Title of Invention

A PROCESS FOR THE PREPARATION OF PEPTIDE SALTS

Abstract Process for the production of a pharmaceutical preparation for parenteral administration to mammals, characterized in that an easily soluble acid addition salt of a basic peptide LHRH antagonist is reacted in the presence of a suitable diluent with a mixed bed ion exchanger or a mixture of an acidic and basic ion exchanger with formation of the free basic peptide, the ion exchanger is subsequently separated off and the free basic peptide is then reacted with an inorganic or organic acid with formation of the desired poorly soluble acid addition salt of the peptide, suitable pharmaceutical excipients, vehicles and/or bulking agents are then added and the diluent is subsequently removed.
Full Text peptide salts
The invention relates to a novel process for the
preparation of peptide salts, in particular of poorly
soluble peptide salts, and their use for the production
of medicaments. The invention furthermore relates to
pharmaceutical preparations which contain at least one
peptide salt prepared according to the invention, and
to their preparation.
In the international patent application No.
PCT/EP94/03904, the preparation of a poorly soluble
peptide by reaction of an aqueous solution of the acid
salt with an acetic acid solution of the basic peptide
with precipitation of the poorly soluble acid addition
salt of the peptide is described. For example, the
preparation of the LHRH antagonist cetrorelix embonate
is described.
The present invention relates to a novel process for
the preparation of peptide salts, characterized in that
an acid addition salt of a basic peptide (starting
peptide salt) (1) is reacted in the presence of a
suitable diluent with a mixed bed ion exchanger or a
mixture of an acidic and basic ion exchanger with
formation of the free basic peptide, the ion exchanger
is subsequently separated off and the free basic
peptide is then reacted with an inorganic or organic
acid with formation of the desired acid addition salt
of the peptide (final peptide salt) (2) and the diluent
is subsequently removed.
The expression "basic peptide" here means poly(amino
acids), even in the sense of a substructure within a
larger overall structure, which contain basic amino
acids, such as arginine, pyridylalanine or lysine, or
Ala10-NH2, detirelix Ac-D-Nal(2)1-D-Phe(4-Cl)2-D-Trp3-Ser4-Tyr5-D-
hArg(Et2)6-Leu7-Arg8-Pro9-D-Ala10-NH2, ramorelix (Stoeckemann
and Sandow, J. Cancer Res. Clin. Oncol. 1993, 119, 457) Ac-D-
Nal(2)1-D-Cpa2-D-Trp3-Ser4-Tyr5-D-Ser(Rha)6-Leu7-Arg8-Pro9-
Azagly10-NH2 and RS-68439 (diacetate salt of Detirelix) Ac-D-
Nal(2)1-D-Cpa2-D-Trp3-Ser4-Tyr5-D-hArg(Et2)6-Leu7-Arg8-Pro9-D-
Ala10-NH2, diacetate (salt). The structures of the peptides mentioned
are found in: Behre et al., GnRH antagonists: an overview,
Proceedings of the 2nd World Conference on Ovulation Induction, The
Parthenon Publishing Group Ltd.; Kutscher et al., Angew, Chem.
1997,109,2240.
The acid addition salts of the peptides employed as starting materials
are preferably easily soluble salts such as the acetates, hydrochlorides,
sulfates.
According to the novel process, the starting peptide salt is completely
or partly dissolved in a diluent or suspended therein. A diluent is then
added. Solvent and diluent can be identical or different. Possible
solvents and diluents are, for example: water, ethanol, methanol,
propanol, isopropanol, butanol, acetone, dimethyl ketone, methyl
ethyl ketone, dimethyl-acetamide, kimethylformamide, N-
methylpyrollidone [sic], acetonitrile, pentane, hexane, heptane and
mixtures thereof. Ethanol, isopropanol or acetone [sic] are preferred.
A water content of 1 to 60%, preferably of 5 to 50%, is likewise to be
preferred.
The mixed bed ion exchanger, i.e. a mixture of an acidic and a basic
ion exchanger, is added to the solution or suspension of the starting
peptide salt. A possible ion exchanger is, for example, Amberlite®.
The amount of ion exchanger depends on the number of
basic groups per peptide. The amount is determined by-
addition until a constant pH is obtained. For example,
10 g of Amberlite MB-3 are needed for 1 g of
cetrorelix.
The pHs of the base solutions during the preparation of
the bases is [sic] adjusted to 7.5 to 13, depending on
the active compound salt employed, in particular in the
case of peptide salts with amino acids having a basic
reaction, but in particular in the case of salts of
LHRH antagonists (for example cetrorelix, D-63153,
abarelix, ganirelix, ramorelix which can be present,
for example, as acetate) , depending on the active
compound used.
The temperature should not exceed 25-30°C in order to
avoid breakdown of the peptide. The reaction time for
the preparation of the free base customarily takes
place [sic] within a few minutes, for example 20 min
starting from cetrorelix acetate. It can also be
longer, for example approximately 1 h, starting from
cetrorelix embonate. After achieving a constant pH,. the
reaction should be discontinued, since decomposition
products, for example, can form on account of the
basicity of the solutions.
The ion exchanger is then removed from the reaction
mixture. Removal can be carried out by screening,
filtering off, centrifuging or column filtration.
The clear to turbid solution of the free peptide base,
which is unstable, should be reacted as rapidly as
possible with the acid with formation of the desired
acid addition salt. The acid can be added as a solid
substance or in solution or as a suspension. In exactly
the same way, the solution of the free peptide base can
take place to the acid [sic].
The reaction times are in the range from a few minutes
to a few hours. For example, for the formation of
cetrorelix embonate the reaction time is 1.5 h.
The reaction solution, which is customarily clear, is
then sterile-filtered. The solvent can then be removed
with obtainment of the pure peptide salt.
Alternatively, before the removal of the solvent
excipients, additives or vehicles can be added to the
solution. The addition of the excipients can take place
as a solid before sterile filtration or after sterile
filtration as a sterile-filtered solution.
Suitable excipients are, for example, mannitol,
sorbitol, xylitol, soluble starch.
According to the invention, the following salts can be
prepared by addition of the appropriate acid: acetate,
adipate, ascorbate, alginate, benzoate, benzene-
sulfonate, bromide, carbonate, citrate, chloride,
dibutylphosphate, dihydrogencitrate, dioctylphosphate,
dihexadecylphosphate, fumarate, gluconate, glucuronate,
glutamate, hydrogencarbonate, hydrogentartrate, hydro-
chloride, hydrogencitrate, iodide, lactate, alpha-
lipoic acid, malate, maleate, malonate, pamoate,
(embonate), palmitate, phosphate, salicylate, stearate,
succinate, sulfate, tartrate, tannate, oleate, octyl-
phosphate
The invention is illustrated by the example below,
without being restricted thereto.
Example 1:
46.47 g of D-20761 were added in portions to 1193 g of
water and dissolved with stirring. (= solution 1).
Solution 1 was then diluted with 32 61 g of 96% ethanol
with stirring. (= solution 2). Solution 2 was filtered
through a glass fiber prefilter after dilution and the
filtrate was mixed with 390 g of Amberlite MB3 (mixed
bed ion exchanger formed of strongly acidic cations
[sic] and anion exchangers) (= mixture 1) with
stirring. 316.8 g of mannitol was [sic] dissolved in
1267 g of water with stirring. (= solution 3) . After
stirring for 15 min, the pH of the supernatant solution
of mixture 1 was measured, and after stirring for a
further 5 minutes the pH was measured again. The
solution was then separated from the Amberlite MB3
through a fine-mesh sieve after a pH of 12.5 had been
reached. (= solution 4).
4162 g of the solution 4 was mixed with 5.34 g of
embonic acid with stirring. This mixture was vigorously
stirred for a further 1.5 h and the somewhat turbid
solution was then filtered through a glass fiber
prefilter. In this solution was a pH of 8.4 [sic] ( =
solution 5) . The measured pHs were measured with a
ground-glass electrode using a viscous electrolyte
liquid. The pHs were only to be regarded as relative
values, since the solutions or suspensions measured
contain ethanol and thus indicated an apparently higher
values [sic].
3333 g of the solution 5 were sterile-filtered in the
reaction apparatus, which was adjusted to RT, and 528 g
of the solution 3 were sterile filtered to give the
solution 5, which was adjusted to RT, with stirring.
(= solution 6).
Solution 6 was warmed to 40°C and the ethanol/water
mixture was then distilled off down to vacuo. (= suspension 1) . The cetrorelix embonate
suspension 1 was cooled to RT and diluted to 3000 g
with sterile-filtered water for injection with
stirring. (= suspension 2). 3.0 g each of the ready-to-
use suspension 2, adjusted to RT, were then dispensed
into 10 ml injection vials, provided with freeze-drying
stoppers and transferred to the freeze-drying unit.
The injection vials were frozen in a freeze-drying unit
at a plate temperature of -40°C. Drying was carried out
by means of a drying program at a plate temperature
increasing from -40°C to 20°C. The FD unit with [sic]
was flooded with sterile-filtered nitrogen, the
injection vials in the unit were sealed and the crimp
caps were then put on and rolled.
After the freeze drying, the sealed injection vials
were sterilized by gamma radiation at 12 kGy (min) -
15 kGy. The latter is optional.
1 140.0(7) mg injection vial contains 34.07 mg of
cetrorelix embonate corresponding to 3 0 mg of
cetrorelix.
and 106 mg of mannitol. For reconstitution, 2 ml of
water for injection are used. The suspension obtained
can be administered by i.m. or s.c. administration.
Biological action:
The cetrorelix embonate (2:1) lyophilizate (30 mg)
obtained according to example 1 is resuspended in 2 ml
of water for injection and can then be administered
parenterally, preferably subcutaneously (s.c.) or
intramuscularly (i.m.).
In the case of s.c. administration, a bioavailability
of about 30-50% (100% = intravenously administered
cetrolix acetate) is found for the cetrorelix embonate
(2:1). A particular advantage of cetrorelix embonate
(2:1) lyophilizate is the small or no 'burst effect' in
the patient. The duration of action is dose-dependent
and is 2-8 weeks or longer at a dose of 30-150 mg. The
cetrorelix embonate (2:1) lyophilizate according to the
invention has already been investigated on humans in
clinical phase I.
Fig. 1 shows the course of the cetrorelix plasma
concentration as a function of time (in hours)
beginning from the i.m. administration of 60 mg of
cetrorelix embonate (2:1) lyophilizate according to
example 1 in humans. No burst effect (about 100 ng/ml)
is determinable. Duration of action more than
700 hours. From 150 h after administration constant
plasma level of about 2 ng/ml. The bioavailability was
about 40%.
The areas of application of the peptide salts according
to the invention are, for example, BPH, myoma and
endometriosis.
We Claim:
1. Process for the production of a pharmaceutical preparation for
parenteral administration to mammals, characterized in that an
easily soluble acid addition salt of a basic peptide LHRH
antagonist is reacted in the presence of a suitable diluent with a
mixed bed ion exchanger or a mixture of an acidic and basic ion
exchanger with formation of the free basic peptide, the ion
exchanger is subsequently separated off and the free basic peptide
is then reacted with an inorganic or organic acid with formation of
the desired poorly soluble acid addition salt of the peptide, suitable
pharmaceutical excipients, vehicles and/or bulking agents are then
added and the diluent is subsequently removed.
2. Process as claimed in claim 1, wherein a salt of cetrorelix [N-Ac-
D-Nal(2)1, D-Cpa2, D-Pal(3)3, D-Cit6, D-Ala10]GnRH Ac-D-
Nal(2)1-D-Cpa2-D-Pal(3)3-Ser4-Tyr5-D-Cit6-Leu7-Arg8-Pro9-D-
Ala10-NH2, teverelix [N-Ac-D-Nal(2)1, D-Cpa2, D-Pal(3)3, D-Hci6,
Lys(iPr)8, D-Ala10]GnRH Ac-D-Nal(2)1-D-Cpa2-D-Pal(3)3-Ser4-
Tyr5-D-Hci6-Leu7-Lys(iPr)8-Pro9-D-Ala10-NH2 abarelix Ac-D-
Nal(2)1-D-Cpa2-D-Pal(3)3-Ser4-N-Me-Tyr5-D-Asn6-Leu7-
Lys(iPr)8-Pro9-D-Ala10-NH2, ganirelix [N-Ac-D-Nal(2)1, D-Phe(4-
Cl)2, D-Pal(3)3, D-hArg(Et2)6, hArg(Et2)8, D-Ala10]GnRH Ac-D-
Nal(2)1-D-Phe(4-Cl)2-D-Pal(3)3-Ser4-Tyr5-D-hArg(Et2)6-Leu7-
hArg(Et2)8-Pro9-D-Ala10-NH2, prazarelix (Azaline B) [N-Ac-D-
Nal(2)1, D-Cpa2, D-Pal(3)3, Aph(Atz)5, D-Aph(Atz)6, Lys(iPr)8,
D-Ala10]GnRH Ac-D-Nal(2)1-D-Cpa2-D-Pal(3)3-Ser4-Aph(Atz)5-
D-Aph(Atz)6-Leu7-Lys(iPr)8-Pro9-D-Ala10-NH2, iturelix (Antide)
[N-Ac-D-Nal(2)1, D-Cpa2, D-Pal(3)3, Lys(Nic)5, D-Lys(Nic)6,
Lys(iPr)8, D-Ala10]GnRH Ac-D-Nal(2)1-D-Cpa2-D-Pal(3)3-Ser4-
Lys(Nic)5-D-Lys(Nic)6-Leu7-Lys(iPr)8-Pro9-D-Ala10-NH2, A-
75998 Ac-D-Nal(2)1-D-Cpa2-D-Pal(3)3-Ser4-N-Me-Tyr5-D-
Lys(Nic)6-Leu7-Lys(iPr)8-Pro9-D-Ala10-NH2, detirelix Ac-D-
Nal(2)1-D-Phe(4-Cl)2-D-Trp3-Ser4-Tyr5-D-hArg(Et2)6-Leu7-Arg8-
Pro9-D-Ala10-NH2, ramorelix Ac-D-Nal(2)1-D-Cpa2-D-Trp3-Ser4-
Tyr5-D-Ser(Rha)6-Leu7-Arg8-Pro9-Azagly10-NH2 or RS-68439
(diacetate salt of Detirelix) Ac-D-Nal(2)1-D-Cpa2-D-Trp3-Ser4-
Tyr5-D-hArg(Et2)6-Leu7-Arg8-Pro9-D-Ala10-NH2, diacetate (salt) is
employed as the starting peptide salt.
3. Process as claimed in claim 1 or 2, wherein the acid is embonic
acid, stearic acid or salicylic acid.
4. Process as claimed in one of claims 1 to 3, wherein the molar ratio
of cetrorelix to embonic acid is 2:1.
5. Process as claimed in one of claims 1 to 4, wherein the diluent is
removed by freeze drying.

Process for the production of a pharmaceutical preparation for parenteral
administration to mammals, characterized in that an easily soluble acid
addition salt of a basic peptide LHRH antagonist is reacted in the presence
of a suitable diluent with a mixed bed ion exchanger or a mixture of an
acidic and basic ion exchanger with formation of the free basic peptide, the
ion exchanger is subsequently separated off and the free basic peptide is then
reacted with an inorganic or organic acid with formation of the desired
poorly soluble acid addition salt of the peptide, suitable pharmaceutical
excipients, vehicles and/or bulking agents are then added and the diluent is
subsequently removed.

Documents:

102-kolnp -2003-granted-abstract.pdf

102-kolnp -2003-granted-claims.pdf

102-kolnp -2003-granted-correspondence.pdf

102-kolnp -2003-granted-description (complete).pdf

102-kolnp -2003-granted-drawings.pdf

102-kolnp -2003-granted-examination report.pdf

102-kolnp -2003-granted-form 1.pdf

102-kolnp -2003-granted-form 18.pdf

102-kolnp -2003-granted-form 2.pdf

102-kolnp -2003-granted-form 26.pdf

102-kolnp -2003-granted-form 3.pdf

102-kolnp -2003-granted-form 5.pdf

102-kolnp -2003-granted-form 6.pdf

102-kolnp -2003-granted-gpa.pdf

102-kolnp -2003-granted-reply to examination report.pdf

102-kolnp -2003-granted-specification.pdf

102-kolnp -2003-granted-translated copy of priority document.pdf


Patent Number 236052
Indian Patent Application Number 102/KOLNP/2003
PG Journal Number 38/2009
Publication Date 18-Sep-2009
Grant Date 17-Sep-2009
Date of Filing 28-Jan-2003
Name of Patentee ZENTARIS GMBH
Applicant Address WEISMULLERSTRASSE 45, 60314 FRANKFURT, MAIN
Inventors:
# Inventor's Name Inventor's Address
1 DAMM, MICHAEL DIEBURGER STR. 106, 63322 RODERMARK
2 SALONEK, WALDEMAR MARGARETE-MASSIAS-STR.8, 69124 HEIDELBERG
3 ENGEL, JURGEN ERLENWEG 3, 63755 ALZENAU
4 BAUER, HORST ROHRENSTRASSE 12A, 91217 HERSBRUCK
5 STACH, GABRIELE BURGHOLZER STRASSE 4, 60433 FRANKFURT
PCT International Classification Number C07K 1/107
PCT International Application Number PCT/EP2001/09219
PCT International Filing date 2001-08-09
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 100 40 700.5 2000-08-17 Germany