RU2259830C2 - Pharmaceutical composition for treatment of diseases associated with loss of osseous mass - Google Patents

Abstract

FIELD: medicine, chemical-pharmaceutical industry.

SUBSTANCE: invention relates to new applying EP₄ receptors agonist for treatment and/or prophylaxis of diseases associated with loss of osseous mass. Agonists of EP₄ receptors show high effectiveness in treatment of diseases associated with loss of osseous mass, among the, as osteoporosis of different genesis. Agonists of EP₄ receptors involve prostaglandin skeleton base.

EFFECT: valuable medicinal properties of pharmaceutical composition.

16 cl, 3 tbl, 5 ex

Description

Technical field

The present invention relates to a pharmaceutical composition for the treatment of diseases related to bone loss containing an EP ₄ receptor agonist as an active ingredient.

In more detail, the invention relates to a pharmaceutical composition for the treatment and / or prevention of diseases associated with bone loss, such as primary osteoporosis, secondary osteoporosis, bone metastasis, hypercalcemia, Paget's disease, bone loss and osteonecrosis, etc .; and a pharmaceutical composition for accelerating osteogenesis, such as osteogenesis after fracture, osteogenesis after bone transplantation, osteogenesis after surgery for an artificial joint, osteogenesis after spinal fusion and osteogenesis after other bone regeneration operations, etc .; and to a pharmaceutical composition for the accelerated treatment of these diseases, containing an EP ₄ receptor agonist as an active ingredient.

State of the art

In about 120-150 days, the bone goes through a cycle including bone resorption by osteoclasts, osteogenesis of osteoblasts, and a resting period (called remodeling). In a healthy adult, both bone resorption and osteogenesis are strictly controlled, so that there is a slight change in total bone mass. However, in a patient suffering from a disease associated with bone loss, the balance between bone resorption and osteogenesis is impaired and bone loss and bone degradation occur. The degree of bone loss reaches 20-30%. Such a patient is prone to bone fractures, which sometimes cause bed rest, body deformation, or, in the worst case, death, when the fractures occur in dangerous places such as the hip joint, etc.

A typical disease associated with bone loss is osteoporosis. Osteoporosis is a systemic disease for which, as the main symptom, bone loss and bone degradation are characteristic.

The causes of osteoporosis can be different, and the most common cause is believed to be age, especially postmenopausal hormonal imbalance in women. Therefore, at present, the main treatment for osteoporosis is the introduction of estrogen, vitamin D or calcitonin. However, with hormone treatments there is a risk of cancer stimulation (especially breast cancer, uterine cancer, etc.), so such treatment is unsafe.

In addition, bisphosphonate was administered as a second-generation drug. However, the problem is that cessation of administration may cause complications.

In addition, even if with the mentioned treatment it is possible to delay the aggravation of osteoporosis, it is difficult to regenerate the bone, the mass of which once decreased.

On the other hand, prostaglandin E ₂ (short for PGE ₂ ) is known as a metabolite in the arachidonic acid cascade. It is known that it has various activities, such as cytoprotective activity, uterine contractile activity, pain inducing effect, stimulating effect on gastrointestinal motility, an awakening effect, inhibitory effect on acid secretion in the stomach, hypotensive activity, diuretic activity, etc. .d.

PGE ₂ is believed to be related to bone resorption (J. Dent Res. 59 (10), 1635 (1980); Nature 266.6455 (1977)).

In addition, there is a patent application stating that macroscopically or microscopically observed bone growth and an increase in bone mass in a dog that was administered PGE ₁ or PGE ₂ orally or intravenously. However, the dose of each compound was 1000 μg / kg / day, which is a very high dose for PG, so it is doubtful that each named PG plays a significant role. Due to such a high dose, it may be difficult to obtain the drug in practice.

A recent study proved the existence of various subtypes of PGE ₂ receptor, which have a physical purpose that distinguishes them from each other. Currently, four receptor subtypes are known; they are named EP ₁ , EP ₂ , EP ₃ , EP ₄ (Ngishi M. et al., J. Lipid Mediators Cell Signaling 12, 379-391 (1995)). By studying the role of each receptor subtype and finding a compound that can form a specific connection with each receptor and cannot form a connection with any other receptors, it became possible to obtain a tool that gives fewer side effects.

It has recently been discovered that an EP ₂ receptor agonist and an EP ₄ antagonist are useful in the treatment of osteoporosis (WO98 / 279769) and accelerated bone resorption (GB2 330307), respectively.

In addition, in the application EP-855389 describes that the compound of formula (IA) was used as a conjugator for the EP ₄ receptor. This patent application shows that the compound of formula (IA) is useful for the treatment and prevention of osteogenesis disorders. However, there is no description or suggestion that the EP ₄ receptor agonist or EP ₄ antagonist is related to osteogenesis or its role in treatment.

As mentioned above, there is an assumption that PGE ₂ or PGE ₁ belong to bone metabolites, and for some of their subtypes this assumption is indeed confirmed. However, it is not clear which subtypes of them, or which particular antagonist or receptor agonist, are applicable.

SUMMARY OF THE INVENTION

The authors of the present invention and others synthesized various compounds that can selectively bind to each subtype of the PGE receptor, and examined them to find compounds that affect osteogenesis as a means for treating diseases associated with bone loss.

Until now, it was believed that the EP ₄ antagonist is applicable for the treatment of diseases associated with bone loss due to its inhibitory effect on bone resorption, and the EP ₄ receptor agonist is not applicable for such treatment due to the stimulating effect on bone resorption, which can cause bone loss. However, unexpectedly, the authors of the present invention found that the EP ₄ receptor agonist in low doses causes an increase in bone mass and bone density, and thereby realized the present invention.

In addition, it was experimentally confirmed that the compounds of formulas (IA) to (IJ), listed below and which are agonists of the EP ₄ receptor, do exert an agonistic effect of the EP ₄ receptor.

The present invention relates to a pharmaceutical composition for the treatment and / or prophylaxis of diseases related to bone loss containing an EP ₄ receptor agonist as an active ingredient.

According to the present invention, it was first confirmed that the EP ₄ receptor agonist has activity in increasing bone mass and bone density and stimulating osteogenesis.

In the present invention, the term "disease associated with bone loss" and means a disease due to conditions such as reduced bone density, degradation of bone tissue, etc., and includes, for example:

1) primary osteoporosis (for example, primary age-related osteoporosis, primary postmenopausal osteoporosis, primary osteoporosis due to oophorectomy, etc.);

2) secondary osteoporosis (e.g. glucocorticoid-induced osteoporosis; hyperthyroidism-induced osteoporosis; immobility-induced osteoporosis; heparin-induced osteoporosis; immunosuppression-induced osteoporosis; osteoporosis; osteoporosis; inflammatory disease; osteoporosis; inflammatory disease; d.);

3) bone diseases such as bone metastasis, hypercalcemia, Paget's disease, bone loss (alveolar bone loss, jaw bone rarefaction, idiopathic bone loss in children, etc.), osteonecrosis, etc.

In addition to treating the aforementioned diseases, the present invention also includes a pharmaceutical composition for accelerating osteogenesis after bone surgery (e.g., osteogenesis after fractures, osteogenesis after bone transplantation, osteogenesis after surgery for an artificial joint, osteogenesis after spinal fusion and osteogenesis after other operations for bone regeneration and etc.), or stimulating bone treatment, or alternative bone transplantation treatment.

According to the present invention, an EP ₄ receptor agonist is each compound that can selectively form a binding to an EP ₄ receptor subtype of a PGE ₂ receptor, and cannot or does not form a binding to any other EP ₁ , EP ₂ , EP ₃ receptor subtype, and has an affinity for EP ₄ .

Therefore, such compounds also include EP ₄ agonists, which will be found in the future, as well as those currently known. In addition, such EP ₄ agonists include both compounds lacking the PG skeleton and those with the PG skeleton.

These compounds can selectively form a bond with the EP ₄ receptor, and they generally do not induce pain that EP ₁ can cause, uterine relaxation that EP ₂ can cause, and uterine contractions that can cause EP ₃ , so that they are not having no influence on the above actions.

As shown in the experiments mentioned later, the EP ₄ agonist has a strong stimulating effect on osteogenesis, so that it is useful for treating the aforementioned diseases associated with bone loss, such as osteoporosis.

Preferred compounds used in the present invention are as follows: for example, compounds selected from EP ₄ agonists consisting of compounds of formula (IA)

where R ^1A means hydroxy, C 1-4 alkyloxy, or NR ^6A R ^7A (where each R ^6A and R ^7A independently means hydrogen or C 1-4 alkyl),

R ^2A means hydrogen or hydroxy-group,

R ^3A means:

(i) C1-8alkyl, C2-8alkenyl or C2-8alkynyl;

(ii) phenyl or C3-7cycloalkyl;

(iii) C1-8 alkyl, C2-8 alkenyl or C2-8 alkynyl substituted with phenyl or C3-7 cycloalkyl (provided that when R ^2A is hydrogen, the alkyl, alkenyl and alkynyl in (i) and (ii) may be substituted with one hydroxy group).

The symbol --- means double or single bond. Provided that its 8-epirequilibrium compound is included.

Compounds of Formula (IB)

Where R ^1B means hydroxy, C 1-6 alkyloxy, or NR ^6B R ^7B (where each R ⁶³ and R ⁷⁸ independently means hydrogen or C 1-6 alkyl),

R ^2B means hydrogen or hydroxy-group,

R ^3B means a single bond or C1-6 alkylene,

R ^4B means:

(i) C1-8 alkyl, C2-8 alkenyl or C2-8 alkynyl substituted with one to three groups selected from C1-balkyloxy or halogen;

(ii) phenyloxy or C3-7cycloalkyloxy;

(iii) furyl, furyloxy, thienyl, thienyloxy, naphthyl, naphthyloxy, phthalanyl or phthalanyloxy;

(iv) phenyl, phenyloxy, C3-7cycloalkyl or C3-7cycloalkyloxy substituted with one to three groups selected from the following group:

C1-6alkyl

C2-6 alkenyl,

C2-6 alkynyl,

C1-6 alkyloxy,

C1-6alkyloxy-C1-6alkyl,

C1-6alkyloxy-C1-6alkyloxy,

C2-6 alkenyloxy-C1-6 alkyl,

C1-6alkyl substituted with one to three hydroxy groups,

C1-6alkyl substituted with one to three halogen atoms,

C1-6alkylthio

C1-6alkylthio-C1-6alkyl,

C1-6alkylthio-C1-6alkyloxy,

C2-6alkenylthio-C1-6alkyl,

C1-6 alkylsulfonyl,

halogen,

trihalomethyl,

cyano

nitro

amino,

hydroxy

C3-7cycloalkyl,

C3-7cycloalkyloxy,

C3-7cycloalkyl-C1-6alkyl,

C3-7cycloalkyloxy-C1-6alkyl,

phenyl,

phenyloxy

phenyl-C1-6alkyl,

phenyl-C2-6 alkenyl,

phenyl-C2-6 alkynyl,

phenyloxy-C1-6alkyl,

phenyloxy-C2-6 alkenyl,

phenyloxy-C2-6 alkynyl,

furil

furyloxy

furyl-C1-6alkyl,

furyloxy-C1-6alkyl,

thienyl

thienyloxy

thienyl C1-6alkyl or

thienyloxy-C1-6alkyl,

(the phenyl, furyl, thienyl, or cycloalkyl mentioned above may be substituted by one to three groups selected from C1-6alkyl, C1-6alkyloxy, C1-6alkyloxy-C1-6alkyl, nitro, halogens, trihalomethyls, amino and hydroxy groups); or

(v) furyl, furyloxy, thienyl, thienyloxy, naphthyl, naphthyloxy, phthalanyl or phthalanyloxy substituted with one to three groups selected from the following groups:

C1-6alkyl

C2-6 alkenyl,

C2-6 alkynyl,

C1-6 alkyloxy,

C1-6alkyloxy-C1-6alkyl,

C1-6alkyloxy-C1-6alkyloxy,

C2-6alkenyloxy-C1-6alkyl,

C1-6alkyl substituted with one to three hydroxy groups,

C1-6alkyl substituted with one to three halogen atoms,

C1-6alkylthio

C1-6alkylthio-C1-6alkyl,

C1-6alkylthio-C1-6alkyloxy,

C2-6alkenylthio-C1-6alkyl,

C1-6 alkylsulfonyl,

halogen,

trihalomethyl,

cyano

nitro

amino,

hydroxy

C3-7cycloalkyl,

C3-7cycloalkyloxy,

C3-7cycloalkyl-C1-6alkyl,

C3-7cycloalkyloxy-C1-6alkyl,

phenyl,

phenyloxy

phenyl C1-6alkyl,

phenyl-C2-6 alkenyl,

phenyl-C2-6 alkynyl,

phenyloxy-C1-6alkyl,

phenyloxy-C2-6 alkenyl,

phenyloxy-C2-6 alkynyl,

furil

furyloxy

furyl-C1-6alkyl,

furyloxy-C1-6alkyl,

thienyl

thienyloxy

thienyl C1-6alkyl or

thienyloxy-C1-6alkyl.

(The phenyl, furyl, thienyl, or cycloalkyl mentioned above may be substituted with one to three groups selected from C1-6alkyl, C1-6alkyloxy, C1-6alkyloxy-C1-6alkyl, nitro, halogen, trihalomethyl, amino, and hydroxy groups),

R ^5B means hydrogen or C1-6 alkyl,

the symbol ----- means double or single bond.

Provided that when R ^2B is hydrogen, the C1-6 alkylene represented by R ^3B may be substituted with one hydroxy group and that its 8-epi-equilibrium compound is included)

Compounds of formula (IC)

where R ^1C means a hydroxy group, C 1-6 alkyloxy, or NR ^6C R ^7C (where each R ^6C and R ^7C independently mean hydrogen or C 1-4 alkyl),

R ^2C means oxo group, halogen or O-COR ^8C (in which R ^8C is C1-4 alkyl, phenyl, or phenyl (C1-4 alkyl)),

R ^3C means hydrogen or hydroxy group,

each R ^4Ca and R ^{4Cb is} independently hydrogen or ^{C1-4 alkyl} ,

R ^5C means phenyl substituted by the following groups:

(i) one to three groups selected from groups

C1-4alkyloxy-C1-4alkyl,

C2-4alkenyloxy-C1-4alkyl,

C2-4alkynyloxy-C1-4alkyl,

C3-7cycloalkyloxy-C1-4alkyl,

C3-7cycloalkyl (C1-4alkyloxy) -C1-4alkyl,

phenyloxy-C1-4alkyl,

phenyl-C-1-4alkyloxy-C1-4alkyl,

C1-4alkylthio-C1-4alkyl,

C2-4alkenylthio-C1-4alkyl,

C2-4alkynylthio-C1-4alkyl,

C3-7cycloalkylthio-C1-4alkyl,

C3-7cycloalkyl (C1-4alkylthio) -C1-4alkyl,

phenylthio-C1-4alkyl, or

phenyl-C1-4alkylthio-C1-4alkyl;

(ii) C1-4alkyloxy-C1-4alkyl and C1-4alkyl groups,

C1-4alkyloxy-C1-4alkyl and C1-4alkyloxy,

C1-4alkyloxy-C1-4alkyl and hydroxy,

C1-4alkyloxy-C1-4alkyl and halogen,

C1-4alkylthio-C1-4alkyl and C1-4alkyl

C1-4alkylthio-C1-4alkyl and C1-4alkyloxy,

C1-4alkylthio-C1-4alkyl and hydroxy, or

C1-4alkylthio-C1-4alkyl and halogen;

(iii) haloalkyls or hydroxy-C1-4alkyls; or

(iv) C1-4 alkyls and hydroxy groups;

the symbol ----- means a single or double bond.

Provided that when R ^2C is O-COR ^8C , the bond between atoms at 8 and 9 positions is double.

Compounds of Formula (ID)

where A ^D is C2-8 alkylene, C2-8 alkenylene, C1-4 alkylene-phenylene or C2-4 alkenylene-phenylene,

R ^1D means a hydroxy group, C 1-6 alkyloxy, C 1-6 alkyloxy-C 1-6 alkyloxy, HO-C 1-6 alkyloxy, or NR ^6D R ^7D (wherein each R ^6D and R ^{7D is} independently hydrogen or C 1-4 alkyl),

R ^2D is oxo, halogen, or R ^8D —COO— (where R ^8D is hydrogen, C 1-4 alkyl, phenyl, phenyl (C 1-4 alkyl), C 1-4 alkyloxy, HOOC-C 1-4 alkyl, C 1-4 alkyloxy-carbonyl-C 1 -4alkyl, HOOC-C2-4alkenyl or C1-4alkyloxycarbonyl-C2-4alkenyl),

R ^3D means hydrogen or hydroxy-group,

R ^4D means alkylene,

R ^5D means phenyl substituted by the following groups:

(i) one to three groups selected from groups

C1-4alkyloxy-C1-4alkyl,

C2-4alkenyloxy-C1-4alkyl,

C2-4alkynyloxy-C1-4alkyl,

C3-7cycloalkyloxy-C1-4alkyl,

C3-7cycloalkyl (C1-4alkyloxy) -C1-4alkyl,

phenyloxy-C1-4alkyl,

phenyl-C1-4alkyloxy-C1-4alkyl,

C1-4alkylthio-C1-4alkyl,

C2-4alkenylthio-C1-4alkyl,

C2-4alkynylthio-C1-4alkyl,

C3-7cycloalkylthio-C1-4alkyl,

C3-7cycloalkyl (C1-4alkylthio) -C1-4alkyl,

phenylthio-C1-4alkyl or

phenyl-C1-4alkylthio-C1-4alkyl;

(ii) C1-4alkyloxy-C1-4alkyl and C1-4alkyl groups,

C1-4alkyloxy-C1-4alkyl and C1-4alkyloxy,

C1-4alkyloxy-C1-4alkyl and hydroxy,

C1-4alkyloxy-C1-4alkyl and halogens,

C1-4alkylthio-C1-4alkyl and C1-4alkyl

C1-4alkylthio-C1-4alkyl and C1-4alkyloxy,

C1-4alkylthio-C1-4alkyl and hydroxy, or

C1-4alkylthio-C1-4alkyl and halogens;

(iii) halo C1-4 alkyl or hydroxy C1-4 alkyl; or

(iv) C1-4 alkyls and hydroxy groups;

the symbol --- means a single or double bond.

Provided that when R ^2D corresponds to the formula R9-COO-, R ^1D is C1-6alkyloxy, C1-6alkyloxy-C1-6alkyloxy or HO-C1-6alkyloxy, the atom bond at the 8th and 9th positions is a double bond.

More preferred compounds used as EP ₄ agonists in the present invention are as follows: (in parentheses under each formula is the published patent or application number and example number). For example, prostaglandin derivatives of the formula (IE)

(WOOO / 03980, example 3);

formulas (IF)

(EP-985663, example 2);

formulas (IG)

(PCT / JP99 / 04934, example 8-6);

formulas (IH)

(PCT / JP99 / 04934, example 2) and formula (IJ)

(PCT / JP99 / 04934, example 2-11)

or alkyl esters of these compounds, their non-toxic salts and their cyclodextrin clathrate compounds.

[Ester]

The compounds of the present invention depicted by formulas (IA) to (IJ) can be converted into esters by known methods. Conversion to the corresponding esters serves to increase the stability and absorbability of the compounds and is therefore applicable for the preparation of pharmaceutical preparations. An alkyl ester is preferred, a C1-4 alkyl ester is more preferred, and a methyl ester is most preferred.

[Salt]

The compounds of the present invention depicted by formulas (IA) to (IJ) can be converted into the corresponding salts by known methods. Non-toxic and water soluble salts are preferred. Suitable salts are, for example, the following:

salts of alkali metals (potassium, sodium, etc.), salts of alkaline earth metals (calcium, magnesium, etc.), ammonium salts, salts of pharmaceutically acceptable organic amines (tetramethylammonium, triethylamine, methylamine, dimethylamine, cyclopentylamine, benzylamine, phenethylamine , piperidine, monoethanolamine, diethanolamine, tri (hydroxymethyl) amine, lysine, arginine, n-methyl-d-glucamine, etc.). The compounds of the present invention depicted by formulas (IA) to (IJ) can be converted to the corresponding hydrates by known methods.

[Clathrates]

The compounds of the present invention depicted by formulas (IA) to (IJ) and their esters can be converted to the corresponding cyclodextrin clathrates by the method described in specification GB 1351238 or GB 1419221 using α-, β- or γ-cyclodextrins or a mixture thereof. Conversion to the corresponding cyclodextrin clathrates enhances the stability and solubility of compounds in water and is therefore useful when used for pharmaceutical preparations.

[Method for producing compounds of the present invention]

Each of the compounds of formulas (IA) to (IJ) and the method for its preparation are described in the following patent applications.

The compound of formula (IA): EP-855389;

The compound of formula (IB): EP-985663;

Compound of formula (IC): WO00 / 03980;

The compound of formula (ID): application No. PCT / JP99 / 04934;

Compound of formula (IE): WO00 / 03980;

The compound of formula (IF): EP-985663;

The compound of formula (IG): application No. PCT / JP99 / 04934;

The compound of formula (IH): application No. PCT / JP99 / 04934;

The compound of formula (IJ): application No. PCT / JP99 / 04934.

BEST MODE FOR CARRYING OUT THE INVENTION

The following reference examples and examples are intended to illustrate, but not limit the scope of the present invention.

Example 1: osteogenesis stimulating effect

Beagle / CSK dogs of 6 months of age were used (n = 3-6 in the group). Each α-cyclodextrin clathrate (containing the active ingredient in the form of α-cyclodextrin clathrate in an amount of 25% w / w) methyl ester (11α, 15β, 13E) -9-oxo-11,15-dihydroxy-16- (3- methoxymethylphenyl) -17,18,19,20-tetranor-5-thiaprost-13-enoic acid (methyl ester of the compound of formula (IE)), (11α, 15α, 13E) -9-hydroxy-11,15-dihydroxy-16 - (3-methoxymethylphenyl) -17,18,19,20-tetranor-3,7-dithiaprost-13-enoic acid (compounds of formula (IF)), (9β, 11α, 15α, 5Z, 13E) -9-chlorine -11,15-dihydroxy-16 (3-ethoxymethylphenyl) -17,18,19,20-tetranorprost-5,13-dienoic acid (compounds of formula (IG)), (11α, 15α, 13E) -9-ok si-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranorprost-13-enoic acid (methyl ester of the compound of formula (IH)) and (11α, 15α, 5Z, 13E) -9-hydroxy-11,15-dihydroxy-16- (3-ethoxymethylphenyl) -17,18,19,20-tetranorprost-5,13-dienoic acid (compounds of formula (1J)) were dissolved in physiological saline and administered to each animal in effective doses of 2, 5, 10 and 25 mcg / kg / day, continuously 24 hours a day for 4 weeks. Each test solution was injected into the femoral vein through a fixed catheter at a rate of 0.2 ml / kg / h. An equal volume of saline was administered to the animal from the control group. After completion of administration, each animal was sacrificed for autopsy.

(1) Determination of bone surface area

The separated femur was fixed with a buffered solution of 10% formalin and then cut into circular sections 10 mm thick perpendicular to the longitudinal axis at a distance of 25 mm on the central side of trochlea sulcus. Images of the bone surface from the side closest to the pineal gland were obtained using a camera connected to a computer at a constant distance, and then the resulting digital image was analyzed to determine the bone surface area. The results are shown in Table 1.

Table 1 Test solution Dose (mcg / kg / day) Bone Surface Area (mm ² ) Non-therapeutic control n = 6 0 68.93 ± 6.64 Compound (IE) according to the invention n = 6 2 77.96 ± 6.94 Compound (IE) according to the invention n = 6 10 190.69 ± 19.03 *** Compound (IE) according to the invention n = 4 25 502.76 ± 54.81 *** The compound (IF) according to the invention n = 3 5 365.29 ± 73.63 Compound (IG) according to the invention n = 3 10 140.10 ± 28.08 Compound (IG) according to the invention n = 3 25 289.38 ± 120.50 The compound (IH) according to the invention n = 3 5 362.99 ± 89.56 The compound (IJ) according to the invention n = 3 5 193.23 ± 11.32

***: p <0.001 (means standard deviation in the Dunnett test)

Discussion:

The group according to the invention, including compounds of formula (IE), at doses of 10 μg / kg / day or higher showed a 2.8-fold increase in bone surface area compared to the control group. A special, more than 7-fold increase in bone surface area was noted in this group at a dose of 25 μg / kg / day.

The group according to the invention, including compounds of formulas (IF), (IH) and (IJ) at a dose of 5 μg / kg / day, also showed a 2.8-5.3-fold increase in bone surface area compared to the control group. In addition, the group according to the invention, including compounds of formula (IG) at a dose of 25 μg / kg / day, also showed a 4.2-fold increase in bone surface area compared to the control group.

(2) Determination of bone density

After taking a side roentgenogram of a 1 cm thick sample used in the aforementioned paragraph (1), the obtained graphic input data were input into a computer to measure the radiation level per unit area at a constant thickness and determine the bone density (Microfocus X-ray System μFX-1000 (Fuji Photo Film Co., Ltd.)). The results are shown in Table 2.

table 2 Test solution Dose (mcg / kg / day) Bone Density (PSL / mm ² ) Non-therapeutic control n = 6 0 5.93 ± 0.72 Compound (IE) according to the invention n = 6 2 5.63 ± 0.70 Compound (IE) according to the invention n = 6 10 8.75 ± 1.19 *** Compound (IE) according to the invention n = 4 25 10.34 ± 1.58 ***

***: p <0.001 (means standard deviation in the Dunnett test)

Discussion:

The group according to the invention, including compounds of formula (IE) at doses of 10 and 25 μg / kg / day, showed a 1.5- and 1.7-fold increase in bone density compared with the control group, respectively. These results confirmed that the present compounds having an affinity for EP4 stimulate a significant increase in bone surface and bone density at a dose of 10 μg / kg / day or higher and have an osteogenesis-stimulating effect.

Example 2: Determination of affinity for EP ₄

The following experiments confirmed that the compounds of formulas (IA) - (IJ) can bind to the E ₄ receptor and that they have an agonistic effect on the EP ₄ receptor.

The agonistic effect of the compounds of formulas (IA) - (IJ) was determined using CHO cells that expressed the EP ₄ receptor (see FEBS Letters 364,339 (1995) with partial changes). It was further confirmed that all compounds cause an increase in cAMP and that they are EP ₄ agonists.

Example 3: osteogenesis stimulating effect

Experimental Method:

Female rats of the SD line of 11 weeks of age were used (n = 8, average body weight 271 g). An incision was made on the abdominal wall of the rat under anesthesia with pentobarbital for ovariectomy, and then a suture was made on the incision area. In the simulation group, animals were only incised and sutured without ovariectomy.

6 days after ovariectomy, the solution prepared by dissolving the α-cyclodextrin clathrate (containing the active ingredient in the form of α-cyclodextrin clathrate in an amount of 25% w / w) methyl ester (11α, 5α, 13E) -9-oxo- 11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranor-5-thiaprost-13-enoic acid (methyl ester of the compound of formula (IE)) in physiological saline, was injected subcutaneously into the back of the head each animal in a dosage of 30 mcg / 5 ml / kg every 8 hours, only three times a day for 2 months. In the control group and the simulation group, each animal was injected with saline. After completion of the experiment, all animals were killed to perform an autopsy. The density of the selected region of the left-sided femur was determined using equipment for the quantitative analysis of distal bone density (KhST-960A, Norland / Stolatec Co.). The results are shown in the following Table.

Table 3 Group Bone Density (mg / cm ³ ) Value ± St. from. Simulation group 404.1 ± 87.0 Control group 226.3 ± 38.9 ### The compound group (IE) according to the invention 794.0 ± 50.2 ***

###: p <0.001 versus simulation group

***: p <0.001 against the control group (in both: T-test)

Discussion:

From Table 3 it is clear that the group of compounds according to the invention showed a significant increase in bone density compared with the control group (without administration).

[Toxicity]

The toxicity of the compounds of formula (IE) of the present invention is very low, therefore, these compounds are safe for use as medications. For example, the maximum tolerated dose of a compound of formula (IE) i.v. the rat was 30 mg / kg body weight or more.

Industrial application

[Pharmaceutical Application]

The EP ₄ agonist and used in the present invention has a stimulating osteogenesis effect, so that it is applicable for the treatment and / or prevention of diseases associated with bone loss. In the present invention, a disease associated with bone loss means a disease that is observed in diseases associated with bone loss, accompanied by a decrease in bone density or degradation of bone tissue, etc. For example, such diseases include:

1) primary osteoporosis (for example, age-related osteoporosis, primary postmenopausal osteoporosis, osteoporosis due to ovariectomy, etc.);

2) secondary osteoporosis (e.g. glucocorticoid-induced osteoporosis; hyperthyroidism-induced osteoporosis; immobility-induced osteoporosis; heparin-induced osteoporosis; immunosuppression-induced osteoporosis; osteoporosis; osteoporosis; inflammatory disease; osteoporosis; inflammatory disease; d.);

3) bone diseases such as metastatic bone, hypercalcemia, Paget's disease, bone loss (alveolar bone rarefaction, jaw bone rarefaction, idiopathic bone rarefaction in children, etc.), osteonecrosis, etc.

In addition, the present invention includes the acceleration of osteogenesis after surgery for bone (for example, osteogenesis after fractures, osteogenesis after bone transplantation, osteogenesis after surgery for an artificial joint, osteogenesis after spinal fusion and osteogenesis after other operations for bone regeneration, etc. ), stimulating bone treatment and alternative treatment related to bone transplantation.

For the purpose described above, an EP ₄ agonist, its ester, its non-toxic salt, or its cyclodextrin clathrate used in the present invention can usually be administered locally by oral administration or parenteral administration. Conversion to a prodrug serves to reduce irritation and improve absorption capacity and stability, etc.

Doses to be administered are determined depending on age, weight, symptoms, desired therapeutic effect, route of administration, duration of treatment, etc. For an adult, human dosages / doses are generally between 1 μg and 100 mg when administered orally up to several times a day, and between 0.1 μg and 10 mg, preferably between 0.2 mg and 5 mg, with parenteral administration (preferably when administered through the skin by the appropriate subcutaneous method or into a vein) up to several times a day.

As mentioned above, the doses to be used depend on various conditions. Therefore, there are cases in which dosages lower or greater than the ranges set above can be used.

The compounds of the present invention can be administered as solid compositions, liquid compositions, or other compositions for oral administration, or as injections, skin ointments, a patch or suppositories, etc. for parenteral administration.

Solid compositions for oral administration include tablets, pills, capsules, dispersible powders, granules, etc.

Capsules include hard capsules and soft capsules.

Such solid compositions contain one or more active compounds mixed with at least one inert diluent, such as lactose, mannitol, glucose, hydroxypropyl cellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, magnesium metaaluminosilicate.

Compositions may also contain, in accordance with normal practice, additional substances other than inert diluents: for example, lubricants such as magnesium stearate, disintegrants such as calcium gluconate cellulose, and dissolution aids such as glutamic acid, aspartic acid . Tablets or pills can optionally be coated with a film of a gastro- or enteric substance, such as sugar, gelatin, hydroxypropyl cellulose or hydroxypropyl cellulose phthalate, etc., or coated with two or more films. Further, the coating may include localized within the capsule absorbent substances, such as gelatin.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, syrups, elixirs, etc. Such liquid compositions contain one or more active compounds in an inert diluent (s) commonly used in the art (for example, purified water, ethyl alcohol). In addition to inert diluents, such compositions may also contain adjuvants, such as wetting agents, suspending agents, sweeteners, flavoring agents, flavoring agents and preservatives.

Other compositions for oral administration include spray compositions that can be prepared by known methods and which contain one or more active compounds. Compositions for spraying may contain additional substances other than inert diluents: for example, stabilizers such as sodium hydrogen sulfate, stabilizers to maintain isotonia, isotonic buffer such as sodium chloride, sodium citrate, citric acid. To prepare such compositions for spraying, you can use the method described, for example, in US patent No. 2868691 or 3095355.

Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Aqueous solutions or suspensions contain distilled water for injection and physiological saline. Non-aqueous solutions or suspensions contain propylene glycol, polyethylene glycol, a vegetable oil such as olive oil, an alcohol such as ethyl alcohol, POLYSORBAT80 (registered trademark), etc. Such compositions may contain additional diluents: for example, preservatives, wetting agents, emulsifiers, dispersants, stabilizers, auxiliary agents, such as dissolution aids (e.g. glutamic acid, aspartic acid). They can be sterilized, for example, by filtration through a bacteria-retaining filter, by combined sterilization of adjuvants in the compositions or by irradiation. They can also be made in the form of sterile solid compositions which can be dissolved in sterilized water or some other sterile injectable diluent immediately before use.

Other compositions for parenteral administration include topical fluids, ointments, liniments, patches, suppositories, etc. that contain one or more active compounds and can be prepared by known methods.

An ointment, in addition to a base, such as white petrolatum, can include pH adjusters, a surfactant, preservatives, emulsifiers, dispersants, stabilizers, dissolution aids, etc.

Example 1, release form: lyophilized form

The following components were mixed in a conventional manner and the solution was sterilized in a conventional manner; 1 ml portions were placed in ampoules and lyophilized in a conventional manner to obtain 100 ampoules, each containing 0.2 mg of active ingredient.

- α-cyclodextrin methyl ester (11α, 15α, 13E) -9-oxo-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranor-5-thiaprost-13-eno acids - 80 mg (active ingredient: 20 mg);

- mannitol - 5 g;

- distilled water - 100 ml.

Example 2, release form: ointment

In a conventional manner, the following components were mixed and placed every 10 g of the mixture in a test tube to obtain 100 ointment tubes, where each contains 0.2 mg of the active ingredient in 1 g of the mixture.

- α-cyclodextrin methyl ester (11α, 15α, 13E) -9-oxo-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranor-5-thiaprost-13-eno acids -800 mg (active ingredient: 200 mg);

- white petroleum jelly - 1 kg.

Claims (18)

1. The use of an EP ₄ receptor agonist for the treatment and / or prevention of diseases associated with bone loss. 2. The use according to claim 1, where the disease associated with bone loss is primary osteoporosis. 3. The use according to claim 2, where the primary osteoporosis is senile osteoporosis, postmenopausal primary osteoporosis, osteoporosis due to ovariectomy. 4. The use of claim 1, wherein the disease associated with bone loss is secondary osteoporosis. 5. The use according to claim 4, where the secondary osteoporosis is osteoporosis induced by glucocorticoids, osteoporosis induced by hyperthyroidism, osteoporosis induced by immobility, osteoporosis induced by heparin, osteoporosis induced by immunosuppression, osteoporosis due to renal failure, osteoporosis or rheumatoid osteoporosis. 6. The use according to claim 1, where the disease associated with bone loss is metastasis to the bone, hypercalcemia, Paget's disease, bone loss, osteonecrosis. 7. The use according to claim 1 or 6, where diseases associated with bone loss are alveolar bone loss, rarefaction of the jaw bone, idiopathic bone loss in children. 8. The use according to claim 1, where the treatment of diseases associated with bone loss is osteogenesis after fracture, osteogenesis after bone transplantation, osteogenesis after surgery for an artificial joint, osteogenesis after spinal fusion, osteogenesis after bone regeneration and alternative bone transplantation treatment . 9. The use according to claim 1, where the EP ₄ agonist is a compound having a prostaglandin skeleton. 10. The use according to claim 9, where the compound having the skeleton of the prostaglandin is a compound of formula (IA)

where R ^1A represents hydroxy, C 1-4 alkyloxy, or NR ^6A R ^7A , where each R ^6A and R ^7A independently represents hydrogen or C 1-4 alkyl, R ^2A represents hydrogen or hydroxy, R ^3A represents: (i) C1-8 alkyl, C2-8 alkenyl, or C2-8 alkynyl; (ii) phenyl or C3-7 cycloalkyl; (iii) C1-8 alkyl, C2-8 alkenyl or C2-8 alkynyl substituted with phenyl or C3-7 cycloalkyl (provided that when R ^2A is hydrogen, alkyl, alkenyl and alkynyl in (i) and (ii) can be substituted with one hydroxy, the symbol ----- means a double or single bond, provided that its 8-epirequilibrium compound is included), or its non-toxic salt, or its cyclodextrin clathrate. 11. The use according to claim 9, where the compound having a prostaglandin skeleton is a compound of formula (IB)

where R ^1B represents hydroxy, C1-6 alkyloxy, or NR ^6B R ^7B , where each R ^6B and R ^7B independently represents hydrogen or C1-6 alkyl, R ^2B represents hydrogen or hydroxy, R ^3B represents a single bond or C1-6 alkylene, R ^4B represents: (i) C1-8 alkyl, C2-8 alkenyl or C2-8 alkynyl substituted with one to three groups selected from C1-6 alkyloxy or halogen; (ii) phenyloxy or C3-7 cycloalkyloxy; (iii) furyl, furyloxy, thienyl, thienyloxy, naphthyl, naphthyloxy, phthalanyl or phthalanyloxy; (iv) phenyl, phenyloxy, C3-7 cycloalkyl or C3-7 cycloalkyloxy substituted with one to three groups selected from the following group: C1-6 alkyl C2-6 alkenyl, C2-6 alkynyl, C1-6 alkyloxy C1-6 alkyloxy-C1-6 alkyl, C1-6 alkyloxy-C1-6 alkyloxy, C2-6 alkenyloxy-C1-6 alkyl, C1-6 alkyl substituted with one to three hydroxy, C1-6 alkyl substituted with one to three halogen atoms, C1-6 alkylthio, C1-6 alkylthio-C1-6 alkyl, C1-6 alkylthio-C1-6 alkyloxy, C2-6 alkenylthio-C1-6 alkyl, C1-6 alkylsulfonyl, halogen, trihalomethyl, cyano nitro amino, hydroxy C3-7 cycloalkyl, C3-7 cycloalkyloxy, C3-7 cycloalkyl-C1-6 alkyl, C3-7 cycloalkyloxy-C1-6 alkyl, phenyl, phenyloxy phenyl C1-6 alkyl, phenyl-C2-6 alkenyl, phenyl-C2-6 alkynyl, phenyloxy-C1-6 alkyl, phenyloxy-C2-6 alkenyl, phenyloxy-C2-6 alkynyl, furil furyloxy furyl C1-6 alkyl, furyloxy-C1-6 alkyl, thienyl thienyloxy thienyl C1-6 alkyl or thienyloxy-C1-6 alkyl, (the phenyl, furyl, thienyl, or cycloalkyl mentioned above may be substituted by one to three groups selected from C1-6 alkyl, C1-6 alkyloxy, C1-6 alkyloxy-C1-6 alkyl, nitro, halogen, trihalomethyl, amino and hydroxy) ; or (v) furyl, furyloxy, thienyl, thienyloxy, naphthyl, naphthyloxy, phthalanyl or phthalanyloxy substituted with one to three groups selected from the following groups: C1-6 alkyl C2-6 alkenyl, C2-6 alkynyl, C1-6 alkyloxy C1-6 alkyloxy-C1-6 alkyl, C1-6 alkyloxy-C1-6 alkyloxy, C2-6 alkenyloxy-C1-6 alkyl, C1-6 alkyl substituted with one to three hydroxy, C1-6 alkyl substituted with one to three halogen atoms, C1-6 alkylthio, C1-6 alkylthio-C1-6 alkyl, C1-6 alkylthio-C1-6 alkyloxy, C2-6 alkenylthio-C1-6 alkyl, C1-6 alkylsulfonyl, halogen, trihalomethyl, cyano nitro amino, hydroxy C3-7 cycloalkyl, C3-7 cycloalkyloxy, C3-7 cycloalkyl-C1-6 alkyl, C3-7 cycloalkyloxy-C1-6 alkyl, phenyl, phenyloxy phenyl C1-6 alkyl, phenyl-C2-6 alkenyl, phenyl-C2-6 alkynyl, phenyloxy-C1-6 alkyl, phenyloxy-C2-6 alkenyl, phenyloxy-C2-6 alkynyl, furil furyloxy furyl C1-6 alkyl, furyloxy-C1-6 alkyl, thienyl thienyloxy thienyl C1-6 alkyl or thienyloxy-C1-6 alkyl (the phenyl, furyl, thienyl, or cycloalkyl mentioned above may be substituted by one to three groups selected from C1-6 alkyl, C1-6 alkyloxy, C1-6 alkyloxy-C1-6 alkyl, nitro, halogen, trihalogenomethyl, amino and hydroxy) , R ^5B represents hydrogen or C1-6 alkyl, the symbol ----- represents a double or single bond, (provided that when R ^2B is hydrogen, the C1-6 alkylene represented by R ^3B can be substituted with one hydroxy and that its 8-epirequilibrium compound is included), or its non-toxic salt, or its cyclodextrin clathrate. 12. The use according to claim 9, where the compound having the skeleton of a prostaglandin is a compound of the formula (1C)

where R ^1C represents hydroxy, C1-6 alkyloxy or NR ^6C R ^7C (where R ^6C and R ^7C independently represent hydrogen or C1-4 alkyl), R ^2C represents oxo, halogen or O-COR ^8C , where R ^8C represents C1-4 alkyl, phenyl or phenyl (C1-4 alkyl), R ^3C represents hydrogen or hydroxy, R ^4Ca and R ^4Cb independently represent hydrogen or C1-4 alkyl, R ^5C represents phenyl substituted with the following groups: (i) one to three groups selected from groups C1-4 alkyloxy-C1-4 alkyl, C2-4 alkenyloxy-C1-4 alkyl, C2-4 alkynyloxy-C1-4 alkyl, C3-7 cycloalkyloxy-C1-4 alkyl, C3-7 cycloalkyl (C1-4 alkyloxy) C1-4 alkyl, phenyloxy-C1-4 alkyl, phenyl-C1-4 alkyloxy-C 1-4 alkyl, C1-4 alkylthio-C1-4 alkyl, C2-4 alkenylthio-C1-4 alkyl, C2-4 alkynylthio-C1-4 alkyl, C3-7 cycloalkylthio-C1-4 alkyl, C3-7 cycloalkyl (C1-4 alkylthio) C1-4 alkyl, phenylthio-C1-4 alkyl or phenyl C1-4 alkylthio C1-4 alkyl; (ii) C1-4 alkyloxy C1-4 alkyl and C1-4 alkyl, C1-4 alkyloxy-C1-4 alkyl and C1-4 alkyloxy, C1-4 alkyloxy-C1-4 alkyl and hydroxy, C1-4 alkyloxy-C1-4 alkyl and halogen, C1-4 alkylthio-C1-4 alkyl and C1-4 alkyl, C1-4 alkylthio-C1-4 alkyl and C1-4 alkyloxy, C1-4 alkylthio-C1-4 alkyl and hydroxy or C1-4 alkylthio-C1-4 alkyl and halogen; (iii) haloalkyl or hydroxy-C1-4 alkyl; or (iv) C1-4 alkyl and hydroxy; the symbol ---- means a single or double bond, (provided that when R ^2C is O-COR ^8C , the bond between the atoms in positions 8 and 9 is double) or its non-toxic salt or its cyclodextrin clathrate. 13. The use according to claim 9, where the compound having the skeleton of a prostaglandin means a compound of the formula (ID)

where A ^D is C2-8 alkylene, C2-8 alkenylene, C1-4 alkylene phenylene or C2-4 alkenylene phenyl, R ^1D is hydroxy, C1-6 alkyloxy, C1-6 alkyloxy-C1-6 alkyloxy, HO-C1- 6 alkyloxy or NR ^6D R ^7D (where R ^6D and R ^{7D are} independently hydrogen or C1-4 alkyl), R ^2D is oxo, halogen, or R ^8D -COO- (where R ^8D is hydrogen, C1-4 alkyl, phenyl, phenyl (C1-4 alkyl), C1-4 alkyloxy, HOOC-C1-4 alkyl, C1-4 alkyloxycarbonyl-C1-4 alkyl, HOOC-C2-4 alkenyl or C1-4 alkyloxycarbonyl-C2-4 alkenyl ), R ^3D is hydrogen or hydroxy, R ^4D represents alkylene, R ^5D represents phenyl substituted with the following groups: (i) one to three groups selected from the groups: C1-4 alkyloxy-C1-4 alkyl, C2-4 alkenyloxy-C1-4 alkyl, C2-4 alkynyloxy-C1-4 alkyl, C3-7 cycloalkyloxy-C1-4 alkyl, C3-7 cycloalkyl (C1-4 alkyloxy) -C1-4 alkyl, phenyloxy-C1-4 alkyl, phenyl C1-4 alkyloxy C1-4 alkyl, C1-4 alkylthio-C1-4 alkyl, C2-4 alkenylthio-C1-4 alkyl, C2-4 alkynylthio-C1-4 alkyl, C3-7 cycloalkylthio-C1-4 alkyl, C3-7 cycloalkyl (C1-4 alkylthio) -C1-4 alkyl, phenylthio-C1-4 alkyl or phenyl C1-4 alkylthio C1-4 alkyl; (ii) C1-4 alkyloxy-C1-4 alkyl and C1-4 alkyl, C1-4 alkyloxy-C1-4 alkyl and C1-4 alkyloxy, C1-4 alkyloxy-C1-4 alkyl and hydroxy, C1-4 alkyloxy-C1-4 alkyl and halogen, C1-4 alkylthio-C1-4 alkyl and C1-4 alkyl, C1-4 alkylthio-C1-4 alkyl and C1-4 alkyloxy, C1-4 alkylthio-C1-4 alkyl and hydroxy, or C1-4 alkylthio-C1-4 alkyl and halogen; (iii) halo C1-4 alkyl or hydroxy C1-4 alkyl, or (iv) C 1-4 alkyl and hydroxy; the symbol ---- means a single or double bond, (provided that when R ^2D corresponds to the formula R ^8D —COO—, R ^1D is C1-6 alkyloxy, C1-6 alkyloxy-C1-6 alkyloxy or HO-C1-6 alkyloxy, the bond between the atoms in positions 8 and 9 is a double bond), or a non-toxic salt thereof or a cyclodextrin clathrate thereof. 14. The use according to claim 9, where the EP ₄ agonist is (11α, 15α, 13E) -9-oxo-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranor- 5-thiaprost-13-enoic acid, (11α, 15α, 13E) -9-oxo-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranor-3,7-dithiaprost-13-enoic acid, (9β, 11α, 15α, 5Z, 13E) -9-chloro-11,15-dihydroxy-16- (3-ethoxymethylphenyl ) -17,18,19,20-tetranorprost-5,13-dienoic acid, (11α, 15α, 13E) -9-oxo-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranorprost-13-enoic acid or (11α, 15α, 5Z, 13E) -9-oxo-11,15-dihydroxy-16- (3-ethoxymethylphenyl) -17,18,19,20-tetranorprost-5,13-dienoic acid or their non-toxic salts, methyl esters or α-cyclodextrin clathrates. 15. The use of the compounds of formula (I)

(11α, 15α, 13E) -9-oxo-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranor-5-thiaprost-13-enoic acid or its non-toxic salt, complex methyl ester or α-cyclodextrin clathrate for the preparation of a medicament for the prevention and / or treatment of diseases associated with bone loss. 16. The use of (11α, 15α, 13E) -9-oxo-11,15-dihydroxy-16- (3-methoxymethylphenyl) -17,18,19,20-tetranor-5-thiaprost-13 methyl ester α-cyclodextrin clathrate -enoic acid for the preparation of a medicament for the prevention and / or treatment of diseases associated with bone loss. Priority on points: 11.24.1999 according to claims 1-8, 10-16; 11/22/2000 according to claim 9.