AU2001265848A1 - Process for the preparation of pharmaceutically acceptable salts of (SS,RS)-S-adenosyl-L-methionine - Google Patents
Process for the preparation of pharmaceutically acceptable salts of (SS,RS)-S-adenosyl-L-methionineInfo
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- AU2001265848A1 AU2001265848A1 AU2001265848A AU2001265848A AU2001265848A1 AU 2001265848 A1 AU2001265848 A1 AU 2001265848A1 AU 2001265848 A AU2001265848 A AU 2001265848A AU 2001265848 A AU2001265848 A AU 2001265848A AU 2001265848 A1 AU2001265848 A1 AU 2001265848A1
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- same
- pharmaceutically acceptable
- process according
- acid
- methionine
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- Granted
Links
- 238000000034 method Methods 0.000 title claims description 28
- 150000003839 salts Chemical class 0.000 title claims description 16
- 238000002360 preparation method Methods 0.000 title claims description 14
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 17
- 239000002253 acid Substances 0.000 claims description 16
- 229920005989 resin Polymers 0.000 claims description 13
- 239000011347 resin Substances 0.000 claims description 13
- 239000006166 lysate Substances 0.000 claims description 9
- 238000000746 purification Methods 0.000 claims description 8
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 7
- 239000001117 sulphuric acid Substances 0.000 claims description 7
- 235000011149 sulphuric acid Nutrition 0.000 claims description 7
- 238000001223 reverse osmosis Methods 0.000 claims description 6
- 238000001471 micro-filtration Methods 0.000 claims description 5
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- 150000007522 mineralic acids Chemical class 0.000 claims description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 13
- 239000000243 solution Substances 0.000 description 12
- 239000000047 product Substances 0.000 description 11
- VFNGKCDDZUSWLR-UHFFFAOYSA-N disulfuric acid Chemical compound OS(=O)(=O)OS(O)(=O)=O VFNGKCDDZUSWLR-UHFFFAOYSA-N 0.000 description 6
- 230000009089 cytolysis Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 241001123227 Saccharomyces pastorianus Species 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 241000235646 Cyberlindnera jadinii Species 0.000 description 2
- 239000004470 DL Methionine Substances 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 235000003534 Saccharomyces carlsbergensis Nutrition 0.000 description 2
- 239000000919 ceramic Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 235000006109 methionine Nutrition 0.000 description 2
- FFEARJCKVFRZRR-UHFFFAOYSA-N methionine Chemical compound CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- CHRJZRDFSQHIFI-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;styrene Chemical group C=CC1=CC=CC=C1.C=CC1=CC=CC=C1C=C CHRJZRDFSQHIFI-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 238000005684 Liebig rearrangement reaction Methods 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 241000775010 Saccharomyces pastorianus CBS 1513 Species 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- JXLHNMVSKXFWAO-UHFFFAOYSA-N azane;7-fluoro-2,1,3-benzoxadiazole-4-sulfonic acid Chemical compound N.OS(=O)(=O)C1=CC=C(F)C2=NON=C12 JXLHNMVSKXFWAO-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000012267 brine Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 229920006026 co-polymeric resin Polymers 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- AXZAYXJCENRGIM-UHFFFAOYSA-J dipotassium;tetrabromoplatinum(2-) Chemical compound [K+].[K+].[Br-].[Br-].[Br-].[Br-].[Pt+2] AXZAYXJCENRGIM-UHFFFAOYSA-J 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006200 ethylation reaction Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- -1 for example Chemical class 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 1
- 235000015497 potassium bicarbonate Nutrition 0.000 description 1
- 239000011736 potassium bicarbonate Substances 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 229910001487 potassium perchlorate Inorganic materials 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 239000013557 residual solvent Substances 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Description
PROCESS FOR THE PREPARATION OF PHARMACEUTICALLY ACCEPTABLE SALTS OF (SS,RS) -S-ADENOSYL-L-METHIONINE
The present invention relates to a process for the preparation of pharmaceutically acceptable salts of (SS,RS) -S-adenosyl-L-methionine (hereinafter referred to as (SS,RS)-SAMe) .
In particular, the invention relates to a process for the preparation of pharmaceutically acceptable salts of (SS,RS) -SAMe, wherein the salified (RS) -(+) -S-adenosyl-L- methionine diastereoisomer (hereinafter referred to .as (RS) - (+) -SAMe) is produced in amounts lower than .or equal to 3% with respect to the salified (SS) -(+) -S-adenosyl-L- methionine diastereoisomer (hereinafter referred to as (SS)-(+)-SAMe) .
As it is known, (SS,RS)-SAMe is a physiological methyl donor involved in enzymatic trans ethylation reactions, that is present in all living organisms and has therapeutical effects on chronic hepatic diseases, adiposis, lipaemia, atherosclerosis and it is desirable, therefore, to produce it in high amounts.
It is also known, (J. . Cornforth, J.A.C.S., 1977, 99,
7292-7300; Stolowitz et al . , J.A.C.S., 1981, 103, 6015-
6019) that the products containing (SS,RS)-SAMe consist of a mixture of two diastereoisomers: (RS) - (+) -SAMe and (SS)-
(+)-SAMe, having the following structural formulae:
SS RS
Moreover, it was demonstrated (De La Haba et al., J.A.C.S., 1959, 81, 3975-3980) that only one of the two diastereoisomers, i.e. (SS) - (+) -SAMe, is enzymatically active for the transmethylation and spontaneously racemises, 'thereby giving rise to the formation of the inactive diastereoisomer (RS) - (+) -SAMe in a percentage equal to about 20% ( u et al . , Biochemistry 1983, 22, 2828-2832) .
The Applicant, in fact, has noted that in all the commercially available products based on (SS, RS) -SAMe, the inactive diastereoisomer (RS) - (+) -SAMe is present in percentages equal to at least 20%; it was also noted that said percentages increase in time even up to 40% and more.
This observation clearly confirms that the diasteroisomer mixture is unstable in time, which, on the other side, had already been noted in relation with the product in solution (G. L. Creason et al., Phytochemistry, vol. 24, N. 6, 1151-1155, 1985; H. C. Uzar, Liebigs Ann. Chem. 1989, 607-610) . The demand for (SS,RS)-SAMe derivatives wherein the percentage of the active (SS) - (+) -SAMe diastereoisomer is clearly higher with respect to the inactive (RS) - (+) -SAMe isomer and wherein said percentage turns out to be stable in time, is particularly felt in the field. It was also found that there is an obstacle to the use of (SS,RS)-SAMe and the pharmaceutically acceptable salts thereof at the industrial level because of their thermal instability, even at room temperature, and of the complexity of the preparation and purification processes thereof.
Several processes for the purification of (SS,RS)-SAMe and for the production of the pharmaceutically acceptable salts thereof are known.
However, the known purification processes, besides providing the use of strong acid resins (JP 13680/1971) or
chelate-type resins (JP 20998/1978) or particular and expensive reactants, such as picric or. picolmic acid' (US 3707536 and US 3954726), bring anyhow to the partial racemisation of the sulphur chiral center of (SS,RS)-SAMe and, therefore, lead to final products containing the inactive diastereoisomer in amounts higher than 20%.
Purification processes that use weak acid resins are also known (JP 14299/1981, FR-A-2531714, EP-A-0141914) , which allow, however, to obtain just a partial separation of (SS,RS)-SAMe and, therefore, an insufficient purity degree for pharmaceutical purposes.
Even if the realization of some of the above- identified processes enables to obtain a higher purity, the partial racemization implies, at any event, that at least 20% of the inactive diastereoisomer should be present; in some cases moreover (FR-2531714) , in order to extract the product from the cells, there is provided the use of potassium bicarbonate, • with subsequent precipitation of potassium perchlorate, which brings about problems firstly in the separation and then in the disposal of the product. In EP-A-0141914, the lysis of the cells of the yeast containing (SS,RS)-SAMe is carried out in the presence of .an organic solvent (for example, ethyl acetate, acetone, etc.) by using, moreover, chromatographic columns based on 100-200 mesh resins, with high investment and maintaining costs. The use of solvents for the extraction of (SS,RS)-SAMe necessarily implies the employment of antideflagrant plants and a recovery, distillation and solvent recovery systems, besides the necessary drying of the exhausted mycelium, in order to avoid that it is discharged with the residual solvent, all these factors clearly bringing about additional investment and operation costs.
According to a first aspect, the present invention relates to a process for the preparation of
pharmaceutically acceptable salts of (SS, RS) -SAMe, wherein the salified (RS) - (+) -SAMe diastereoisomer is present in amounts lower than or equal to 3% with respect to the salified (SS) - (+) -SAMe diastereoisomer, which, at a temperature higher than or equal to 0-12 °C, comprises:
- the purification of (SS, RS) -S-adenosyl-L-methionine from enriched yeast, which shall contain at least 6 g/1 thereof, which comprises:
(a) - the adjustment of the pH value to 1.2-3.5; (b) - the preparation of an aqueous lysate of (SS,RS)- SAMe from the enriched yeast;
(c) - the microfiltration of the resulting lysate;
(d) - the absorption of the resulting microfiltrate on a weak acid resin, by eluting with a 0.1-2 N inorganic acid solution;
(e) - the decolouration of the resulting eluate;
- the concentration of the decolourised eluate, by reverse osmosis, from 30 to 70% by volume;
- the addition of stoichiometric amounts of at least one pharmaceutically acceptable acid salt to the concentrated eluate, so as to obtain the corresponding pharmaceutically acceptable salt of (SS, RS) -SAMe.
According to a preferred aspect, the so obtained pharmaceutically acceptable salt of (SS,RS)-SAMe can be subjected to lyophilization.
According to another preferred aspect, the process of the invention is carried out at a temperature of 2-5 °C.
According to a further preferred aspect, the pH value in step (a) is 1-2, whereas the preparation of the lysate in step (b) can take place by passing the yeast through a breaking-cells equipment, then proceeding with the microfiltration of the so obtained yeast, for example on a ceramic membrane.
The enriched yeast on which (SS,RS)-SAMe is purified preferably contains at least 8-10 g/1 of (SS, RS) -SAMe; the
pharmaceutically acceptable acid is selected, preferably, from sulphuric acid and paratoluensulphonic acid.
It can be noted that the process of the invention allows to use resin/product ratios equal to, for example, 10-20 liters of resin pro kg of absorbed pro'duct, which are advantageous with respect to what has been disclosed in JP 20998/1978. -, ''
The process of the invention allows to produce salts of (SS,RS)-SAMe wherein, even at room temperature, it is possible to detect a percentage of the (SS) - (+) -SAMe diastereoisomer equal to at least 97 with respect to the (RS) - (+) -SAMe diastereoisomer which is present, accordingly, in percentages lower than or equal to 3.
The process of the invention allows moreover to exclude the use of organic solvents in the preparation of the lysate, with remarkable advantages with respect to the purification steps of the pharmaceutically acceptable salts of (SS,RS) -SAMe, as well as ecological and environmental advantages . It is furthermore possible to obtain a higher yield and purity of the pharmaceutically acceptable salts of (SS,RS)-SAMe with respect to those obtainable by known processes; a purity equal to at least 98% in (SS,RS)-SAMe and a yield equal to at least 90 are obtained, in fact, with respect to the fermented product.
Thanks to its particular conditions, the process of the invention allows to avoid the degradation of (SS,RS)- SAMe during the preparation of the lysate and allows to obtain a lysis with a yield higher than 98% and with a content of by-products, the main product of which being 5- deacyl-5-methylthioadenosine, lower than 1%.
(SS, RS) -SAMe, suitably salified as above described, can be produced, for example, by fermenting a suitable microorganism, such as Saccharomyces pastorianus (ex Saccharomyces carlsbergensis CBS 1513) , Saccharomyces
cerevisiae (IFO 2044) , Torulopsis utilis and Candida utilis .
The yeast containing (SS,RS)-SAMe can be enriched by the processes known in the field, such as for example, the Schlenk method described in "Journal of Biological Chemistry", vol. 29, page 1037, (1987), which was modified only in optimizing the use of DL methionine and which was conducted at a maximum temperature of 27,5°C for about 20 hours . The (SS,RS) -SAMe-enriched yeast (which, in order to be advantageously employed in the realization of the present invention, indicatively contains at least 6 g/1 of (SS,RS) -SAMe, undergoes, upon adjustment of the pH value to 1.2-3.5, a cellular lysis process, by passing the yeast, preferably, through a cell-breaking equipment.
The resulting lysate, after being subjected to microfiltration, for example on a ceramic membrane such as Kerasep® K09A, is adsorbed on a weak acid carboxylic resin, preferably of the cationic type, such as Rohm and Haas® IRC86, preferably until saturation (about 150 g/1), and eluted with a solution of an inorganic acid such as, for example, 0.1-2 N sulphuric acid, hydrochloric acid, etc.
The decolouration of the resulting eluate takes then place, for example by means of a copolymer resin with a styrene-divinylbenzene unit, such as Resindion® 825L.
The resulting eluate containing (SS,RS)-SAMe is concentrated, by reverse osmosis, from 30 to 70%, preferably from 40 to 50% by volume. The so obtained concentrate is added with stoichiometrically equivalent amounts of an acid or a mixture of pharmaceutically acceptable acids, such as those indicated above. The so obtained products can be used for possible preparations in solution or can be subjected to lyophilisation, when one
wishes to use them in the solid form.
The following examples illustrate the invention without limiting it. EXAMPLE 1 1000 kg of yeast obtained by fermentation of Saccharomyces carlsbergensis were enriched with (SS,RS)- SAMe according to the Schlenk method, modified as follows. The yeast was added with 100 kg of yeast cream (which, upon dilution with 100 1 of deionised water has a 2.2 g/1 titer) , 2 kg of DL methionine, 12 kg of hydrated glucose and 1.5 kg of citric acid, keeping under stirring at 27 °C + 0,5°C for 22 hours, aerating through emission of sterile filtered air at a flow of 0.6 1/1/m, thereby obtaining 9 g/1 of (SS,RS) -SAMe. After adjusting pH at 1.2 by means of H2S04, lysis was carried out, at a temperature of 12 °C, by the " Constant Cell Disruption System" produced by Constant System Ltd. , a pressure-type cell-breaking system with a cooling system. The solution was then cooled by using first cold water and then brine, until the solution was brought to a temperature of about 2°C.
The obtained mixture was then conveyed to a microfitration plant, endowed with cartridges of the type Verind A-10 HFM 180 SM, for separating the exhausted solid from the enriched liquid. The panel was washed with 2000 1 of demineralised water at 2°C. The filtration yield was 98%.
The enriched solution was passed through the IRC 86 resin (Rohm and Haas®) , a carboxylic resin and eluted with 1 N sulphuric acid, still keeping the temperature at about 2°C.
The collected eluate was decoulorised by using a Resindion® 825L resin. The enriched solution was concentrated by reverse osmosis until a 40% concentration of (SS,RS)-SAMe was obtained. Corresponding stoichiometric amounts of sulphuric acid and paratoluensulphonic acid
were then added to give the disulphate paratoluensulphonate of (SS, RS) -SAMe . The final yield of (SS,RS)-SAMe disulphate paratoluensulphate was 90%.
The content of (RS) - (+) -SAMe disulphate paratoluen.sulphonate in the diastereoisomer mixture of
(SS,RS)-SAMe disulphate paratoluensulphonate, analyzed by
HPLC, turned out to be 1%. The relevant data are reported in the following, in the table concerning sample N. 4.
EXAMPLE 2 1000 kg of yea-st, obtained by fermentation of Saccharomyces carlsbergensis enriched with (SS,RS)-SAMe according to the method described in EXAMPLE 1, with an activity equal to 8.2 g/kg, were lysated by a cell- breaking system at a temperature of 12 °C and at a 2 pH. After adding 500 1 of water to the resulting solution, the microfiltration and the subsequent steps were carried out, analogously to what described in EXAMPLE 1, washing with
2000 1 of cold demineralized water (about 5°C) . 7,5 kA of
(SS,RS)-SAMe were obtained which, after being concentrated by reverse osmosis, were salified obtaining a 91,4% yield of (SS,RS)-SAMe disulphate paratoluensulphonate (lysis yield: 99%; purification yield: 98%) . The time elapsing from the end of the fermentation to the concentration by reverse osmosis was 32 hours. The relevant data are reported in the following, in the table concerning sample N. 5. i
EXAMPLE 3
The solution obtained by the process of EXAMPLE 1, after absorption on IRC 86 (Rohm and Haas®) resin, was eluted with 1 N sulphuric acid.
The obtained solution was concentrated up to 20% and then added with sulphuric acid and paratoluensulphonic acid in a stoichiometric amount, thereafter it was further concentrated until a 40% solution was obtained. 14.09 kg
of (SS,RS)-SAMe disulphate paratoluensulphonate were obtained, with a transformation yield of 97.8%. The relevant data are reported in the following, in the table relating to sample N. 6.
EXAMPLE 4 (comparative)
3 samples of SAMIR® [ (SS, RS) -SAMe] , produced by Knoll Farmaceutici S.p.A., were analyzed by HPLC. The values measured for each sample are as follows:
SAMPLE 1 - 100 mg of SAMIR® (vials) batch 045-021; expiration date 06/20Q0.
Peak No. 5, corresponding to (SS) - (+) -SAMe, indicates a percentage of 58%, whereas peak No. 6, corresponding to (RS) - (+) -SAMe, indicates a percentage of 42%.
SAMPLE 2 - 200 mg of SAMIR® (tablets); batch 121; expiration date 05/2002.
Peak No. 5, corresponding to (SS) - (+) -SAMe, indicates a percentage of 60%, whereas peak No. 6, corresponding to
(RS) - (+) -SAMe, indicates a percentage of 40%.
SAMPLE 3 - 400 mg of SAMIR® (tablets); batch 040; expiration date 10/2002.
Peak No. 3, correspond ng to (SS) - (+) -SAMe, indicates a percentage of 59%, whereas peak No. 4, corresponding to (RS) - (+) -SAMe, indicates a percentage of 41%.
EXAMPLE 5
The products obtained according to the process of the invention in EXAMPLES 1-3, samples 4-6 respectively, were analyzed, similarly to what has been described in example 4, after four months from the date of their production.
The measured values for each sample were as follows :
SAMPLE 4 (EXAMPLE 1); batch 003/R.
Peak No. 7, corresponding 'to (SS) - (+) -SAMe, indicates a percentage of 99%, whereas peak No. 8, corresponding to (RS) - (+) -SAMe, indicates a percentage of 1%.
SAMPLE 5 (Example 2); KF = 2.3%; titre = 102.6%; batch
001/R.
Peak No. 7, corresponding to (SS) - (+) -SAMe, indicates a percentage of 98%, whereas peak No. 8, corresponding to (RS) - (+) -SAMe, indicates a percentage of 2%.
SAMPLE 6 (EXAMPLE 3); KF = 1.39%; titre = 102.7; batch 004/R.
Peak No. 7, corresponding to (SS) - (+) -SAMe, indicates a percentage of 98%, whereas peak No. 6, corresponding to (RS) - (+) -SAMe, indicates a percentage of 2%.
Claims (8)
1. A process for the preparation of pharmaceutically acceptable salts of (SS,RS) -SAMe, wherein the salified (RS) - (+) -SAMe diastereoisomer is present in amounts lower than or equal to 3% with respect to the salified (SS)-(+)- SAMe diastereoisomer, which, at a temperature of 0-12 °C, comprises:
- the purification of (SS, RS) -S-adenosyl-L-methionine from enriched yeast, which shall contain at least 6 g/1 thereof, which comprises:
(a) - the adjustment of the pH value to 1.2-3.5;
(b) - the preparation of an aqueous lysate of- (SS,RS)- SAMe from the enriched yeast;
(c) - the microfiltration of the resulting lysate; (d) - the absorption of the resulting icrofiltrate on a weak acid resin, by eluting with a 0.1-2 N inorganic acid solution;
(e) - the decolouration of the resulting eluate; the concentration of the decolourised eluate by reverse osmosis, from 30 to 70% by volume;
- the addition of stoichiometric amounts of at least a pharmaceutically acceptable acid to the concentrated eluate, so as to obtain the corresponding pharmaceutically acceptable salt of (SS, RS) -SAMe .
2. Process according to claim 1, wherein the pharmaceutically acceptable salt of (SS,RS)-SAMe is subjected to lyophilization.
3. Process according to claim 1 or 2, wherein the pH value in step (a) is 1-2.
4. Process according to any of the preceding claims, wherein the preparation of the lysate in step (b) takes place by passing the yeast through a cell-breaking equipment .
5. Process according to any of the preceding claims, wherein the temperature is 2-5 °C.
6. Process according to any of the preceding claims, wherein the enriched yeast contains at least 8-10 g/1 of
(SS,RS) -S-adenosyl-L-methionine.
7. Process according to any of the preceding claims, i wherein the pharmaceutically acceptable acid is selected from sulphuric acid and paratoluensulphonic acid.
8. A pharmaceutically acceptable salt of- (SS,RS)-S- adenosyl-L-methionine obtainable by the process according to any of the preceding claims .
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IT2000MI001158A IT1318535B1 (en) | 2000-05-25 | 2000-05-25 | PROCESS FOR THE PREPARATION OF PHARMACEUTICALLY ACCEPTABLE SALTS OF (SS, RS) -S-ADENOSYL-METHIONINE. |
| ITMI2000A001158 | 2000-05-25 | ||
| PCT/EP2001/003633 WO2001090130A1 (en) | 2000-05-25 | 2001-03-30 | Process for the preparation of pharmaceutically acceptable salts of (ss-rs)-s-adenosyl-l-methionine |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| AU2001265848A1 true AU2001265848A1 (en) | 2002-02-21 |
| AU2001265848B2 AU2001265848B2 (en) | 2006-03-23 |
| AU2001265848B8 AU2001265848B8 (en) | 2006-05-04 |
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| AU6584801A Pending AU6584801A (en) | 2000-05-25 | 2001-03-30 | Process for the preparation of pharmaceutically acceptable salts of (ss-rs)-s-adenosyl-l-methionine |
| AU2001265848A Expired AU2001265848B8 (en) | 2000-05-25 | 2001-03-30 | Process for the preparation of pharmaceutically acceptable salts of (SS,RS)-S-adenosyl-L-methionine |
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| AU6584801A Pending AU6584801A (en) | 2000-05-25 | 2001-03-30 | Process for the preparation of pharmaceutically acceptable salts of (ss-rs)-s-adenosyl-l-methionine |
Country Status (14)
| Country | Link |
|---|---|
| US (2) | US20020010147A1 (en) |
| EP (1) | EP1283845B1 (en) |
| JP (1) | JP4777588B2 (en) |
| KR (1) | KR100737193B1 (en) |
| AT (1) | ATE256695T1 (en) |
| AU (2) | AU6584801A (en) |
| CA (1) | CA2407559C (en) |
| DE (1) | DE60101575T2 (en) |
| DK (1) | DK1283845T3 (en) |
| ES (1) | ES2208610T3 (en) |
| IT (1) | IT1318535B1 (en) |
| PT (1) | PT1283845E (en) |
| TR (1) | TR200302297T4 (en) |
| WO (1) | WO2001090130A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2002307018A1 (en) | 2001-03-30 | 2002-10-15 | Board Of Regents | Biosynthesis of s-adenosylmethionine in a recombinant yeast strain |
| CN100398659C (en) * | 2002-09-16 | 2008-07-02 | 上海青瑞生物医药技术有限公司 | Method for preparing optically pure (s, s)-adenosylmethionine |
| US20050272687A1 (en) * | 2004-06-08 | 2005-12-08 | Hebert Rolland F | Stable S-adenosyl-l-methionine |
| US20090012036A1 (en) * | 2005-05-24 | 2009-01-08 | Hebert Rolland F | Stable S-adenosyl-L-methionine |
| PL1731596T3 (en) * | 2005-06-09 | 2013-08-30 | Gnosis Spa | Dried, freeze-dried and/or microencapsulated Saccharomyces cerevisiae cells with a high content of (S)-(+)-S-adenosyl-L-methionine |
| ITRM20050344A1 (en) * | 2005-06-30 | 2007-01-01 | Luca Maria De | SALTS OR COMPLEXES OF METHYL-DONOR SUBSTANCES WITH PHYTIC ACID OR ITS DERIVATIVES AND RELATIVE METHOD OF SYNTHESIS. |
| CN1955306B (en) * | 2005-10-28 | 2011-03-16 | 上海医药工业研究院 | Method of saccharomycetes bio-synthesing, of producing optical pure adenosine methionine and its culture solution |
| ITMI20060629A1 (en) * | 2006-03-31 | 2007-10-01 | Daniele Giovannone | ORAL SOLID COMPOSITIONS BASED ON S-ADENOSYLMETIONINE AND PROCESS FOR THEIR ACHIEVEMENT |
| WO2007129701A1 (en) | 2006-05-10 | 2007-11-15 | Mitsubishi Gas Chemical Company, Inc. | Method of producing dry yeast containing s-adenosyl-l-methionine and composition for oral intake |
| US20070265211A1 (en) * | 2006-05-12 | 2007-11-15 | Matthias Rath | Novel composition and method effective in inhibiting the atherogenic process |
| ITMI20071374A1 (en) * | 2007-07-10 | 2009-01-11 | Gnosis Spa | STABLE STABLE OF S-ADENOSYLMETHIONINE AND PROCESS FOR THEIR ACHIEVEMENT. |
| US20100004191A1 (en) * | 2008-07-01 | 2010-01-07 | Rolland F Hebert | Compositions of S-adenosyl-L-methionine. |
| JP6479681B2 (en) | 2013-01-16 | 2019-03-06 | ヘーベルト サム−イー エルエルシー | Stable indole-3-propionate salts of S-adenosyl-L-methionine |
| ITMI20130426A1 (en) * | 2013-03-20 | 2014-09-21 | Gnosis Spa | S-ADENOSYLMETHIONINE STERILE HIGH-ISOMER CONTENT ACTIVE FOR INJECTABLE SOLUTIONS AND PROCEDURE TO OBTAIN IT |
| CN106349311B (en) * | 2016-08-23 | 2018-04-13 | 北京金阳利康医药有限公司 | The method that S Ademetionines are extracted from yeast fermentation broth |
| IT201700074957A1 (en) * | 2017-07-04 | 2019-01-04 | Gnosis Spa | SALT OF (SS) -ADENOSYL METHIONINE WITH ESAFOSPHATE INOSITOL AND PROCEDURE TO OBTAIN IT |
| CN110396120B (en) * | 2019-02-13 | 2020-05-12 | 山东惠仕莱生物科技有限公司 | Method for extracting and separating S-adenosylmethionine from S-adenosylmethionine fermentation liquor |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IE39517B1 (en) * | 1973-06-27 | 1978-10-25 | Bioresearch Sas | Double salts of s-adenosyl-l-methhionine |
| JPS56145299A (en) * | 1980-04-11 | 1981-11-11 | Kanegafuchi Chem Ind Co Ltd | Purification of s-adenosyl-l-methionine |
| JPS5929700A (en) * | 1982-08-13 | 1984-02-16 | Nippon Zeon Co Ltd | Purification of s-adenosyl-l-methionine |
| IT1169773B (en) * | 1983-08-24 | 1987-06-03 | Bioresearch Spa | PROCESS FOR THE PRODUCTION OF STABLE SALTS OF SULPHO-ADENOSYL-L-METHIONINE |
| IT1173990B (en) * | 1984-05-16 | 1987-06-24 | Bioresearch Spa | STABLE SALTS OF SULPHO-ADENOSYL-METHIONINE (SAME) PARTICULARLY SUITABLE FOR PARENTERAL USE |
-
2000
- 2000-05-25 IT IT2000MI001158A patent/IT1318535B1/en active
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2001
- 2001-03-30 DE DE60101575T patent/DE60101575T2/en not_active Expired - Lifetime
- 2001-03-30 DK DK01943206T patent/DK1283845T3/en active
- 2001-03-30 WO PCT/EP2001/003633 patent/WO2001090130A1/en active IP Right Grant
- 2001-03-30 AT AT01943206T patent/ATE256695T1/en active
- 2001-03-30 JP JP2001586317A patent/JP4777588B2/en not_active Expired - Lifetime
- 2001-03-30 TR TR2003/02297T patent/TR200302297T4/en unknown
- 2001-03-30 KR KR1020027015667A patent/KR100737193B1/en not_active Expired - Lifetime
- 2001-03-30 EP EP01943206A patent/EP1283845B1/en not_active Expired - Lifetime
- 2001-03-30 AU AU6584801A patent/AU6584801A/en active Pending
- 2001-03-30 PT PT01943206T patent/PT1283845E/en unknown
- 2001-03-30 AU AU2001265848A patent/AU2001265848B8/en not_active Expired
- 2001-03-30 CA CA002407559A patent/CA2407559C/en not_active Expired - Lifetime
- 2001-03-30 ES ES01943206T patent/ES2208610T3/en not_active Expired - Lifetime
- 2001-04-11 US US09/829,906 patent/US20020010147A1/en not_active Abandoned
-
2002
- 2002-05-13 US US10/142,876 patent/US6958233B2/en not_active Expired - Lifetime
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