CA2222103C - Inositolglycans having insulin-like action - Google Patents
Inositolglycans having insulin-like action Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/04—Disaccharides
Abstract
Inositolglycans having insulin-like action Compounds of the formula I
A-Z-R (I) are described, where A is the radical H - P(O)(OH) -, H - P(S)(OH) -, HO--P(S)(OH) -, HS-P(S)(OH) -, (C1-C4) - alkyl - P(O)(OH) -, (C1-C4) - alkyl -P(S)(OH)-, S(O)2(OR1) -, S(O)(OR1)-, NH2 - C(O) -, R1R2N -, R1R2N - C(O) -NH -, R1O - SO2 - NH -, (C1-C4) - alkyl - SO2 -, (C1-C4) - alkyl - S(O) - or R1 - S -, Z is 2 to 6 substituted or unsubstituted sugar radicals and R is substituted or unsubstituted inositol.
The compounds of the formula I are suitable for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
A-Z-R (I) are described, where A is the radical H - P(O)(OH) -, H - P(S)(OH) -, HO--P(S)(OH) -, HS-P(S)(OH) -, (C1-C4) - alkyl - P(O)(OH) -, (C1-C4) - alkyl -P(S)(OH)-, S(O)2(OR1) -, S(O)(OR1)-, NH2 - C(O) -, R1R2N -, R1R2N - C(O) -NH -, R1O - SO2 - NH -, (C1-C4) - alkyl - SO2 -, (C1-C4) - alkyl - S(O) - or R1 - S -, Z is 2 to 6 substituted or unsubstituted sugar radicals and R is substituted or unsubstituted inositol.
The compounds of the formula I are suitable for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
Description
Hoechst Aktiengesellschaft HOE 96/F 321 Dr. MS/St Description Inositolglycans having insulin-like action The invention relates to inositolglycans having insulin-like action, which are suitable for the treatment of diabetes mellitus.
It is known that the metabolic action of insulin also causes the formation of low molecular weight compounds which also have insulin-like action (US 4,446,064).
A
number of inositoiglycan compounds have already been proposed which have insulin-like action (WO 96/14075, JP 6/293790, JP 4/120089) Diabetes type II, noninsulin-dependent diabetes, is accompanied by insulin resistance of the peripheral tissue, such as muscle or fatty tissue. The glucose utilization, which is thereby reduced, is caused by a lack of insulin stimulation of glucose transport and subsequent metabolic processes.
In attempting to find further active compounds having insulin-like action, it has now been found that the compounds according to the invention have in vitro insulin-like action, exhibit good serum stability and also have insulin-like action on insulin-resistant tissues and are thus suitable for the treatment of diabetes mellitus.
The invention therefore relates to inositolglycans having insulin-like action, of the formula I
A-Z-R (I) and/or physiologically tolerable salts of the compound of the formula I and/or stereoisomeric forms of the compound of the formula I, where A is the radical 1) H - P(O)(OH) - , 2) H - P(S)(OH) - , 3) HO-P(S)(OH) -, 4) HS-P(S)(OH) -, 5) (Cl-C4) - alkyl - P(O)(OH) - , 6) (Cl-C4) - alkyl - P(S)(OH) - , 7) S(O)2(OR') - , 8) S(O)(OR') -, 9) NH2 - C(O) -, 10) R'R2N -, 11) R1R2N - C(O) - NH -, 12) R'O - S02 - NH -, 13) (CI-C4) - alkyl - SO2 - , 14) (Cl-C4) - alkyl - S(O) - or 15) R' - S -in which R' and R2 independently of one another are hydrogen or (Cl-C4) - alkyl, Z is 1) 2 to 6 sugar radicals, 2) 2 to 6 sugar radicals, mono- to hexasubstituted independently of one another by 2.1 methyl, 2.2 sugar radical 2.3 disugar radical 2.4 - S02 - OH, 2.5 - C(O) - NR' R2, 2.6 - C(O) - (CI-C4) - alkyl, 2.7 - P(O)(H)OH, 2.8 - P(O)(OH)2, 2.9 - P(S)(H)OH, 2.10 - P(S)(OH)2, 2.11 - P(S)(SH)(OH), 2.12 - P(O)(OH) - 0- CH2 - CH2 -NR1 R2 or 2.13 the glycosidic bond between the 2 to 6 sugar radicals is replaced one to six times by- CH2 - or - S-, and in which R' and R2 independently of one another are a hydrogen atom or (CI-C4) - alkyl, R is 1) inositol, 2) inositol phosphate, 3) inositol thiophosphate, 4) inositol cyclophosphate, 5) inositol cyclothiophosphate, 6) a radical from the group defined under R 2) to 5) mono- or disubstituted independently of one another by 6.1 phosphate or 6.2 thiophosphate, 7) a radical from the group defined under R 2) to 5) monosubstituted by 7.1 a cyclophosphate radical or 7.2 a cyclothiophosphate radical, or 8) inositol, where two adjacent OH groups are substituted by 8.1 -CH2-S02-NH-.
Preferred compounds of the formula I are those wherein A is 1) H - P(O)(OH) -, 2) S(0)2(OR') - or 3) NH2 - C(O) -Z is 1) 2 to 6 sugar radicals which originate from the group consisting of 1.1 mannose, 1.2 glucose, 1.3 gluconic acid, 1.4 galactonic acid, 1.5 mannonic acid, 1.6 glucosamine, 1.7 fructose or 1.8 galactose, 2) 2 to 6 sugar radicals which originate from the group defined under Z1.1 to 1.8 and are mono- to hexasubstituted independently of one another by 2.1 methyl, 2.2 mannose, 2.3 glucosamine, 2.4 dimannose or 2.5 mannose-glucosamine and the glycosidic bond of the two sugars mannose and glucosamine is between the carbon atoms 1-3, 1-2 or 1-6 of the two sugars, and R is 1) inositol, 2) inositol phosphate, 3) inositol thiophosphate, 4) inositol cyclothiophosphate or 5) inositol cyclophosphate.
Particularly preferred compounds of the formula I are those wherein A is H - P(O)(OH) -, Z is 1) 2 to 4 sugar radicals which originate from the group consisting of 1.1 mannose or 1.2 glucosamine or 2) 2 to 4 sugar radicals which originate from the group consisting of 2.1 mannose or 2.2 glucosamine, monosubstituted by mannose and R is 1) inositol, 2) inositol phosphate or 3) inositol cyclophosphate.
Sugar radicals are understood as meaning compounds which are derived from aldoses and ketoses having 3 to 7 carbon atoms, which can belong to the D or L
series; these also include amino sugars or uronic acids. Examples which may be mentioned are glucose, mannose, fructose, galactose, ribose, erythrose, glyceraldehyde, sedoheptulose, glucosamine, galactosamine, glucuronic acid, galacturonic acid, gluconic acid, galactonic acid or mannonic acid.
By disugar, saccharides are meant which consist of two sugar units. Tri-, tetra-, penta- or hexasugars are formed by acetal-like linkage of 3 to 6 sugars. The linkages can in this case occur in the a- or R-form. The linkages between the sugars preferably take place via carbon atom 1 and carbon atom 6, carbon atom 1 and carbon atom 2, and carbon atom 1 and carbon atom 4 of the respective sugars.
The a-form of the linkage between the sugars is preferred.
The linkage of A to Z takes place, for example, via one of the oxygen atoms of Z or via one of the carbon atoms of Z, preferably via the carbon atom of the CH2 group of Z. The linkage of the radicals for A 10) to 12) preferably takes place via a carbon atom of Z, the other linkages of A preferably take place via an oxygen atom of Z.
The linkage of R to Z takes place analogously to the linkages of the di-, tri-, tetra-, penta- or hexasugars. Furthermore, the linkage from R to Z can also be replaced one or more times by -CH2 - or -S-.
If the sugar is substituted, the substitution preferably takes place on the hydrogen atom of an OH group of the sugars.
The term insulin-resistant tissue is understood as meaning, for example, rat fat cells which no longer contain any insulin receptor.
The compounds according to the invention can contain one or more phosphate groups which can also be additionally derivatized by a phosphate protective group.
Phosphate protective groups are, for example, phenyl, benzyl or hydroxypropylnitrile (Houben Weyl, Methoden der Organischen Chemie [Methods of Organic Chemistry], Volume 12/1 or Volume 12/2; Teilheimer, Synthetic Methods of Organic Chemistry, Vol 45).
Physiologically tolerable salts of the compound of the formula I are in particular understood as meaning pharmaceutically utilizable or nontoxic salts. Such salts are formed, for example, from compounds of the formula I which contain acidic groups, e.g. phosphates or sulfates, with alkali metals or alkaline earth metals, such as, for example, Na, K, Mg and Ca, and also with physiologically tolerable organic amines, such as, for example, triethylamine and tris(2-hydroxyethyl)amine. Compounds of the formula I which contain basic groups, e.g. an amino group, form salts with inorganic acids, such as, for example, hydrochloric acid, sulfuric acid or phosphoric acid and with organic carboxylic or sulfonic acids, such as, for example, acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid and p-toluenesulfonic acid. Compounds in which basic and acidic groups are present in equal number form internal salts and are not dependent on a third salt component.
The invention furthermore relates to a process for the preparation of the compound of the formula I, which comprises synthesizing the inositolglycan stepwise from protected sugar and inositol precursors, then adding the radical A and removing from the compound obtained one or more protective groups introduced temporarily for the protection of other functions and converting the compound of the formula I
thus obtained, if appropriate, into its physiologically tolerable salt.
The synthesis of the di- to polysugars is carried out by known processes (H.
Paulsen, Angew. Chem. Int. Ed. 21 (1982) p. 155). The trichloroacetimidate method is preferably used for the synthesis of oligosaccharides (R. R. Schmidt, Angew.
Chem. Int. Ed. 25 (1986) 212 - 235; T. Ogawa, Tetrahedron Lett. 31 (1990) 2439 -2442).
The synthesis of the phosphates is carried out with the aid of the H-phosphate and the phosphoramidite method.(W. Bannwath et al., Helvetica Chemica Acta, 70 (1987), pages 175-186; L.A. Slotin, Synthesis (1977), pages 737-752) The following are essentially possible protective groups for the hydroxyl groups of the sugars: benzyl, acetyl, benzoyl, pivaloyl, trityl, tert-butyidimethylsilyl, benzylidene, cyclohexylidene or isopropylidene protective groups.
The compounds of the formula I and their physiologically tolerable salts are primarily used as active compounds for pharmaceutical preparations for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
The invention therefore also relates to a pharmaceutical preparation which comprises at least one compound of the formula I and/or at least one of its physiologically tolerable salts in dissolved, amorphous and/or crystalline form -preferably in amorphous and/or crystalline form.
The pharmaceutical preparation is preferably a solution or suspension for injection having a pH of approximately 3.0 to 9.0, preferably of approximately 5.0 to 8.5, which contains a suitable isotonicizing agent, a suitable preservative and, if appropriate, a suitable buffer, and if appropriate also a depot principle, all in sterile aqueous solution or suspension. All of the preparation constituents apart from the active compound forms the preparation excipient. Suitable isotonicizing agents are, for example, glycerol, glucose, mannitol, NaCI, calcium or magnesium compounds such as, for example, CaCi2 or MgCI2. Suitable preservatives are, for example, phenol, m-cresol, benzyl alcohol and/or p-hydroxybenzoic acid esters.
Buffer substances which can be used, in particular for adjusting a pH of approximately 5.0 to 8.5, are, for example, sodium acetate, sodium citrate or sodium phosphate. Otherwise, physiologically acceptable dilute acids (typically HCI) or alkalis (typically NaOH) are also suitable for adjusting the pH.
For the purpose of varying the profile of action of the preparation according to the invention, modified (cf. EP-B 132 769 and EP-B 132 770) and/or unmodified insulins, preferably bovine, porcine or human insulin, in particular human insulin, can also be admixed.
The pharmaceutical preparation is prepared by bringing at least one compound of the formula I and/or at least one of its physiologically tolerable salts, if appropriate together with modified and/or unmodified insulin or insulin derivatives, into a suitable administration form using a physiologically acceptable excipient and also, if appropriate, using suitable additives and auxiliaries.
The invention is now explained in greater detail by means of the following examples.
Example 1:
Preparation of the compound B
The synthesis course can be inferred from the reaction scheme at the end of Example 1.
Synthesis of compound 3 60 g (94 mmol) of 1(T. G. Mayer, B. Kratzer, R. R. Schmidt, Angew. Chem. 106 (1994) 2289- 93) and 21.2 g (42.4 mmol) of 2 (A. Termin, R. R. Schmidt, Liebigs Ann. Chem. (1989) 789- 795) are dissolved in 200 ml of dry methylene chloride and 400 ml of dry n-heptane. After addition of 70 g of dried molecular sieve (0.4 nm), the mixture is stirred at room temperature for 15 minutes. 5ml of 0.05 M
trimethylsilyltrifluormethane-sulfonic acid in methylene chloride (described as catalyst solution in the following procedures) are then added. After 15 minutes, 300 ml of n-heptane/ethyl acetate (1:1) are added and the mixture is filtered through silica gel. The silica gel is washed with n-heptane/ethyl acetate (1:1) and the filtrate is then concentrated. After purification by means of flash chromatography, 41.0 g (99%) of 3 are obtained as a colorless oil. TLC: n-heptane/ethyl acetate (1:1), Rf =
0.6, MS: (M + Li)+ = 981.1, calculated C55H67N3O Si, M= 974.21.
Synthesis of imidate 4 41 g (42.0 mmol) of 3 are dissolved in 400 ml of tetrahydrofuran (THF) and 9 ml of acetic acid. After addition of 70 ml of 1 M TBAF/THF solution, the mixture is allowed to stand at room temperature for 8 hours. The acetic acid is removed by freezing (16 h at -30 C) and the filtrate is purified by means of flash chromatography after concentration. Yield 34.1 g (94%) of deprotected compound. This is dissolved in 300 ml of dry methylene chloride. After addition of 50 ml of trichloroacetonitrile and 20 g of potassium carbonate, the mixture is stirred at room temperature for 4 hours.
It is filtered through silica gel, the silica gel is washed with n-heptanelethyl acetate (1:1) and the filtrate is concentrated. Crude yield: 42.1 g. TLC: n-heptane/ethyl acetate (2:1), Rf = 0.5. 1 H- NMR (CDCI3): characteristic signals for imidate;
6= 8,78, for NH and 5.63 for the anomer (f3- imidate).
Synthesis of trisaccharide 6 33.2 g (33.0 mmol) of 4 and 13.3 g (25.0 mmol) of 5 (R. Aneja, S. G. Aneja, A.
Parra, Tetrahedron, Asymmetry 6 (1995) 17- 18; C. J. J. Elie, R. Verduyn, C.
E.
Dreef, D. M. Braunts, G. A. van der Marel, J. H van Boom, Tetrahedron, 46 (1990) 8243- 54) are dissolved in 120 ml of dry methylene chloride and 360 ml of dry n-heptane. After addition of 100 g of molecular sieve, the mixture is stirred at room temperature for 15 minutes. It is cooled to -20 C under argon and then treated with ml of catalyst solution. After 30 minutes, the mixture is allowed to thaw to room temperature. For working up, it is filtered through silica gel and the silica gel is 15 washed with n-heptane/ethyl acetate (1:1). The filtrate is concentrated and the crude product (46.2 g) is dissolved in 150 ml of methylene chloride. After addition of 400 ml of methanol and 15 ml of 1 M NaOMe/MeOH solution, the mixture is allowed to stand at room temperature for 16 hours. The solution is treated with 1 ml of water and concentrated. The oil obtained is dissolved in 50 ml of ethyl acetate, diluted 20 with 200 ml of n-heptane/ethyl acetate (1:1) and filtered through silica gel. After concentration, the residue is purified by flash chromatography. Yield 32.5 g (97%) of white foam as an anomer mixture. TLC: n-heptane/ethyl acetate (2:1), Rf =0.5.
MS:
(M +Li)+ = 1336.7; calculated C80H87N3015, M = 1330.59.
Synthesis of trisaccharide 7 The anomer mixture 6 can only be easily separated chromatographically as the derivative 7. 32.4 g (24.4 mmol), of 6 are dissolved in 200 ml of methylene chloride.
After addition of 500 ml of 0.5 M HCI/MeOH (from 17.5 ml of AcCI in 500 ml of MeOH) and 20 ml of ethylene glycol, the mixture is allowed to stand at room temperature for 17 hours. After concentration, the residue is purified by flash chromatography. The product obtained (26.9 g, 88%) is dissolved in 300 ml of methylene chloride. 3.0 g of imidazole and 4.6 g of TBDMSCI are added. After hours at room temperature, the mixture is diluted with 500 ml of n-heptane/ethyl acetate (2:1) and filtered through silica gel. The silica gel is washed with n-heptane/ethyl acetate (2:1) and the filtrate is concentrated. The crude product obtained is purified by flash chromatography. Yield 21.1 g(72 r6) of 7 and 6.3 g 5 (22%) of alpha product. TLC: n-heptane/ethyl acetate (2:1), Rf =0.5 for 7 and Rf =0.3 for the alpha product. MS: (M +Li)+ = 1370.6; calculated C$oH93N3O15Si , M = 1364.71.
Synthesis of trisaccharide 8 10 21.2 g (15.5 mmol) of 7 are dissolved in 60 ml of methylene chloride and 180 ml of dimethoxypropane. 250 mg of TsOH are added and the mixture is allowed to stand at room temperature for 1 hour. After addition of 2 mi of trimethylamine, it is concentrated and 23.1 g of crude product are obtained. This is dissolved in 150 ml of THF and treated with 30 ml of 1 M TBAF/THF solution. After 16 hours, it is concentrated and purified by flash chromatography. Yield: 20.0 g (99%) of 8 as a white foam. TLC: n-heptane/ethyl acetate (2:1), Rf= 0.4. MS: (M + Li)+ =
1296.7;
calculated C77H83N3015, M= 1290.51.
Synthesis of tetrasaccharide 10 20.0 g (15.4 mmol) of 8 and 15.0 g (23.5 mmol) of 9 (T. G. Mayer, B. Kratzer, R. R.
Schmidt, Angew. Chem. 106 (1994) 2289- 93) are reacted analogously to the procedure for compound 6 and 20.3 g (76%) of tetrasaccharide 10 are obtained as a white foam. TLC: n-heptane/ethyl acetate (2:1), Rf= 0,6, MS: (M + Li)+ = 1770, calculated C107H15N3020, M= 1763.09.
Synthesis of pentasaccharide 12 20.3 g(11.8 mmol) of 10 and 12.0 g (20.3 mmol) of 11(T. G. Mayer, B. Kratzer, R. R.
Schmidt, Angew. Chem. 106 (1994) 2289- 93) are reacted analogously to the procedure for compound 6 and 19.4 g (80%) of deacylated product are obtained.
This product is dissolved in 200 mi of methylene chloride and treated with 3.4 g (50.0 mmol) of imidazole. 6.0 g (40.0 mmol) of TBDMSCI are added and the mixture is stirred at room temperature for 15 hours. After addition of 5 ml of methanol, it is allowed to stand for 10 minutes, then diluted with 200 ml of n-heptane/ethyl acetate (1:1) and filtered through silica gel. The silica gel is additionally washed with 200 ml of n-heptane/ethyl acetate (1:1), and the filtrate is concentrated and purified by means of flash chromatography. Yield: 19.4 g (95%) of 12 as a white foam. TLC:
n-heptane/ethyl acetate (2:1), Rf= 0,7. MS: (M + Li)+ = 2226; calculated C1331-1151 N3025S1, M= 2219.75 Synthesis of hexasaccharide 14 19.4 g (8.9 mmol) of 12 and 14.0 g (20.,0 mmol) of 13 (T. G. Mayer, B.
Kratzer, R. R.
Schmidt, Angew. Chem. 106 (1994) 2289- 93) are dissolved in 100 ml of dry methylene chloride and 300 ml of dry n-heptane. After addition of 40 g of molecular sieve (0.4 nm), the mixture is stirred at room temperature for 15 minutes. 10 ml of catalyst solution are added and the mixture is stirred for a further 15 minutes. For working up, it is filtered through silica gel and the silica gel is washed with n-heptane/ethyl acetate (1:1). The filtrate is concentrated and 33 g of crude product are obtained. This is dissolved in 200 ml of methylene chloride and treated with 500 ml of 0.5 M HCI in methanol. After two hours at room temperature, the mixture is concentrated and concentrated several times with methylene chloride. The intermediate obtained is dissolved in 200 ml of methylene chloride and treated with 3.4 g (50 mmol) of imidazole and 6.0 g of (40 mmol) of TBDMSCI. After 16 hours (h), the mixture is worked up analogously to compound 12, Yield 20.2 g (85%) of 14 as a white foam. TLC: n-heptane/ethyl acetate (2:1), Rf= 0,3. MS: (M+Li)+= 2669;
calculated C161H177N3030Si, M= 2662.26 Synthesis of compound 15 30.0 g of triazole are dissolved in 800 ml of dry THF.13.5 ml of phosphorus oxychloride are added dropwise at 10 C. 60 ml of triethylamine are then added dropwise and the mixture is stirred at room temperature for 15 minutes. The precipitate is filtered and washed with a little dry THF. The filtrate is added to 19.1 g (7.2 mmol) of 14. The solution is concentrated to 100 ml. After 15 minutes, it is diluted with 500 ml of ethyl acetate and washed twice with 100 ml of water.
The organic phase is dried over magnesium sulfate, filtered and concentrated.
After flash chromatography, 19.0 g (97%) of cyclic phosphate derivative are obtained as a white foam. TLC: methylene chloride/methanol/33% NH3 (100/7/1), Rf= 0.3.
MS: (M+2Li-H)+ = 2737; calculated C161 H174N3032PSi, M= 2724.22.
The cyclic phosphate is dissolved in 350 ml of THF and 100 ml of TBAF (1 M in THF) are added. After 20 hours, the mixture is concentrated and the residue is purified by flash chromatography. Yield 18.1 g (99%) of 15 as a white foam. TLC: methylene chloride/methanol/33% NH3 (100/7/1), Rf= 0,3 (runs identically to the starting material) MS: (M+2Li-H)+ = 2622; calculated C155H162N3032P, M= 2609.96.
Synthesis of compound 16 14 g of phosphorous acid is concentrated four times with pyridine and then taken up in 200 ml of dry pyridine. 16 ml of pivaloyl chloride are added dropwise at 10 C. This reaction solution is allowed to stand at room temperature for 15 minutes. 18.1 g (6.9 mmol) of 15 are introduced into the reaction solution described above.
After 1 hour, it is diluted with 200 ml of toluene and 150 ml of methylene chloride/methanol/33% NH3 (30/10/3). After concentrating, residual pyridine is distilled out a further three times with toluene. The residue is suspended in 200 ml of methylene chloride/methanol (20:1). The nonsoluble constituents are filtered and washed twice with 50 ml of methylene chloride/methanol (20:1). The filtrate is concentrated and purified by flash chromatography. Yield 16.9 g (91 %) of protected final product. TLC: methylene chloride/methanol/33% NH3 (100/7/1), Rf = 0.25.
MS: (M+3Li-2H)+ = 2691; calculated C155H163N3034P2, M = 2673,94. For deprotection, 600 ml of ammonia are condensed at -78 C. 4.7 g (204 mmol) of sodium are dissolved therein. This solution is diluted with 300 ml of dry THF
and 16.9 g (6.3 mmol) of protected final product dissolved in 100 ml of dry THF
are then slowly added dropwise at a reaction temperature of -78 C. After a reaction time of 15 minutes (blue color must not disappear), the mixture is treated cautiously with 10 g of ammonium chloride. When the blue color has disappeared, the mixture is diluted cautiously with 100 ml of water and 300 ml of methanol. It is allowed to thaw and then concentrated to around 150 ml. This solution is diluted with 2 I of methylene chioride/methanol/33% NH3 (3/3/1) and added to a flash silica gel column (700 ml of silica gel). It is eluted with 3 1 of methylene chloride/methanol/33% NH3 (3/3/2) and then with 3 I(3/3.5/3). The product elutes if the mixture is then chromatographed using n-butanol/ ethanol/water/33% NH3 (2/2/2/1).Yield 5.5 g (78%) of 16 as a white solid. TLC: (2/2/2/1), Rf = 0,4.
MS:
(M+H)+= 1116,5; calculated C36H63NO34P2, M= 1115,83. 31P- NMR (D20) a= 16,3 ppm for cyclic phosphate and 7.9 for H-phosphate.
OBn Ac0 Bn0 O O
BnO + HO OTBDMS
BnO BnO
Oy CC13 N3 TMS-Trf Ac0 BnO
p BnO OBn BnO 0 BnO
1. TBAF, AcO H
2. C13CCN, K2CO3 Ac0 Bnp p BnO OBn BnO p O O~NH
BnO N3 4 cci, BnO O r n HO OBn 1.
2.NaOMe HO Bn0 p Bn0 OBn BnO p p BnO O OBn Bn0 N3 O p OBn HO BnOp BnO OBn BnO p O BnO O OBn BnO
N3 O p OBn 1. HCI / MeOH
10 2. TBDMSCI, imidazole TBDMSO Bn0 OBn BnO
BnO p BnO O OBn O Hp OBn +
BnO
TBDMSO Bnpp 7 BnO OBn BnO p O
BnO OBn0 O OB OBn n HO
1. dimethoxypropane, TsOH
2. TBAF
HO BnOp BnO OBn BnO p BnO O OBn O O p OBn Bn0 BnO
Ac00 1. B BnO
~ CC13 NH
2. NaOMe BnO HOO
BnO
Bn0 0 Bn0 0 Bn0 OBn Bn0 Bn0 O
O O O OBn BnO N 0 OBn 3 nll AcO Ac0 O
1. Bn0 Bn0 11 y NH
2. NaOMe Bn0 Bn0 3. TBDMSCI, imidazole Bn0 0 BnO .0 BnO
0 BnO0 Bn0 OBn Bn0 Bn0 0 0 0 O Bn0 N j,-"~OBn OBn '12 3 II
It is known that the metabolic action of insulin also causes the formation of low molecular weight compounds which also have insulin-like action (US 4,446,064).
A
number of inositoiglycan compounds have already been proposed which have insulin-like action (WO 96/14075, JP 6/293790, JP 4/120089) Diabetes type II, noninsulin-dependent diabetes, is accompanied by insulin resistance of the peripheral tissue, such as muscle or fatty tissue. The glucose utilization, which is thereby reduced, is caused by a lack of insulin stimulation of glucose transport and subsequent metabolic processes.
In attempting to find further active compounds having insulin-like action, it has now been found that the compounds according to the invention have in vitro insulin-like action, exhibit good serum stability and also have insulin-like action on insulin-resistant tissues and are thus suitable for the treatment of diabetes mellitus.
The invention therefore relates to inositolglycans having insulin-like action, of the formula I
A-Z-R (I) and/or physiologically tolerable salts of the compound of the formula I and/or stereoisomeric forms of the compound of the formula I, where A is the radical 1) H - P(O)(OH) - , 2) H - P(S)(OH) - , 3) HO-P(S)(OH) -, 4) HS-P(S)(OH) -, 5) (Cl-C4) - alkyl - P(O)(OH) - , 6) (Cl-C4) - alkyl - P(S)(OH) - , 7) S(O)2(OR') - , 8) S(O)(OR') -, 9) NH2 - C(O) -, 10) R'R2N -, 11) R1R2N - C(O) - NH -, 12) R'O - S02 - NH -, 13) (CI-C4) - alkyl - SO2 - , 14) (Cl-C4) - alkyl - S(O) - or 15) R' - S -in which R' and R2 independently of one another are hydrogen or (Cl-C4) - alkyl, Z is 1) 2 to 6 sugar radicals, 2) 2 to 6 sugar radicals, mono- to hexasubstituted independently of one another by 2.1 methyl, 2.2 sugar radical 2.3 disugar radical 2.4 - S02 - OH, 2.5 - C(O) - NR' R2, 2.6 - C(O) - (CI-C4) - alkyl, 2.7 - P(O)(H)OH, 2.8 - P(O)(OH)2, 2.9 - P(S)(H)OH, 2.10 - P(S)(OH)2, 2.11 - P(S)(SH)(OH), 2.12 - P(O)(OH) - 0- CH2 - CH2 -NR1 R2 or 2.13 the glycosidic bond between the 2 to 6 sugar radicals is replaced one to six times by- CH2 - or - S-, and in which R' and R2 independently of one another are a hydrogen atom or (CI-C4) - alkyl, R is 1) inositol, 2) inositol phosphate, 3) inositol thiophosphate, 4) inositol cyclophosphate, 5) inositol cyclothiophosphate, 6) a radical from the group defined under R 2) to 5) mono- or disubstituted independently of one another by 6.1 phosphate or 6.2 thiophosphate, 7) a radical from the group defined under R 2) to 5) monosubstituted by 7.1 a cyclophosphate radical or 7.2 a cyclothiophosphate radical, or 8) inositol, where two adjacent OH groups are substituted by 8.1 -CH2-S02-NH-.
Preferred compounds of the formula I are those wherein A is 1) H - P(O)(OH) -, 2) S(0)2(OR') - or 3) NH2 - C(O) -Z is 1) 2 to 6 sugar radicals which originate from the group consisting of 1.1 mannose, 1.2 glucose, 1.3 gluconic acid, 1.4 galactonic acid, 1.5 mannonic acid, 1.6 glucosamine, 1.7 fructose or 1.8 galactose, 2) 2 to 6 sugar radicals which originate from the group defined under Z1.1 to 1.8 and are mono- to hexasubstituted independently of one another by 2.1 methyl, 2.2 mannose, 2.3 glucosamine, 2.4 dimannose or 2.5 mannose-glucosamine and the glycosidic bond of the two sugars mannose and glucosamine is between the carbon atoms 1-3, 1-2 or 1-6 of the two sugars, and R is 1) inositol, 2) inositol phosphate, 3) inositol thiophosphate, 4) inositol cyclothiophosphate or 5) inositol cyclophosphate.
Particularly preferred compounds of the formula I are those wherein A is H - P(O)(OH) -, Z is 1) 2 to 4 sugar radicals which originate from the group consisting of 1.1 mannose or 1.2 glucosamine or 2) 2 to 4 sugar radicals which originate from the group consisting of 2.1 mannose or 2.2 glucosamine, monosubstituted by mannose and R is 1) inositol, 2) inositol phosphate or 3) inositol cyclophosphate.
Sugar radicals are understood as meaning compounds which are derived from aldoses and ketoses having 3 to 7 carbon atoms, which can belong to the D or L
series; these also include amino sugars or uronic acids. Examples which may be mentioned are glucose, mannose, fructose, galactose, ribose, erythrose, glyceraldehyde, sedoheptulose, glucosamine, galactosamine, glucuronic acid, galacturonic acid, gluconic acid, galactonic acid or mannonic acid.
By disugar, saccharides are meant which consist of two sugar units. Tri-, tetra-, penta- or hexasugars are formed by acetal-like linkage of 3 to 6 sugars. The linkages can in this case occur in the a- or R-form. The linkages between the sugars preferably take place via carbon atom 1 and carbon atom 6, carbon atom 1 and carbon atom 2, and carbon atom 1 and carbon atom 4 of the respective sugars.
The a-form of the linkage between the sugars is preferred.
The linkage of A to Z takes place, for example, via one of the oxygen atoms of Z or via one of the carbon atoms of Z, preferably via the carbon atom of the CH2 group of Z. The linkage of the radicals for A 10) to 12) preferably takes place via a carbon atom of Z, the other linkages of A preferably take place via an oxygen atom of Z.
The linkage of R to Z takes place analogously to the linkages of the di-, tri-, tetra-, penta- or hexasugars. Furthermore, the linkage from R to Z can also be replaced one or more times by -CH2 - or -S-.
If the sugar is substituted, the substitution preferably takes place on the hydrogen atom of an OH group of the sugars.
The term insulin-resistant tissue is understood as meaning, for example, rat fat cells which no longer contain any insulin receptor.
The compounds according to the invention can contain one or more phosphate groups which can also be additionally derivatized by a phosphate protective group.
Phosphate protective groups are, for example, phenyl, benzyl or hydroxypropylnitrile (Houben Weyl, Methoden der Organischen Chemie [Methods of Organic Chemistry], Volume 12/1 or Volume 12/2; Teilheimer, Synthetic Methods of Organic Chemistry, Vol 45).
Physiologically tolerable salts of the compound of the formula I are in particular understood as meaning pharmaceutically utilizable or nontoxic salts. Such salts are formed, for example, from compounds of the formula I which contain acidic groups, e.g. phosphates or sulfates, with alkali metals or alkaline earth metals, such as, for example, Na, K, Mg and Ca, and also with physiologically tolerable organic amines, such as, for example, triethylamine and tris(2-hydroxyethyl)amine. Compounds of the formula I which contain basic groups, e.g. an amino group, form salts with inorganic acids, such as, for example, hydrochloric acid, sulfuric acid or phosphoric acid and with organic carboxylic or sulfonic acids, such as, for example, acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid and p-toluenesulfonic acid. Compounds in which basic and acidic groups are present in equal number form internal salts and are not dependent on a third salt component.
The invention furthermore relates to a process for the preparation of the compound of the formula I, which comprises synthesizing the inositolglycan stepwise from protected sugar and inositol precursors, then adding the radical A and removing from the compound obtained one or more protective groups introduced temporarily for the protection of other functions and converting the compound of the formula I
thus obtained, if appropriate, into its physiologically tolerable salt.
The synthesis of the di- to polysugars is carried out by known processes (H.
Paulsen, Angew. Chem. Int. Ed. 21 (1982) p. 155). The trichloroacetimidate method is preferably used for the synthesis of oligosaccharides (R. R. Schmidt, Angew.
Chem. Int. Ed. 25 (1986) 212 - 235; T. Ogawa, Tetrahedron Lett. 31 (1990) 2439 -2442).
The synthesis of the phosphates is carried out with the aid of the H-phosphate and the phosphoramidite method.(W. Bannwath et al., Helvetica Chemica Acta, 70 (1987), pages 175-186; L.A. Slotin, Synthesis (1977), pages 737-752) The following are essentially possible protective groups for the hydroxyl groups of the sugars: benzyl, acetyl, benzoyl, pivaloyl, trityl, tert-butyidimethylsilyl, benzylidene, cyclohexylidene or isopropylidene protective groups.
The compounds of the formula I and their physiologically tolerable salts are primarily used as active compounds for pharmaceutical preparations for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
The invention therefore also relates to a pharmaceutical preparation which comprises at least one compound of the formula I and/or at least one of its physiologically tolerable salts in dissolved, amorphous and/or crystalline form -preferably in amorphous and/or crystalline form.
The pharmaceutical preparation is preferably a solution or suspension for injection having a pH of approximately 3.0 to 9.0, preferably of approximately 5.0 to 8.5, which contains a suitable isotonicizing agent, a suitable preservative and, if appropriate, a suitable buffer, and if appropriate also a depot principle, all in sterile aqueous solution or suspension. All of the preparation constituents apart from the active compound forms the preparation excipient. Suitable isotonicizing agents are, for example, glycerol, glucose, mannitol, NaCI, calcium or magnesium compounds such as, for example, CaCi2 or MgCI2. Suitable preservatives are, for example, phenol, m-cresol, benzyl alcohol and/or p-hydroxybenzoic acid esters.
Buffer substances which can be used, in particular for adjusting a pH of approximately 5.0 to 8.5, are, for example, sodium acetate, sodium citrate or sodium phosphate. Otherwise, physiologically acceptable dilute acids (typically HCI) or alkalis (typically NaOH) are also suitable for adjusting the pH.
For the purpose of varying the profile of action of the preparation according to the invention, modified (cf. EP-B 132 769 and EP-B 132 770) and/or unmodified insulins, preferably bovine, porcine or human insulin, in particular human insulin, can also be admixed.
The pharmaceutical preparation is prepared by bringing at least one compound of the formula I and/or at least one of its physiologically tolerable salts, if appropriate together with modified and/or unmodified insulin or insulin derivatives, into a suitable administration form using a physiologically acceptable excipient and also, if appropriate, using suitable additives and auxiliaries.
The invention is now explained in greater detail by means of the following examples.
Example 1:
Preparation of the compound B
The synthesis course can be inferred from the reaction scheme at the end of Example 1.
Synthesis of compound 3 60 g (94 mmol) of 1(T. G. Mayer, B. Kratzer, R. R. Schmidt, Angew. Chem. 106 (1994) 2289- 93) and 21.2 g (42.4 mmol) of 2 (A. Termin, R. R. Schmidt, Liebigs Ann. Chem. (1989) 789- 795) are dissolved in 200 ml of dry methylene chloride and 400 ml of dry n-heptane. After addition of 70 g of dried molecular sieve (0.4 nm), the mixture is stirred at room temperature for 15 minutes. 5ml of 0.05 M
trimethylsilyltrifluormethane-sulfonic acid in methylene chloride (described as catalyst solution in the following procedures) are then added. After 15 minutes, 300 ml of n-heptane/ethyl acetate (1:1) are added and the mixture is filtered through silica gel. The silica gel is washed with n-heptane/ethyl acetate (1:1) and the filtrate is then concentrated. After purification by means of flash chromatography, 41.0 g (99%) of 3 are obtained as a colorless oil. TLC: n-heptane/ethyl acetate (1:1), Rf =
0.6, MS: (M + Li)+ = 981.1, calculated C55H67N3O Si, M= 974.21.
Synthesis of imidate 4 41 g (42.0 mmol) of 3 are dissolved in 400 ml of tetrahydrofuran (THF) and 9 ml of acetic acid. After addition of 70 ml of 1 M TBAF/THF solution, the mixture is allowed to stand at room temperature for 8 hours. The acetic acid is removed by freezing (16 h at -30 C) and the filtrate is purified by means of flash chromatography after concentration. Yield 34.1 g (94%) of deprotected compound. This is dissolved in 300 ml of dry methylene chloride. After addition of 50 ml of trichloroacetonitrile and 20 g of potassium carbonate, the mixture is stirred at room temperature for 4 hours.
It is filtered through silica gel, the silica gel is washed with n-heptanelethyl acetate (1:1) and the filtrate is concentrated. Crude yield: 42.1 g. TLC: n-heptane/ethyl acetate (2:1), Rf = 0.5. 1 H- NMR (CDCI3): characteristic signals for imidate;
6= 8,78, for NH and 5.63 for the anomer (f3- imidate).
Synthesis of trisaccharide 6 33.2 g (33.0 mmol) of 4 and 13.3 g (25.0 mmol) of 5 (R. Aneja, S. G. Aneja, A.
Parra, Tetrahedron, Asymmetry 6 (1995) 17- 18; C. J. J. Elie, R. Verduyn, C.
E.
Dreef, D. M. Braunts, G. A. van der Marel, J. H van Boom, Tetrahedron, 46 (1990) 8243- 54) are dissolved in 120 ml of dry methylene chloride and 360 ml of dry n-heptane. After addition of 100 g of molecular sieve, the mixture is stirred at room temperature for 15 minutes. It is cooled to -20 C under argon and then treated with ml of catalyst solution. After 30 minutes, the mixture is allowed to thaw to room temperature. For working up, it is filtered through silica gel and the silica gel is 15 washed with n-heptane/ethyl acetate (1:1). The filtrate is concentrated and the crude product (46.2 g) is dissolved in 150 ml of methylene chloride. After addition of 400 ml of methanol and 15 ml of 1 M NaOMe/MeOH solution, the mixture is allowed to stand at room temperature for 16 hours. The solution is treated with 1 ml of water and concentrated. The oil obtained is dissolved in 50 ml of ethyl acetate, diluted 20 with 200 ml of n-heptane/ethyl acetate (1:1) and filtered through silica gel. After concentration, the residue is purified by flash chromatography. Yield 32.5 g (97%) of white foam as an anomer mixture. TLC: n-heptane/ethyl acetate (2:1), Rf =0.5.
MS:
(M +Li)+ = 1336.7; calculated C80H87N3015, M = 1330.59.
Synthesis of trisaccharide 7 The anomer mixture 6 can only be easily separated chromatographically as the derivative 7. 32.4 g (24.4 mmol), of 6 are dissolved in 200 ml of methylene chloride.
After addition of 500 ml of 0.5 M HCI/MeOH (from 17.5 ml of AcCI in 500 ml of MeOH) and 20 ml of ethylene glycol, the mixture is allowed to stand at room temperature for 17 hours. After concentration, the residue is purified by flash chromatography. The product obtained (26.9 g, 88%) is dissolved in 300 ml of methylene chloride. 3.0 g of imidazole and 4.6 g of TBDMSCI are added. After hours at room temperature, the mixture is diluted with 500 ml of n-heptane/ethyl acetate (2:1) and filtered through silica gel. The silica gel is washed with n-heptane/ethyl acetate (2:1) and the filtrate is concentrated. The crude product obtained is purified by flash chromatography. Yield 21.1 g(72 r6) of 7 and 6.3 g 5 (22%) of alpha product. TLC: n-heptane/ethyl acetate (2:1), Rf =0.5 for 7 and Rf =0.3 for the alpha product. MS: (M +Li)+ = 1370.6; calculated C$oH93N3O15Si , M = 1364.71.
Synthesis of trisaccharide 8 10 21.2 g (15.5 mmol) of 7 are dissolved in 60 ml of methylene chloride and 180 ml of dimethoxypropane. 250 mg of TsOH are added and the mixture is allowed to stand at room temperature for 1 hour. After addition of 2 mi of trimethylamine, it is concentrated and 23.1 g of crude product are obtained. This is dissolved in 150 ml of THF and treated with 30 ml of 1 M TBAF/THF solution. After 16 hours, it is concentrated and purified by flash chromatography. Yield: 20.0 g (99%) of 8 as a white foam. TLC: n-heptane/ethyl acetate (2:1), Rf= 0.4. MS: (M + Li)+ =
1296.7;
calculated C77H83N3015, M= 1290.51.
Synthesis of tetrasaccharide 10 20.0 g (15.4 mmol) of 8 and 15.0 g (23.5 mmol) of 9 (T. G. Mayer, B. Kratzer, R. R.
Schmidt, Angew. Chem. 106 (1994) 2289- 93) are reacted analogously to the procedure for compound 6 and 20.3 g (76%) of tetrasaccharide 10 are obtained as a white foam. TLC: n-heptane/ethyl acetate (2:1), Rf= 0,6, MS: (M + Li)+ = 1770, calculated C107H15N3020, M= 1763.09.
Synthesis of pentasaccharide 12 20.3 g(11.8 mmol) of 10 and 12.0 g (20.3 mmol) of 11(T. G. Mayer, B. Kratzer, R. R.
Schmidt, Angew. Chem. 106 (1994) 2289- 93) are reacted analogously to the procedure for compound 6 and 19.4 g (80%) of deacylated product are obtained.
This product is dissolved in 200 mi of methylene chloride and treated with 3.4 g (50.0 mmol) of imidazole. 6.0 g (40.0 mmol) of TBDMSCI are added and the mixture is stirred at room temperature for 15 hours. After addition of 5 ml of methanol, it is allowed to stand for 10 minutes, then diluted with 200 ml of n-heptane/ethyl acetate (1:1) and filtered through silica gel. The silica gel is additionally washed with 200 ml of n-heptane/ethyl acetate (1:1), and the filtrate is concentrated and purified by means of flash chromatography. Yield: 19.4 g (95%) of 12 as a white foam. TLC:
n-heptane/ethyl acetate (2:1), Rf= 0,7. MS: (M + Li)+ = 2226; calculated C1331-1151 N3025S1, M= 2219.75 Synthesis of hexasaccharide 14 19.4 g (8.9 mmol) of 12 and 14.0 g (20.,0 mmol) of 13 (T. G. Mayer, B.
Kratzer, R. R.
Schmidt, Angew. Chem. 106 (1994) 2289- 93) are dissolved in 100 ml of dry methylene chloride and 300 ml of dry n-heptane. After addition of 40 g of molecular sieve (0.4 nm), the mixture is stirred at room temperature for 15 minutes. 10 ml of catalyst solution are added and the mixture is stirred for a further 15 minutes. For working up, it is filtered through silica gel and the silica gel is washed with n-heptane/ethyl acetate (1:1). The filtrate is concentrated and 33 g of crude product are obtained. This is dissolved in 200 ml of methylene chloride and treated with 500 ml of 0.5 M HCI in methanol. After two hours at room temperature, the mixture is concentrated and concentrated several times with methylene chloride. The intermediate obtained is dissolved in 200 ml of methylene chloride and treated with 3.4 g (50 mmol) of imidazole and 6.0 g of (40 mmol) of TBDMSCI. After 16 hours (h), the mixture is worked up analogously to compound 12, Yield 20.2 g (85%) of 14 as a white foam. TLC: n-heptane/ethyl acetate (2:1), Rf= 0,3. MS: (M+Li)+= 2669;
calculated C161H177N3030Si, M= 2662.26 Synthesis of compound 15 30.0 g of triazole are dissolved in 800 ml of dry THF.13.5 ml of phosphorus oxychloride are added dropwise at 10 C. 60 ml of triethylamine are then added dropwise and the mixture is stirred at room temperature for 15 minutes. The precipitate is filtered and washed with a little dry THF. The filtrate is added to 19.1 g (7.2 mmol) of 14. The solution is concentrated to 100 ml. After 15 minutes, it is diluted with 500 ml of ethyl acetate and washed twice with 100 ml of water.
The organic phase is dried over magnesium sulfate, filtered and concentrated.
After flash chromatography, 19.0 g (97%) of cyclic phosphate derivative are obtained as a white foam. TLC: methylene chloride/methanol/33% NH3 (100/7/1), Rf= 0.3.
MS: (M+2Li-H)+ = 2737; calculated C161 H174N3032PSi, M= 2724.22.
The cyclic phosphate is dissolved in 350 ml of THF and 100 ml of TBAF (1 M in THF) are added. After 20 hours, the mixture is concentrated and the residue is purified by flash chromatography. Yield 18.1 g (99%) of 15 as a white foam. TLC: methylene chloride/methanol/33% NH3 (100/7/1), Rf= 0,3 (runs identically to the starting material) MS: (M+2Li-H)+ = 2622; calculated C155H162N3032P, M= 2609.96.
Synthesis of compound 16 14 g of phosphorous acid is concentrated four times with pyridine and then taken up in 200 ml of dry pyridine. 16 ml of pivaloyl chloride are added dropwise at 10 C. This reaction solution is allowed to stand at room temperature for 15 minutes. 18.1 g (6.9 mmol) of 15 are introduced into the reaction solution described above.
After 1 hour, it is diluted with 200 ml of toluene and 150 ml of methylene chloride/methanol/33% NH3 (30/10/3). After concentrating, residual pyridine is distilled out a further three times with toluene. The residue is suspended in 200 ml of methylene chloride/methanol (20:1). The nonsoluble constituents are filtered and washed twice with 50 ml of methylene chloride/methanol (20:1). The filtrate is concentrated and purified by flash chromatography. Yield 16.9 g (91 %) of protected final product. TLC: methylene chloride/methanol/33% NH3 (100/7/1), Rf = 0.25.
MS: (M+3Li-2H)+ = 2691; calculated C155H163N3034P2, M = 2673,94. For deprotection, 600 ml of ammonia are condensed at -78 C. 4.7 g (204 mmol) of sodium are dissolved therein. This solution is diluted with 300 ml of dry THF
and 16.9 g (6.3 mmol) of protected final product dissolved in 100 ml of dry THF
are then slowly added dropwise at a reaction temperature of -78 C. After a reaction time of 15 minutes (blue color must not disappear), the mixture is treated cautiously with 10 g of ammonium chloride. When the blue color has disappeared, the mixture is diluted cautiously with 100 ml of water and 300 ml of methanol. It is allowed to thaw and then concentrated to around 150 ml. This solution is diluted with 2 I of methylene chioride/methanol/33% NH3 (3/3/1) and added to a flash silica gel column (700 ml of silica gel). It is eluted with 3 1 of methylene chloride/methanol/33% NH3 (3/3/2) and then with 3 I(3/3.5/3). The product elutes if the mixture is then chromatographed using n-butanol/ ethanol/water/33% NH3 (2/2/2/1).Yield 5.5 g (78%) of 16 as a white solid. TLC: (2/2/2/1), Rf = 0,4.
MS:
(M+H)+= 1116,5; calculated C36H63NO34P2, M= 1115,83. 31P- NMR (D20) a= 16,3 ppm for cyclic phosphate and 7.9 for H-phosphate.
OBn Ac0 Bn0 O O
BnO + HO OTBDMS
BnO BnO
Oy CC13 N3 TMS-Trf Ac0 BnO
p BnO OBn BnO 0 BnO
1. TBAF, AcO H
2. C13CCN, K2CO3 Ac0 Bnp p BnO OBn BnO p O O~NH
BnO N3 4 cci, BnO O r n HO OBn 1.
2.NaOMe HO Bn0 p Bn0 OBn BnO p p BnO O OBn Bn0 N3 O p OBn HO BnOp BnO OBn BnO p O BnO O OBn BnO
N3 O p OBn 1. HCI / MeOH
10 2. TBDMSCI, imidazole TBDMSO Bn0 OBn BnO
BnO p BnO O OBn O Hp OBn +
BnO
TBDMSO Bnpp 7 BnO OBn BnO p O
BnO OBn0 O OB OBn n HO
1. dimethoxypropane, TsOH
2. TBAF
HO BnOp BnO OBn BnO p BnO O OBn O O p OBn Bn0 BnO
Ac00 1. B BnO
~ CC13 NH
2. NaOMe BnO HOO
BnO
Bn0 0 Bn0 0 Bn0 OBn Bn0 Bn0 O
O O O OBn BnO N 0 OBn 3 nll AcO Ac0 O
1. Bn0 Bn0 11 y NH
2. NaOMe Bn0 Bn0 3. TBDMSCI, imidazole Bn0 0 BnO .0 BnO
0 BnO0 Bn0 OBn Bn0 Bn0 0 0 0 O Bn0 N j,-"~OBn OBn '12 3 II
BnO
Bn00 1. Bn0 Bn0 0 y CC13 Bn0 13 Bn00 NH
B B O 2. HCI/MeOH
TBDMSO 0.0 B B O 3. TBDMSCI, imidazole BnO O
Bn0 O
Bn0 O Bn00 BnO OBn Bn0 O 0 O Bn0 OH OBn BnO HO OBn 1. tris(1,2,4-triazole) phosphate BnO Bn0 O 2. TBAF
Bn0 Bn0 HO O
Bn0 .O
BnO
Bn0 00 B Bn0 O Bn0 0 BnO OBn Bn0 "Llo Bn0 0 0 0OBn Bn0 0 OBn 0jPIOH
Bn00 1. Bn0 Bn0 0 y CC13 Bn0 13 Bn00 NH
B B O 2. HCI/MeOH
TBDMSO 0.0 B B O 3. TBDMSCI, imidazole BnO O
Bn0 O
Bn0 O Bn00 BnO OBn Bn0 O 0 O Bn0 OH OBn BnO HO OBn 1. tris(1,2,4-triazole) phosphate BnO Bn0 O 2. TBAF
Bn0 Bn0 HO O
Bn0 .O
BnO
Bn0 00 B Bn0 O Bn0 0 BnO OBn Bn0 "Llo Bn0 0 0 0OBn Bn0 0 OBn 0jPIOH
BnO BnO
BnO
Bn0 HO p -O
BnO
BnO
Bn0 p =0 BnO
BnO
O
I Bn0 0 BnO OBn BnO p BnO O
O 0 OBn Bn0 N ~--OBn 15 p P-OH
1. P(OH)3, PivCl 2. Na, NH3(l), NH4C1 HO Hp p O HO
IP~O
H OO
HO HO
HO
HO p =0 HO
HO
p HpO
HO OH
OH
~ p O OH
H2N p HO O
16=B
BnO
Bn0 HO p -O
BnO
BnO
Bn0 p =0 BnO
BnO
O
I Bn0 0 BnO OBn BnO p BnO O
O 0 OBn Bn0 N ~--OBn 15 p P-OH
1. P(OH)3, PivCl 2. Na, NH3(l), NH4C1 HO Hp p O HO
IP~O
H OO
HO HO
HO
HO p =0 HO
HO
p HpO
HO OH
OH
~ p O OH
H2N p HO O
16=B
Example 2:
The compounds A, C, D, E, F, G, H and I are prepared analogously to the methods according to Example 1. Table 1 shows the structural formulae, empirical formula and mass spectrum.
Compound Empirical formula Mass spectrum (M+H)+
C36 H63 N O34 P2 1116.5 HO Ho 0 HO
O HO
H-P'O O O
HO HO
HO
HO O
O
HO
HO
O O
HO HO
~,O OH HO~P
HO O
O O
O
A HO O OH
HZN HO HO
C36H63NO34P2 1116.5 HO HO
HO
O HO
H_p---o 60 I HO
HO HO
HO
HO
HO O
HO OH
HO OH
B HZN O
Compound Empirical formula Mass spectrum (M+H)+
5 C36H63NO33P2S 1132.7 HO HO O
DOHO
HO HO
HO
HO
HO
O
HO O
HO OH
15 0 0 oH
HO OH
C HO.~P' S
The compounds A, C, D, E, F, G, H and I are prepared analogously to the methods according to Example 1. Table 1 shows the structural formulae, empirical formula and mass spectrum.
Compound Empirical formula Mass spectrum (M+H)+
C36 H63 N O34 P2 1116.5 HO Ho 0 HO
O HO
H-P'O O O
HO HO
HO
HO O
O
HO
HO
O O
HO HO
~,O OH HO~P
HO O
O O
O
A HO O OH
HZN HO HO
C36H63NO34P2 1116.5 HO HO
HO
O HO
H_p---o 60 I HO
HO HO
HO
HO
HO O
HO OH
HO OH
B HZN O
Compound Empirical formula Mass spectrum (M+H)+
5 C36H63NO33P2S 1132.7 HO HO O
DOHO
HO HO
HO
HO
HO
O
HO O
HO OH
15 0 0 oH
HO OH
C HO.~P' S
20 C36H65NO35P2 1134.4 HO HO
HO
O HO
HO
HO HO
O
HO
HO
OH
O
OH
HO 0-P,, Compound Empirical formula Mass spectrum (M+H)+
C40H75N3040P4 1362.8 HO HO O
HO
O HO
HO HO
HO
O
HZN~O-PO O
/ HO O
HO HO o HO"iO" VNHZ
O
HO OH
HO p HO O
E HO oH
HO O
C36H65NO35P2 1134.4 HO HO
HO
O HO
Ho HO
HO
HO O
O
HO
HO O
n HO HOO OH HOOH
HO O
O HO
IL OH
F HO O
HO
HO
O HO
HO
HO HO
O
HO
HO
OH
O
OH
HO 0-P,, Compound Empirical formula Mass spectrum (M+H)+
C40H75N3040P4 1362.8 HO HO O
HO
O HO
HO HO
HO
O
HZN~O-PO O
/ HO O
HO HO o HO"iO" VNHZ
O
HO OH
HO p HO O
E HO oH
HO O
C36H65NO35P2 1134.4 HO HO
HO
O HO
Ho HO
HO
HO O
O
HO
HO O
n HO HOO OH HOOH
HO O
O HO
IL OH
F HO O
HO
Compound Empirical formula Mass spectrum (M+H)+
C18H33N0j9P2 630.3 HO-i -O
HO
H O
HO OH O
HO 0 HO ~
O O P~
~ O
HO
O
OH
G HO HO
C18H34NO17P 568.2 HO'P-O
HO
H O
HO OH
HO
O
::S:~ OH
H HO HO
C18H33N0j9P2 630.3 HO-i -O
HO
H O
HO OH O
HO 0 HO ~
O O P~
~ O
HO
O
OH
G HO HO
C18H34NO17P 568.2 HO'P-O
HO
H O
HO OH
HO
O
::S:~ OH
H HO HO
Compound Empirical formula Mass spectrum (M+H)+
O 1334.6 II
HO-i-O HOO
H HHO
O HOO
HHo O HOO
HHO
O OH OH
HO HOO HO 5~ O O HO 101 HO O HZN HO O P~OH
Ho HO
Example 3 Pharmaceutical activity The biological activity of the compounds of the formula I according to the invention is determined with the aid of isolated fat cells from the rat.
The preparation of fat cells from the rat was carried out as follows:
White fatty tissue of the epididymis (Wistar rat, 160 - 180 g, no food restriction) is digested with collagenase and the resulting isolated fat cells are separated off by means of filtration of undigested tissue and washed by flotation several times using Krebs-Ringer-Henseleit buffer (KRH buffer).
A) Lipogenesis This test determines the insulin-stimulable conversion of glucose into toluene-soluble products (triglycerides, phospholipids, fatty acids), which promotes glucose transport and triglyceride (glycerol-3-P synthesis, esterification)/phospholipid/fatty acid synthesis including the insulin signal transmission cascade. At a glucose concentration of 2.5 mM in the test, the esterification (and not the glycerol-synthesis including glucose transport) is rate-determining for the stimulation of lipogenesis.
200 NI (3 x105 cells/ml) of rat fat cells in KRH buffer are incubated at 37 C
for 90 min with 100 NI of D-[3-3H] glucose (25 mM, 0.4 pCi) in the presence or absence of insulin (10 ng/ml) or compound of the formula I according to the invention (final volume 1 ml). By addition of a toluene-soluble scintillation cocktail (10 ml), the cells are disrupted and the lipids are separated off from water-soluble products and the incubation medium. After phase separation, the radioactivity incorporated in lipid products is determined directly without removal of the aqueous phase by scintillation measurement ([3H]Iipid [dpmx10-3]). A control value (incubation under identical conditions but without cells) is subtracted from this radioactivity. The lipogenesis rate is linear up to 120 min. The maximum stimulation factor - this is the ratio of the incubation result with insulin to the incubation results without insulin - is set at 100%. The percentage data under %Insma;' are parts of the maximum stimulation factor defined in this way. The term EC50 indicates the concentration of the compound of the formula I at which 50% of the maximum stimulation to be achieved by the respective compound of the formula I is to be observed.
B) Glucose transport Rat fat cells in 100 NI of KRH buffer (titer 5 x 105 cells/mi) are incubated with gentle shaking at 37 C for 15 min with insulin or with the compound of the formula I
according to the invention. After addition of 15 NI of 2-[3H]-deoxyglucose (0.3 mM, 0.2 pCi), the incubation is continued at room temperature. After specified times (0-20 min), 100 Ni of the test batch are removed and transferred to a reaction vessel (contents 400 NI), in which 250 NI of dinonyl phthalate have been initially introduced.
After centrifugation (15,000 x g, 1 min), the cells are separated off from the oily layer of the incubation medium underneath the oily layer by cutting the tube at the level of 5 the oily layer and transferring to a scintillation vessel. After addition of 5 ml of water-soluble scintillation fluid, the radioactivity is determined. This total cell-associated radioactivity is corrected for [3H]-deoxyglucose which has passively diffused into the cells and is included in the cell interstices by subtraction of a control value (incubation in the presence of the glucose transport inhibitor 10 cytochalasin B). The initial (stimulated) glucose transport rate is linear up to 15 min. The maximum stimulation factor - this is the ratio of the incubation result with insulin to the incubation result without insulin - is set at 100%.
The terms "%lnsmax" and"EC50" are as defined under A) lipogenesis.
Table 2 Compound Upogenesis Glucose transport Formulae %Insmu EC50[NM] %Insnu EC5014,iM]
HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- A 59 7 4GIuN(31-6(L)inositol-1,2-(cyclic)-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Manal-6Mana1- B 77 8.2 41 8.1 4GIuNp1-6(D)inositol-1,2-(cyclic)-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- C 64 7 29 10 4GIuNp1-6(D)inositol-1,2-(cyclic)-thiophosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- D 33 20 18 18 3GIuN(31-6(L)inositol-3-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1(6-EtNP)- E 48 60 34 30 6Mana1(2-EtNP)-4GIuNp1-6(D)inositol-1,2-(cyclic)-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- F 53 15 26 25 4GIuNa1-6(L)inositol-2-phosphate HO-PO(H)O-6Mana 1-4GIuNa 1-6(L)inositol-1,2-(cyclic)- G 25 15 9 15 phosphate HO-PO(H)O-6Mana1-4GIuNa1-6(L)inositol H 15 25 2 10 F HO-PO(H)O-6Mana1-6-Mana1-6Mana1-6Mana1- I 18 20 14 50 3GIuN(31-6(L)inositol-3-phosphate
O 1334.6 II
HO-i-O HOO
H HHO
O HOO
HHo O HOO
HHO
O OH OH
HO HOO HO 5~ O O HO 101 HO O HZN HO O P~OH
Ho HO
Example 3 Pharmaceutical activity The biological activity of the compounds of the formula I according to the invention is determined with the aid of isolated fat cells from the rat.
The preparation of fat cells from the rat was carried out as follows:
White fatty tissue of the epididymis (Wistar rat, 160 - 180 g, no food restriction) is digested with collagenase and the resulting isolated fat cells are separated off by means of filtration of undigested tissue and washed by flotation several times using Krebs-Ringer-Henseleit buffer (KRH buffer).
A) Lipogenesis This test determines the insulin-stimulable conversion of glucose into toluene-soluble products (triglycerides, phospholipids, fatty acids), which promotes glucose transport and triglyceride (glycerol-3-P synthesis, esterification)/phospholipid/fatty acid synthesis including the insulin signal transmission cascade. At a glucose concentration of 2.5 mM in the test, the esterification (and not the glycerol-synthesis including glucose transport) is rate-determining for the stimulation of lipogenesis.
200 NI (3 x105 cells/ml) of rat fat cells in KRH buffer are incubated at 37 C
for 90 min with 100 NI of D-[3-3H] glucose (25 mM, 0.4 pCi) in the presence or absence of insulin (10 ng/ml) or compound of the formula I according to the invention (final volume 1 ml). By addition of a toluene-soluble scintillation cocktail (10 ml), the cells are disrupted and the lipids are separated off from water-soluble products and the incubation medium. After phase separation, the radioactivity incorporated in lipid products is determined directly without removal of the aqueous phase by scintillation measurement ([3H]Iipid [dpmx10-3]). A control value (incubation under identical conditions but without cells) is subtracted from this radioactivity. The lipogenesis rate is linear up to 120 min. The maximum stimulation factor - this is the ratio of the incubation result with insulin to the incubation results without insulin - is set at 100%. The percentage data under %Insma;' are parts of the maximum stimulation factor defined in this way. The term EC50 indicates the concentration of the compound of the formula I at which 50% of the maximum stimulation to be achieved by the respective compound of the formula I is to be observed.
B) Glucose transport Rat fat cells in 100 NI of KRH buffer (titer 5 x 105 cells/mi) are incubated with gentle shaking at 37 C for 15 min with insulin or with the compound of the formula I
according to the invention. After addition of 15 NI of 2-[3H]-deoxyglucose (0.3 mM, 0.2 pCi), the incubation is continued at room temperature. After specified times (0-20 min), 100 Ni of the test batch are removed and transferred to a reaction vessel (contents 400 NI), in which 250 NI of dinonyl phthalate have been initially introduced.
After centrifugation (15,000 x g, 1 min), the cells are separated off from the oily layer of the incubation medium underneath the oily layer by cutting the tube at the level of 5 the oily layer and transferring to a scintillation vessel. After addition of 5 ml of water-soluble scintillation fluid, the radioactivity is determined. This total cell-associated radioactivity is corrected for [3H]-deoxyglucose which has passively diffused into the cells and is included in the cell interstices by subtraction of a control value (incubation in the presence of the glucose transport inhibitor 10 cytochalasin B). The initial (stimulated) glucose transport rate is linear up to 15 min. The maximum stimulation factor - this is the ratio of the incubation result with insulin to the incubation result without insulin - is set at 100%.
The terms "%lnsmax" and"EC50" are as defined under A) lipogenesis.
Table 2 Compound Upogenesis Glucose transport Formulae %Insmu EC50[NM] %Insnu EC5014,iM]
HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- A 59 7 4GIuN(31-6(L)inositol-1,2-(cyclic)-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Manal-6Mana1- B 77 8.2 41 8.1 4GIuNp1-6(D)inositol-1,2-(cyclic)-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- C 64 7 29 10 4GIuNp1-6(D)inositol-1,2-(cyclic)-thiophosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- D 33 20 18 18 3GIuN(31-6(L)inositol-3-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1(6-EtNP)- E 48 60 34 30 6Mana1(2-EtNP)-4GIuNp1-6(D)inositol-1,2-(cyclic)-phosphate HO-PO(H)O-6Mana1(Mana1-2)-2Mana1-6Mana1- F 53 15 26 25 4GIuNa1-6(L)inositol-2-phosphate HO-PO(H)O-6Mana 1-4GIuNa 1-6(L)inositol-1,2-(cyclic)- G 25 15 9 15 phosphate HO-PO(H)O-6Mana1-4GIuNa1-6(L)inositol H 15 25 2 10 F HO-PO(H)O-6Mana1-6-Mana1-6Mana1-6Mana1- I 18 20 14 50 3GIuN(31-6(L)inositol-3-phosphate
Claims (15)
1. A compound of the formula I
A-Z-R (I) and/or a physiologically tolerable salt of the compound of the formula I
and/or a stereoisomeric form of the compound of the formula I, where A is the radical 1) H - P(O)(OH) -,
A-Z-R (I) and/or a physiologically tolerable salt of the compound of the formula I
and/or a stereoisomeric form of the compound of the formula I, where A is the radical 1) H - P(O)(OH) -,
2) H - P(S)(OH) -,
3) HO-P(S)(OH) -,
4) HS-P(S)(OH) -,
5) (C1-C4) - alkyl - P(O)(OH)-,
6) (C1-C4) - alkyl - P(S)(OH) -,
7) S(O)2(OR1) -,
8) S(O)(OR1) -,
9) NH2 - C(O) -,
10) R1R2N -,
11) R1R2N - C(O) - NH -,
12) R1O-SO2-NH-,
13) (C1-C4) - alkyl - SO2 -,
14) (C1-C4) - alkyl - S(O) - or
15) R1- S -, in which R1 and R2 independently of one another are hydrogen or (C1-C4) - alkyl, Z is 1) 2 to 6 sugar radicals, 2) 2 to 6 sugar radicals, mono- to hexasubstituted independently of one another by 2.1 methyl, 2.2 sugar radical 2.3 disugar radical 2.4 - SO2 - OH, 2 5 - C(O) - NR1R2, 2.6 - C(O) - (Cl-C4) - alkyl, 27 - P(O)(H)OH, 2.8 - P(O)(OH)2, 2.9 - P(S)(H)OH, 2 10 - P(S)(OH)2, 2.11 - P(S)(SH)(OH), 2 12 - P(O)(OH) - O- CH2 - CH2 -NR1R2 or 2.13 the glycosidic bond between the 2 to 6 sugar radicals is replaced one to six times by- CH2- or - S -, and in which R1 and R2 independently of one another are a hydrogen atom or (C1-C4) - alkyl, R is 1) inositol, 2) inositol phosphate, 3) inositol thiophosphate, 4) inositol cyclophosphate, 5) inositol cyclothiophosphate, 6) a radical from the group defined under R 2) to 5) mono- or disubstituted independently of one another by 6.1 phosphate or 6.2 thiophosphate, 7) a radical from the group defined under R 2) to 5) monosubstituted by 7.1 a cyclophosphate radical or 7.2 a cyclothiophosphate radical, or 8) inositol, where two adjacent OH groups are substituted by 8.1 -CH2-SO2-NH-2. A compound of the formula I as claimed in claim 1, wherein A is 1) H - P(O)(OH) -, 2) S(O)2(OR1) - or 3) NH2 - C(O) -Z is 1) 2 to 6 sugar radicals which are selected from the group consisting of 1.1 mannose, 1.2 glucose, 1.3 gluconic acid, 1.4 galactonic acid, 1.5 mannonic acid, 1.6 glucosamine, 1.7 fructose and 1.8 galactose, 2) 2 to 6 sugar radicals which are selected from the group defined under Z1.1 to 1.8 and are mono- to hexasubstituted independently of one another by 2.1 methyl, 2.2 mannose, 2.3 glucosamine, 2.4 dimannose or 2.5 mannose-glucosamine and the glycosidic bond of the two sugars mannose and glucosamine is between the carbon atoms 1-3, 1-2 or 1-6 of the two sugars, and R is 1) inositol, 2) inositol phosphate, 3) inositol thiophosphate, 4) inositol cyclothiophosphate or 5) inositol cyclophosphate.
3. A compound of the formula I as claimed in claim 1, wherein A is H - P(O)(OH) -, Z is 1) 2 to 6 sugar radicals which are selected from the group consisting of 1.1 mannose and 1.2 glucosamine or 2) 2 to 6 sugar radicals which are selected from the group consisting of 2.1 mannose and 2.2 glucosamine, monosubstituted by mannose and R is 1) inositol, 2) inositol phosphate or 3) inositol cyclophosphate.
4. A process for the preparation of a compound of the formula I as claimed in any one of claims 1 to 3, which comprises synthesizing the inositolglycan stepwise from protected sugar and inositol precursors, then adding the radical A and removing from the compound obtained one or more protective groups introduced temporarily for the protection of other functions and converting the compound of the formula I thus obtained, if appropriate, into its physiologically tolerable salt.
5. A pharmaceutical preparation, comprising at least one compound of the formula I as claimed in any one of claims 1 to 3 in dissolved, amorphous or crystalline form and a pharmaceutical excipient.
6. A pharmaceutical preparation as claimed in claim 5, comprising an amount of at least one modified or unmodified insulin or insulin derivative.
7. A process for the production of a pharmaceutical preparation as claimed in claim 5 or 6, which comprises bringing at least one compound of the formula I into a suitable administration form using a physiologically acceptable excipient.
8. The process according to claim 7 further comprising using additives or auxiliaries.
9. The process as claimed in claim 7 or 8 wherein at least one modified or unmodified insulin or insulin derivative is added.
10. A use of at least one compound of the formula I as claimed in any one of claims 1 to 3 for the production of a pharmaceutical preparation for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
11. A use of at least one compound of the formula I as claimed in any one of claims 1 to 3 for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
3. A compound of the formula I as claimed in claim 1, wherein A is H - P(O)(OH) -, Z is 1) 2 to 6 sugar radicals which are selected from the group consisting of 1.1 mannose and 1.2 glucosamine or 2) 2 to 6 sugar radicals which are selected from the group consisting of 2.1 mannose and 2.2 glucosamine, monosubstituted by mannose and R is 1) inositol, 2) inositol phosphate or 3) inositol cyclophosphate.
4. A process for the preparation of a compound of the formula I as claimed in any one of claims 1 to 3, which comprises synthesizing the inositolglycan stepwise from protected sugar and inositol precursors, then adding the radical A and removing from the compound obtained one or more protective groups introduced temporarily for the protection of other functions and converting the compound of the formula I thus obtained, if appropriate, into its physiologically tolerable salt.
5. A pharmaceutical preparation, comprising at least one compound of the formula I as claimed in any one of claims 1 to 3 in dissolved, amorphous or crystalline form and a pharmaceutical excipient.
6. A pharmaceutical preparation as claimed in claim 5, comprising an amount of at least one modified or unmodified insulin or insulin derivative.
7. A process for the production of a pharmaceutical preparation as claimed in claim 5 or 6, which comprises bringing at least one compound of the formula I into a suitable administration form using a physiologically acceptable excipient.
8. The process according to claim 7 further comprising using additives or auxiliaries.
9. The process as claimed in claim 7 or 8 wherein at least one modified or unmodified insulin or insulin derivative is added.
10. A use of at least one compound of the formula I as claimed in any one of claims 1 to 3 for the production of a pharmaceutical preparation for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
11. A use of at least one compound of the formula I as claimed in any one of claims 1 to 3 for the treatment of diabetes mellitus or noninsulin-dependent diabetes.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19649350.1 | 1996-11-28 | ||
| DE19649350A DE19649350A1 (en) | 1996-11-28 | 1996-11-28 | Inositol glycans with an insulin-like effect |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2222103A1 CA2222103A1 (en) | 1998-05-28 |
| CA2222103C true CA2222103C (en) | 2007-08-28 |
Family
ID=7813041
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002222103A Expired - Fee Related CA2222103C (en) | 1996-11-28 | 1997-11-25 | Inositolglycans having insulin-like action |
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|---|---|
| US (1) | US6004938A (en) |
| EP (1) | EP0845475B1 (en) |
| JP (1) | JP4299378B2 (en) |
| CN (1) | CN1136224C (en) |
| AR (1) | AR018246A1 (en) |
| AT (1) | ATE290541T1 (en) |
| AU (1) | AU728637B2 (en) |
| BR (1) | BR9706041C1 (en) |
| CA (1) | CA2222103C (en) |
| CZ (1) | CZ294886B6 (en) |
| DE (2) | DE19649350A1 (en) |
| DK (1) | DK0845475T3 (en) |
| ES (1) | ES2237784T3 (en) |
| HK (1) | HK1008786A1 (en) |
| HU (1) | HUP9702242A3 (en) |
| PL (1) | PL188604B1 (en) |
| PT (1) | PT845475E (en) |
| RU (1) | RU2178794C2 (en) |
| TR (1) | TR199701432A2 (en) |
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| US7312196B2 (en) * | 1997-01-08 | 2007-12-25 | Amylin Pharmaceuticals, Inc. | Formulations for amylin agonist peptides |
| GB9814039D0 (en) * | 1998-06-29 | 1998-08-26 | Univ London | Materials and methods relating to the prevention or treatment of ischaemia-reperfusion injury |
| US6953781B2 (en) | 2000-05-12 | 2005-10-11 | Rodaris Pharmaceuticals Limited | Compounds and their uses |
| US6759390B2 (en) | 2000-05-12 | 2004-07-06 | Manuel Martin-Lomas | Compounds and their uses |
| US6716826B2 (en) | 2000-05-12 | 2004-04-06 | Rodaris Pharmaceuticals Limited | Compounds and their uses |
| EP1409498B1 (en) * | 2000-05-12 | 2008-03-26 | Rodaris Pharmaceuticals Limited | Inositol phosphoglycan derivatives and their medical uses |
| US6939857B2 (en) | 2000-05-12 | 2005-09-06 | Rodaris Pharmaceuticals Limited | Compounds and their uses |
| CA2433183A1 (en) * | 2000-05-12 | 2001-11-15 | Rodaris Pharmaceuticals Limited | Inositolglycans and their uses |
| DE60133137D1 (en) * | 2000-05-12 | 2008-04-17 | Rodaris Pharmaceuticals Ltd | INOSITOLPHOSPHOGLYCANDERIVATE AND ITS MEDICAL USE |
| GB0015627D0 (en) * | 2000-06-26 | 2000-08-16 | Rademacher Group Limited | Phosphoglycan messengers and their medical uses |
| FR2828206B1 (en) * | 2001-08-03 | 2004-09-24 | Centre Nat Rech Scient | USE OF LYSYL OXIDASE INHIBITORS FOR CELL CULTURE AND TISSUE ENGINEERING |
| EP1503768B1 (en) * | 2002-04-29 | 2015-10-28 | NormOxys, Inc. | Inositol pyrophosphates, and use thereof |
| US7943594B2 (en) * | 2002-07-10 | 2011-05-17 | Massachusetts Institute Of Technology | Solid-phase and solution-phase synthesis of glycosylphosphatidylinositol glycans |
| CN100369610C (en) * | 2002-10-09 | 2008-02-20 | 中国药科大学 | Plant extract for preventing and treating diabetes, and preparing method and pharmaceutic use thereof |
| US20070135389A1 (en) * | 2004-07-06 | 2007-06-14 | Claude Nicolau | Tumor eradication by inositol-tripyrophosphate |
| US7745423B2 (en) * | 2004-07-06 | 2010-06-29 | NormOxys, Inc | Calcium/sodium salt of inositol tripyrophosphate as an allosteric effector of hemoglobin |
| US20060106000A1 (en) * | 2004-07-06 | 2006-05-18 | Claude Nicolau | Use of inositol-tripyrophosphate in treating tumors and diseases |
| FR2878850B1 (en) * | 2004-12-02 | 2008-10-31 | Cis Bio Internat Sa | DERIVATIVES OF INOSITOL-1-PHOSPHATE |
| EP2114414B1 (en) * | 2006-12-21 | 2013-11-06 | Trustees Of Tufts College | Synthetic lipophilic inositol glycans for treatment of cancer and glucose-metabolism disorders |
| EP2111226A4 (en) * | 2006-12-29 | 2010-02-10 | Normoxys Inc | Cyclitols and their derivatives and their therapeutic applications |
| EP2152085A4 (en) * | 2007-05-01 | 2012-05-23 | Normoxys Inc | Erythropoietin complementation or replacement |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4446064A (en) * | 1980-12-19 | 1984-05-01 | The University Of Virginia Alumni Patents Foundation | Insulin mediator substance |
| DE3326472A1 (en) * | 1983-07-22 | 1985-02-14 | Hoechst Ag, 6230 Frankfurt | NEW INSULIN DERIVATIVES, METHOD FOR THE PRODUCTION AND USE THEREOF AND PHARMACEUTICAL AGENTS FOR TREATING THE DIABETES MELLITUS |
| DE3326473A1 (en) * | 1983-07-22 | 1985-01-31 | Hoechst Ag, 6230 Frankfurt | PHARMACEUTICAL AGENT FOR TREATING THE DIABETES MELLITUS |
| US4906468A (en) * | 1986-04-11 | 1990-03-06 | The Rockefeller University | Insulin activity messengers, their antibodies, and thereof |
| YU66892A (en) * | 1991-08-20 | 1995-10-24 | Hoechst Ag. | PHOSPHOINOSYTOLGLICAN - A PEPTID WITH ACTION AS INSULIN |
| EP0545198B1 (en) * | 1991-11-29 | 1999-09-15 | Hoechst Aktiengesellschaft | Peptides with insulin like activity |
| JPH06293790A (en) * | 1993-04-08 | 1994-10-21 | Kyowa Hakko Kogyo Co Ltd | Physiologically active substance, inositol glucan |
| US5652221A (en) * | 1994-11-07 | 1997-07-29 | The University Of Virginia Patent Foundation | Method of treating defective glucose metabolism using synthetic insulin substances |
-
1996
- 1996-11-28 DE DE19649350A patent/DE19649350A1/en not_active Withdrawn
-
1997
- 1997-11-13 DE DE59712220T patent/DE59712220D1/en not_active Expired - Fee Related
- 1997-11-13 PT PT97119835T patent/PT845475E/en unknown
- 1997-11-13 AT AT97119835T patent/ATE290541T1/en not_active IP Right Cessation
- 1997-11-13 EP EP97119835A patent/EP0845475B1/en not_active Expired - Lifetime
- 1997-11-13 DK DK97119835T patent/DK0845475T3/en active
- 1997-11-13 ES ES97119835T patent/ES2237784T3/en not_active Expired - Lifetime
- 1997-11-25 CA CA002222103A patent/CA2222103C/en not_active Expired - Fee Related
- 1997-11-26 CN CNB971229589A patent/CN1136224C/en not_active Expired - Fee Related
- 1997-11-26 HU HU9702242A patent/HUP9702242A3/en unknown
- 1997-11-26 AR ARP970105573A patent/AR018246A1/en active IP Right Grant
- 1997-11-26 AU AU45382/97A patent/AU728637B2/en not_active Ceased
- 1997-11-26 TR TR97/01432A patent/TR199701432A2/en unknown
- 1997-11-26 US US08/979,865 patent/US6004938A/en not_active Expired - Fee Related
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- 1997-11-27 JP JP32550897A patent/JP4299378B2/en not_active Expired - Fee Related
- 1997-11-27 CZ CZ19973775A patent/CZ294886B6/en not_active IP Right Cessation
- 1997-11-27 RU RU97120547/04A patent/RU2178794C2/en not_active IP Right Cessation
- 1997-11-28 PL PL97323407A patent/PL188604B1/en not_active IP Right Cessation
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| AU728637B2 (en) | 2001-01-11 |
| HK1008786A1 (en) | 1999-05-21 |
| JP4299378B2 (en) | 2009-07-22 |
| MX9709207A (en) | 1998-05-31 |
| RU2178794C2 (en) | 2002-01-27 |
| AR018246A1 (en) | 2001-11-14 |
| DE19649350A1 (en) | 1998-06-04 |
| CZ377597A3 (en) | 1998-06-17 |
| HUP9702242A3 (en) | 1999-01-28 |
| PL188604B1 (en) | 2005-03-31 |
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| ES2237784T3 (en) | 2005-08-01 |
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| BR9706041C1 (en) | 2000-06-06 |
| EP0845475A1 (en) | 1998-06-03 |
| TR199701432A2 (en) | 1998-06-22 |
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