the Random Hexamer Procedure.

Gene expression analysis was performed with 50ng to 200ng cDNA per reaction on

SDS7900HT Real.time PCR System (Applied Biosystems, Carlsbad, USA) using TaqMan probes (Applied Biosystems, IGFBP-Rn00565713_m1, Cyp26a1 Rn00590308_m1, PPIA Rn00690933_m1) and the Fast Blue qPCR

MasterMix Plus (Eurogentec, Liege, Belgium; # RT-QP2X-03 + FB). Cyclophilin A (PPIA) was used as an endogenous control for relative quantification. The calculation of the relative expression levels was carried out according to the comparative delta delta CT method.

Results:

18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-one (compound A) and 18-methyl-6a, 7a, 15β, 16β-dimethylene-19-nor-20- Spirox-4-en-3-one (compound B) showed dose-dependent local efficacy by weight gain in the IUS-bearing

Uterine horn (Figs. 1/8 and 2/8).

In the tested release range (for Compound A: 0.6 -10 pg per animal and day or for Compound B: 1-45 pg / animal and day) both showed progestins

Surprisingly, with the exception of the 10pg / animal and day dose of Compound A, no LH depression and thus no systemic side effect (Figs 3/8 and 4/8).

The pharmacokinetic profile of 18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-one and 18-methyl-6a, 7a, 15β, 16β-dimethylene-19- nor-20-Spirox-4-en-3-one showed a very rapid degradation rate in all in vitro metabolism studies (liver) as well as in all animal species examined in vivo.

In the local application by means of IUS (rods) in the rat showed

Compound A at 4 to 7 fold higher potency in induction of gene expression than levonorgestrel at equal release rates (Figure 3/4). This higher local potency also supports the possibility of achieving faster and stronger local gestagenic effects on the uterus without causing systemic side effects.

Thus, these progestins can be dosed with local activity so without the side effects described for levonorgestrel occur in women. Also for humans, very rapid degradation rates were also found in vitro (liver). Rapid in vitro degradation in the liver can also be implicated in rapid in vivo degradation, resulting in a very low systemic exposure of 18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-ene 3-one and 18-methyl-6a, 7a, 15β, 16β-dimethylene-19-nor-20-spirox-4-en-3-one is calculated after application by an IUS. The expected substance concentrations (Css = concentration at steady state) are calculated from the release rate from the IUS divided by the disappearance rate (clearance). Using a 20 μg dose per day and woman equivalent to that of Mirena results in a more than 30-fold lower systemic exposure to 18-methyl-15β, 16β-methylene-19-nor-20-spirox -4- en-3-οη and 18-methyl-6a, 7a, 15SS, 16ss-dimethylene-19-nor-20-Spirox-4-en-3-one in comparison with Mirena ^®.

Example 2

The effect on the human androgen receptor (hAR) was determined by means of

Transactivation analyzes examined. For this purpose, the test substances were in

different concentrations to cells that express the human androgen receptor stable and can be detected via a reporter gene activation of the An d rogre reze pto rs. Method:

For transactivation studies, PC3 (human prostate carcinoma) cells stably transfected with hAR and reporter gene MTV-luc were used. When

Culture medium was RPMI medium (without L-glutamine; without phenol red) # E15-49 PAA L-glutamine 200mM # 25030-024 Gibco BRL 100U / 100g / ml penicillin

/ Streptomycin Gibco # 15140-122 used with 10% fetal calf serum (FCS). The cultivation of the cells was carried out at 37 ° C and 5% C0 ₂ . The test medium was the same as the culture medium except that the 10% FCS was replaced by 5% activated charcoal-treated FCS (CCS). Cells were seeded in wells of a 96 well plate ("CulturPlate" from Packard # 6005180) with 2x104 cells / well / 200μΙ assay medium The cells were incubated with different concentrations of the test substances and 80μΙ substrate with the "steadylite HTS Reporter Gene Assay System" of Measure Perkin Elmer. Results:

The results show Compound A (18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-one) and Compound B (18-methyl-6a, 7a, 15β, 16β- bis-methylene-19-nor-20-spirox-4-en-3-one) have a more than 10-fold higher EC50 in the transactivation of the hAR than levonorgestrel: while the EC50 values for Compound A: 6.9 nM and for Compound B: 56 nM, levonorgestrel has an EC50 of only 0.5 nM. This> 10-fold dissociation with levonorgestrel means that at

Applications of compounds with no systemic androgenic effects is to be expected, even if in local uterine applications systemic

Drug levels should occur as they are observed for levonorgestrel in Mirena ^® applications.

Example 3

The amount of active ingredient (A) or (B) released was determined by reversed-phase liquid chromatography with UV detection in a 1% 2-hydroxypropyl-β-cyclodextrin (2-HPBCD) solution.

For a rod coated with a PTFPMS membrane, the in vitro release rates reported in Figures 7/8 and 8/8 were determined.

Claims

claims

1. Intrauterine Use of 18-Methyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-ones of the General Formula (1)

Formula (1) wherein R ⁶ and R ^{7 is} a hydrogen atom or together an α-methylene group.

2. Intrauterine system containing 18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-one of the general formula (1)

The intrauterine system of claim 2 containing 18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-ene-3-one.

4. Intrauterine system according to claim 2 containing 18-methyl-6a, 7a, 15ß, 16ß-dimethylene-19-nor-20-spirox-4-en-3-one.

5. Intrauterine system according to claim 3, characterized in that from the system a daily dose of 1 -200 ug 18-methyl-15ß, 16ß-methylene-19-nor-20-spirox-4-en-3-one is released ,

6. An intrauterine system according to claim 5, characterized in that from the system a daily dose of 1 -100 ug 18-methyl-15ß, 16ß-methylene-19-nor-20-spirox-4-en-3-one is released ,

7. An intrauterine system according to claim 6, characterized in that from the system a daily dose of 2-50 ug 18-ethyl-15ß, 16ß-methylene-19-nor-spirospox-4-en-3-one one is released ,

8. An intrauterine system according to claim 4, characterized in that from the system a daily dose of 2-500 micrograms of 18-methyl-6a, 7a, 15ß, 16ß-dimethyl-19-nor-20-spirox-4-ene-3 -on is released.

9. Intrauterine system according to claim 8, characterized in that from the system a daily dose of 2-200 mg of 18-methyl-6a, 7a, 15β, 16β-dimethyl-19-nor-20-spirox-4-ene-3 -on is released.

10. An intrauterine system according to claim 9, characterized in that from the system a daily dose of 5-100 micrograms of 18-methyl-6a, 7a, 15ß, 16ß-dimethyl-19-nor-20-spirox-4-ene-3 -on is released.

1 1. Intrauterine system according to any one of claims 2-10 for use in contraception.

12. An intrauterine system according to any one of claims 2-10 for use in gynecological therapy with the exception of the therapy of menorrhagia and other uterine bleeding.

13. Use of 18-methyl-15β, 16β-methylene-19-nor-20-spirox-4-en-3-one in contraception and gynecological therapy with the exception of the therapy of menorrhagia and other uterine bleeding.