U.S. patent application number 12/240002 was filed with the patent office on 2009-04-30 for solid oral compositions based on s-adenosyl methionine and/or nadh and process for obtaining them.
Invention is credited to CARLO DE ANGELIS, DANIELE GIOVANNONE.
Application Number | 20090110729 12/240002 |
Document ID | / |
Family ID | 37327762 |
Filed Date | 2009-04-30 |
United States Patent
Application |
20090110729 |
Kind Code |
A1 |
GIOVANNONE; DANIELE ; et
al. |
April 30, 2009 |
SOLID ORAL COMPOSITIONS BASED ON S-ADENOSYL METHIONINE AND/OR NADH
AND PROCESS FOR OBTAINING THEM
Abstract
This invention relates to solid oral compositions based on SAMe
and/or NADH or their salts in association with calcium oxide and/or
calcium hydroxide and a process for obtaining them. This invention
also relates to a method for stabilising a solid oral composition
based on SAMe and/or NADH or their salts, making use of calcium
oxide, calcium hydroxide optionally in association with malic acid,
glutamic acid, xylitol, calcium sulphate hemihydrate, magnesium
oxide and/or mixtures thereof. This invention also relates to the
use of SAMe or its salts in association with calcium oxide and/or
calcium hydroxide with the possible further addition of melatonin
and/or 1-theanine and/or 1-tryptophan and/or 5-hydroxytryptophan
for the treatment of depressive states.
Inventors: |
GIOVANNONE; DANIELE;
(FROSINONE, IT) ; DE ANGELIS; CARLO; (FONTANA
LIRI, IT) |
Correspondence
Address: |
DYKAS, SHAVER & NIPPER, LLP
P.O. BOX 877
BOISE
ID
83701-0877
US
|
Family ID: |
37327762 |
Appl. No.: |
12/240002 |
Filed: |
September 29, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
PCT/IT2006/000610 |
Aug 8, 2006 |
|
|
|
12240002 |
|
|
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|
Current U.S.
Class: |
424/474 ;
514/44R; 514/46 |
Current CPC
Class: |
A61K 9/2095 20130101;
A61K 9/282 20130101; A61K 31/7076 20130101; A61K 31/52 20130101;
A61K 31/7084 20130101; A61P 25/24 20180101; A61K 9/2009 20130101;
A61K 9/0007 20130101 |
Class at
Publication: |
424/474 ; 514/46;
514/44 |
International
Class: |
A61K 9/28 20060101
A61K009/28; A61K 31/7076 20060101 A61K031/7076; A61P 25/24 20060101
A61P025/24; A61K 31/7084 20060101 A61K031/7084 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 31, 2006 |
IT |
MI2006A000629 |
Claims
1. A composition comprising SAMe and/or NADH or its salts in
association with calcium oxide and optionally pharmaceutically
acceptable excipients.
2. A composition according to claim 1, in which the said SAMe is
S-adenosyl methionine paratoluene sulphonate, S-adenosyl
methionine-1,4-butene disulphonate, S-adenosyl methionine sulphate,
S-adenosyl methionine tosylate.
3. A composition according to claim 1, in which the SAMe is present
in a quantity varying from approximately 30 to approximately 90% by
weight with respect to the weight of the composition.
4. A composition according to claim 3, in which the SAMe is present
in a quantity varying from approximately 50 to 85% by weight with
respect to the weight of the composition.
5. A composition according to claim 1, in which the NADH is present
in a quantity varying from approximately 1 to approximately 90% by
weight with respect to the weight of the composition.
6. A composition according to claim 5, in which the NADH is present
in a quantity varying from approximately 5 to 50% by weight with
respect to the weight of the composition.
7. A composition according to claim 1, in which the calcium oxide
is present in a quantity varying from approximately 1 to
approximately 40% by weight with respect to the weight of the
composition.
8. A composition according to claim 1, in which the calcium oxide
is present in a quantity varying from approximately 2 to
approximately 20% by weight with respect to the weight of the
composition.
9. A composition according to claim 1, comprising at least one
further active ingredient preferably selected from 1-melatonin,
1-theanine and/or 1-tryptophan and/or 5-hydroxytryptophan or
mixtures thereof.
10. A composition according to claim 1, in which at least one of
the pharmaceutically acceptable excipients is calcium sulphate
hemihydrate and/or glucono-delta-lactone.
11. A composition according to claim 1, in the form of a direct
mixture, tablet, capsule, granulate or powder.
12. A composition according to claim 1, in the form of a tablet,
preferably an ordinary, coated, film-coated and/or gastroresistant
tablet.
13. A composition according to claim 1, characterised in that it is
in the form of a gastroresistant tablet.
14. A process for the preparation of a tablet according to claim 1,
comprising the following stages: a) mixing of the SAMe and/or NADH
and/or their salts with calcium oxide and pharmaceutically
acceptable excipients, b) precompression and subsequent granulation
of the mixture obtained in stage a), c) mixing of the granulate
obtained in stage b) with pharmaceutically acceptable excipients
such as calcium sulphate hemihydrate, xylitol, malic acid, glutamic
acid, glucono-delta-lactone, magnesium oxide, hydrogenated fatty
acids, precipitated silica, magnesium stearate, saccharose,
glycerol behenate, d) compression of the mixture obtained in stage
c), with the optional addition of sweeteners and/or flavourings, e)
optional coating of the tablet obtained in stage d) with
hydrogenated fatty acids, f) optional aqueous phase film-coating of
the tablet obtained in stage e).
15. A process according to claim 14 in which the SAMe is S-adenosyl
methionine paratoluene sulphonate, S-adenosyl methionine-1,4-butene
disulphonate, S-adenosyl methionine sulphate, S-adenosyl methionine
tosylate.
16. A process according to claim 14, in which the said calcium
oxide is calcium oxide alone.
17. A process according to claim 14, in which the absolute moisture
content is less than approximately 50%-60% and the temperature is
maintained around 20.degree. C. and 25.degree. C., in particular
around 20.degree. C.
18. A process according to claim 14, in which in stage c) at least
one further active ingredient preferably selected from 1-melatonin,
1-theanine and/or 1-tryptophan and/or 5-hydroxytryptophan and/or
mixtures thereof may be added to the mixture.
19. A process according to claim 14, in which the coating mentioned
in stage e) is applied using hydrogenated fatty acids in a quantity
of between 0.4 and 1.5% by weight with respect to the weight of the
tablet.
20. A process according to claim 14, in which the aqueous phase
film-forming mentioned in stage f) is performed using a varnish
preferably selected from gum Lac and/or its salts (Shellac.TM.),
methacrylic acid, cellulose acetophthalates, titanium dioxide,
talc, triethyl citrate, PVP K30, curcumin, lutein,
hydroxypropylcellulose, hydroxypropylmethylcellulose and/or
mixtures thereof.
21. A process according to claim 20, in which the varnish lies
within a range between approximately 1.0 and approximately 5.0% by
weight with respect to the composition.
22. Compositions which can be obtained through the process in
claims 14.
23. A method for the treatment of depressive states which comprises
the administration of SAMe or its salts in association with calcium
oxide to a patient in such a need.
24. A method according to claim 23 in which magnesium oxide is
further added to the said calcium oxide.
25. A method according to claim 23, in which the said SAMe is
S-adenosyl methionine paratoluene sulphonate, S-adenosyl
methionine-1,4-butene disulphonate, S-adenosyl methionine sulphate,
S-adenosyl methionine tosylate.
26. A method according to claim 23, in which the said calcium oxide
is calcium oxide alone.
27. A method for stabilising a composition based on SAMe or its
salts comprising use of the mixture of SAMe and its salts with
calcium oxide.
28. A method for stabilising a composition based on NADH or its
salts comprising use of the mixture of NADH and its salts with
calcium oxide.
29. A method according to claim 27, in which SAMe or its salts is
present in a quantity of between approximately 30 and approximately
90% by weight calculated in relation to the weight of the
composition.
30. A method according to claim 27, in which SAMe or its salts is
present in a quantity of between approximately 50 and approximately
85% by weight calculated in relation to the weight of the
composition.
31. A method according to claim 28, in which NADH or its salts is
present in a quantity of between approximately 1 and approximately
90% by weight calculated in relation to the weight of the
composition.
32. A method according to claim 28, in which NADH or its salts is
present in a quantity of between approximately 5 and approximately
50% by weight calculated in relation to the weight of the
composition.
33. A method according to claim 27 in which the calcium oxide is
present in a quantity which varies from approximately 1 to
approximately 40% by weight with respect to the weight of the
composition.
34. A method according to claim 33, in which the calcium oxide is
present in a quantity which varies from approximately 2 to
approximately 20% by weight with respect to the weight of the
composition.
35. A method according to claim 33, comprising the addition of
pharmaceutically acceptable excipients.
Description
[0001] This application is a continuation-in-part of PCT
application IT2006/000610 filed Aug. 8, 2006, which claims priority
benefit of Italian application Serial No. MI2006A000629, file Mar.
31, 2006, the contents of which are hereby incorporated by
referenced into the present disclosure as if fully put forth
therein.
[0002] S-adenosyl methionine (SAMe) is a physiological donor of
methyl groups present in all living organisms and is involved in
enzyme transmethylation reactions.
[0003] This substance therefore has a very important biological
role and is essentially used in clinical practice as an
antidepressant.
[0004] By "SAMe" is meant both the racemic mixture and the
individual diastereoisomers (RS)-(+)-S-adenosyl-L-methionine
[(RS)-(+)-SAMe)] and (SS)-(+)-S-adenosyl-L-methionine
[(SS)-(+)-SAMe)], as well as mixtures other than the racemic
mixture.
[0005] The difficulty of using S-adenosyl methionine as a drug
and/or dietetic is however known because it is extremely unstable
at temperatures above 0.degree. C. or in the presence of moisture,
through both degradation of the active ingredient, understood to be
the sum of the two diastereoisomers, and through the conversion of
active (SS)-(+)-S-adenosyl-L-methionine to inactive
(RS)-(+)-S-adenosyl-L-methionine (racemisation of the
substance).
[0006] Italian Patent no. 829906 describes a process for the
preparation of pharmaceutically acceptable salts of
(SS,RS)-S-adenosyl-L-methionine with quantities of inactive
diastereoisomer (RS)-(+)-S-adenosyl-L-methionine of 3% or less with
respect to the active diastereoisomer
(SS)-(+)-S-adenosyl-L-methionine of 97% or more. The same applies
with regard to the need to use racemic mixtures with a high
percentage of the active S,S diastereoisomer as this is the only
one which is pharmacologically active. However, the patent confirms
that although more than 97% of active S,S diastereoisomer is
obtained at ambient temperature, the racemic mixture is unstable
over time, with conversion of the (SS)-(+)-S-adenosyl-L-methionine
into (RS)-(+)-S-adenosyl-L-methionine in a relatively short
time.
BRIEF DESCRIPTION OF THE DRAWINGS
[0007] FIG. 1 represents a graph illustrating the water absorption
capacity of several materials over time.
[0008] FIG. 2 illustrates the water absorbency of several materials
at a varying Relative Humidity.
[0009] U.S. Pat. Nos. 13,627, 663,943, 98,102 and 354,263 describe
a method for stabilising pharmaceutically acceptable salts of
S-adenosyl methionine comprising S-adenosyl methionine paratoluene
sulphonate, S-adenosyl methionine-1,4-butene disulphonate,
S-adenosyl methionine sulphate, S-adenosyl methionine tosylate with
a group of substances comprising chitosan, dextrin,
carboxymethylcellulose, fumaric acid, azelaic acid and tryptophan.
In particular the first of these patents indicates that it is
important to have a product with the highest amount of S,S
diastereoisomer which is the most stable possible over time because
the R,S diastereoisomer is not only inactive but has a
pharmacological effect which opposes that of the S,S. However, U.S.
Pat. Nos. 13,627 and 98,102 describe methods for stabilising
S-adenosyl methionine salts using the abovementioned substances in
a percentage by weight with respect to the active ingredient which
is very much higher than 50%, and adding them in reconstituted
aqueous solution to S-adenosyl methionine salts, with final
lyophilisation. This gives rise to high production costs and very
low yields because the % of ions in the final product falls from
approximately 50% to approximately 25%.
[0010] Racemisation of the S-adenosyl methionine is linked to three
basic parameters: [0011] 1. The nature of S-adenosyl-L-methionine
salt formation. [0012] 2. The residual moisture content in the
powder after drying. [0013] 3. The temperature at which the product
is stored.
[0014] The rate of racemisation of SAMe as a salt of S-adenosyl
methionine paratoluene sulphonate differs from the racemisation of
SAMe in the form of S-adenosyl methionine-1,4-butene disulphonate
salt, or S-adenosyl methionine sulphate or as S-adenosyl methionine
tosylate.
[0015] Although they have different pH for the same residual
moisture content, these four salts have very different stabilities
and racemisation. The reason for this has to be sought in the
mechanisms of diastereoisomer degradation and conversion in the
various salts.
[0016] It is known that the drier the starting material the more
stable the product will be.
[0017] The same consideration applies to rate of racemisation.
Theoretically, with zero moisture content, the conversion rate of
the S,S diastereoisomer at a given storage temperature is at a
minimum.
[0018] It is also known that the rate of degradation and therefore
also racemisation is associated with the thermal energy of the
material. This is reflected in the fact that the higher the storage
temperature for the material, the more rapidly it degrades and
racemises.
[0019] If not formulated on the basis of specific procedures and
using specific measures, formulations based on S-adenosyl
methionine reflect the abovementioned instability and racemisation
of the active ingredient, (conversion of the active S,S
diastereoisomer into the inactive R,S diastereoisomer), with
obvious adverse repercussions for the preservation and storage of
the material, even for short periods of time.
[0020] U.S. Pat. Nos. 3,954,726 and 4,057,672 describe relatively
stable salts of S-adenosyl methionine, that is up to 25.degree. C.
and 45.degree. C., respectively. U.S. Pat. No. 4,465,672 also
describes stable salts of S-adenosyl methionine with 5 mols of a
sulphonic acid with a pK of less than 2.5.
[0021] In this latter United States patent, the process of
preparing the product comprises preparation of a concentrated
aqueous solution of an impure salt of SAMe, purification of the
solution and its elution with a dilute aqueous solution of the
preselected sulphonic acid, titration of the resulting eluate,
concentration and lyophilisation or spraying. Because of the high
instability of SAMe and its derivatives the use of an aqueous
environment makes the limitations of this process obvious, and even
if residual moisture content is successfully contained it is still
unsuitable because of the properties of the inactive
ingredient.
[0022] Also these patents do not describe the rate of conversion of
the active S,S enantiomer at various operating and storage
temperatures for the product. Up to now no methods for stabilising
the active (SS)-(+)-S-adenosyl-L-methionine diastereoisomer in
acceptable percentages in solid oral formulations, particularly
tablets, are known. The only known concept is the need to keep
moisture content, impurities and the active
(SS)-(+)-S-adenosyl-L-methionine diastereoisomer under strict
control, protecting the tablets by either compression or
film-forming.
[0023] NADH is an active ingredient normally used as an energising
agent and antioxidant. Currently known compositions based on NADH,
such as those for example described in U.S. Pat. Nos. 5,332,727 and
7,034,011 are based on stabilising the active ingredient through
association with other antioxidants.
[0024] There has therefore hitherto been felt a need to identify a
simple and economic process which will make it possible to obtain a
product based on SAMe and/or NADH, with the removal of moisture and
low hygroscopic properties, with as a consequence increased
stability in terms of both the active ingredient and reduced
racemisation in favour of stabilisation of the reduced (S,S)
enantiomer and NADH.
[0025] Surprisingly it has been found that the addition of calcium
oxide and/or calcium hydroxide brings about improved stability of
both the SAMe, regarded as the sum of the two S,S and R,S
diastereoisomers, and the (S,S) diastereoisomer and the NADH,
through reducing the water content of the SAMe and the NADH and by
reducing its hygroscopic properties, further favoring synergistic
antidepressant action through the provision of calcium.
[0026] Calcium oxide and/or hydroxide directly mixed with atomised
SAMe and/or NADH powder, or with solid formulations based on SAMe
and/or NADH, are successful in removing water through a chemical
reaction with the powder or the preparation itself.
[0027] In fact no other excipients which succeed in removing
moisture in direct mixture with the powder and/or preparations of
SAMe and/or NADH over time at relatively lower temperatures
(15-20.degree. C.), reaching values of close to zero, are
known.
[0028] The main reason is due to the highly hygroscopic nature of
the SAMe which is even greater than that of substances which are
well known as excellent desiccants such as silica gel, calcium
chloride and others. This means that by mixing SAMe with excipients
having a moisture content of close to zero, the residual water in
mixtures and/or preparations based on SAMe is the same in absolute
terms as that present in the initial SAMe powder. As a consequence
there is only a percentage reduction in moisture content in the
preparations through the dilution effect, but the same percentage
by weight of water with respect to the weight of SAMe used. For
this reason, in a direct mixture and/or SAMe preparations, it has
never hitherto been possible to achieve higher stability of the
active ingredient, and therefore a reduced racemisation rate, than
that of the starting material, but at the limit this stability can
be achieved.
[0029] Calcium oxide is instead a natural desiccant with very high
reactivity in relation to water. It reacts with it and changes to a
calcium hydroxide, eliminating it permanently in preparations.
CaO+2H.sub.2O.fwdarw.Ca(OH).sub.2
[0030] FIG. 1 shows the rate of absorption of H.sub.2O with
different absorbent substances including calcium oxide.
[0031] It will be seen that calcium oxide absorbs slowly but
constantly up to 28% of its weight.
[0032] FIG. 2 shows the absorption capacity for water vapour of
various desiccants as the environmental humidity (RH) varies.
[0033] In this case it will be seen that calcium oxide absorbs
approximately 28% of water in a highly reactive way in an
environment with a very low relative humidity.
[0034] Table 1 summarises the absorbent capacities of various
desiccants under different relative humidity and temperature
conditions.
TABLE-US-00001 TABLE 1 Properties of adsorbents Montmorillonite
Property Molecular sieve Silica gel clay CaO CaSO.sub.4 Adsorption
capacity at low Excellent Poor Slight Excellent Good concentrations
of H.sub.2O Absorption ratio Excellent Good Good Poor Good Capacity
for water @77.degree. F. High High Medium High Low 40% RH
Separation by molecular Yes No No No No dimensions Adsorption
capacity at high Excellent Poor Poor Good Good temperatures
[0035] Specifically the shape of the two FIGS. 1 and 2 and the
summary values in Table 1 demonstrate that calcium oxide is the
only substance which is consistently capable of removing the very
small quantities of residual moisture content of SAMe or the
relatively high moisture content of NADH, or its salts
(approximately 1-1.5% K.F./approximately 5-7% K.F.) by chemical
conversion purely by physical contact, reducing it to values close
to zero.
[0036] This therefore reduces the second instability factor in
SAMe, or its salts, because of the high rate of racemisation of its
active S,S diastereoisomer. Table 2 provides moisture content
values for five lots of starting material of SAMe (S-adenosyl
methionine paratoluene sulphonate) with its corresponding analysis
prior to mixing with calcium oxide and storage at 20.degree. C. for
21 days, and the relative accelerated stability at 53.degree. C.
for 5 days.
TABLE-US-00002 TABLE 2 stress test 5 days at 53.degree. C. Moisture
Moisture Total Moisture content % content % K.F. % S, S SAMe titre
% impurities % content % SAMe Total Lot K.F. t = 0 t = 21 days at
20.degree. C. t = 0 t = 0 t = 0 K.F. % S, S titre % impurities %
001 1.15 1.13 80.87 52.96 0.66 1.09 56.21 51.19 5.17 002 1.08 1.05
80.02 51.98 0.73 1.05 56.31 50.84 5.54 003 1.06 1.03 80.21 52.76
1.03 1.03 56.12 50.11 4.55 004 1.09 1.09 79.82 52.23 0.94 0.99
55.79 49.58 4.34 005 1.04 1.12 81.54 52.29 1.04 1.00 55.28 49.99
5.02
[0037] Table 3 shows moisture content values for five lots of
starting material of SAMe (S-adenosyl methionine paratoluene
sulphonate) with its corresponding analysis after mixing with
calcium oxide and storage at 20.degree. C. for 21 days, and the
relative accelerated stability at 53.degree. C. for 5 days.
TABLE-US-00003 TABLE 3 Stress test 5 days at 53.degree. C. Moisture
Moisture content Total Moisture content % K.F. % K.F. t = 21 days %
S, S SAMe titre % impurities content % Total Lot t = 0 at
20.degree. C. t = 0 t = 0 % t = 0 K.F. % S, S SAMe titre %
impurities % 001 0.98 0.63 80.67 50.22 0.66 0.43 66.47 50.09 3.17
002 1.16 0.55 80.32 50.02 0.73 0.41 65.43 50.00 2.78 003 1.00 0.70
80.11 50.16 1.03 0.39 66.56 49.81 2.65 004 1.04 0.59 79.99 50.23
0.94 0.35 65.79 49.98 2.89 005 0.95 0.61 81.23 50.19 1.04 0.38
67.25 49.87 3.02
[0038] Table 4 shows moisture content values for five lots of
starting material of SAMe (S-adenosyl methionine-1,4-butene
disulphonate) with corresponding analysis prior to mixing with
calcium oxide and storage at 20.degree. C. for 21 days, and the
relative accelerated stability at 53.degree. C. for 5 days.
TABLE-US-00004 TABLE 4 Stress test 5 days at 53.degree. C. Moisture
Moisture content Total Moisture content % K.F. % K.F. t = 21 days %
S, S SAMe titre % impurities % content % SAMe Total Lot t = 0 at
20.degree. C. t = 0 t = 0 t = 0 K.F. % S, S titre % impurities %
001 2.03 2.03 84.58 51.34 0.44 2.09 59.43 50.94 4.06 002 2.01 2.31
85.34 51.54 0.56 2.21 60.02 50.93 4.23 003 1.98 1.99 83.89 52.34
0.45 2.00 60.32 51.03 4.05 004 1.89 1.99 84.82 52.02 0.67 1.96
59.49 51.72 4.63 005 1.94 2.02 85.34 51.78 0.64 1.93 58.98 50.79
4.47
[0039] Table 5 shows moisture content values for five lots of
starting material of SAMe (S-adenosyl methionine-1,4-butene
disulphonate) with corresponding analysis after mixing with calcium
oxide and storage for 23.degree. C. for 21 days, and the relative
accelerated stability at 53.degree. C. for 5 days.
TABLE-US-00005 TABLE 5 Stress test 5 days at 53.degree. C. Moisture
Moisture content Total Moisture content % % K.F. t = 21 days % S, S
SAMe titre % impurities % content Total Lot K.F. t = 0 at
20.degree. C. t = 0 t = 0 t = 0 % K.F. % S, S SAMe titre %
impurities % 001 1.94 1.33 84.21 50.01 0.49 0.78 70.34 50.00 2.03
002 1.89 1.45 85.02 49.78 0.50 0.87 70.02 50.01 1.98 003 1.87 1.27
83.49 50.12 0.49 0.93 71.32 49.89 2.00 004 1.80 1.38 84.54 50.34
0.57 0.81 71.89 50.04 2.13 005 1.84 1.40 85.25 50.08 0.53 0.88
70.94 50.00 1.35
[0040] Table 6 shows moisture content values for five lots of
starting material of NADH with corresponding analysis prior to
mixing with calcium oxide and storage at 20.degree. C. for 21 days,
and the relative accelerated stability at 53.degree. C. for 5
days.
TABLE-US-00006 TABLE 6 Stress test 5 days at 53.degree. C. NADH
NADH Moisture Moisture content (sodium salt) Total Moisture (sodium
salt) content % K.F. % K.F. t = 21 days titre % impurities %
content % titre % Total Lot t = 0 at 20.degree. C. t = 0 t = 0 K.F.
t = 0 impurities % 001 6.45 6.34 92.43 1.66 6.09 82.19 7.17 002
6.38 6.32 91.98 1.73 6.05 83.84 7.54 003 6.66 6.34 92.73 1.33 6.23
83.11 8.55 004 7.09 6.87 92.23 1.44 6.54 84.58 7.34 005 5.94 5.76
92.45 1.64 5.00 83.99 7.02
[0041] Table 7 shows moisture content values for five lots of
starting material of NADH with corresponding analysis after mixing
with calcium oxide and storage at 20.degree. C. for 21 days, and
the relative accelerated stability at 53.degree. C. for 5 days.
TABLE-US-00007 TABLE 7 Stress test 5 days at 53.degree. C. NADH
NADH Moisture Moisture content (sodium salt) Total Moisture (sodium
salt) content % K.F. % K.F. t = 21 days titre % impurities %
content % titre % Total Lot t = 0 at 20.degree. C. t = 0 t = 0 K.F.
t = 0 impurities % 001 6.21 3.20 84.53 (*) 1.50 3.49 81.19 2.45 002
6.33 4.32 85.32 (*) 1.48 3.45 82.34 3.54 003 6.44 4.01 83.93 (*)
1.44 3.23 80.56 2.67 004 7.23 4.39 84.23 (*) 1.54 3.54 82.56 3.14
005 6.87 3.98 83.95 (*) 1.43 3.10 82.49 3.02 (*) Lower titre
because mixed with 10% of calcium oxide.
[0042] From the data shown in Tables 2, 3, 4, 5, 6, 7 it will be
seen that the mixture of calcium oxide in combination with SAMe
(S-adenosyl methionine paratoluene sulphonate and S-adenosyl
methionine-1,4-butene disulphonate) or with NADH causes the
stability of the material at 53.degree. C. for 5 days to increase
with permanent removal of approximately 40% of the moisture content
when the mixture is stored for 21 days at 20.degree. C., and
approximately 60% after the stress test at 53.degree. C. for 5
days.
[0043] Thus, one object of this invention relates to compositions
comprising SAMe and/or NADH, or their salts, in association with
calcium oxide and/or calcium hydroxide, and optionally
pharmaceutically acceptable excipients.
[0044] According to this invention, by "SAMe" is meant both the
racemic mixture and the individual (RS)-(+)-S-adenosyl-L-methionine
[(RS)-(+)-SAMe)] and (SS)-(+)-S-adenosyl-L-methionine
[(SS)-(+)-SAMe)] diastereoisomers, including the mixtures other
than the racemic mixture.
[0045] In particular, the compositions according to this invention
contain SAMe, or its salts, in a quantity of between 30 and 90% by
weight, preferably between 50 and 85% by weight, with respect to
the weight of the composition, in association with calcium oxide
and/or calcium hydroxide in a quantity of between 1 and 40% by
weight, preferably between 2 and 20% by weight, with respect to the
weight of the composition.
[0046] In particular, the compositions according to this invention
contain NADH, or its salts, in a quantity between 1 and 90% by
weight, preferably between 5 and 50% by weight, with respect to the
weight of the composition, in association with calcium oxide and/or
calcium hydroxide in a quantity of between 1 and 40% by weight,
preferably between 2 and 20% by weight, with respect to the weight
of the composition.
[0047] Preferably the said SAMe, or its salts, is S-adenosyl
methionine paratoluene sulphonate, S-adenosyl methionine-1,4-butene
disulphonate, S-adenosyl methionine sulphate, S-adenosyl methionine
tosylate.
[0048] Preferably, the NADH is present in the form of its
pharmaceutically acceptable salts.
[0049] Preferably the said calcium oxide and/or calcium hydroxide
is calcium oxide alone, calcium hydroxide alone, or a mixture
thereof.
[0050] The pharmaceutically acceptable excipients used according to
this invention are preferably selected from calcium sulphate
hemihydrate, magnesium oxide, malic acid, glutamic acid,
glucono-delta-lactone, xylitol and/or their mixtures.
[0051] Compositions according to this invention may optionally
comprise at least one further active ingredient, preferably
selected from melatonin, 1-theanine and/or 1-tryptophan and/or
5-hydroxytryptophan and/or their mixtures.
[0052] The compositions according to this invention may be in the
form of a direct mixture, tablets, capsules, granules and/or
powder. In this invention by direct mixture is meant a mixture of
atomised powder of SAMe and/or NADH, or their salts, in association
with calcium oxide and/or calcium hydroxide alone, without the
addition of other excipients.
[0053] Preferably, the compositions according to this invention are
in the form of tablets, more preferably in the form of ordinary,
coated, film-coated and/or gastroresistant tablets.
[0054] In this invention, by ordinary tablet is meant a tablet
obtained by direct compression or compression after granulation
without coating; by coated tablet is meant a tablet coated with
non-gastroresistant substances; by film-coated tablet is meant a
coated tablet which is further covered with water-based varnishes,
which varnishes may have a gastroresistant action.
[0055] Thus, the compositions according to this invention may be
film-coated with water-based varnishes preferably selected from gum
Lac (Shellac.TM.) and/or its salts, methacrylic acid, cellulose
acetophthalates, titanium dioxide, talc, triethyl citrate, PVP K30,
curcumin, lutein, hydroxypropylcellulose,
hydroxypropylmethylcellulose and/or mixtures thereof.
[0056] By gastroresistant tablets according to this invention are
meant tablets capable of passing unchanged through the gastric
barrier.
[0057] The said film coating with varnishes, when provided through
Shellac.TM., salts, cellulose acetophthalates and/or other coatings
which are insoluble in an acid environment, may render the
compositions according to the invention resistant to passage
through the stomach. The varnishes according to this invention may
be present in a quantity varying from 1.0 to 1.98% by weight with
respect to the composition.
[0058] The compositions according to this invention have
approximately 60% less moisture content (KF) than the compositions
based on SAMe known hitherto and are approximately 12 times less
hygroscopic than shown in Table 6 above.
TABLE-US-00008 TABLE 8 Known tablets Known tablets based based on
SAMe on SAMe SAMe/CaO SAMe/CaO SAMe 400 mg SAMe 400 mg tablets
tablets tablets tablets (Example 1) (Example 1) KF % T = 0 KF % T =
24 h* KF % T = 0 KF % T = 24 h* Lot 01 1.24 3.76 0.45 0.76 Lot 02
1.21 3.87 0.51 0.68 Lot 03 1.10 3.98 0.52 0.70 Lot 04 1.33 3.75
0.43 0.64 Lot 05 1.39 3.76 0.57 0.74 at 40.degree. C. -75Rh KF
(moisture content according to the Karl Fischer method) T =
time
[0059] The compositions according to this invention are preferably
intended for the treatment of depressive states.
[0060] A further object of this invention is a process for the
preparation of solid compositions for oral use comprising SAMe
and/or NADH, or their salts, in association with calcium oxide
and/or calcium hydroxide which comprises the following stages:
[0061] a) mixing of the SAMe, or its salts, with calcium oxide and
pharmaceutically acceptable excipients, [0062] b) precompression
and subsequent granulation of the mixture obtained in stage a),
[0063] c) mixing of the granulate obtained in stage b) with
pharmaceutically acceptable excipients such as calcium sulphate
hemihydrate, xylitol, malic acid, glutamic acid, magnesium oxide,
hydrogenated fatty acids, precipitated silica, magnesium stearate,
saccharose, glycerol behenate, [0064] d) compression of the mixture
obtained in stage c), with the optional addition of sweeteners
and/or flavourings, [0065] e) optional coating of the tablet
obtained in stage d) with hydrogenated fatty acids, [0066] f)
optional aqueous phase film-forming on the tablet obtained in stage
e).
[0067] The process according to this invention is carried out in an
environment in which the relative humidity lies below 20% and the
temperature is held between 18 and 25.degree. C., preferably around
20.degree. C.
[0068] Granulation according to this invention is preferably
carried out using a rotating blade granulator fitted with a
stainless mesh having holes of between 1.2 mm and 3.2 mm in
diameter.
[0069] SAMe, or its salts, is used in a quantity varying from 30 to
90% by weight, preferably from 50 to 85% by weight, with respect to
the weight of the composition.
[0070] NADH, or its salts, is used in a quantity varying from 1 to
90% by weight, preferably from 5 to 50% by weight, with respect to
the weight of the composition.
[0071] In particular, the pharmaceutically acceptable excipients
used in the process according to the invention are preferably
selected from calcium sulphate hemihydrate, magnesium oxide,
calcium carbonate, malic acid, glutamic acid, xylitol, saccharose,
anhydrous microcrystalline cellulose, hydrogenated fatty acids,
magnesium stearate, glycerol behenate, precipitated silica.
[0072] More particularly, in step a) the active ingredient is
preferably mixed with calcium oxide from approximately 1.0 to
approximately 10% by weight and/or magnesium stearate from
approximately 0.5 to approximately 5% by weight and/or precipitated
silica from approximately 0.5 to approximately 2.0% by weight
calculated with respect to the active ingredient.
[0073] In stage c), the granulate obtained in b) is preferably
mixed with magnesium hydroxide from approximately 1.0 to 10.0% by
weight and/or microcrystalline cellulose from approximately 1.0 to
approximately 20.0% by weight and/or hydrogenated fatty acids from
approximately 1.0 to approximately 10% by weight and/or malic acid
from approximately 1 to approximately 10% by weight and/or glutamic
acid from approximately 1 to approximately 10% by weight and/or
glucono-delta-lactone from approximately 1 to approximately 10% by
weight, magnesium stearate from approximately 0.5 to approximately
5% by weight and/or glycerol behenate from approximately 1.0 to
approximately 5.0% calculated with respect to the active
ingredient.
[0074] Optionally, in said stage c) of the process according to the
invention at least one further active ingredient preferably
selected from melatonin, 1-theanine and/or 1-tryptophan and/or
5-hydroxytryptophan and/or their mixtures may be added to the
mixture for the treatment of depressive states.
[0075] At stage e) coating with hydrogenated fatty acids,
preferably molten hydrogenated vegetable fatty acids, may be
performed using conventional processes known in the art, with if
appropriate the addition of surfactants which are miscible in the
oily liquid.
[0076] According to this invention the coating mentioned in stage
e) may be performed using hydrogenated fatty acids, preferably
molten hydrogenated vegetable fatty acids, in a quantity of between
approximately 0.4 and approximately 1.5% by weight with respect to
the weight of the composition.
[0077] The said stage h) in the process according to this
invention, makes it possible to reduce the hygroscopic nature of
the tablet obtained in stage g) by approximately twelve times,
bringing about appreciable advantages in any subsequent stage of
aqueous phase film-forming.
[0078] Aqueous phase film-forming (stage i) may be carried out
using a substance or varnish preferably selected from gum Lac
(Shellac.TM.) and/or its salts, methacrylic acid, cellulose
acetophthalates, titanium dioxide, talc, triethyl citrate, PVP K30,
curcumin, lutein, hydroxypropylcellulose,
hydroxypropylmethylcellulose and/or mixtures thereof.
[0079] In particular the said film-forming may be carried out using
substances preferably selected from gum Lac (Shellac.TM.) and/or
its salts.
[0080] A further object of this invention is the use of SAMe or its
salts in association with calcium and magnesium for the preparation
of pharmaceutical, dietetic and/or nutritional/pharmaceutical
compositions for the treatment of depressive states.
[0081] Yet a further object of this invention is a method for
stabilising SAMe and/or NADH, preferably the (S,S) enantiomer, or
its salts, which comprises the use of calcium oxide and/or calcium
hydroxide in the percentages indicated above.
EXAMPLES
Example 1
TABLE-US-00009 [0082] TABLETS OF 400 mg SAMe ion/tablet Composition
based on SAMe sulphate p-toluene sulphonate A. SAMe sulphate
p-toluene sulphonate 800.00 mg B Calcium oxide 70.00 mg C.
Magnesium hydroxide 80.00 mg D. Saccharose 100.00 mg E. Calcium
carbonate 80.00 mg F. Magnesium stearate 20.00 mg G. Malic acid
40.00 mg E. Hydrogenated fatty acid 50.00 mg Total weight of core
1240.00 mg F. Hydrogenated vegetable fatty acids 4.00 mg G. Shellac
.RTM. 30.00 mg H. PVP K 30 6.0 mg I. Titanium dioxide 5.00 mg L.
Talc 10.00 mg M. Triethyl citrate 5.00 mg N. Curcumin 0.050 mg
Total weight of tablet 1300.50 mg
1.1. Mixing
[0083] The working environment was conditioned to a temperature of
20.degree. C. and a relative humidity value of approximately 20%
RH. A, B, C, D, E and G and 50% of F were then transferred to the
mixer in the quantities indicated above, leaving them with stirring
for approximately 30 minutes. At the end of this operation the
resulting mixture was transferred to dry containers, always
controlling moisture content and temperature.
1.2. Precompression
[0084] Precompression of the mixture was effected using a rotary
machine equipped with round punches of 25.0 mm. The hardness of the
tablets produced had to be regulated to subsequently produce a
granulate having good flow characteristics.
1.3 Granulation
[0085] The tablets produced during the first processing stage were
granulated on a 1000-1500 .mu.m mesh, again in a
humidity-controlled environment.
1.4 Mixing
[0086] The granulate obtained in stage 1.3 was transferred into the
mixer, adding magnesium stearate and leaving it with stirring for
approximately 30 minutes. At the end of this operation the
resulting mixture was transferred into dry containers.
1.5 Compression
[0087] Final compression of the granulate was carried out using a
rotary machine equipped with oblong punches of 21.0.times.9.8 mm
adjusting the weight to 1240 mg/tablet and the compression force to
at least 25 KP. The tablets produced had a hardness of between 25
and 35 Kp.
[0088] Friability: .ltoreq.1.0%; disaggregation time: .ltoreq.15
minutes (measured using the method described in U.S.P. 24.sup.th
ed.)
[0089] Moisture content according to K.F..ltoreq.1.50%
[0090] Stability tests on uncoated tablets were performed at only
40.degree. C. and 75% RH for six months and for a single lot
because this is not a finished product. The samples were stored in
alu/alu blisters.
TABLE-US-00010 TABLE 9 Lot 001 - cores of 400 mg ion/tablet
qualitative/quantitative composition in Example 1) Moisture content
% Lot (T/t).sup.1 (K. Fischer) S, S % AD.sup.2 (%) MTAD.sup.3 (%)
SAMe.sup.4 001 0.66 79.9 0.21 0.43 409.98 (20/0) 001A 0.56 75.7
0.33 0.67 409.58 (40/1) 001B 0.44 72.5 0.54 0.78 407.02 (40/3) 001C
0.35 70.3 0.76 0.98 404.78 (40/6) .sup.1Temperature (.degree.
C.)/time (months); .sup.2adenosine; .sup.3methylthioadenosine;
.sup.4SAMe sulphate p-toluene sulphonate (mg/tablet);
[0091] The data in Table 9 show that the tablets have optimum
stability.
1.6: Tablet Coating
[0092] The tablets resulting from the preceding processing stages
were coated in a bowl with a mixture of hydrogenated fatty acids
(4.0 mg/tablet).
[0093] Hydrogenated fatty acid melting at 70.degree. C. was placed
in a glass container of 2.0 litres and the temperature of the
mixture was raised to approximately 75.degree. C. obtaining a
homogeneous fused mass.
[0094] After the bowl had been preheated to approximately
65.degree. C., approximately 250 kg of tablets were added and
allowed to heat up to 60.degree. C. The cores were then protected
by causing the previously prepared fused mass to adhere to the
moving tablets. The cores so treated were again left at 60.degree.
C. for approximately 3 minutes, until the waxy layer had been
completely cleaned from the basket of the bowl.
1.7: Film-Forming on the Tablets
[0095] Shellac.TM. and PVP were dissolved in a container of
suitable size until a solution of 20% w/v was obtained, and
triethyl citrate was added slowly with constant stirring.
[0096] In another steel container again fitted with a stirrer,
talc, titanium dioxide and curcumin were dispersed in 4.0 l of
deionised water. The resulting suspension was poured into the
Shellac.TM. solution, washing the container with approximately 1.0
l of deionised water, subsequently diluting with a further 4.0 l of
deionised water.
[0097] During the first coating stage the temperature of the cores
was held at 54.degree. C. for approximately 40 minutes, and this
was then reduced in regular steps down to a value of 45.degree. C.
in the final stage.
[0098] After coating of the protected cores was complete, they were
allowed to dry for a further 10 minutes, again at 45.degree. C.
Finally reduction in the temperature to 42-43.degree. C. was
awaited so that emptying of the bowl could begin, taking care to
store the tablets in suitable envelopes which were impermeable to
moisture. No increase in percentage water content was observed in
the tablets produced in this way. All the checks specified by the
quality specifications were also carried out on these.
Example 2
TABLE-US-00011 [0099] TABLETS OF 400 mg SAMe ion/tablet
Compositions based on SAMe sulphate p-toluene sulphonate A. SAMe
sulphate p-toluene sulphonate 800.00 mg B. L-melatonin 2.00 mg C
Calcium oxide 70.00 mg D. Magnesium hydroxide 100.00 mg E. Calcium
sulphate hemihydrate 100.00 mg F. Calcium carbonate 160.00 mg G.
Magnesium stearate 20.00 mg H. Malic acid 40.00 mg I. Hydrogenated
fatty acid 40.00 mg Total weight of core 1332.00 mg L. Hydrogenated
vegetable fatty acids 4.00 mg M. Shellac .RTM. 30.00 mg N. PVP K 30
6.0 mg O. Titanium dioxide 5.00 mg P. Talc 10.00 mg Q. Triethyl
citrate 5.00 mg R. Curcumin 0.050 mg Total weight of tablet 1302.50
mg
[0100] The quantities relate to the preparation of a standard
industrial lot of 250.00 kg of tablets.
[0101] The tablets were prepared in the manner described in Example
1 using the components and quantities indicated above.
TABLE-US-00012 TABLE 10 Lot 002 - cores of 400 mg/ion/tablet
(qualitative/quantitative composition in Example 2) Moisture Lot
content % AD.sup.2 MTAD.sup.3 L-melatonin (T/t).sup.1 (K. Fischer)
S, S % (%) (%) SAMe.sup.4 mg 002 0.71 81.2 0.29 0.39 413.11 2.04
(20/0) 002A 0.50 76.8 0.35 0.58 410.21 2.03 (40/1) 002B 0.52 73.0
0.49 0.65 411.54 2.03 (40/3) 002C 0.42 71.0 0.79 0.83 409.40 2.01
(40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
[0102] The data in Table 10 indicate that the tablets have optimum
stability.
Example 3
TABLE-US-00013 [0103] TABLETS OF 400 mg SAMe ion/tablet Composition
based on SAMe sulphate p-toluene sulphonate A. SAMe sulphate
p-toluene sulphonate 800.00 mg B. L-theanine 200.00 mg C Calcium
oxide 70.00 mg D. Magnesium hydroxide 100.00 mg E. Xylitol 50.00 mg
F. Calcium carbonate 100.00 mg G. Microcrystalline cellulose 60.00
mg H. Magnesium stearate 20.00 mg I. Malic acid 40.00 mg L.
Hydrogenated fatty acid 40.00 mg Total weight of core 1480.00 mg M.
Hydrogenated vegetable fatty acids 4.00 mg N. Shellac .RTM. 30.00
mg O. PVP K 30 6.0 mg P. Titanium dioxide 5.00 mg Q. Talc 10.00 mg
R. Triethyl citrate 5.00 mg S. Hydroxypropylmethylcellulose 10.00
mg T. Curcumin 0.050 mg Total weight of tablet 1550.05 mg
[0104] The quantities relate to the preparation of a standard
industrial lot of 250.00 kg of tablets.
[0105] The tablets were prepared in the manner described in Example
1 using the components and quantities indicated above.
TABLE-US-00014 TABLE 11 Lot 003 - cores of 400 mg ion/tablet
(qualitative/quantitative composition in Example 3) Moisture
content % AD.sup.2 MTAD.sup.3 Lot (T/t).sup.1 (K. Fischer) S, S %
(%) (%) SAMe.sup.4 L-theanine 003 0.59 80.4 0.23 0.34 411.32 204.54
(20/0) 003A 0.53 76.6 0.32 0.61 410.54 203.54 (40/1) 003B 0.45 73.4
0.45 0.72 410.02 203.01 (40/3) 003C 0.37 71.3 0.69 0.88 407.56
201.92 (40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
[0106] The data in Table 11 show that the tablets have optimum
stability.
Example 4
TABLE-US-00015 [0107] TABLETS OF 400 mg SAMe ion/tablet Composition
based on SAMe sulphate p-toluene sulphonate A. SAMe sulphate
p-toluene sulphonate 800.00 mg B Calcium oxide 70.00 mg C.
Magnesium hydroxide 100.00 mg D. Calcium carbonate 150.00 mg E.
Magnesium stearate 20.00 mg F. Malic acid 40.00 mg G. Hydrogenated
fatty acid 40.00 mg Total weight of core 1220.00 mg H. Hydrogenated
vegetable fatty acids 8.00 mg I. Hydroxypropylmethylcellulose 30.00
mg L. PVP K 30 6.0 mg M Titanium dioxide 5.00 mg N. Talc 10.00 mg
O. Triethyl citrate 5.00 mg P. Curcumin 0.050 mg Total weight of
tablet 1284.05 mg
[0108] The quantities relate to the preparation of a standard
industrial lot of 250.00 kg of tablets.
[0109] The tablets were prepared in the manner described in Example
1 using the components and quantities indicated above.
Example 5
TABLE-US-00016 [0110] TABLETS OF 400 mg SAMe ion/tablet Composition
based on SAMe sulphate p-toluene sulphonate A. SAMe sulphate
p-toluene sulphonate 800.00 mg B. Folic acid 3.00 mg C Calcium
oxide 70.00 mg D. Magnesium hydroxide 100.00 mg E. Calcium
carbonate 100.00 mg F. Calcium sulphate 100.00 mg G. Magnesium
stearate 20.00 mg H. Malic acid 40.00 mg I. Hydrogenated fatty acid
40.00 mg Total weight of core 1273.00 mg L. Hydrogenated vegetable
fatty acids 8.00 mg M. Hydroxypropylmethylcellulose 30.00 mg N. PVP
K 30 6.0 mg O Titanium dioxide 5.00 mg P. Talc 10.00 mg Q. Triethyl
citrate 5.00 mg R. Curcumin 0.050 mg Total weight of tablet 1284.05
mg
[0111] The quantities relate to the preparation of a standard
industrial lot of 250.00 kg of tablets.
[0112] The tablets were prepared in the manner described in Example
1 using the components and quantities indicated above.
TABLE-US-00017 TABLE 12 Lot 004 - cores of 400 mg ion/tablet
(qualitative/quantitative composition in Example 5) Moisture Lot
content % AD.sup.2 MTAD.sup.3 Folic acid (T/t).sup.1 (K. Fischer)
S, S % (%) (%) SAMe.sup.4 mg 004 0.59 80.11 0.33 0.23 410.89 3.23
(20/0) 004A 0.53 75.4 0.45 0.55 410.43 3.24 (40/1) 004B 0.45 72.8
0.55 0.67 409.76 3.21 (40/3) 004C 0.37 69.6 0.79 0.99 408.67 3.19
(40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
[0113] The data in Table 12 indicate that the tablets have optimum
stability.
Example 6
TABLE-US-00018 [0114] TABLETS OF 400 mg SAMe ion/tablet Composition
based on SAMe sulphate p-toluene sulphonate A. SAMe sulphate
p-toluene sulphonate 800.00 mg B. Folic acid 3.00 mg C. Melatonin
2.00 mg C Calcium oxide 70.00 mg D. Magnesium hydroxide 100.00 mg
E. Calcium carbonate 100.00 mg F. Calcium sulphate 100.00 mg G.
Magnesium stearate 20.00 mg H. Malic acid 40.00 mg I. Hydrogenated
fatty acid 40.00 mg Total weight of core 1275.00 mg
[0115] The quantities relate to the preparation of a standard
industrial lot of 250.00 kg of tablets.
[0116] The tablets were prepared in the manner described in Example
1 using the components and quantities indicated above.
TABLE-US-00019 TABLE 13 Lot 005 - cores of 400 mg ion/tablet
(qualitative/quantitative composition in Example 6). Moisture
content % AD.sup.2 MTAD.sup.3 Folic acid L-melatonin Lot
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg mg 005 0.53
81.3 0.29 0.40 415.12 3.12 2.21 (20/0) 005A 0.50 76.2 0.38 0.59
414.21 3.03 2.12 (40/1) 005B 0.41 73.2 0.51 0.73 413.34 3.02 2.04
(40/3) 005C 0.29 69.2 0.83 1.09 412.21 3.00 2.08 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
[0117] The data in Table 13 reveal that the tablets have optimum
stability.
Example 7
TABLE-US-00020 [0118] TABLETS OF 5.5 mg of NADH/tablet as sodium
salt Compositions based on NADH without calcium oxide QUANTITY
DESCRIPTION OF COMPONENTS PER UNIT Active ingredient A) NADH mg
5.50 Excipients (core) B) Microcrystalline cellulose mg 7.00 C)
Mannitol mg 26.0 D) Glycerol behenate mg 2.00 E) "Light" magnesium
oxide* mg 8.0 F) Magnesium stearate mg 0.50 G) Calcium carbonate mg
1.00 Total weight of core mg 50.00 Excipients (coating) H) Schellac
mg 2.00 I) Povidone (PVP) mg 0.20 L) Titanium dioxide mg 0.10 M)
Anhydrous colloidal silica mg 0.20 N) Talc mg 0.20 O) Triethyl
citrate mg 0.15 Overall weight of the coated tablets mg 52.85 *=
Magnesium oxide light is a better lubricant than the heavy form
Example 8
TABLE-US-00021 [0119] TABLETS OF 5.5 mg of NADH/tablet as sodium
salt Composition based on NADH with calcium oxide QUANTITY
DESCRIPTION OF COMPONENTS PER UNIT Active ingredient A) NADH mg
5.50 Excipients (core) B) Microcrystalline cellulose mg 7.00 C)
Mannitol mg 20.0 D) Glycerol behenate mg 2.00 E) Calcium oxide mg
6.00 F) "Light" magnesium oxide* mg 8.00 G) Magnesium stearate mg
0.50 H) Calcium carbonate mg 1.00 Total weight of core mg 50.00
Excipients (coating) I) Schellac mg 2.00 L Povidone (PVP) mg 0.20
M) Titanium dioxide mg 0.10 N) Anhydrous colloidal silica mg 0.20
O) Talc mg 0.20 P) Triethyl citrate mg 0.15 Overall weight of the
coated tablets mg 52.85 *= Magnesium oxide light is a better
lubricant than the heavy form
[0120] The quantities relate to the preparation of a standard
industrial lot of 20.00 kg of tablets
Experimental Part
Stability Tests on the Finished Product
[0121] Stability at 40.degree. C. 75% RH (STRESS TEST) and at
ambient temperature over a long period (SHELF LIFE) for the
compositions in Examples 1, 2, 3, 4, 5, 6, 7, 8 obtained according
to the process according to the invention were evaluated for
changes in appearance (essentially change in colour), titre of SAMe
sulphate p-toluene sulphonate and NADH and other active ingredients
(mg/tablet), increase in degradation purities, moisture content
(K.F.) and % of the active (SS)-(+)-S-adenosyl-L-methionine
diastereoisomer; the presence of any degradation products, which
can be substantially identified as adenosine and
methylthioadenosine and oxidised NADH, expressed as a percentage
with respect to the mg of SAMe-toluene sulphonate per tablet and
reduced NADH, was further checked by HPLC.
Stress Test
[0122] The tablets were prepared in stoppered glass bottles and
enclosed in such a way as to reproduce the conditions of final
packaging (generally aluminium/aluminium blister).
[0123] The samples so prepared were stored for six months in a
stove thermostatted to a temperature of 40.+-.2.degree. C. and 75%
RH.
[0124] Nine samples from three different lots were used for the 400
mg tablets (Examples 1, 2, 3, 4, 5, 6), and each sample from each
lot was sampled after 0, 1, 3 and 6 months.
[0125] The following tables (14-37) report the results of the
stress test.
TABLE-US-00022 TABLE 14 Lot 006 - tablets of 400 mg ion/tablet
(Example 1) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 006 0.73 78.4 0.24 0.41
411.98 (20/0) 006 0.59 74.2 0.36 0.63 409.45 (40/1) 006B 0.54 71.5
0.59 0.73 409.02 (40/3) 006C (40/6) 0.43 68.9 0.87 0.91 405.71
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00023 TABLE 15 Lot 007 - tablets of 400 mg ion/tablet
(Example 1) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 007 0.61 79.2 0.31 0.55
412.32 (20/0) 007A 0.62 75.4 0.39 0.69 411.88 (40/1) 007B 0.57 73.1
0.52 0.72 410.67 (40/3) 007C (40/6) 0.49 70.1 0.77 0.89 408.65
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00024 TABLE 16 Lot 008 - tablets of 400 mg ion/tablet
(Example 1) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 008 0.81 77.9 0.34 0.49
408.54 (20/0) 008A 0.76 73.4 0.53 0.59 407.58 (40/1) 008B 0.61 71.1
0.74 0.74 407.04 (40/3) 008C (40/6) 0.55 68.8 0.88 0.84 404.21
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00025 TABLE 17 Lot 009 - tablets of 400 mg ion/tablet
(EXAMPLE 2) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 009 0.54 80.3
0.34 0.33 412.13 2.02 (20/0) 009A 0.50 77.4 0.39 0.45 410.54 2.01
(40/1) 009B 0.43 72.5 0.54 0.67 410.01 2.00 (40/3) 009C 0.32 70.3
0.84 0.93 408.44 1.98 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00026 TABLE 18 Lot 010 - tablets of 400 mg ion/tablet
(EXAMPLE 2) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 010 0.61 80.0
0.52 0.53 410.54 2.03 (20/0) 010A 0.57 75.4 0.55 0.58 408.65 2.03
(40/1) 010B 0.51 72.3 0.67 0.69 408.56 2.00 (40/3) 010C 0.48 70.0
0.86 0.98 406.98 1.95 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00027 TABLE 19 Lot 011 - tablets of 400 mg ion/tablet
(EXAMPLE 2) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 011 0.75 78.3
0.24 0.34 412.21 2.00 (20/0) 011A 0.55 75.8 0.35 0.55 410.29 2.02
(40/1) 011B 0.50 73.1 0.44 0.77 409.65 1.98 (40/3) 011C 0.47 71.3
0.75 0.97 407.65 1.95 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00028 TABLE 20 Lot 012 - tablets of 400 mg ion/tablet
(Example 3) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 012 0.66 80.3
0.34 0.54 414.43 205.65 (20/0) 003A 0.61 75.4 0.43 0.66 413.43
203.54 (40/1) 012B 0.58 72.2 0.54 0.76 411.32 203.32 (40/3) 012C
0.43 70.2 0.64 0.89 410.98 202.46 (40/6) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00029 TABLE 21 Lot 013 - tablets of 400 mg ion/tablet
(Example 3) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 013 0.73 79.5
0.25 0.53 412.45 203.01 (20/0) 003A 0.64 76.1 0.38 0.64 412.01
202.83 (40/1) 013B 0.55 72.5 0.65 0.72 410.52 202.01 (40/3) 013C
0.47 69.9 0.79 0.96 409.74 201.21 (40/6) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00030 TABLE 22 Lot 014 - tablets of 400 mg ion/tablet
(Example 3) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 014 0.62 79.2
0.35 0.44 412.22 202.01 (20/0) 003A 0.60 76.4 0.45 0.55 411.01
201.43 (40/1) 014B 0.57 72.9 0.67 0.76 410.52 200.01 (40/3) 014C
0.47 70.7 0.85 0.93 409.44 198.21 (40/6) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00031 TABLE 23 Lot 015 - tablets of 400 mg ion/tablet
(Example 4) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 018 0.63 79.4 0.43 0.52
412.54 (20/0) 018A 0.52 74.7 0.44 0.69 411.58 (40/1) 018B 0.41 71.5
0.58 0.78 49.78 (40/3) 018C (40/6) 0.31 68.9 0.72 0.99 407.75
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00032 TABLE 24 Lot 016 - tablets of 400 mg ion/tablet
(Example 4) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 016 0.56 79.2 0.33 0.49
410.54 (20/0) 001A 0.46 75.9 0.39 0.67 410.11 (40/1) 016B 0.42 72.9
0.50 0.69 409.67 (40/3) 016C (40/6) 0.39 70.7 0.86 0.87 408.65
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00033 TABLE 25 Lot 017 - tablets of 400 mg ion/tablet
(Example 4) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 017 0.69 78.9 0.35 0.49
413.54 (20/0) 017A 0.59 75.4 0.45 0.69 412.58 (40/1) 017B 0.56 72.9
0.59 0.79 409.02 (40/3) 017C (40/6) 0.49 71.7 0.87 0.96 407.59
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00034 TABLE 26 Lot 018 - tablets of 400 mg ion/tablet
(Example 5) Moisture content % AD.sup.2 MTAD.sup.3 Folic acid Lot
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 018 0.69 80.4
0.29 0.36 412.45 3.13 (20/0) 018A 0.56 75.7 0.35 0.58 411.98 3.04
(40/1) 018B 0.50 73.2 0.54 0.87 410.71 3.01 (40/3) 018C 0.38 70.3
0.66 1.05 407.37 3.09 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00035 TABLE 27 Lot 019 - tablets of 400 mg ion/tablet
(Example 5) Moisture Lot content % AD.sup.2 MTAD.sup.3 Folic acid
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 019 0.59 80.1
0.55 0.33 410.00 3.10 (20/0) 019A 0.53 75.4 0.65 0.45 410.02 3.03
(40/1) 019B 0.45 72.8 0.87 0.61 408.43 3.06 (40/3) 019C 0.37 69.6
1.01 0.79 40627 3.07 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00036 TABLE 28 Lot 020 - tablets of 400 mg ion/tablet
(Example 5) Moisture content % AD.sup.2 MTAD.sup.3 Folic acid Lot
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 020 0.49 80.8
0.23 0.33 414.89 3.00 (20/0) 020A 0.50 75.8 0.37 0.51 412.29 2.89
(40/1) 020B 0.37 72.3 0.51 0.63 409.76 2.98 (40/3) 020C 0.28 69.1
0.63 0.87 408.63 2.78 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00037 TABLE 29 Lot 021 - tablets of 400 mg ion/tablet
(Example 6) Moisture content % Folic L- Lot (K. S, AD.sup.2
MTAD.sup.3 acid melatonin (T/t).sup.1 Fischer) S % (%) (%)
SAMe.sup.4 mg mg 021 0.65 78.9 0.21 0.49 415.12 3.19 2.11 (20/0)
021A 0.53 76.1 0.34 0.57 414.21 3.23 2.02 (40/1) 021B 0.41 72.1
0.50 0.63 413.34 3.03 2.04 (40/3) 021C 0.26 69.0 0.81 0.94 412.21
3.00 2.01 (40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00038 TABLE 30 Lot 022 - tablets of 400 mg ion/tablet
(Example 6) Moisture content % Folic L- Lot (K. S, AD.sup.2
MTAD.sup.3 acid melatonin (T/t).sup.1 Fischer) S % (%) (%)
SAMe.sup.4 mg mg 022 0.76 80.2 0.25 0.42 412.34 3.32 2.11 (20/0)
022A 0.64 75.9 0.27 0.54 411.21 3.23 2.10 (40/1) 022B 0.59 72.4
0.43 0.77 410.12 3.12 2.09 (40/3) 022C 0.46 70.1 0.55 0.90 408.91
3.08 2.05 (40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00039 TABLE 31 Lot 023 - tablets of 400 mg ion/tablet
(Example 6) Moisture content % Folic L- Lot (K. S, AD.sup.2
MTAD.sup.3 acid melatonin (T/t).sup.1 Fischer) S % (%) (%)
SAMe.sup.4 mg mg 023 0.53 81.0 0.22 0.47 411.87 3.05 2.13 (20/0)
023A 0.50 76.0 0.33 0.69 409.27 3.03 2.12 (40/1) 023B 0.41 73.7
0.55 0.73 405.34 3.08 2.07 (40/3) 023C 0.29 69.9 0.74 0.87 404.71
3.03 2.04 (40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00040 TABLE 32 Lot 001 - tablets of 5.5 mg (Example 7)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 001 3.30 1.41 5.43 (20/0) 001A 3.89 2.03 4.98 (40/1)
001B 3.84 2.43 3.21 (40/3) 001C 3.63 3.61 2.21 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00041 TABLE 33 Lot 002 - tablets of 5.5 mg (Example 7)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 002 3.10 1.55 5.23 (20/0) 002A 3.87 2.22 4.34 (40/1)
002B 3.99 2.67 3.00 (40/3) 002C 3.77 3.89 2.02 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00042 TABLE 34 Lot 003 - tablets of 5.5 mg (Example 7)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 003 3.90 1.21 5.33 (20/0) 003A 3.65 2.23 4.58 (40/1)
003B 3.44 2.5 3.31 (40/3) 003C 3.93 3.81 2.51 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00043 TABLE 35 Lot 001 - tablets of 5.5 mg (Example 8)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 001 2.10 1.31 5.40 (20/0) 001A 1.99 1.43 5.32 (40/1)
001B 1.80 1.53 5.21 (40/3) 001C 1.33 1.81 5.00 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00044 TABLE 36 Lot 002 - tablets of 5.5 mg (Example 8)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 002 2.00 1.23 5.52 (20/0) 002A 1.36 1.32 5.34 (40/1)
002B 1.45 1.57 5.12 (40/3) 002C 1.27 1.99 4.89 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00045 TABLE 37 Lot 003 - tablets of 5.5 mg (Example 8)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 003 2.90 1.29 5.35 (20/0) 003A 1.65 1.56 5.21 (40/1)
003B 1.44 1.99 4.98 (40/3) 003C 1.12 2.31 4.67 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
[0126] From the stability data at 40.degree. C. and 75% RH (STRESS
TEST) it will be seen that all the lots examined after six months
had suffered degradation equal to approximately 2.5% of both SAMe
and the other active ingredients with a reduction of approximately
10% in the active (SS)-(+)-S-adenosyl-L-methionine
diastereoisomers;
[0127] From the stability data at 40.degree. C. and 75% RH (STRESS
TEST) it will be seen that all the lots of NADH examined containing
calcium oxide had undergone approximately 50% less degradation than
the lots without calcium oxide after six months.
Shelf Life
[0128] The tablets were packed in stoppered glass bottles and
enclosed in such a way as to reproduce the conditions of final
packaging (generally aluminium/aluminium blister).
[0129] The samples were selected in the same way and in the same
quantities as described for the stress test and kept in an
environment thermostatted to a temperature of 25.+-.2.degree. C.
and a humidity of 60% RH.
[0130] Nine samples originating from three different lots were used
for the 400 mg tablets (Examples 1, 2, 3, 4, 5, 6, 7, 8) and each
sample from each lot was sampled after 0, 3, 6, 12 months.
[0131] The following tables (38-61) show the results for SHELF
LIFE.
TABLE-US-00046 TABLE 38 Lot 024 - tablets of 400 mg ion/tablet
(Example 1) Moisture content Lot (T/t).sup.1 % (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 024 0.65 79.4 0.32 0.28
413.48 (20/0) 024A 0.56 75.3 0.44 0.34 413.23 (25/3) 024B 0.52 72.5
0.59 0.65 411.89 (25/6) 024C 0.44 69.9 0.83 0.79 409.76 (25/12)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00047 TABLE 39 Lot 025 - tablets of 400 mg ion/tablet
(Example 1) Moisture content Lot (T/t).sup.1 % (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 006 0.69 78.9 0.27 0.46
410.67 (20/0) 025 0.65 74.6 0.39 0.67 408.78 (25/3) 025B 0.56 73.5
0.65 0.74 409.02 (25/6) 025C 0.34 70.4 0.79 0.89 405.32 (25/12)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00048 TABLE 40 Lot 026 - tablets of 400 mg ion/tablet
(Example 1) Moisture content Lot (T/t).sup.1 % (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 026 0.78 78.7 0.20 0.47
411.65 (20/0) 006 0.65 74.9 0.39 0.60 409.43 (25/3) 026B 0.54 72.5
0.69 0.70 408.02 (25/6) 026C 0.48 68.45 0.88 0.94 404.43 (25/12)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00049 TABLE 41 Lot 027 - tablets of 400 mg ion/tablet
(Example 2) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 027 0.70 80.0
0.41 0.20 410.24 2.12 (20/0) 010A 0.64 75.7 0.54 0.47 408.65 2.04
(25/3) 027B 0.55 72.7 0.69 0.58 405.56 2.05 (25/6) 027C 0.43 70.4
0.83 0.85 406.58 1.99 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00050 TABLE 42 Lot 028 - tablets of 400 mg ion/tablet
(Example 2) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 010 0.64 80.4
0.33 0.35 413.44 2.07 (20/0) 028A 0.54 76.73 0.45 0.54 412.35 2.09
(25/3) 028B 0.50 73.9 0.67 0.56 408.46 2.04 (25/6) 028C 0.37 73.0
0.85 0.67 406.58 2.02 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00051 TABLE 43 Lot 029 - tablets of 400 mg ion/tablet
(Example 2) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 029 0.64 78.7
0.33 0.45 408.43 2.13 (20/0) 029A 0.57 75.3 0.34 0.54 407.55 2.12
(25/3) 029B 0.51 72.5 0.54 0.56 404.45 2.05 (25/6) 029C 0.39 71.2
0.67 0.76 403.23 1.99 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00052 TABLE 44 Lot 030 - tablets of 400 mg ion/tablet
(Example 3) Moisture content % AD.sup.2 MTAD.sup.3 Lot (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 030 0.71 79.76
0.22 0.34 413.49 209.35 (20/0) 030A 0.61 74.7 0.33 0.46 412.33
203.54 (25/3) 030B 0.55 73.2 0.51 0.66 410.32 202.32 (25/6) 030C
0.49 71.4 0.69 0.79 404.98 200.32 (25/12) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00053 TABLE 45 Lot 031 - tablets of 400 mg ion/tablet
(Example 3) Moisture content % AD.sup.2 MTAD.sup.3 Lot (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 031 0.62 80.4
0.37 0.43 412.43 205.21 (20/0) 031A 0.56 74.4 0.40 0.54 410.45
204.54 (25/3) 031B 0.58 71.2 0.50 0.65 407.78 203.23 (25/6) 031C
0.49 68.5 0.61 0.79 407.21 201.34 (25/12) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00054 TABLE 46 Lot 032 - tablets of 400 mg ion/tablet
(Example 3) Moisture content % AD.sup.2 MTAD.sup.3 Lot (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 032 0.63 81.5
0.44 0.24 409.99 203.65 (20/0) 032A 0.65 75.5 0.43 0.46 406.78
202.45 (25/3) 032B 0.59 73.4 0.64 0.56 406.54 203.00 (25/6) 032C
0.50 70.0 0.84 0.75 404.21 201.23 (25/12) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00055 TABLE 47 Lot 033 - tablets of 400 mg ion/tablet
(Example 4) Moisture Lot content % AD.sup.2 (T/t).sup.1 (K.
Fischer) S, S % (%) MTAD.sup.3 (%) SAMe.sup.4 033 0.74 79.9 0.39
0.29 411.23 (20/0) 033A 0.64 74.4 0.44 0.38 409.45 (25/3) 033B 0.59
73.5 0.63 0.57 406.02 (25/6) 033C 0.34 70.6 0.88 0.89 404.23
(25/12) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00056 TABLE 48 Lot 034 - tablets of 400 mg ion/tablet
(Example 4) Moisture Lot content % AD.sup.2 (T/t).sup.1 (K.
Fischer) S, S % (%) MTAD.sup.3 (%) SAMe.sup.4 034 0.59 78.3 0.25
0.39 410.23 (20/0) 034A 0.60 73.4 0.35 0.57 408.58 (25/3) 034B 0.53
70.9 0.49 0.88 404.32 (25/6) 034C 0.39 68.5 0.68 0.90 402.12
(25/12) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00057 TABLE 49 Lot 035 - tablets of 400 mg ion/tablet
(Example 4) Moisture Lot content % AD.sup.2 (T/t).sup.1 (K.
Fischer) S, S % (%) MTAD.sup.3 (%) SAMe.sup.4 035 0.59 78.7 0.38
0.39 408.56 (20/0) 035A 0.49 74.9 0.49 0.57 409.65 (25/3) 035B 0.50
72.0 0.65 0.68 404.73 (25/6) 035C 0.36 70.2 0.97 0.87 402.12
(25/12) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00058 TABLE 50 Lot 036 - tablets of 400 mg ion/tablet
(Example 5) Moisture Lot content % AD.sup.2 MTAD.sup.3 Folic acid
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 036 0.70 80.4
0.47 0.37 413.00 3.05 (20/0) 036A 0.58 74.4 0.56 0.40 410.45 3.03
(25/3) 036B 0.42 72.0 0.78 0.66 408.99 3.06 (25/6) 036C 0.39 69.8
0.89 0.72 404.67 3.01 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00059 TABLE 51 Lot 037 - tablets of 400 mg ion/tablet
(Example 5) Moisture Lot content % AD.sup.2 MTAD.sup.3 Folic acid
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 037 0.69 78.7
0.49 0.39 411.30 3.05 (20/0) 037A 0.63 74.5 0.64 0.55 408.57 3.01
(25/3) 037B 0.59 71.8 0.81 0.67 405.98 3.00 (25/6) 037C 0.48 69.2
1.00 0.89 402.56 2.89 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00060 TABLE 52 Lot 038 - tablets of 400 mg ion/tablet
(Example 5) Moisture Lot content % AD.sup.2 MTAD.sup.3 Folic acid
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 038 0.70 81.2
0.52 0.31 410.99 3.11 (20/0) 038A 0.63 75.4 0.60 0.43 407.32 3.08
(25/3) 038B 0.58 73.2 0.76 0.68 405.89 3.03 (25/6) 038C 0.49 70.6
0.80 0.93 401.34 3.01 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00061 TABLE 53 Lot 039 - tablets of 400 mg ion/tablet
(Example 6) Moisture content Folic L- Lot % (K. S, AD.sup.2
MTAD.sup.3 acid melatonin (T/t).sup.1 Fischer) S % (%) (%)
SAMe.sup.4 mg mg 039 0.63 81.4 0.29 0.43 410.43 3.03 2.06 (20/0)
039A 0.53 74.7 0.39 0.65 406.89 3.05 2.07 (25/3) 039B 0.57 72.7
0.58 0.79 403.69 3.00 2.03 (25/6) 039C 0.42 70.9 0.79 0.89 401.34
2.89 2.02 (25/12) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00062 TABLE 54 Lot 040 - tablets of 400 mg ion/tablet
(Example 6) Moisture content Folic L- Lot % (K. S, AD.sup.2
MTAD.sup.3 acid melatonin (T/t).sup.1 Fischer) S % (%) (%)
SAMe.sup.4 mg mg 040 0.63 78.8 0.35 0.40 408.88 3.10 2.05 (20/0)
040A 0.58 74.5 0.45 0.67 404.47 3.07 2.02 (25/3) 040B 0.48 72.5
0.60 0.70 403.34 3.03 2.07 (25/6) 040C 0.37 69.3 0.78 0.89 400.45
3.00 2.00 (25/12) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00063 TABLE 55 Lot 041 - tablets of 400 mg ion/tablet
(Example 6) Moisture content Folic L- Lot % (K. S, AD.sup.2
MTAD.sup.3 acid melatonin (T/t).sup.1 Fischer) S % (%) (%)
SAMe.sup.4 mg mg 041 0.73 81.6 0.42 0.38 410.48 3.15 2.10 (20/0)
023A 0.70 75.3 0.43 0.49 407.56 3.09 2.12 (25/3) 041B 0.58 72.4
0.58 0.70 406.65 3.08 2.08 (25/6) 041C 0.49 70.4 0.73 0.88 402.39
3.05 2.03 (25/12) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
TABLE-US-00064 TABLE 56 Lot 001 - tablets of 5.5 mg (Example 7)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 001 3.30 1.41 5.43 (20/0) 001A 3.34 1.53 5.23 (25/1)
001B 3.54 1.73 5.11 (25/3) 001C 3.23 2.21 4.65 (25/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00065 TABLE 57 Lot 002 - tablets of 5.5 mg (Example 7)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 002 3.10 1.55 5.23 (20/0) 002A 3.02 1.65 5.02 (25/1)
002B 3.00 1.87 4.70 (25/3) 002C 3.17 2.79 4.45 (25/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00066 TABLE 58 Lot 003 - tablets of 5.5 mg (Example 7)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 003 3.90 1.21 5.33 (20/0) 003A 3.75 1.43 5.21 (25/1)
003B 3.84 1.50 5.11 (25/3) 003C 3.34 2.61 4.87 (25/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00067 TABLE 59 Lot 001 - tablets of 5.5 mg (Example 8)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 001 2.10 1.31 5.40 (20/0) 001A 1.87 1.33 5.38 (25/1)
001B 1.89 1.43 5.31 (25/3) 001C 1.43 1.51 5.20 (25/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00068 TABLE 60 Lot 002 - tablets of 5.5 mg (Example 8)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 002 2.00 1.23 5.52 (20/0) 002A 1.76 1.30 5.44 (25/1)
002B 1.85 1.47 5.42 (25/3) 002C 1.57 1.67 5.29 (25/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
TABLE-US-00069 TABLE 61 Lot 003 - tablets of 5.5 mg (Example 8)
Moisture content % Lot (T/t).sup.1 (K. Fischer) NAD.sup.2 (%)
NADH.sup.3 003 2.90 1.29 5.35 (20/0) 003A 1.75 1.36 5.29 (25/1)
003B 1.84 149 5.12 (25/3) 003C 1.62 1.78 5.07 (25/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2Oxidised NADH;
.sup.3NADH sodium salt (mg/tablet);
[0132] From the stability data at 25.degree. C. and 60% RH (SHELF
LIFE) it will be seen that all the lots examined after twelve
months had suffered very little degradation of the SAMe with a
reduction of approximately 10% in the active
(SS)-(+)-S-adenosyl-L-methionine diastereoisomer;
[0133] From the stability data at 25.degree. C. and 60% RH (SHELF
LIFE) it will be seen that all the lots of NADH examined which
contained calcium oxide had undergone approximately 50% less
degradation than the lots without calcium oxide after six
months.
COMPARATIVE EXAMPLES
[0134] The following three comparative examples reproducing the
formulation of examples 1, 2, and 3 without the presence of calcium
oxide have been introduced.
EXAMPLES
Example 1A
TABLE-US-00070 [0135] TABLETS OF 400 mg SAMe ion/tablet Composition
based on SAMe sulphate p-toluene sulphonate A. SAMe sulphate
p-toluene sulphonate 800.00 mg B Calcium oxide absent 00.00 mg C.
Magnesium hydroxide 150.00 mg D. Saccharose 100.00 mg E. Calcium
carbonate 80.00 mg F. Magnesium stearate 20.00 mg G. Malic acid
40.00 mg E. Hydrogenated fatty acid 50.00 mg Total weight of core
1240.00 mg F. Hydrogenated vegetable fatty acids 4.00 mg G. Shellac
.RTM. 30.00 mg H. PVP K 30 6.0 mg I. Titanium dioxide 5.00 mg L.
Talc 10.00 mg M. Triethyl citrate 5.00 mg N. Curcumin 0.050 mg
Total weight of tablet 1300.50 mg
[0136] Stability tests on uncoated tablets were performed at only
40.degree. C. and 75% RH for six months and for a single lot
because this is not a finished product. The samples were stored in
alu/alu blisters.
TABLE-US-00071 TABLE 9A Lot 001A - cores of 400 mg ion/tablet
(qualitative/quantitative composition in Example 1A) Moisture
content % Lot (T/t).sup.1 (K. Fischer) S, S % AD.sup.2 (%)
MTAD.sup.3 (%) SAMe.sup.4 001 0.69 79.6 0.25 0.49 412.32 (20/0)
001A 0.66 73.2 0.65 0.97 409.23 (40/1) 001B 0.87 63.4 1.98 3.23
386.32 (40/3) 001C 0.79 53.2 3.32 9.56 365.67 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
[0137] The data in Table 9A show that the tablets have non good
stability.
Example 2A
TABLE-US-00072 [0138] TABLETS OF 400 mg SAMe ion/tablet
Compositions based on SAMe sulphate p-toluene sulphonate A. SAMe
sulphate p-toluene sulphonate 800.00 mg B. L-melatonin 2.00 mg C
Calcium oxide absent 00.00 mg D. Magnesium hydroxide 170.00 mg E.
Calcium sulphate hemihydrate 100.00 mg F. Calcium carbonate 160.00
mg G. Magnesium stearate 20.00 mg H. Malic acid 40.00 mg I.
Hydrogenated fatty acid 40.00 mg Total weight of core 1332.00 mg L.
Hydrogenated vegetable fatty acids 4.00 mg M. Shellac .RTM. 30.00
mg N. PVP K 30 6.0 mg O. Titanium dioxide 5.00 mg P. Talc 10.00 mg
Q. Triethyl citrate 5.00 mg R. Curcumin 0.050 mg Total weight of
tablet 1302.50 mg
[0139] The quantities relate to the preparation of a standard
industrial lot of 250.00 kg of tablets.
[0140] The tablets were prepared in the manner described in Example
1 using the components and quantities indicated above.
TABLE-US-00073 TABLE 10A Lot 002A - cores of 400 mg/ion/tablet
(qualitative/quantitative composition in Example 2A) Moisture Lot
content % AD.sup.2 MTAD.sup.3 L-melatonin (T/t).sup.1 (K. Fischer)
S, S % (%) (%) SAMe.sup.4 mg 002 0.87 80.0 0.34 0.69 412.21 2.00
(20/0) 002A 0.91 66.5 0.76 1.38 403.45 2.01 (40/1) 002B 0.92 63.2
2.02 4.75 381.64 2.01 (40/3) 002C 0.90 51.0 3.79 10.73 360.32 1.98
(40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
[0141] The data in Table 10A indicate that the tablets have non
good stability.
Example 3A
TABLE-US-00074 [0142] TABLETS OF 400 mg SAMe ion/tablet Composition
based on SAMe sulphate p-toluene sulphonate A. SAMe sulphate
p-toluene sulphonate 800.00 mg B. L-theanine 200.00 mg C Calcium
oxide absent 00.00 mg D. Magnesium hydroxide 170.00 mg E. Xylitol
50.00 mg F. Calcium carbonate 100.00 mg G. Microcrystalline
cellulose 60.00 mg H. Magnesium stearate 20.00 mg I. Malic acid
40.00 mg L. Hydrogenated fatty acid 40.00 mg Total weight of core
1480.00 mg M. Hydrogenated vegetable fatty acids 4.00 mg N. Shellac
.RTM. 30.00 mg O. PVP K 30 6.0 mg P. Titanium dioxide 5.00 mg Q.
Talc 10.00 mg R. Triethyl citrate 5.00 mg S.
Hydroxypropylmethylcellulose 10.00 mg T. Curcumin 0.050 mg Total
weight of tablet 1550.05 mg
[0143] The quantities relate to the preparation of a standard
industrial lot of 250.00 kg of tablets.
[0144] The tablets were prepared in the manner described in Example
1 using the components and quantities indicated above.
TABLE-US-00075 TABLE 11A Lot 003A - cores of 400 mg ion/tablet
(qualitative/quantitative composition in Example 3A) Moisture
content % AD.sup.2 MTAD.sup.3 Lot (T/t).sup.1 (K. Fischer) S, S %
(%) (%) SAMe.sup.4 L-theanine 003 0.98 78.2 0.34 0.74 408.39 203.53
(20/0) 003A 0.93 72.3 0.75 1.34 399.74 202.54 (40/1) 003B 1.05 62.3
2.45 4.72 378.32 202.11 (40/3) 003C 1.07 51.9 4.69 12.78 354.67
200.62 (40/6) .sup.1Temperature (.degree. C.)/time (months);
.sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe sulphate
p-toluene sulphonate (mg/tablet);
[0145] The data in Table 11A show that the tablets have non good
stability.
Experimental Part
Stability Tests on the Finished Product
[0146] Stability at 40.degree. C. 75% RH (STRESS TEST) and at
ambient temperature over a long period (SHELF LIFE) for the
compositions in Examples 1A, 2A, 3A, obtained according to the
process according to the invention were evaluated for changes in
appearance (essentially change in colour), titre of SAMe sulphate
p-toluene sulphonate, increase in degradation purities, moisture
content (K.F.) and % of the active (SS)-(+)-S-adenosyl-L-methionine
diastereoisomer; the presence of any degradation products, which
can be substantially identified as adenosine and
methylthioadenosine, expressed as a percentage with respect to the
mg of SAMe-toluene sulphonate per tablet, was further checked by
HPLC.
Stress Test
[0147] The tablets were prepared in stoppered glass bottles and
enclosed in such a way as to reproduce the conditions of final
packaging (generally aluminium/aluminium blister).
[0148] The samples so prepared were stored for six months in a
stove thermostatted to a temperature of 40.+-.2.degree. C. and 75%
RH.
[0149] Nine samples from three different lots were used for the 400
mg tablets (Examples 1A, 2A, 3A,), and each sample from each lot
was sampled after 0, 1, 3 and 6 months.
[0150] The following tables (14A-22A) report the results of the
stress test.
TABLE-US-00076 TABLE 14A Lot 006A - tablets of 400 mg ion/tablet
(Example 1A) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 006 0.99 78.1 0.35 0.81
410.23 (20/0) 006 1.16 71.2 0.85 1.37 404.45 (40/1) 006B 1.18 60.1
2.54 4.63 379.34 (40/3) 006C 1.29 50.8 4.82 11.98 350.78 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00077 TABLE 15A Lot 007A- tablets of 400 mg ion/tablet
(Example 1A) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 007 1.00 77.3 0.43 0.79
411.01 (20/0) 007A 0.96 71.8 0.85 1.65 406.29 (40/1) 007B 1.09 62.0
2.33 2.98 380.11 (40/3) 007C 1.09 52.1 4.01 10.90 355.99 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00078 TABLE 16A Lot 008A- tablets of 400 mg ion/tablet
(Example 1A) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 008 1.02 75.7 0.33 0.85
410.11 (20/0) 008A 1.03 70.3 0.95 1.47 403.98 (40/1) 008B 1.23 61.4
2.98 4.45 376.29 (40/3) 008C 1.56 51.1 5.02 13.54 345.87 (40/6)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00079 TABLE 17A Lot 009A - tablets of 400 mg ion/tablet
(EXAMPLE 2A) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 009 0.99 78.1
0.41 0.88 409.55 2.03 (20/0) 009A 1.21 68.4 0.96 1.65 401.33 2.01
(40/1) 009B 0.98 59.8 2.23 4.45 376.64 2.11 (40/3) 009C 1.11 52.3
3.99 11.23 351.34 1.98 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00080 TABLE 18A Lot 010A - tablets of 400 mg ion/tablet
(EXAMPLE 2A) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 010 0.87 77.3
0.46 0.83 409.59 2.12 (20/0) 010A 1.11 67.2 1.06 1.55 402.66 2.08
(40/1) 010B 1.18 59.4 2.55 4.67 381.23 2.01 (40/3) 010C 1.31 51.2
3.67 10.45 367.34 2.03 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00081 TABLE 19A Lot 011A - tablets of 400 mg ion/tablet
(EXAMPLE 2A) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 011 1.07 76.8
0.57 0.89 409.00 2.10 (20/0) 011A 1.14 67.5 1.33 1.72 400.87 2.04
(40/1) 011B 1.23 58.9 2.87 4.87 376.29 2.04 (40/3) 011C 1.38 53.5
4.88 11.89 362.54 2.03 (40/6) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00082 TABLE 20A Lot 012A - tablets of 400 mg ion/tablet
(Example 3A) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 012 1.21 76.7
0.36 0.84 407.00 202.72 (20/0) 003A 1.18 69.1 0.88 1.54 397.64
201.39 (40/1) 012B 1.35 63.4 2.75 4.87 379.98 200.41 (40/3) 012C
1.57 52.7 4.43 11.68 361.82 198.42 (40/6) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00083 TABLE 21A Lot 013A - tablets of 400 mg ion/tablet
(Example 3A) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 013 1.04 77.3
0.39 0.89 408.34 203.56 (20/0) 003A 1.07 68.4 0.85 1.57 399.84
202.49 (40/1) 013B 1.24 62.3 2.45 4.38 383.67 201.83 (40/3) 013C
1.37 51.9 4.23 9.87 370.52 199.27 (40/6) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00084 TABLE 22A Lot 014A - tablets of 400 mg ion/tablet
(Example 3A) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 014 1.00 75.3
0.53 0.92 406.23 205.52 (20/0) 003A 1.02 67.3 1.03 1.71 395.29
203.67 (40/1) 014B 1.13 61.2 2.82 3.89 371.28 202.61 (40/3) 014C
1.29 50.6 4.65 10.21 356.72 201.56 (40/6) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
Shelf Life
[0151] The tablets were packed in stoppered glass bottles and
enclosed in such a way as to reproduce the conditions of final
packaging (generally aluminium/aluminium blister).
[0152] The samples were selected in the same way and in the same
quantities as described for the stress test and kept in an
environment thermostatted to a temperature of 25.+-.2.degree. C.
and a humidity of 60% RH.
[0153] Nine samples originating from three different lots were used
for the 400 mg tablets (Examples 1A, 2A, 3A,), and each sample from
each lot was sampled after 0, 3, 6, 12 months.
[0154] The following tables (38A-46A) show the results for SHELF
LIFE.
TABLE-US-00085 TABLE 38A Lot 024A - tablets of 400 mg ion/tablet
(Example 1A) Moisture Lot content % (T/t).sup.1 (K. Fischer) S, S %
AD.sup.2 (%) MTAD.sup.3 (%) SAMe.sup.4 024 0.99 78.1 0.35 0.81
410.23 (20/0) 024A 1.02 68.2 0.59 1.24 404.21 (25/3) 024B 1.05 64.1
0.78 1.75 399.32 (25/6) 024C 1.14 60.2 1.83 2.88 396.65 (25/12)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00086 TABLE 39A Lot 025A - tablets of 400 mg ion/tablet
(Example 1A) Moisture Lot content % MTAD.sup.3 (T/t).sup.1 (K.
Fischer) S, S % AD.sup.2 (%) (%) SAMe.sup.4 006 1.00 77.3 0.43 0.79
411.01 (20/0) 025 1.02 67.2 0.65 1.36 407.67 (25/3) 025B 1.15 65.5
0.87 1.97 401.87 (25/6) 025C 1.19 61.3 1.78 2.85 396.34 (25/12)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00087 TABLE 40A Lot 026A - tablets of 400 mg ion/tablet
(Example 1A) Moisture Lot content % MTAD.sup.3 (T/t).sup.1 (K.
Fischer) S, S % AD.sup.2 (%) (%) SAMe.sup.4 026 1.02 75.7 0.33 0.85
410.11 (20/0) 006 1.09 66.4 0.83 1.54 405.45 (25/1) 026B 1.23 63.4
1.08 2.03 400.43 (25/6) 026C 1.49 59.9 1.99 2.77 395.87 (25/12)
.sup.1Temperature (.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00088 TABLE 41A Lot 027A - tablets of 400 mg ion/tablet
(Example 2A) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 027 0.99 76.7
0.46 0.83 409.59 2.12 (20/0) 010A 1.12 68.2 0.93 1.52 401.43 2.08
(25/3) 027B 0.98 62.6 1.59 2.54 397.53 2.07 (25/6) 027C 1.04 52.6
2.21 4.95 391.28 2.04 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00089 TABLE 42A Lot 028A - tablets of 400 mg ion/tablet
(Example 2A) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 010 0.87 75.8
0.46 0.83 409.59 2.12 (20/0) 028A 1.02 66.6 0.97 1.59 404.67 2.06
(25/3) 028B 0.99 61.4 1.74 2.83 398.93 2.03 (25/6) 028C 1.24 53.6
2.23 5.54 395.68 2.02 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00090 TABLE 43A Lot 029A - tablets of 400 mg ion/tablet
(Example 2A) Moisture Lot content % AD.sup.2 MTAD.sup.3 L-melatonin
(T/t).sup.1 (K. Fischer) S, S % (%) (%) SAMe.sup.4 mg 029 1.07 76.8
0.57 0.89 409.00 2.10 (20/0) 029A 1.06 67.7 1.42 1.93 403.28 2.06
(25/3) 029B 1.09 61.2 1.91 2.78 397.81 2.06 (25/6) 029C 1.26 52.6
2.43 4.76 393.98 2.04 (25/12) .sup.1Temperature (.degree. C.)/time
(months); .sup.2adenosine; .sup.3methylthioadenosine; .sup.4SAMe
sulphate p-toluene sulphonate (mg/tablet);
TABLE-US-00091 TABLE 44A Lot 030A - tablets of 400 mg ion/tablet
(Example 3A) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 030 1.21 76.7
0.36 0.84 407.00 202.72 (20/0) 030A 1.32 68.7 1.23 1.43 402.23
203.33 (25/3) 030B 1.52 63.2 1.91 2.63 397.37 202.11 (25/6) 030C
1.43 53.8 2.53 4.77 394.88 201.09 (25/12) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00092 TABLE 45A Lot 031A - tablets of 400 mg ion/tablet
(Example 3A) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 031 1.04 77.3
0.39 70.89 408.34 203.56 (20/0) 031A 1.11 66.9 1.41 1.63 403.83
202.23 (25/3) 031B 1.34 621 1.98 2.93 398.44 201.61 (25/6) 031C
1.27 54.0 2.71 4.79 395.98 200.18 (25/12) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet);
TABLE-US-00093 TABLE 46A Lot 032A - tablets of 400 mg ion/tablet
(Example 3A) Moisture Lot content % AD.sup.2 MTAD.sup.3 (T/t).sup.1
(K. Fischer) S, S % (%) (%) SAMe.sup.4 L-theanine 032 1.00 75.3
0.53 0.92 406.23 205.52 (20/0) 032A 1.31 65.6 1.37 1.88 402.99
204.65 (25/3) 032B 1.30 57.1 1.79 2.79 396.47 203.51 (25/6) 032C
1.43 50.0 2.55 4.67 390.38 202.58 (25/12) .sup.1Temperature
(.degree. C.)/time (months); .sup.2adenosine;
.sup.3methylthioadenosine; .sup.4SAMe sulphate p-toluene sulphonate
(mg/tablet).
Results
[0155] The stability at 40.degree. C. 75% RH (STRESS TEST) and at
temperature over a long period (SHELF LIFE) for the compositions of
Examples 1A, 2A and 3A (without calcium oxide) is lower than the
stability for the compositions of Examples 1, 2 and 3 (with calcium
oxide), valued at the same conditions (STRESS TEST and SHELF
LIFE).
* * * * *