The present invention relates to an inositol phosphate containing pharmaceutical composition with improved bio¬ availability of inositol phosphate and the use of at least one aliphatic ester or ether compound with the formula I or II below for the preparing of an inositol phosphate containing medicament with improved bioavailabi¬ lity of inositol phosphate at non-parenteral administra¬ tion.

Hydrophilic and ionized drugs most often encounter rather poor penetration of the epithelial barriers to the capillaries of the portal circulation as most of this type of substances are transported by passive diffusion. Inositol phosphates belong to this group.

Normally it is the unionized fraction of the drug that partition across the lipid membranes and this fraction is most often small over the pH-range encountered in the gastrointestinal tract.

It is known from the literature, for example M. Sekine et al., (1984), J.Pharm. Dyn. 7, 856-863 that a combination of specific glyceryl-containing compounds and some anti¬ biotics render an improved absorption of the antibiotic after rectal administration. However the amount of the glyceryl containing compound is very high resulting in an inconvienient formulation with potential side effects.

Nothing has been described in the literature about the use of an aliphatic ester or ether compound with formula I or II together with inositol phosphate for improvement of the bioavailability of the inositol phosphate.

At oral administration the properties of inositol phosphates per se result in limitations in respect of the uptake of the compounds from the intestine. In order to optimize the effect of these substances in the body it is desirable that as large a portion as possible of the added amount can be utilized effectively. Thereby the added amount can be reduced which is advantageous for the patient for example when the drug D-myo-inositol-1,2,6- trisphosphate (IP-.) is used.

According to the present invention it has now quite surprisingly been possible to meet the above desire and bring about an inositol phosphate containing pharma¬ ceutical composition which comprises an aliphatic ester or ether compound with formula I or II below for improve¬ ment of the bioavailability of the inositol phosphate in mammals including man at non-parenteral administration.

The expression bioavailability stands for the measurement of how large a portion of an administered drug that occurs in the blood stream when the way of administration of the drug is non-parenteral. The term thus shows the amount of the drug that has been able to penetrate membrane barriers after for example oral administration, topical administration or intraperitoneal administration.

The aliphatic ester or ether compound with 3 to 18 carbon atoms can be defined by the formula I.

CH_OR CHOR CH ₂ OR_ wherein R R and/or R_ independently are

i) hydrogen; ii) a straight or branched alkyl with 1 to 15 carbon atoms such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl and pentadecyl; iii) a cycloalkyl with 3 to 15 carbon atoms such as cyclopropyl, cyclopentyl and/or cyclohexyl; iv) an alkenyl with 2 to 14 carbon atoms such as vinyl, allyl, propenyl, octadienyl, decenyl, dodecenyl and/or tetradecenyl; v) a cycloalkenyl with 5 to 14 carbon atoms such as cyclopentenyl, cyclopentadienyl, cyclohexenyl and/or cyclohexadienyl; vi) a straight or branched acyl radical of a saturated aliphatic carboxylic acid with 1 to 15 carbon atoms such as formyl, acetyl, propionyl, butyryl, iso- butyryl, valeryl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, pivaloyl, lauroyl and/or myristoyl; vii) a straight or branched acyl radical of an unsatu¬ rated aliphatic carboxylic acid with 1 to 15 carbon atoms such as acryloyl, propiolyl, oleoyl, aleoyl, fumaroyl and/or citraconoyl; viii)an acyl radical of a saturated cyclic carboxylic acid such as cyclopentanoyl or ix) an acyl radical of an unsaturated cyclic carboxylic acid such as furoyl.

The above groups (ii) to (ix) are unsubstituted or substituted with hydroxy; oxo; alkoxy; carboxy; esterified carboxy; amine, substituted amine; acyloxy or acylamine.

In one preferred embodiment of the invention where a compound with formula I is used R and R are

i) hydrogen or ii) a straight or branched alkyl selected from the group of methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl and/or octyl

and R is

i) a straight or branched alkyl selected from the group of butyl, hexyl, octyl and/or decyl or ii) a straight or branched acyl radical of a saturated aliphatic carboxylic acid with 1 to 15 carbon atoms such as butanoyl, hexanoyl, octanoyl and/or decanoyl.

Preferred compounds in this group are 1,2-dihydroxy- propylbutanoate, 2,3-dihydroxypropylbutanoate, 1,3-di-hydroxypropylbutanoate, 1,2-dihydroxypropy1- hexanoate, 2,3-dihydroxypropylhexanoate, 1,3-di- hydroxypropylhexanoate, 1,2-dihydroxypropyloxtanoate, 1,3-dihydroxypropyloctanoate, 2,3-dihydroxypropy1- octanoate, 1,2-dihydroxypropyldecanoate, 1,3-dihydroxy¬ propyloctanoate and 2,3-dihydroxypropyldecanoate.

The aliphatic ester or ether compound can also have the following formula:

a) CH ₂ OR ₁ CH ₂ OR ₂ , b) CH.CHOR ₁ CH ₂ OR ₂ , c) CH OR- CCHOR ) CH OR- wherein n is an integer between 1 and 4 or d) C((CH ₂ OR ₁ ) ₂ CH ₂ OR ₂ CH ₂ OR ₃ )

wherein R , R and/or R independently are as defined above.

In one embodiment of this type of the invention where the compound has the formula (a) or (b) , R- and/or R„ are

i) hydrogen; ii) a straight or branched alkyl selected from the group of methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl and/or octyl or iii) a straight or branched acyl radical of a saturated, aliphatic carboxylic acid with 1 to 15 carbon atoms, such as butanoyl, hexanoyl, octanoyl and/or decanoyl.

In another embodiment of this type of the invention where the compound has the formula (c) or (d) , R and/or R are

i) hydrogen; ii) a straight or branched alkyl selected from the group of methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl and/or octyl or

iii) a straight or branched acyl radical of a saturated, aliphatic carboxylic acid with 1 to 15 carbon atoms, such as butanoyl, hexanoyl, octanoyl and/or decanoyl

and R_ is

i) a straight or branched alkyl selected from the group of butyl, hexyl, octyl and/or decyl or ii) a straight or branched acyl radical of a saturated aliphatic carboxylic acid with 1 to 15 carbon atoms such as butanoyl, hexanoyl, octanoyl and/or decanoyl.

The aliphatic ester or ether compound can also have the formula II, ' R4.COOR5-. wherein R4. is

i) hydrogen; ii) a straight or branched alkyl with 1 to 15 carbon atoms such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl and/or pentadecyl; iii) a cycloalkyl with 3 to 15 carbon atoms such as cyclopropyl, cyclopentyl and/or cyclohexyl or iv) an alkenyl with 2 to 14 carbon atoms such as vinyl, allyl, propenyl, octadionyl, decenyl, dodecenyl and/or tetradecenyl;

the above groups (i) to (iv) being unsubstituted or substituted with hydroxy; oxo; alkoxy; carboxy; esteri- fied carboxy; amine; substituted amine; acyloxy or acylamine, and wherein R is

i) hydrogen; ii) a cation such as sodium and/or potassium or iii) ' R4. as defined above

In one preferred embodiment of the invention where the compound is used, R is selected from the group consisting of propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl and decyl and R _g is

i) hydrogen; ii) a cation such as sodium and/or potassium or iii) a lower alkyl such as methyl, ethyl or propyl.

According to the invention the amount of the aliphatic ester or ether compound in the composition can be fairly low. From the view of the patient this is very favourable as no side effects or other pharmacological effects from these compounds are observed.

One way of expressing the amount of the compound needed is the weight per kg body weight. The final formulation can contain 0.2 to 200 mg of the compound per kg body weight, for example preferably 0.2 to 50 mg of the compound per kg body weight and most preferably 0.2 to 20 mg of the compound per kg body weight.

Another way of expressing the amount of the compound needed is the weight per weight of active substance i.e. the amount of inositol phosphate. The final formulation can contain 0.1 to 100 mg of the compound per mg inositol phosphate, for example preferably 0.5 to 10 mg of the compound per mg inositol phosphate and most preferably 0.5 to 5 mg of the compound per mg inositol phosphate.

The expression inositol phosphate comprises phosphate or phosphates of different isomers of inositol for example myo-inositol, allo-inositol, cis-inositol, chiro- inositol, scyllo-inositol, muco-inositol, neo-inositol or epi-inositol. Most often the inositol isomer is myo- inositol.

The degree of phosphorylation can vary between one phosphate group/inositol moiety to six phosphate groups/inositol moiety.

Preferably inositol trisphosphate isomers are used according to the invention and most preferably D-myo- inositol-1,2,6-trisphosphate (IPO is used.

According to the invention the inositol phosphate is most often present in an salt form or in a form where only a few of the negative charges are protonated. The salt can contain one or more cations in different combinations. Examples of cations are sodium and potassium ions.

The pharmaceutical composition according to the invention may be administered orally, topically, rectally or by inhalation spray in dosage forms or formulations comprising conventional, non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. The preferred administration route is oral administration.

The pharmaceutical composition for oral use can be present in different forms such as capsules, granules, tablets, troches, lozenges, aqueous suspensions, dispensible powders, emulsions, syrups or elixirs. When the composition is present in liquid form capsules are

preferably utilized. At the use of granules, these preferably have a size of 0.15-2 mm. Either the granules can consist of the pharmaceutical composition per se or of the composition and suitable fillers. When the pharma¬ ceutical composition is used in a tablet form, the tablets can have a weight of 50-1500 mg, preferably 50-800 mg and most preferably 100-500 mg.

Formulations for oral use include tablets which contain the active ingredient in a mixture with non-toxic pharma¬ ceutically acceptable excipients. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium chloride, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, potato starch, or alginic acid; binding agents, for example, starch, gelatin or acacia; and lubricating agents, for example, magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed.

Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil.

For the rectal application of the composition of this invention, typical dosage forms include suppositories, rectal gelatin capsules (solutions and suspensions) , and enemas or micro-enemas (solutions and suspensions) . Thus, in a typical suppository formulation, any one of the compounds of this invention is combined with any pharma¬ ceutically acceptable suppository base such as cocoa butter, esterified fatty acids, glycerinated gleatin, and various water-soluble or dispersible bases like poly¬ ethylene glycols and polyoxyethylene sorbitan fatty acid ester. Various additives like salicylates or surfactant materials may be incorporated.

For topical use, creams, ointments, gels, solutions or the like containing the compositions are employed according to methods recognized in the art.

Naturally, the therapeutic dosage range for the compounds of the present invention will vary with the size and needs of the patient and the particular pain or disease symptom being treated.

The administration of the pharmaceutical composition according to the invention can be in a combined dosage form or in separate dosage forms.

As mentioned above the invention also relates to the use of at least one aliphatic ester or ether compound with formula I och II for the preparing of an inositol phos¬ phate containing medicament with improved bioavailability of inositol phosphate in mammals including man at non- parenteral administration.

As mentioned above the aliphatic ester or ether compound can have the following formulas:

A. CH ₂ OR ₁ CHOR ₂ CH ₂ OR ₃

B. CH ₂ OR ₁ CH ₂ OR ₂

C. CH CHOR ₁ CH ₂ OR ₂

D. CH 2_OR1_ (CHOR2_)nCH2_OR3.

E. C((CH ₂ OR ₁ ) ₂ CH ₂ OR ₂ CH ₂ OR ₃ )

wherein R , R , R , R , R-. and n are as defined above.

Preferably the compound is selected from the group of dihydroxypropy1 butanoate, dihydroxypropy1 hexanoate, dihydroxypropyl octanoate and dihydroxypropyl decanoate.

Preferably the inositol phosphate is inositol trisphosphate and most preferably the inositol trisphospate is D-myo-inositol-l,2,6-trisphosphate.

Another embodiment of the invention relates to pharmaceutical compositions comprising a aliphatic ester or ether compound with anyone of formulas A-F above and other phosphorus-containing inositol derivatives. For example different types of phosphonates of inositol encounter the need of improved bioavailability after non-parenteral administration. Still another embodiment of the invention relates to pharmaceutical compositions comprising an aliphatic ester or ether compound with anyone of formulas A-F and derivatives of inositol phos¬ phates and inositol phosphonates. The derivatives could be compounds where the phosphate or phosphonate group or

groups are further esterified or compounds where the hydroxyl groups on the inositol moiety are etherified or esterified, such as alkanoyl and carbamoyl derivatives. A further embodiment of the invention is related to composi¬ tions comprising an aliphatic ester or ether compound with anyone of formulas A-F above and inositol derivatized with other charged groups such as carboxyl and sulphonyl. The aliphatic ester or ether compound used in these embodi¬ ments belong to the same groups as disclosed above.

The invention also relates to the improvement of the penetration of cellmembranes in a wider aspect than in the above definition of bioavailability. Under certain circumstances there is a need to be able to obtain an improved ability of different isomers of inositol phos¬ phates or derivatives thereof to cross cell membranes.

Such an example is when it is desirable to let the inositol phosphate or its derivative influence different intracellular metabolic pathways. This can be the case for example when new drugs based on inositolphosphate compounds are being developed. Another example is when the knowledge of different cellular signal systems is further explored. For example it is known that some iso- meric structures of inositol phosphates exist intra¬ cellularly. In many cases it is advantageous to be able to add these substances from the extracellular space in order to clarify in more detail how they effect the be¬ haviour of the cells. This knowledge can then be used in order to develop agonistic or antagonistic compounds effecting the cellular response.

The combination of an aliphatic ester or ether compound as disclosed above and an inositol phosphate or its derivative will facilitate the penetration across cellular membranes.

Another area is when it is desirable to facilitate the transport of nucleosides across cellular membranes. The combination of an aliphatic ester or ether compound as described above and a nucleoside will enhance the pene¬ tration of cellular membranes.

Furthermore the invention can be used for the pharma¬ ceuticals which suffer from low bioavailability due to the same reasons as inositol phosphates and their deriva¬ tives. One such application is a pharmaceutical composi¬ tion comprising an aliphatic ester or ether compound as defined above and a bisphosphonate or a derivative there¬ of useful for the treatment of bone disorders such as osteoporosis and other conditions characterized by ab¬ normal calcium metabolism.

The bisphosphonate or the derivative thereof could be selected from the group of hydroxy ethane bisphosphonate, amino ethane bisphosphonate, dichloromethylene bisphos¬ phonate, methylene bisphosphonate, and preferably:

l-hydroxy-3-aminopropane-l,1-bisphosphonate l-hydroxy-6-aminohexane-l,1-bisphosphonate 4-amino-l-hydroxybutylidene-l,1-bisphonate N-methyl-4-amino-l-hydroxy butylidene-l,1-bisphosphonate 3-amino-l-hydroxypropylidene-l,1-bisphosphonate 1-hydroxy ethylidene-l,1-bisphosphonate 4-(hydroxy methylene-bisphosphonate)piperidine

Another application is a pharmaceutical composition com¬ prising an aliphatic ester or ether compound as defined above and a bisphosphonate or a derivative thereof useful for the treatment of cardiovascular conditions such as increased cholesterol levels and atherosclerosis. The bisphosphonate or the derivative thereof could be selected from the group of alkyl bisphosphonate, alkenyl bisphos- phonates, alkynyl bisphosphonates, phenyl alkyl bisphos- phonates and phenoxyalkyl bisphosphonates.

Still another application is a pharmaceutical composition comprising an aliphatic ester or ether compound as de¬ fined above and a specific bisphosphonate useful as anti¬ microbial agents. The bisphosphonate in this application is selected from the group of alkyl ethenylidene bisphos¬ phonates such as tetraisopropyl ethenylidene bisphosphonate and diethyl dibutyl ethenylidene bisphosphonate. The in¬ vention can also be used for other pharmaceuticals suffering from low bioavailability such as antiviral agents. One such application is a pharmaceutical composition comprising an aliphatic ester or ether compound as defined above and a charged phosphorus-containing pharmaceutical or a biologi¬ cally active compound such as:

phosphonoformiate, phosphonoacetate, 3-azido thymidine-5-phosphate, 9-(2-phosphonyl methoxy ethyl) adenine, S-9-(3-hydroxy-2-phosphonyl methoxy propyl)- -adenine/cytosine,

2 ' ,3 '-didehydro-2,3-dideoxy thy idine, (2R, 5R)-9-(2,5-dihydro-5-phosphonomethoxyl)-2- -furanyl adenosine and derivatives thereof. The composi¬ tions are useful for the treatment of CMV-infections, HIV-infections, herpes simplex and hepatitis B.

The invention can also be used for other pharmaceuticals and biologically active compounds with low and limited bioavailability such as phosphorylated and phosphonylated sugar residues, for example hydroxy phosphonyl glucose- amine derivatives, purine and pyrimidine nucleotides and derivatives thereof for example 2 '-deoxy-5-fluorouridine 5'-phosphate and oligonucleotides and derivatives thereof. Those skilled in the art will realize that the list of compounds is not exhaustive and the invention is also applicable to other pharmaceuticals and biologically active compounds comprising a phosphate or phosphonate moiety.

Preferably, the above aliphatic ester or ether compounds contain 3 to 18 carbon atoms.

In some embodiments of the invention also an aromatic ester or ether compound with 3 to 18 carbon atoms could be contemplated to be utilized.

The invention will be explained further in connection with the embodiment examples below, however without limiting it thereto.

Example 1 shows the manufacturing of a pharmaceutical composition comprising a mixture of dihydroxypropyl octanoate and IP . Example 2 and 3 illustrate the determination of the bioavailability of two different dihydroxypropyl esters and IP after oral administration.

Example 1

The pentasodium salt of D-myo-inositol-1,2,6-trisphos- phate (IP ₃ ) 2.0 g, was mixed with 4,1 g of 2,3-di- hydroxypropyl octanoate to form an composition consisting of 4 moles of the 2,3-dihydroxypropyl octanoate compound/IP -moiety.

Example 2

The bioavailability of a composition comprising a mixture of 2,3-dihydroxypropyl octanoate and D-myo-inositol- -1,2,6-trisphosphate (IP ₃ ) (obtained according to Example 1) after oral administration was determined in an experiment with pigs. Five pigs were given an in¬ travenous injection of the sodium salt of IP and blood samples were collected 0.25 hr, 0.5 hr, 1 hr, 1,5 hrs, 2 hrε, 4 hrs, 6 hrs and 8 hrs after the in¬ jection. The blood samples were assayed for the con¬ tent of IP_ and these measurements were used for the calculation of the reference data as 100 % of the substance is introduced into the blood stream after an intravenous injection. Another five pigs were given the composition comprising the mixture of 2,3-dihydroxy- propyl octanoate and IP as an intraduodenal dosing. The animals were treated with Atropin and anaesthetized with Hypnodil /Sedaperone® whereafter a laparotomy was carried out. Blood samples were collected at the time intervals mentioned above and the concentration of IP was determined in each sample. By calculating the area under the curve (AUC) from a graph where the concen¬ tration versus time is shown and comparing the obtained value with a similar calculation from the data after intravenous injection of the substance the bioavailabi¬ lity for this specific formulation is obtained.

Still another five pigs were given the penta-sodium salt of IP_ orally. Blood samples were collected at the same time intervals as above and the concentration of IP was determined. Following a similar procedure the AUC was determined.

The following concentrations of IP were obtained after intravenous injection of the sodium salt, after oral ad¬ ministration of the composition comprising 2, 3-dihydroxy¬ propyl octanoate and IP- and after oral administration of the sodium salt respectively.

time intravenous cone. IP (μM) oral formu¬ injection, oral formulation, lation, sodium salt composition com¬ sodium salt of IP. prising 2,3-di- of IP- hydroxypropyl octanoate and IP-

0 0 0

0.25 53.7 24.0

0.5 40.0 20.2 0.6

1.0 21.6 13.2 0.5

1.5 12.1 11.2 0.5

2.0 9.3 8.6 0.6

4.0 2.6 2.8 0.4

6.0 1.1 1.1 0.4

8.0 0.6 0.5 0

Normaliz ;ed

AUC (%) 100 58

The bioavailability of IP_ at the use of the oral formulation consisting of 2,3-dihydroxypropyl octanoate and IP_ was 58 %. Irritancy test showed no toxic effects on the intestinal barriers.

Example 3

The bioavailability of a composition comprising a mixture of 2,3-dihydroxypropyl butanoate and IP- after oral ad¬ ministration was determined with a method similar to the description in Example 2. Five pigs were given an intravenous injection of the penta-sodium salt of IP while another five animals were given a formulation consisting of a mixture of 2,3-dihydroxypropyl butanoate and IP as and intraduodenal dosing. Still another group of five pigs were given the penta-sodium salt of IP orally. Blood samples were collected and the concentra¬ tion of IP were determined.

The following concentrations of IP were obtained after intravenous injection of the sodium salt, after oral ad¬ ministration of the composition comprising 2,3-dihydroxy- propyl butanoate and IP and after oral administration of the sodium salt respectively.

time intra- cone IP-(μM) oral (hrs) venous, oral formula- formulation, injection tion, composi- sodium salt sodium salt tion comprising of IP. of IP, 2,3-dihydroxy propylbutanoate and IP_

0 0 0

0.25 53.7 2.8

0.5 40.0 4.2 0.6

1.0 21.6 4.0 0.5

1.5 12.1 3.8 0.5

2.0 9.3 1.7 0.6

4.0 2.6 0.8 0.4

6.0 1.1 0.6 0.4

8.0 0.6 0.1 0

Normalized

AUC (%) 100 23

The bioavailability of IP at the use of the oral formulation consisting of the 2,3-dihydroxypropyl butanoate and IP- was 23 %. Irritancy test showed no toxic effects on the intestinal barriers.