WO1992000744A1

WO1992000744A1 - THE USE OF INOSITOLMONOPHOSPHATE FOR THE PREPARING OF A MEDICAMENT EFFECTIVE AS A NEUROPEPTIDE η-ANTAGONIST

Info

Publication number: WO1992000744A1
Application number: PCT/SE1991/000466
Authority: WO
Inventors: Lars Edvinsson; Bo Fallgren
Original assignee: Perstorp Ab
Priority date: 1990-07-04
Filing date: 1991-06-27
Publication date: 1992-01-23
Also published as: AU8225391A; SE9002342D0; SE467340B

Abstract

The present invention relates to the use of a compound of formula (I): C6H6(OH)5 (OPO3H2), for the preparing of a medicament for preventing, alleviating or combatting different conditions in mammals including man.

Description

THE USE OF INOSITOLMONOPHOSPHATE FOR THE PREPARING OF A MEDICAMENT EFFECTIVE AS A NEUROPEPTIDE Y-ANTAGONIST

The present invention relates to the use of a compound having the formula (I): C₆H₆ (OH)₅ (OPO₃H₂) (I) in acid or salt form for the preparing of a medicament for preventing, alleviating or combatting different conditions in mammals including man.

Inositolphosphates are a group of compounds occurring in different plants, as constituents in cell membranes, etc. The hexaphosphoiylated myo-inositol, phytic acid, is present in different grains at varying amounts. It is known that lower inositol phosphate derivatives are formed when the grains are

germinated. The end products are inositol and inorganic phosphate.

Some other inositolphosphates exist in the blood of birds, especially the inositol pentaphosphate. This compound regulates the affinity for oxygen to hemoglobin in the eiythrocytes.

Lower inositolphosphates linked to glycerol moities are constituents of plasma membranes in cells.

This is valid for very specific isomers of inositoltrisphosphates, inositolbisphosphates and inositolmonophosphates. These inositolphosphates are released within the cells and regulate different metabolic pathways. From C.A., vol. 33 (1939), abstr. no. 7351, no 3/4, the use of phosphates including inositolphosphates as an antirachitic diet has been reported. No reference is made to specific inositolphosphates.

The US-patent no. 4.473.563 discloses the extra corporal treatment of erythrocytes to incorporate therein inositolphosphates to improve the oxygen supply. Then erythrocytes are separated from the blood, which has been pumped out of the body for that purpose. After complicated treatment of erythrocytes the latter are reintroduced into the blood. There is no disclosure of administering inositolphosphates directly to the body.

In the US-patent no. 2.723.938 the use of inositolphosphates is disclosed for stabilizing dispersions of an aqueous suspension of penicillin. This ensures that brief simple manual shaking will restore a state of complete and uniform dispersion of the penicillin after prolonged storage.

From the European Patent no. 179.439 a pharmaceutical composition comprising as a pharmaceutically active ingredient at least one isomer of inositoltrisphosphate is known. In said patent the effect of this pharmaceutical composition is shown for different areas such as platelet aggregation.

Neuropeptide Y (NPY) is a peptide present in the central and peripheral nervous system. The peptide co-exists with noradrenaline in many neurons and as a neurotransmittor per se or synergistically together with noradrenaline. NPY-containing fibres are numerous around arteries especially for arteries in the heart but also arteries in the respiratory tract, the gastrointestinal tract and genitourinary tract. NPY is also present in the cerebrum area with effects on e.g. blood pressure and release of different hormones. NPY belongs to the pancreatic polypeptide (PP) family which includes also for example peptide YY (PYY).

In man and animals abnormal NPY-levels are associated with the following diseases or conditions:

Diseases pertaining to the vascular system, the heart, blood vessels and the peripheral circulation such as vasospasm, angina, hemorrhage, high blood pressure, cardiac hypertrophy, myocardial infarction;

Diseases pertaining to the renal system such as abnormal renal conditions like impaired flow of fluid and renal failure; Cerebral diseases and diseases related to the central nervous system such as stroke and conditions associated with stroke, cerebral vasospasm and hemorrhage and depression;

Abnormal drink and food intake such as obesity;

Diabetes or complications of diabetes;

Inflammatory diseases such as arthritis:

Respiratory diseases such as asthma;

Regulation of hormone release from e.g. the pituitary.

The invention relates to the use of a compound of formula (I) for the preparing of a medicament effective as an NPY-antagonist.

In other preferred embodiments of the invention the medicament is intended to be used for preventing, alleviating or combatting diseases pertaining to the heart, the blood vessels, the renal sytem and brain injuries.

The invention relates to the use of at least one compound of formula (I) for the preparing of a medicament for preventing, alleviating or combatting for example the following conditions:

Diseases pertaining to the heart, blood bessels or the renal system such as vasospasm, heart failure, cardiac hypertrophy, increased blood pressure, angina, myocardial infarction or conditions related to myocardial infarction, sudden cardiac death, arrythmia. peripheral vascular diseases and abnormal renal conditions such as impaired flow of fluid, abnormal mass transport and renal failure;

Conditions related to increased sympathetic nerve activity for example during or after coronary artery surgery and operations and surgery in the gastrointestinal tract;

Brain injuries and diseases related to the central nervous system such as infarctions in the brain like stroke and conditions related to stroke,

vasospasm and hemorrhage in the brain, depression and dementia;

Diseases related to abnormal gastrointestinal motility and secretion such as different forms of ileus and Crohn's disease;

Abnormal drink and food intake such as obesity, anorexia and metabolic disorders;

Respiratory diseases such as asthma or conditions related to asthma and bronchoconstriction; Diseases related to abnormal hormone release for example leutinizing hormone, growth hormone and prolactin.

Abnormal NPY-levels or abnormal NPY-induced effects are associated with the above mentioned disorders and the compound of formula (I) is acting primarily as an NFY-antagonist or as an antagonist against NPY-induced effects. Peptides which are related to NPY such as PYY and other peptides belonging to the pancreatic polypeptide-fold (PP-fold) family of regulatory peptides also occur abnormally in some of the conditions mentioned above. Under certain circurmstances the effect of the compound of formula (I) could also be directed against properties or effects induced by these peptides. In some conditions the effects induced by for example NPY could be mediated by substances as endothelin and alike. However, the compound might also be effective against the above mentioned conditions by other mechanisms and pathways not related to the described peptides.

The different isomers of the compound of formula (I) are inositol-monophosphates. In one preferred embodiment of the invention the inositol-isomer is myoinositol but also other inositols, i.e. allo-inositol, epi-inositol, chiro-inositol, muco-inositol, cis-inositol, neo-inositol, scyllo-inositol are to be contemplated to be in the scope of the invention.

The different compound of formula (I) can be produced by degradation of inositolphosphates containing more than one phosphate groups/inositol molecule followed by separation and purification. The different isomers could also be produced by synthetic methods, chemically or enzymatically starting with e.g. inositol and a phosphorus source. Furthermore the use of micro-biological methods and hybrid-DNA-techniques are also suitable.

The compound of formula (I) can also exist in salt form or as a mixture of salt and acid. Preferably the salt is a salt of sodium, potassium, calcium, magnesium or zinc or a mixture of two or more thereof.

It is suitable that the medicament used according to the invention exists in unit dosage form. Tablets, granules or capsules are suitable administration forms for such unit dosage. These forms can easily be modified in order to provide controlled absorption of the compound in the intestine. A common pharmaceutically acceptable additive, excipient and carrier can be Included in the medicament. The tablets or granules can also contain a disintegrate, which causes the tablets or granules to disintegrate in the intestine. Other administration forms are e.g. slow release of transdermal administration, nasal, rectal, intra-articular. topical, intraperitoneal and subcutaneous administration. In certain cases, especially in acute situations, it is preferable to use the unit dosage in the form of a solution for intravenous administration.

The medicament according to the present invention can preferably also contain another pharmaceutically active ingredient in addition to the compound of formula (I). The amount of the compound of formula (I) is in the range of 5 % to 95 % especially 15 % to 95 % by weight of the pharmaceutically active ingredients.

The medicament is preferably containing 0.01 g to 1.5 g of the compound of formula (I).

For administration to human patients appropriate dosages can routinely be determined by those skilled in this art by the extension of the results obtained in animals at various dosages.

The preferred dosage for human falls within the range of 0.1 mg to 1.000 mg. especially 0.1 mg to 200 mg of the compound of formula (I)/day/kg body weight.

The compound of formual (I) has a cyclic form. In order to better describe specific compounds of formula (I) used in preferred embodiments of the inventions the following formula (II) is used:

In preferred embodiments of the invention the phsophate group is axial to the ring structure.

In preferred embodiments of the invention the compounds of formula (II) are selected from the group where:

R₃ is phosphate, R₁, R₅, R₈, R₉, R₁₂ are hydroxyl and R₂, R₄, R₆, R₇, R₁₀, R_{1 1} are hydrogen.

Rx is phosphate, R₃, R₅, R₇, R₁₀, R₁₁ are hydroxyl and R₂, R₄, R₆, R₈, R₉, R₁₂ are hydrogen.

R₅ is phosphate, R₁, R₃, R₇, R₁₀ R₁₁ are hydroxyl and R₂, R₄, R₆, R₈, R₉, R₁₂ are hydrogen.

Rx is phosphate, R₃, R₅, R₇, R₉, R₁₂ are hydroxyl and R₂, R₄, R₅, R₈, R₁₀,

R_{1 1} are hydrogen.

R₉ is phosphate, R_1, R₃, R₆, R₇, R₁₂ are hydroxyl and R₂, R₄, R₅, R₈, R₁₀,

R₁₁ are hydrogen.

R₁₂ is phosphate, R₁, R₃, R₆, R₇, R₉ are hydroxyl and R₂, R₄, R₅, R₈, R₁₀,

R_{1 1} are hydrogen.

R3 is phosphate, R₁, R₅, R₈, R₁₀, R₁₂ are hydroxyl and R₂, R₄, R₆, R₇, R₉, R₁₁ are hydrogen.

R₁₀ is phosphate, R₁, R₃, R₅, R₈, R₁₂ are hydroxyl and R₂, R₄, R₆, R₇, R₉, R₁₁ are hydrogen. R₃ is phosphate, R₁, R₆, R₈, R₉, R₁₂ are hydroxyl and R₂, R₄, R₅, R₇, R₁₀, R_{1 1} are hydrogen.

R₆ is phosphate. R₁, R₃, R₈, R₉, R₁₂ are hydroxyl and R₂, R₄, R₅, R₇, R₁₀, R_{1 1} are hydrogen.

R₁ is phosphate, R₃, R₆, R₇, R₁₀, R₁₂ are hydroxyl and R₂, R₄, R₅, R₈, R₉, R_{1 1} are hydrogen.

R₁₂ is phosphate. R₁, R₃, R₆, R₇, R₁₀ are hydroxyl and R₂, R₄, R₅, R₈, R₉, R_{1 1} are hydrogen.

R₁ is phosphate, R₃, R₅, R₈, R₁₀, R_{1 1} are hydroxyl and R₂, R₄, R₆, R₇, R₉,

R₁₂ are hydrogen.

R₅ is phosphate, R₁, R₃, R₈, R₁₀, R_{1 1} are hydroxyl and R₂, R₄, R₆, R₇, R₉,

R₁₂ are hydrogen.

R₈ is phosphate, R₁, R₃, R₅, R₁₀, R_{1 1} are hydroxyl and R₂, R₄, R₆, R₇, R₉,

R₁₂ are hydrogen.

R₁ is phosphate, R₃, R₅, R₇, R₉, R_{1 1} are hydroxyl and R₂, R₄, R₆, R₈, R₁₀,

R₁₂ are hydrogen.

R₅ is phosphate, R₁, R₃, R₇, R₉, R_{1 1} are hydroxyl and R₂, R₄, R₅, R₈, R₁₀,

R₁₂ are hydrogen.

R₉ is phosphate, R₁, R₃, R₅, R₇, R₁₁ are hydroxyl and R₂, R₄, R₆, R₈, R₁₀

R₁₂ are hydrogen.

One of R₁, R₄, R₅, R₈, R₉, R₁₂ is phosphate, the remaining hydroxyl and R₂, R₃, R₆, R₇, R₁₀, R_{1 1} are hydrogen.

The invention is further explained below in connection with embodiment examples of which Example 1 shows the production of myo-inositolmonophosphate (IP₁).

Example 2 teaches the production of a solution of a sodium salt of IP₁.

Example 3 illustrates the counteractive effect of IP₁ against NPY-induced contraction. EXAMPLE 1

A 1.2 gram quantity of the sodium salt of myo-inositol-1,2,3-trisphosphate was dissolved in 900 ml of an aqueous solution of sodium acetate buffer pH 4.6.

50 gram of baker's yeast was added with stirring and incubation was continued at 45°C for 90 mins. The hydrolysis was then stopped by adding 30 ml of ammonia to pH 12. The suspension was centrifuged and the supernatant was collected.

500 ml of the supernatant was passed through an ion-exchange column (Dowex 1, chloride form, 25 mm × 250 mm) and eluted with a linear gradient of sodium chloride (0 to 0.5 M NaCl) at pH 12.

Aliquots of eluted fractions were completely hydrolyzed in order to determine the contents of phosphorus and inositol. The peaks correspond to different inositolphosphates and the largest peak with a phosphorus/inositol ratio of one was further investigated with NMR.

Structural determination showed the product to be myo-inositol-2-monophosphate.

EXAMPLE 2

0.5 gram of a sodium salt of myo-inositol-2-monophosphate and 0.77 gram NaCl were dissolved in 98.73 ml of water for injection to form a solution suitable for injection into a person or an animal.

EXAMPLE 3

The basilar arteries from guinea pigs were dissected free and were then cut into cylindrical segments (2-3 mm long; 0.2 - 0.3 mm outer diameter) before the experiments started. Each segment was mounted on two L-shaped metal prongs, where one was connected to a force displacement transducer to continuous recording of the tension and the other to a displacement device.

The mounted specimens were immersed in tissue baths (37°C) containing a buffer solution of the following composition (mM) and pH 7.4: sodium chloride, 119; sodium hydrogen carbonate, 15; potassium chloride 4.6;

magnesium chloride, 1.2; sodium dihydrogen phosphate, 1.2; calcium chloride, 1.5; glucose, 11. A tension of 2 mM was applied to the arterial segments and the contractile capacity of each vessel segment was examined by exposure to a potassiumrich (60 mM) buffer solution. The maximum contraction obtained in this way was given the value 100 %.

The NPY-induced contraction was measured after dissolving NPY in the above mentioned buffer to a final concentration of 0.3 μM NPY.

The antagonistic properties of myo-inositol-2-monophosphate (IP₁) in this system were evaluated by incubation with different concentrations of IP₁ twenty minutes before the addition of NPY to the tissue bath. The following results were obtained:

Compound Contraction (%)

NPY, no IP₁ 88

NPY + IP₁ (10^-8M) 47

NPY + IP₁ (10^-7M) 28

NPY + IP₁ (10^-6M) 18

IP₁ demonstrates a significant decrease of the vasoconstriction induced by

NPY also in very low concentrations. These effects imply a very potent inhibition of vessel constriction, which is a dominant component in diseases and conditions such as vasospasm, angina, stroke and hypertension.

Claims

1. The use of a compound having the formula (I) C₆H₆ (OH)₅ (OPO₃H₂)

(I) in acid form for the preparing of a medicament effective as a neuropeptide Y- (NPY-) antagonist.

2. The use of a compound of formula (I) for the preparing of a medicament for preventing, alleviating or combatting brain injuries and diseases related to the central nervous system (CNS).

3. The use according to claim 2 wherein the brain injuries are related to stroke, vasospasm and hemorrhage.

4. The use according to claim 2 wherein the CNS diseases are related to depression and dementia.

5. The use of a compound of formula (I) for the preparing of a medicament for preventing, alleviating or combatting diseases pertaining to the heart, blood vessels and the renal system.

6. The use according to claim 5 wherein the diseases pertaining to the heart, blood vessels and renal system are related to vasospasm, heart failure, increased blood pressure, angina and myocardial infarction.

7. The use of a compound of formula (I) for the preparing of a medicament for preventing, alleviating or combatting diseases related to abnormal food and drink intake.

8. The use of a compound of formula (I) for the preparing of a medicament for preventing, alleviating or combatting diseases related to abnormal gastrointestinal motility and secretion.

9. The use according to anyone of claims 1-8 wherein said compound is in salt form.

10 . The use according to claim 9 wherein said salt of the compound of

formula (I) is a salt of sodium, potassium, calcium or zinc.

11. The use according to anyone of claims 1-10 wherein the medicament is in unit dosage forms comprising tablets, granules and solutions.

12. The use according to anyone of claims 1-11 wherein said compound is myo-inositol-2-monophosphate.