AU2008202073A1 - 1-Butyric acid derivatives and their use - Google Patents

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P001 Section 29 Regulation 3.2(2)

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION STANDARD PATENT Application Number: Lodged: Invention Title: 1-Butyric acid derivatives and their use The following statement is a full description of this invention, including the best method of performing it known to us: P111AHAU/1107 -t- 00 Ct 1-Butyric acid derivatives and their use M Description O The invention concerns butyric acid derivatives, 00 10 pharmaceutical compounds containing such derivatives as well as uses of such derivatives for the manufacture of pharmaceutical compounds for the treatment of various illnesses.

Today cancer is one of the most common causes of death, and the number of cancer cases in industrialized countries is constantly on the rise. Above all this is connected with the fact that malignant tumours are an illness associated with advanced age, and thanks to the successful fight against infectious diseases more people now reach this age. Despite all the progress in diagnostic and therapeutic areas, the prospects of a successful cure for the most commonly occurring internal forms of cancers are seldom greater than 20%. At the present time, a cancerous growth can be eradicated or its growth inhibited. A reverse transformation of a tumour cell to a normal cell is not yet achievable. The most important therapeutic measures surgery and irradiation remove the cancer cells from the organism. The common current chemotherapeutic treatments of cancer, cytostatics, only lead to destruction or damage of tumour cells. The effect in most cases is not very selective, so that at the same time severe damage to healthy cells occurs. In general tumour cells have a different metabolism from healthy cells, in particular for glycolysis. Thus a change of the enzyme system involved in glycolysis and a change in NADH transport is typical for tumour cells. Among other things the activity of the enzymes 00 for glycolysis is increased. This enables rapid cell growth under the aerobic conditions typical for tumour cells. For details regarding this, please refer to E. Eigenbrodt et al., Biochemical and Molecular Aspects of Selected Cancers, Vol. 2, p. 311 ff, 1994.

State of the art The use of glucose analogues for the inhibition of glycolysis 00 10 is known from the citation E. Eigenbrodt et al., Biochemical 0 and Molecular Aspects of Selected Cancers, Vol. 2, p. 311 ff, 1994. Other approaches discussed in this source are the use of inhibitors for glycolytic isoenzymes, for example by the formation of suitable complexes or the inhibition of complex formation. The result is that tumour cells are effectively starved. A difficulty with the aforementioned compounds is that many of them are genotoxic and/or insufficiently specific for tumour cells.

In connection with a new agent against inflammatory illnesses as well as autoimmune reactions, U. Mangold et al., Eur. J.

Biochem., 266:1-9, 1999 report that these agents, namely leflunomide derivatives, also affect glycolysis.

The technical problem of the invention The present invention is based on the technical problem of preparing agents that are able to inhibit proliferation, particularly of cancer cells, and thereby inhibit the growth of neoplastic tumours and inflammations as well as exaggerated immune reactions of the body, such as septic shock, autoimmune diseases, transplant rejections as well as acute and chronic inflammation reactions, while in fact exhibiting little to no cytotoxicity toward normal cells in the blood, the immune system, and tissues.

00 For the solution of this technical problem the invention demonstrates the use of a compound of Formula I R1 R3 Formula I R C1 2 R CH -2Hb R2 R4 where a and b can be the same or different and have the values Sof 0 or 1, 00 10 where R1 -Cl-C18-alkyl, -cycloalkyl or -aryl, and 0where R2 -OX1, -SX1, -COO-, -(CH 2 )n-COOX1 or -COOX1 with Xl -Cl-C18-alkyl, -cycloalkyl or -aryl and n 1-8, where R3 -CN, -COO-, -COOX2, -CO-X2, -CO-NHX2 with X2 -Cl-C18-alkyl, -cycloalkyl or -aryl, where R4 is -NHY or -CO-NHZ with Y -CO-R (R -Cl-C18-alkyl, -cycloalkyl or -aryl or NHA, with A -Cl-C18-alkyl, -cycloalkyl or -aryl) and Z phenyl, naphthyl, phenyl substituted with -Hal and/or -0- Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl or naphthyl group substituted with -Hal and/or -O-Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl (-Hal -Cl or -Br), in which a and b correspond to the number of remaining carbon valences for C 1 and C 2 where a ring closure via R3 can occur at with the elimination of Xl in R2 and X2 in R3, or a physiologically compatible salt of such a compound for the manufacture of a pharmaceutical compound for treatment and/or prophylaxis of illnesses from the group consisting of "neoplastic tumours, inflammatory illnesses, autoimmune diseases, in particular systemic lupus erythematosus, degenerative diseases of the joints, illnesses of the rheumatic type with cartilage degeneration, all progressive forms of arthritis, in particular rheumatoid and chronic polyarthritis, joint trauma, cartilage loss due to 00 Simmobilization, septic shock, illnesses with impaired leukocyte adhesion, illnesses due to elevated TNF alpha concentrations, cachexia, Crohn's syndrome, psoriasis of the skin, Wegener's granulomatosis, rejection reactions after transplants, in particular in the course of cell therapy or stem cell therapy".

SSome of the substances falling under the aforementioned definition are known and familiar in other contexts. Other 00 10 substances falling under the aforementioned definition, Showever, are new. Therefore the invention includes further compounds according to Formula I R1 R3 Formula I C' C2Hb R2 R4 where a and b can be the same or different and have the values of 0 or 1, where R1 -C1-C18-alkyl, -cycloalkyl or -aryl, and where R2 -OX1, -SX1, -COO-, -(CH 2 )n-COOXl or -COOX1 with X1 -Cl-C18-alkyl, -cycloalkyl or -aryl and n 1-8, where R3 -CN, where R4 is -NHY or -CO-NHZ with Y -CO-R (R -Cl-C18-alkyl, -cycloalkyl or -aryl or NHA, with A -Cl-C18-alkyl, -cycloalkyl or -aryl) and Z phenyl, naphthyl, phenyl substituted with -Hal and/or -0- Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl or naphthyl group substituted with -Hal and/or -O-Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl (-Hal -Cl or -13r), in which a and b correspond to the number of remaining carbon valences for C 1 and C 2 or a physiologically compatible salt of such a compound 00 Finally the invention covers a pharmaceutical composition containing a compound with the formula R1 R3 Formula I R C1H;2-Hb R R2 R4 p where a and b can be the same or different and have the values Sof 0 or 1, Swhere R1 -Cl-C18-alkyl, -cycloalkyl or -aryl, and 00 10 where R2 -OX1, -SX1, -COO-, (CH 2 -COOX1 or -COOX1 with 0 X1 -Cl-C18-alkyl, -cycloalkyl or -aryl and n 1-8, where R3 -CN, -COO-, -COOX2, -CO-X2, -CO-NHX2 with X2 -Cl-C18-alkyl, -cycloalkyl or -aryl, where R4 is -NHY or -CO-NHZ with Y -CO-R (R -Cl-C18-alkyl, -cycloalkyl or -aryl or NHA, with A -Cl-C18-alkyl, -cycloalkyl or -aryl) and Z phenyl, naphthyl, phenyl substituted with -Hal and/or -0- Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl or naphthyl group substituted with -Hal and/or -0-Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl (-Hal -Cl or whereby a and b correspond to the number of remaining carbon valences for C 1 and C 2 where a ring closure via R3 can occur at C with the elimination of Xl in R2 and X2 in R3, or a physiologically compatible salt of such a compound as well as at least one physiologically compatible inactive ingredient and/or vehicle.

It is self-evident that in some cases various stereoisomers of compounds corresponding to Formula I can exist (especially enantiomers and diastereomers), which are all objects of the invention. The term "alkyl" encompasses linear and branched 00 alkyl groups. The term "cycloalkyl" also encompasses cycloalkyl groups with linear or branched alkyl substituents.

The term aryl also includes aryl groups where there may be alkyl substituents that are alkyl or cycloalkyl.

Surprisingly it was found that such 1-butyric acid derivatives of the general Formula I are able, depending on the dose, to inhibit the proliferation of cancer cells in therapeutically relevant concentrations in vitro. In the range of dosages 00 10 investigated no cytotoxic effect was determined. Thus Scompounds of the general Formula I have an antiproliferative effect, that is, the propagation of tissues is slowed down.

For example in chronic polyarthritis, excessive growth of the synovial sheath, which increases almost like a tumour, penetrates into the joint and with the progress of the illness destroys the cartilage and bone. The antiproliferative effect of the compounds corresponding to Formula I of the invention causes an immediate slowing of the growth as well as a decrease of the so-called systemic inflammation activity and in fact for all proliferation processes having to do with the inflammation, wound healing or regeneration. Thus, due to their pharmacological properties, the compounds in accordance with the invention are likewise outstandingly suited for treatment and prevention of the other previously listed illnesses. The aforementioned medical definitions and terms can be found in the Roche Lexikon Medizin (Roche Medical Lexicon), 4 th Edition, Munich 1999.

Furthermore the invention concerns a diagnostic agent containing at least one compound corresponding to Formula I for the detection of illnesses such as those mentioned above, for which purpose a cell or cell culture to be tested is brought into contact with such a compound and evaluated appropriately. Refer to Example 2 for this.

00 O Embodiments of the invention Within the scope of the invention there are various, nonrestrictive embodiments possible. Thus a pharmaceutical composition corresponding to the invention can have several different compounds meeting the aforementioned definition.

Moreover, a pharmaceutical composition corresponding to the invention can additionally contain at least one active Singredient that is different to the compound defined by 00 10 Formula I. Such a case is a combination preparation. The 0 various active ingredients in this can be prepared in a single pharmaceutical form, that is, the active ingredients are mixed together in the administration form. However, it is also possible to prepare the various active ingredients in physically separate administration forms of the same of different types. Preferred active ingredients in this regard are so-called immune modulators like leflunomide (Arava®), Methotrexate or antirheumatic agents.

It is preferable that the compounds according to the invention have the following groups: R1 methyl or ethyl, R2 -OX, -COO-, or COOX with X1 H, methyl or ethyl, R3 -CN, -COOH, -COO-, or COX2, -CO-NHX2 or a ring closure via R3 to with the elimination of X1 in R2 and X2 in R3, R4 is -NHY with Y H or -COR (R methyl, ethyl or -NHA with A H, methyl or ethyl) or CO-NHZ with Z -Br, -Cl, -0-C1 and/or -0-Br substituted phenyl.

Particularly appropriate examples of compounds falling under Formula I are explained below.

Compound 1: R1 methyl, R2= -OH, R3 -CN, R4 -NH 2 a b 0 Compound 2: 00 RI methyl, R2 -OH, R3 =COOH, R4 -NI1 2 a =b =0 Compound 3: R1 methyl, R2 -OH, R3 -CN, R4 =-NHY, a =b =0 Compounds 4 6: Ri methyl, R2= -OH, R3 R4 =-CO-NH-C 6

H

4 F meta)

-CO-NH-C

6

H

3 Br 2 ortho, meta), or C 6

H

4 O(_l para) a b =0 Compound 7: R1 methyl, R2 -OH, R3 -CN, R4 -CO--NH-Z, a =b =0 00 10 Compound 8: R1 methyl, R2 -OH, R3 -CN, R4 -NH 2 a b =0 Compound 9: R1 R2 -COO-methyl, R3 -CN, R4 a b =0 Compound Ri R2 -COO-, R3 -COOH, R4 a 1, b =0 Compound 11: R1 R2 -COO-, R3 -COOH, R4 -NH-CO-NH 2 a =b 1 Compound 12: R1 R2 -COO-, R3 -COOH, R4 NH 2 a =b =1 Compound 13: R1 R2 -CH 2 -COO-methyl, R3 -CN, R4 a 1, b 0 Compound 14: R1 R2 =-OXi, R3 =-CO-X2, R4 NH 2 a b Xl and X2 eliminated Compound R1 R2 -COOH, R3 =-COOH, R4 -NH--CO-NH 2 a =b 1 Compound 16: Ri R2 -OXi, R3 =-CO-NHX2, R4 =NH 2 a b 1, Xl and X2 eliminated Most particularly preferable are those compounds according to Formula I that show (oxo-enol) tautomerism, such as the methyl ester of 4-cyano-4-oxo-butyric acid (Compound 9, earlier: carbomethoxypropionyl cyanide).- 00

OCH,

O ^N 0 0 Counter-ions which come into question for ionic compounds in accordance with Formula I are, for example, Na+, Li+, S 5 cyclohexylammonium or basic amino acids (such as lysine, 0 arginine, ornithine, glutamine). The pharmaceuticals 00 0 manufactured with compounds in accordance with the invention Scan be administered orally, intramuscularly, periarticularly, intraarticularly, intravenously, intraperitonealy, subcutaneously or rectally. The invention concerns the processes for the manufacture of pharmaceuticals which are characterized in that they contain at least one compound according to Formula I with a pharmaceutically suitable and physiologically compatible carrier and perhaps additional active ingredients, additives or auxiliary materials in an appropriate administration form. Suitable solid or liquid galenic preparations or formulations are, for example, granulates, powders, coated tablets, tablets, (micro) capsules, suppositories, syrups, juices, suspensions, emulsions, drops or injectable solutions as well as preparations with a protracted release of the active ingredient, in the manufacture of which typical auxiliary materials such as carriers, disintegrants, binding agents, coatings, swelling agents, lubricants or grease, flavour additives, sweeteners, solubilizers find use. Auxiliary materials named are magnesium carbonate, titanium dioxide, lactose, mannite and other sugars, talc, milk protein, gelatine, starch, cellulose and its derivatives, animal and vegetable oils such as cod-liver oil, sunflower oil, peanut oil or sesame oil, polyethylene glycols and solubilizers such as sterile water and single or multi-functional alcohols such as glycerine.

00 O Preferably the pharmaceuticals are manufactured and administered in dosage units, each unit: containing as the active component a particular dose of the compound corresponding to the invention as represented by Formula I.

For solid dosage units such as tablets, capsules, coated pills or suppositories this dose can consist of 1 to 1000 mg, with to 300 mg preferred, and for injection solutions in ampules the dose can be 0.3 to 300 mg, with 10 to 100 mg preferred.

00 10 For treatment of an adult patient weighing between 50 and 100 Skg, for example weighing 70 kg, daily doses of 20 to 1000 mg of the active ingredient, preferably 100 to 500 mg, are indicated. In some circumstances, however, higher or lower daily doses can also be applied. The administration of the daily dosage can occur as a single application of a single dosage unit, but it can also occur with several smaller dosage units as well as multiple administrations of divided doses at particular intervals.

The invention is explained in more detail in the following sections using examples intended solely to illustrate the embodiments.

Example 1 The compound carbomethoxypropionyl cyanide was synthesized according to the method described by Q. Tang and S. Sen (Tetrahedron Letters 39 1998, pp 2249-2252). Typically 1.5 g mmol) carbomethoxypropionyl chloride was added to a solution of 1.79 g CuCN (20 mmol) in 10 ml acetonitrile. The mixture was refluxed for 30 minutes and after cooling to room temperature concentrated with the rotary evaporator. The residue was dissolved in ether, and the ether solution was filtered. After removal of the solvent a slightly yellow oil remained (yield 0.95 g, 67% theoretical); IR (cm1) 2225, 1727.

00 0 Example 2 The Novikoff hepatoma cells used came from the tumour bank of the Deutsches Krebsforschungszentrum (German Cancer Research Centre) in Heidelberg (Cancer Research 1951, 17, 1010). Each cm 2 culture flask was seeded with 100,000 cells. The substance pertaining to the invention from Example 1, Lcycloserine or dehydrothreonine was dissolved in a solvent Ssuited for use in cell cultures, such as water, dilute 00 10 ethanol, dimethyl sulfoxide or similar and added in increasing concentrations to the culture medium. For example Lcycloserine (Compound 16) or dehydrothreonine (Compound 2) was used in a concentration range from 80 pM to 5000 pM, carbomethoxypropionyl cyanide (Compound 13) in a concentration range of 100 pM to 300 pM. After four days of cultivation the cell count in each flask was measured. The results are shown in Figures 1 and 2. A dosage-dependent inhibition of proliferation can be seen in comparison to the control sample which has no added compound according to the invention.

Example 3 The investigation of the effects of carbomethoxypropionyl cyanide (CMPC) on the metabolism of the Novikoff cells showed that CMPC greatly inhibits the glycolysis rate, as shown in Figure 3.

Claims (8)

1. The use of a compound according to Formula I Formula I RI R3 SC1Ha^.C2Hb SR2 R4 0 where a and b can be the same or different and have the values 00 10 of 0 or 1, 0 where R1 -Cl-C18-alkyl, -cycloalkyl or -aryl, and where R2 -OX1, -SX1, -COO-, -(CH 2 )n-COOX1 or -COOX1 with Xl -Cl-C18-alkyl, -cycloalkyl or -aryl and n 1-8, where R3 -CN, -COO-, -COOX2, -CO-X2, -CO-NHX2 with X2 -Cl-C18-alkyl, -cycloalkyl or -aryl, where R4 is -NHY or -CO-NHZ with Y -CO-R (R Cl-C18-alkyl, -cycloalkyl or -aryl or -NHA, with A Cl-C18-alkyl, -cycloalkyl or -aryl) and Z phenyl, naphthyl, phenyl substituted with -Hal and/or -0- Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl or naphthyl group substituted with -Hal and/or -0-Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl (-Hal -Cl or in which a and b correspond to the number of remaining carbon valences for C' and C 2 in which a ring closure via R3 can occur at with the elimination of X1 in R2 and X2 in R3, or a physiologically compatible salt of such a compound for the manufacture of a pharmaceutical composition for the treatment and/or prophylaxis of illnesses from the group consisting of "neoplastic tumours, inflammatory illnesses, autoimmune diseases, in particular systemic lupus erythematosus, degenerative diseases of the joints, illnesses of the rheumatic type with cartilage degeneration, all progressive forms of arthritis, in particular' rheumatoid and chronic polyarthritis, joint trauma, cartilage loss due to 00 3 immobilization, septic shock, illnesses with impaired Sleukocyte adhesion, illnesses due to elevated TNF alpha concentrations, cachexia, Crohn's syndrome, psoriasis of the skin, Wegener's granulomatosis, rejection reactions after transplants, in particular in the course of cell therapy or stem cell therapy".

2. Compound according to Formula I R1 R3 00 10 Formula I R 1 R3 SC' H b Ci R2 R4 where a and b can be the same or different and have the values of 0 or 1, where R1 -Cl-C18-alkyl, -cycloalkyl or -aryl, where R2 -OX1, -SX1, -COO-, -(CH 2 )n-COOX1 or -COOX1 with Xl -Cl-C18-alkyl, -cycloalkyl or -aryl and n 1-8, where R3 -CN, where R4 is -NHY or -CO-NHZ with Y -CO-R (R -Cl-C18-alkyl, -cycloalkyl or -aryl or NHA, with A -Cl-C18-alkyl, -cycloalkyl or -aryl) and Z phenyl, naphthyl, phenyl substituted with -Hal and/or -0- Hal and/or C1-C18-alkyl, -cycloalkyl or -aryl or naphthyl group substituted with -Hal and/or -0-Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl (-Hal -Cl or -Br), in which a and b correspond to the number of remaining carbon valences for C 1 and C 2 or a physiologically compatible salt of such a compound.

3. Pharmaceutical composition containing a compound according to the formula R1 R3 Formula I C'H^z--CZHb R2 R4 00 t where a and b can be the same or different and have the values of 0 or 1, where R1 -Cl-C18-alkyl, -cycloalkyl or -aryl, where R2 -OX1, -SX1, -COO-, -(CH 2 )n-COOX1 or -COOX1 with M Xl -C1-C18-alkyl, -cycloalkyl or -aryl and n 1-8, 0 where R3 -CN, -COO-, -COOX2, -CO-X2, -CO-NHX2 with 0 X2 -Cl-C18-alkyl, -cycloalkyl or -aryl, 00 10 where R4 is -NHY or -CO-NHZ with SY -CO-R (R Cl-C18-alkyl, -cycloalkyl or -aryl or -NHA, with A -Cl-C18-alkyl, -cycloalkyl or -aryl) and Z phenyl, naphthyl, phenyl substituted with -Hal and/or -0- Hal and/or -Cl-C18-alkyl, -cycloalkyl or -aryl or naphthyl group substituted with -Hal and/or -0-Hal and/or Cl-C18-alkyl, -cycloalkyl or -aryl (-Hal -Cl or in which a and b correspond to the number of remaining carbon valences for C' and C 2 where a ring closure via R3 can occur at C 1 with the elimination of X1 in R2 and X2 in R3, or a physiologically compatible salt of such a compound as well as at least one physiologically compatible auxiliary and/or carrier substance.

4. A pharmaceutical composition according to claim 3 which contains in addition at least one active ingredient differing from the compounds corresponding to Formula I, preferably leflunomide (raaf Methotrexate or antirheumatic agents.

5. The use of a compound or pharmaceutical composition according toone of the claims 1 to 4, where R1 methyl or ethyl, R2 -OX1, -COO-, or -COOX1 with X1 methyl or ethyl, R3 -CN, -COOH, -COO-, -COX2, -CO-NHX2 with a ring closure via R3 to with the elimination of X1 in R2 and X2 in R3, 00 O R4 is -NHY with Y H or -COR (R methyl, ethyl or -NHA with A H, methyl or ethyl) or CO-NHZ with Z -Br, -Cl, -0-C1 and/or -0-Br substituted phenyl.

6. A compound according to Formula I and one of the claims 1 to 4, characterized in that it has an anti- proliferative effect.

S7. A compound according to Formula I and claim 6, 00 10 characterized in that has the effect of inhibiting Sinflammation.

8. A diagnostic agent containing at least one compound according to Formula I and claim 1.