TEPZZ - 7 Z58B - T: European Patent Specification
TEPZZ - 7 Z58B - T: European Patent Specification
TEPZZ - 7 Z58B - T: European Patent Specification
TEPZZ_7¥¥Z58B_T
(11) EP 1 733 058 B1
(12) EUROPEAN PATENT SPECIFICATION
Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent
Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the
Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been
paid. (Art. 99(1) European Patent Convention).
Description
5 [0001] The present invention relates to a process for making iron sucrose complexes.
[0003] One iron formulation, iron dextran, has been associated with significant adverse effects. Such effects are
reported in approximately 26% of patients receiving iron dextran. See, Gupta et al., Kidney Int., 1999 May; 55(5):1891-8.
20 The underlying cause of the immediate severe reactions is unclear. However, known anaphylactic reactions to dextran
have implicated dextran as the cause of the severe reactions to IV iron dextran. IV iron products free of dextran are
thought to decrease or avoid these severe reactions.
[0004] One product that is dextran-free is iron sucrose complex in sucrose (VENOFER®). Product safety reports for
iron sucrose demonstrate a low incidence of adverse effects. One study of 77 patients receiving a total of 757 doses,
25 reported only 4 patients experiencing adverse events related to the administration of iron sucrose. The events reported
were diarrhea, abdominal pain, nausea, constipation, and a transient minty taste. Ten patients in this study had a
documented history of sensitivity reactions to iron dextran that were consistent with anaphylaxis and none experienced
a hypersensitivity reaction with iron sucrose. See, Charytan et al., Am J Kidney Dis. 2001 Feb ; 37(2):300-7
[0005] IN 187116 describes a process for preparing saccharated iron oxide in powder from by (i) filtering a solution
30 of sodium carbonate in distilled water; (ii) filtering a solution ferric chloride in distilled water; (iii) adding the solution of
step (ii) in solution of step (iii) with constant stirring for at least 45 minutes; (iv) adjusting the pH 7 to 7.5 by addition of
sodium carbonate at room temperature; (v) decanting the supernant liquid from the slurry of ferric hydroxide thus formed;
(vi) separating the ferric hydroxide; (vii) mixing powdered sucrose with ferric hydroxide of step (vi); (viii) heating the
mixture and adjusting the pH in the range of 6.5 to 7.5 by 15% sodium hydroxide; and (ix) drying at 85˚ C to obtain
35 powdered ferric hydroxide.
[0006] Iron sucrose complex in sucrose generally contains contaminants including excipients, free sucrose and by-
40 products of the synthesis of the complex which are readily detected by techniques such as gel permeation chromatography
(GPC). The compendial method of analysis for Iron sucrose complex in sucrose is reported in United States Pharma-
copoeia (USP 26).
[0007] A chromatographic method for separating and purifying an iron saccharidic complex product is disclosed in
US Patent Application Publications 2002/0076821 and 2003/0153086. An iron sucrose complex, substantially free of
45 excipients having a molecular weight of less than about 5,000 Daltons, is also disclosed.
[0008] Small variations in molecular structure and composition can determine the difference between an active iron
complex having no adverse effects, and another iron complex that may induce adverse reactions. See, "Raising the Bar
for Quality Drugs", pp. 26-31, Chemical and Engineering News, American Chemical Society, Mar. 19, 2001. There is a
reported correlation between toxicity of iron saccharate complexes and higher molecular weight and the variability of
50 size of the complex. See, Fishbane et al., Semin Dial. 2000 Nov-Dec; 13(6):381-4.
[0009] A composition of iron sucrose complex comprising a narrower molecular weight distribution may yield a safer
and more efficacious therapy. There exists a need for an iron sucrose complex preparative method that results in a
product with narrower molecular weight distribution as compared to existing compositions.
[0010] According to one embodiment of the invention there is provided a process of preparing iron sucrose complex,
containing no more than 15% (wt./wt.) of excipients, according to claim 1.
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[0011] The selected molar ratio of sucrose to ferric hydroxide is from 2:1 to 50:1, preferably from 2:1 to 20:1, more
preferably from 5:1 to 20:1.
[0012] The sodium ions are present in the aqueous reaction mixture in a molar ratio of sodium ions to sucrose in the
range of from 1.0:0.5 to 1:20, preferably from 1:1 to 1:10; more preferably from 1:3 to 1:8.
5 [0013] The pH of the aqueous reaction mixture is in the range from 6.5 to 13, preferably 7 to 13, preferably from 8 to
12, most preferably from 8 to 10.
[0014] The selected temperature of the aqueous reaction mixture is a temperature in the range from 75˚C to 120˚C,
preferably in the range from 95˚C to 120˚C. The selected time interval is in the range from 2 minutes to 40 hours,
preferably in the range from 2 minutes to 300 minutes.
10 [0015] According to some sub-embodiments of the invention the aqueous reaction mixture comprises from 0.2% w/w
to 8% w/w based on the weight of the reaction mixture.
[0016] The aqueous reaction mixture may be concentrated prior to the step of isolating the iron sucrose complex to
reduce the volume of the reaction mixture to a volume that is preferably in the range from 20% to 80% of the original
volume thereof, more preferably in the range from 20% to 70% of the original volume thereof.
15 [0017] According to one embodiments of the invention, the weight average molecular weight of the isolated iron
sucrose complex is in the range from 20,000 to 400,000 Daltons, preferably, in the range from 20,000 to 120,000 Daltons,
more preferably in the range from 30,000 to 60,000 Daltons. According to certain sub-embodiments of the invention,
the weight average molecular weight of the prepared iron sucrose complex is about 35,000, about 40,000, about 45,000,
about 50,000, about 55,000, about 60,000, about 65,000, about 70,000, about 75,000, about 80,000, about 85,000,
20 about 90,000, about 95,000, about 100,000, about 105,000, about 110,000, about 115,000, about 130,000, about
135,000, about 140,000, about 145,000, about 150,000, about 155,000, about 160,000, about 165,000, or about 170,000
Daltons.
[0018] According to one sub-embodiment of the invention, the step of isolating the iron sucrose comprises concentrating
the reaction mixture of step (b) to form a residue comprising iron sucrose complex.
25 [0019] According to another sub-embodiment of the invention, the step of isolating the iron sucrose complex comprises
(i) forming a mixture by adding to the reaction mixture of step (b) at least one water-miscible organic solvent in an
amount sufficient to precipitate iron sucrose complex; and
(ii) collecting the precipitated iron sucrose complex, from the mixture formed in step (i).
30
[0020] According to one embodiment of step of isolating the iron sucrose complex, the step of collecting the precipitated
iron sucrose complex comprises filtration of the mixture formed in step (i).
[0021] According to another embodiment of isolating the iron sucrose complex, the step of collecting the precipitated
iron sucrose complex comprises centrifugation of the mixture formed in step (i).
35 [0022] According to another embodiment of isolating the iron sucrose complex, the step of collecting the precipitated
iron sucrose complex comprises freeze drying of the mixture formed in step (i).
[0023] According to one preferred embodiment of the invention, the ferric hydroxide used to form the iron sucrose
complex according to the invention is prepared by reacting at least one ferric salt, preferably, ferric chloride, ferric nitrate,
or a mixture thereof, with at least one base in a reaction mixture comprising an aqueous medium.
40 [0024] The collected iron sucrose complex is optionally purified, such as by.
[0025] According to one embodiment of purification of the collected iron sucrose complex, the step of separating the
purified precipitated iron sucrose complex comprises filtration of the mixture formed in step (b).
[0026] According to another embodiment of purification of the collected iron sucrose complex, the step of separating
50 the purified precipitated iron sucrose complex comprises centrifugation of the mixture formed in step (b).
[0027] The purified iron sucrose complex is optionally dried.
[0028] According to another embodiment of the invention there is provided a process of preparing an aqueous solution
of sucrose and iron sucrose complex, comprising the steps of:
55 (a) reacting ferric hydroxide and sucrose in an aqueous reaction mixture comprising sodium ions, at a selected
molar ratio of sucrose to ferric hydroxide, for a selected time interval, at a selected temperature, and at a pH in the
range from 6.5 to 13;
(b) isolating iron sucrose complex from the reaction mixture;
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(c) dissolving the isolated iron sucrose complex in an aqueous solvent to form a solution;
(d) forming a mixture by adding to the solution formed in step (c) at least one water-miscible organic solvent in an
amount sufficient to precipitate iron sucrose complex from the solution;
(e) collecting purified iron sucrose complex from the mixture formed in step (d); and
5 (f) dissolving the purified iron sucrose complex, prepared in step (e), in a solution of sucrose in water, preferably in
a solution of sucrose in water containing in the range from 20% to 40% (wt/wt) sucrose in water, more preferably
in a 30% (wt/wt) solution of sucrose in water.
[0029] According to another embodiment of the invention there is provided a process of preparing a co-precipitate
10 comprising iron sucrose complex and sucrose, the process comprising the steps of:
(a) reacting ferric hydroxide and sucrose in an aqueous reaction mixture comprising sodium ions, at a selected
molar ratio of sucrose to ferric hydroxide, for a selected time interval, at a selected temperature, and at a pH in the
range from 6.5 to 13;
15 (b) isolating iron sucrose complex from the reaction mixture;
(c) dissolving the isolated iron sucrose complex in an aqueous solvent to form a solution;
(d) forming a mixture by adding to the solution formed in step (c) at least one water-miscible organic solvent in an
amount sufficient to precipitate iron sucrose complex from the solution;
(e) collecting the purified iron sucrose complex from the mixture formed in step (d);
20 (f) dissolving purified iron sucrose complex product prepared according to step (e) in an aqueous sucrose solution;
(g) forming a mixture by adding to the solution of iron sucrose complex formed in step (f) at least one water-miscible
organic solvent in an amount sufficient to co-precipitate iron sucrose complex and sucrose;
(h) collecting the co-precipitate formed in step (g); and optionally
(i) drying the co-precipitate.
25
[0030] The ratio of purified iron sucrose complex to sucrose in the aqueous sucrose solution in step (a) of the co-
precipitate preparation method is from 1:0.1 to 1:20 by weight (wt/wt), preferably in the range of from 1:0.5 to 1:5 by
weight (wt/wt). The concentration of the sucrose solution employed to produce the co-precipitate is preferably in the
range from 10% to 50% weight/volume of sucrose in water.
30 [0031] According to another embodiment of the invention, another process of preparing a co-precipitate comprising
iron sucrose complex and sucrose is provided. The process comprises the steps of:
(a) providing a reaction mixture comprising a ferric salt dissolved in an aqueous medium;
(b) adding to the reaction mixture a first base in an amount in the range from about 1 to about 2 equivalents based
35 on the amount of ferric salt;
(c) allowing the reaction mixture to equilibrate for a time interval in the range from 10 to 60 minutes;
(d) forming a mixture by adding sucrose to the equilibrated reaction mixture of step (c) in a selected molar ratio to
the amount of ferric salt in the reaction mixture;
(e) heating the mixture formed in step (d) to a first temperature;
40 (f) forming a mixture by adding to the heated mixture formed in step (e) a second base in an amount sufficient to
adjust the pH of the reaction mixture to a selected pH;
(g) heating the mixture formed in step (f) at a second temperature for a selected time interval;
(h) after the selected time interval, cooling the reaction mixture to a temperature in the range of from 20˚ to 30˚C; and
(i) isolating the co-precipitate from the cooled reaction mixture.
45
[0032] The first selected temperature is preferably in the range from 60˚ to 90˚C, more preferably from 60˚ to 80˚C
and the second selected temperature is in the range from 75˚ to 120˚C, preferably in the range from 95˚ to 120˚C.
Suitable bases for use as the first and/or second selected bases include, for example, alkali metal carbonates, e.g.
sodium carbonate; alkali metal bicarbonates, e.g., sodium bicarbonate; alkali metal hydroxides, e.g., sodium hydroxide;
50 water-soluble amines, e.g., trishydroxymethyl-aminomethane; and mixtures thereof.
[0033] The selected molar ratio of sucrose to the ferric salt in step (d) is in the range from 2:1 to 50:1, preferably, in
the range from 2:1 to 20:1.
[0034] The step of isolating the co-precipitate preferably comprises the steps of:
55 (a) forming a mixture by adding to the cooled reaction mixture formed in step (h) at least one water-miscible organic
solvent in an amount sufficient to co-precipitate iron sucrose complex and sucrose;
(b) collecting the co-precipitate formed in step (a); and optionally
(c) drying the co-precipitate.
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[0035] The cooled reaction mixture of step (h) may be optionally concentrated to reduce the volume of the reaction
mixture to a volume in the range preferably from 20% to 80% of the original step (h) reaction mixture volume, more
preferably from 20% to 70% of the original step (h) reaction mixture volume prior to the addition of the water-miscible
organic solvent;
5 [0036] According to another embodiment of the invention there is provided a process of preparing an aqueous solution
of sucrose and iron sucrose complex, comprising the steps of:
(a) reacting ferric hydroxide and sucrose in an aqueous reaction mixture comprising sodium ions, at a selected
molar ratio of sucrose to ferric hydroxide, for a selected time interval, at a selected temperature, and at a pH in the
10 range from 6.5 to 13;
(b) isolating iron sucrose complex from the reaction mixture;
(c) dissolving the isolated iron sucrose complex in an aqueous solvent to form a solution;
(d) forming a mixture by adding to the solution formed in step (c) at least one water-miscible organic solvent in an
amount sufficient to precipitate iron sucrose complex from the solution;
15 (e) collecting the purified iron sucrose complex from the mixture formed in step (d);
(f) dissolving the purified iron sucrose complex product formed in step (e) in an aqueous sucrose solution;
(g) forming a mixture by adding to the iron sucrose complex solution formed in step (f) at least one water-miscible
organic solvent in an amount sufficient to co-precipitate iron sucrose complex and sucrose;
(h) collecting the co-precipitate formed in step (g); and
20 (i) dissolving the collected co-precipitate in water.
[0037] According to another embodiment of the invention there is provided a process of preparing an aqueous solution
of sucrose and iron sucrose complex, comprising the steps of:
25 (a) combining ferric hydroxide and sucrose, in an aqueous reaction mixture, at a selected molar ratio of sucrose to
ferric hydroxide, at a selected temperature and at a pH in the range from 7 to 13, preferably from 8 to 12, more
preferably from 9.5 to 12, most preferably from 10.5 to 12;
(b) maintaining the reaction mixture at the selected temperature for a time interval from 2 to 300 minutes; and
(c) adding to the reaction mixture a selected quantity of sucrose.
30
[0038] The selected temperature is in the range from 75˚ to 120˚C, preferably in the range from 95˚ to 120˚C, and the
selected molar ratio of sucrose to ferric hydroxide in step (a) is in the range from 2:1 to 50:1, preferably, in the range
from 2:1 to 20:1, more preferably, in the range from 5:1 to 20:1.
[0039] The quantity of sucrose added in step (c) is preferably from 1 to 50 times the amount of ferric hydroxide used
35 in step (a), on a mol/mol basis, more preferably from 1 to 20 times the amount of ferric hydroxide used in step (a), on a
mol/mol basis.
[0040] The iron sucrose complexes prepared according to the process of the invention comprise from 1 to 60% ferric
iron (wt/wt), preferably from 1 to 55% ferric iron (wt/wt), more preferably from 1 to 50% ferric iron (wt/wt). According to
some preferred embodiments, the iron sucrose complex prepared according to the process of the invention comprises
40 from 30% to 50% ferric iron (wt/wt), most preferably about 45% ferric iron. According to other preferred embodiments,
the iron sucrose complex prepared according to the process of the invention comprises from 2% to 15% ferric iron
(wt/wt), most preferably about 5% ferric iron (wt/wt).
[0041] According to another embodiment of the invention, a pharmaceutical composition in a solid dosage form is
provided comprising a pharmaceutically acceptable carrier and an iron sucrose complex having a molecular weight in
45 the range from 20,000 to 400,000 Daltons.
[0042] Preferably, the pharmaceutical composition of the invention comprises an iron sucrose complex prepared by
the process according to the present invention.
Definitions
50
[0043] The expression, "substantially free of excipients," used to describe the product iron sucrose complex formed
by the method of the invention means that the product contains 15% (wt/wt) of excipients or less, and correspondingly
contains 85% (wt/wt) or more iron sucrose complex. Preferably the iron sucrose complex formed by the method of the
invention contains 10% (wt/wt) of excipients or less. Most preferably, the iron sucrose complex formed by the method
55 of the invention contains 5% (wt/wt) of excipients or less.
[0044] The term "excipients" as used herein refers to components of the product of a process of the invention that
are other than iron sucrose complex. Examples include, free sucrose, water and solvents, and substances related to
the synthesis process. By "substances related to the synthesis process" is meant the reagents used in the synthesis
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and products of degradation of either the synthesis reagents of the reaction products.
[0045] The expression "weight average molecular weight" unless otherwise indicated, is one expression of the mo-
lecular weight of a substance which comprises a distribution of molecular weights rather than a single molecular weight.
The "weight average molecular weight" is calculated as a summation of the squares of the weights of a fraction of the
5 molecular weight distribution, divided by the total weight of the molecules. The weight average molecular weight may
be determined by gel permeation chromatography (GPC) using refractive index, light scattering, small angle neutron
scattering (SANS), or by sedimentation velocity.
[0046] The expression "alkali metal," as employed herein refers to metals or ions of metals found in Group I of the
periodic table. Preferred alkali metals are lithium, sodium and potassium.
10 [0047] The term "base" as employed herein refers to a chemical species that donates electrons or hydroxide ions
(Arrhenius definition) or that accepts protons (Bro"nsted definition). Bases include strong bases, i.e., bases that are
completely dissociated in aqueous solution and weak bases, i.e., bases that are only partially dissociated in aqueous
solution. Examples of strong bases include sodium hydroxide and potassium hydroxide. Examples of weak bases include
ammonia and alkyl amines.
15 [0048] The expression "water-miscible organic solvent," unless otherwise indicated, refers to an organic solvent which
is soluble in water in all proportions at standard temperature and pressure. Suitable water-miscible organic solvents
include, for example, methanol, ethanol, isopropanol, acetone, acetonitrile, tetrahydrofuran, dioxane, dimethyl forma-
mide, dimethylacetamide, and N-methylpyrrolidinone.
[0049] The term "co-precipitation", used herein refers to simultaneous precipitation of more than one dissolved sub-
20 stance from a solution or suspension.
[0050] The expressions, "aqueous medium" and "aqueous solvent" refer, unless otherwise indicated, to a solvent or
medium that is water, or a mixture of water and one or more water-miscible organic solvents.
[0051] The expression "substantially free of crystalline material" refers, unless otherwise indicated, to a material that
is indistinguishable via X-ray powder diffraction from the same material present as exclusively an amorphous solid.
25 [0052] The expression "solid dosage form" means a solid pharmaceutical preparation in the form of, for example a
tablet, capsule, pill, powder, or granule.
30 [0053]
Fig. 1 shows a gel permeation chromatography (GPC) trace of iron sucrose complex prepared by the process of
the invention, having a weight average molecular weight of 49,000 Daltons.
Fig. 2 shows a GPC trace of iron sucrose complex in 20% aqueous solution, prepared by the process of the invention,
35 wherein the iron sucrose complex has a weight average molecular weight of 50,000 Daltons.
Fig. 3 shows a GPC trace of VENOFER® brand iron sucrose complex in sucrose wherein the iron sucrose complex
has a weight average molecular weight of 46,000 Daltons.
Fig. 4 shows a GPC trace of iron sucrose complex wherein the iron sucrose complex has a weight average molecular
weight of 90,000 Daltons.
40 Fig. 5 shows an X-ray diffractogram of a dried, purified iron sucrose complex prepared in Example 6 which is
amorphous and substantially free of sucrose.
Fig. 6 shows an X-ray diffractogram of a dried, purified iron sucrose complex prepared in Example 8 which contains
a detectable amount of crystalline iron sucrose complex.
[0054] Iron sucrose complexes presently employed in therapy contain significant amounts of contaminants detectable
by GPC analysis. The present invention provides a process for the preparation of iron sucrose complexes that are
substantially free of excipients. The iron sucrose complexes, substantially free of excipients, can be used to formulate
50 a therapeutic iron sucrose composition containing lower levels of contaminants.
A. Ferric Hydroxide
[0055] The term "ferric hydroxide" as employed herein, includes the various forms of ferric hydroxide, including, for
55 example, hydrated ferric oxide, ferric oxy hydroxide, polymeric ferric hydroxide, ferric hydroxide gel, partially neutralized
ferric salts and partially neutralized polymeric ferric salts. The various forms of ferric hydroxide may be expressed
according to Formula I:
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{(FeIII)a[Y]k[X/e]m[O]b[H]d}z I
wherein "a" and "z" represent integers that are independently 1 to about 1000, preferably 1 to about 500; "Y" is a cation
other than FeIII, for example, ammonium or alkyl ammonium; "b," "d," "k" and "m" represent integers that are independently
5 0 to about 1000, preferably 0 to about 500; "X" is an anion, for example, chloride, bromide, iodide, nitrate, sulfate, acetate,
citrate, and other acid anions; "e" represents the equivalent number of the anion X.
10 [0056] The ferric hydroxide utilized as the starting material in the process of the present invention may be prepared
by reacting a ferric salt with at least about one molar equivalent of a base, based on the amount of the ferric salt. A
mixture of ferric salts, and/or a mixture of bases, may be employed. Suitable ferric salts include ferric salts wherein the
anion is an anion of an acid such as, for example, chloride, bromide, iodide, nitrate, sulfate, acetate, citrate and other
acid anions. Preferred ferric salts include, for example ferric chloride and ferric nitrate.
15 [0057] Suitable bases for reaction with the ferric salt include, for example, alkali metal carbonates, alkali metal bicar-
bonates, alkali metal hydroxides, water-soluble amines and mixtures thereof. Preferred bases include sodium carbonate,
sodium bicarbonate, sodium hydroxide, tris-hydroxymethylaminoethane and mixtures thereof.
[0058] The ferric hydroxide may be prepared by (a) providing a reaction mixture comprising a ferric salt dissolved in
an aqueous medium; (b) adding to the reaction mixture a first base in an amount from 1 to 2 equivalents based on the
20 amount of ferric salt; (c) allowing the reaction mixture to equilibrate for a time interval greater than about 10 minutes,
preferably from 10 minutes to 120 minutes, more preferably from 10 minutes to 60 minutes; (d) adding to the equilibrated
reaction mixture a second base in an amount sufficient to adjust the pH of the reaction mixture to a selected pH; and
(e) collecting the ferric hydroxide from the reaction mixture.
[0059] The first and second bases may be the same, or may be different bases. The bases may be added to the
25 reaction mixture in solution or suspension in an aqueous solvent. Alternately the bases may be added neat, i.e., a base
such as sodium carbonate may be added as a dry solid.
[0060] The first base may be added to the reaction mixture batchwise, i.e., all at once, or continuously or semi-
continuously over a time interval at a constant or variable addition rate. A slow continuous addition may be performed
as a titration wherein the pH of the mixture is continuously monitored, preferably using a pH meter. The addition of the
30 base may be stopped when a selected pH, preferably in the range from 2.0 to 2.5, is achieved in the reaction mixture.
The addition rate of the base for slow continuous addition is preferably from 0.02 to 0.2 equivalents of the base per
minute, based on the amount of the ferric salt in the reaction mixture.
[0061] After the addition of the first base to the reaction mixture, the reaction mixture is allowed to equilibrate, with or
without stirring. The temperature is preferably maintained in the range from 20˚ to 30˚C. The pH of the reaction mixture
35 is typically observed to drop to a pH in the range from 1.4 to 1.8 during the time interval when the reaction mixture is
allowed to equilibrate.
[0062] The second base is preferably added to the reaction mixture continuously at a constant addition rate while the
pH of the resulting mixture is monitored. Suitable addition rates are from 0.02 to 0.2 equivalents of the base per minute,
based on the amount of the ferric salt in the reaction mixture.
40 [0063] The addition of the second base to the reaction mixture is continued until the pH of the reaction mixture is in
the range from 3.5 to 9. According to some embodiments, the desired pH is about 4. According to other embodiments,
the desired pH is about 7. According to still other embodiments, the desired pH is about 8.3. Ferric hydroxide forms as
a precipitate in the reaction mixture during the second base addition. The ferric hydroxide precipitate begins to form at
a pH of about 3.
45 [0064] Following complete addition of the second base, the reaction mixture, comprising a suspension of ferric hy-
droxide, is allowed to equilibrate for a time interval from about 5 minutes to about 60 minutes. The ferric hydroxide
precipitate is observed to settle during the equilibration period.
[0065] The ferric hydroxide precipitate may be collected from the reaction mixture by any suitable method, including,
for example, filtration, centrifugation, or decanting. Filtration is preferred. Suitable filtration methods include vacuum
50 filtration, e.g., for example through a Buckner funnel. For larger scale or manufacturing operations, an agitated nutsch
filter is preferred. The resulting filter cake comprising ferric hydroxide is washed with water and then prepared as a slurry
in an aqueous solvent. Suitable aqueous solvents include water and mixtures of water with one or more water-miscible
organic solvents, wherein the water-miscible organic solvent comprises up to about 30% of the aqueous solvent.
[0066] To prepare iron sucrose, a suspension or a slurry of ferric hydroxide, as prepared above, is reacted with sucrose
in a reaction mixture comprising sodium ions. The sucrose is preferably provided as a solution of sucrose in an aqueous
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solvent, wherein the ratio of sucrose to aqueous solvent is preferably in the range from 1:0.1 to 1:4, more preferably in
the range from 1:0.2 to 1:4. The sodium ions are preferably provided by addition of a sodium base, preferably aqueous
sodium hydroxide. The mixture of the ferric hydroxide and sucrose may become basic upon the addition of the sodium
base. The sodium base may be added to the aqueous medium, and the ferric hydroxide may subsequently be added.
5 Alternately, the sodium base may be added to the mixture of the ferric hydroxide and sucrose in the aqueous medium.
[0067] The reaction mixture is optionally cooled to a temperature in the range from 20˚ to 30˚C, prior to the step of
isolating iron sucrose complex from the reaction mixture.
[0068] The reaction of ferric hydroxide and sucrose may be optionally monitored to determine the weight average
molecular weight and purity of the iron sucrose product. Monitoring may be done by removing an aliquot of the reaction
10 mixture and conducting a molecular weight analysis on the aliquot.
[0069] When the reaction of ferric hydroxide and sucrose is complete (as determined by observation of the clarity of
the reaction mixture and by GPC analysis of reaction aliquots) the iron sucrose complex is isolated from the reaction
mixture. Isolation may be achieved by adding one or more water-miscible organic solvents to the reaction mixture to
precipitate the iron sucrose complex. The amount of water-miscible organic solvent added to the reaction mixture is
15 preferably in the range of from 0.3 to 10 times the volume of the reaction mixture to which it is added. The product iron
sucrose complex that precipitates from the reaction mixture after addition of the water-miscible organic solvent is collected
from the reaction mixture. Suitable methods for collecting the product include, for example, filtration, centrifugation and
decanting. The product is preferably collected by filtration. The selection of suitable filtration media, e.g., for example,
a sintered glass funnel or Bucknes funnel, is within the capability of one of ordinary skill in the art.
20 [0070] Alternately, the iron sucrose complex is isolated from the reaction mixture by concentrating the reaction mixture
to form a residue comprising the iron sucrose complex. Concentration of the reaction mixture may be carried out at
reduced pressure or at atmospheric pressure, utilizing concentration techniques such as, lyophilization, distillation or
vacuum centrifugation. The concentration of the reaction mixture is preferably done at a temperature from -80˚ to 105˚C,
more preferably at a temperature from 35˚ to 105˚C. Isolation of the iron sucrose complex by concentrating the reaction
25 mixture at low temperature and low pressures is referred to herein as "freeze drying." Freeze drying may be performed
using a commercially available freeze-drying apparatus by cooling the reaction mixture to below the freezing temperature,
preferably from -80˚ to -50˚C and applying vacuum, preferably a pressure in the range from 10 millitorr to 50 torr, more
preferably in the range from 10 millitorr to 10 torr, most preferably from 10 millitorr to 1 torr.
[0071] The product iron sucrose complex thus obtained, in the form of a filtrate or a residue as described above, may
30 be optionally purified by (a) dissolving the isolated iron sucrose in an aqueous solvent; (b) forming a mixture by adding
at least one water-miscible organic solvent to the solution formed in step (a), the solvent being added in an amount
sufficient to precipitate iron sucrose complex from the mixture formed in step (b); and (c) collecting the precipitated
purified iron sucrose complex.
[0072] The aqueous solvent used to dissolve the filtrate or residue is preferably employed in an amount in the range
35 from 0.2 to 10 times the weight of the filtrate or residue to be dissolved therein, more preferably from 0.3 to 7 times the
weight of the filtrate or residue to be dissolved therein. The pH of the resulting solution is preferably adjusted to a pH in
the range from 8 to 14, preferably from 9 to 13, more preferably from 10 to 14, by the addition of a base such as sodium
hydroxide. Suitable water-miscible organic solvents for the precipitation of purified iron sucrose complex include, for
example, methanol, ethanol, acetone, tetrahydrofuran, dioxane, acetonitrile and mixtures thereof. The amount of the
40 water-miscible organic solvent added to the solution of the residue or filtrate to precipitate iron sucrose complex is
preferably in the range from a 15:1 to a 1:7 ratio of the water-miscible organic solvent to aqueous solvent (wt/wt), more
preferably from a 1:0.3 to a 1:7 ratio or from a 15:1 to a 1:1 ratio (wt/wt) of the water-miscible organic solvent to aqueous
solvent.
[0073] The purified iron sucrose complex thus obtained, is optionally dried. Drying of the purified iron sucrose complex
45 may be carried out under vacuum or at atmospheric pressure, in air or under an inert atmosphere such as, for example,
nitrogen. Preferably the purified product is dried at a temperature from 25˚ to 140˚C.
[0074] The purified iron sucrose complex prepared according to the process of the invention preferably contains no
more than about 10% wt/wt free sucrose. The purified iron sucrose complex prepared according to the process of the
invention preferably no more than about 10% (wt/wt) of sodium salts such as for example, sodium chloride, sodium
50 bromide, sodium iodide, sodium nitrate, sodium sulfate, sodium acetate or sodium citrate, or mixtures thereof.
[0075] The purified iron sucrose complex preferably contains no more than about 15% wt/wt water and solvent. The
purified iron sucrose complex may exist in a hydrated form: [Complex]u[H2O]v; wherein "Complex" is the iron sucrose
complex; "u" is an integer from 1 to 1000, preferably from 1 to 500, and v is a rational number from 0.2 to 50, preferably
from 0.2 to 25, more preferably from 0.2 to 10.
55 [0076] The thus-obtained precipitated purified iron sucrose complex, may be formulated as a parenteral iron formu-
lation. One example of a parenteral iron formulation comprises dissolving the precipitated purified iron sucrose complex,
containing from 20% to 50% ferric iron, in an aqueous sucrose solution, preferably 20% aqueous sucrose, to form a
parenteral iron formulation. The concentration of precipitated purified iron sucrose complex in the sucrose solution is
8
EP 1 733 058 B1
selected such that the composition is suitable as an injectable form of ferric iron. Another example of a parenteral iron
formulation comprises dissolving the precipitated purified iron sucrose complex, containing from 2% to 15% ferric iron,
in water for injection to form a parenteral iron formulation. The concentration of precipitated purified iron sucrose complex
in the aqueous solution is selected such that the composition is suitable as an injectable form of ferric iron.
5 [0077] According to another embodiment of the invention, there is provided a pharmaceutical composition in solid
dosage form comprising a pharmaceutically acceptable carrier and an iron sucrose complex having a molecular weight
in the range from 20,000 to 400,000 Daltons. The pharmaceutical composition of the invention may be formulated for
oral administration and may be in the form of a tablet, capsule, pill, powder, granule or other suitable solid dosage form
with suitable excipients and additives.
10 [0078] For example, the iron sucrose complex formed by the process of the present invention may be combined with
at least one excipient such as a filler, binder, humectant, disintegrating agent, solution retarder, absorption accelerator,
wetting agent absorbent or lubricating agent. According to one embodiment of a solid dosage form of an iron sucrose
complex formed the claimed process, the active agent may be combined with carboxymethylcellulose calcium, magne-
sium stearate, mannitol and starch, and then formed into tablets by conventional tableting methods.
15 [0079] The pharmaceutical composition according the invention comprises an iron sucrose complex that contains
from 1 to 60% ferric iron by weight, preferably from 1 to 50% ferric iron by weight, more preferably from 30 to 50% ferric
iron by weight.
[0080] The pharmaceutical composition according the invention preferably contains from 5 to 200 mg of ferric iron,
more preferably from 10 to 150 mg, most preferably from 25 to 100 mg.
20 [0081] The practice of the invention is illustrated by the following non-limiting examples.
Examples
[0082] The following analytical methods are employed in the Examples that follow.
25
Molecular weight determination for iron sucrose complex
[0083] GPC analyses were performed using a Shirnadzu Class VP, SCL10A-VP, with an LC10AD pump, equipped
with a refractive index detector (Shimadzu RID 10A). The mobile phase employed was an aqueous buffer prepared by
30 dissolving 7.12g of dibasic sodium phosphate dihydrate, 5.52 g of monobasic sodium phosphate, and 0.40 g of sodium
azide in 2 liters of water.
[0084] The separation media consists of two 7.8-mm x 30-cm columns (Waters Ultrahydrogel GPC column containing
packing L39, with pore sizes of 1000A and 120A, respectively) set up in series. The column temperatures were maintained
at 45˚ +/- 2˚ and the flow rate was about 0.5 mL per minute.
35 [0085] Standard solutions (Waters Dextran molecular weight standard kit part # WAT 054392) were prepared by
accurately weighing about 20 mg of each polysaccharide molecular weight standard (5,000- 400,000 Da) into separate
5mL volumetric flasks. Mobile phase (about 4 mL) was added to each flask and the resulting mixture was allowed to
stand at or below 25˚C for a minimum of 12 hours. After the agglomerate particles of each standard solution swelled to
their fullest extent, each standard solution was gently agitated until the polysaccharide dissolved. Chromatograms of
40 freshly prepared standard solutions regularly show a small, unidentified secondary peak following the main peak. Any
standard solutions wherein the secondary peak reached half the height of the main peak were discarded.
[0086] A system suitability test solution was also prepared by dissolving 200 mg of high molecular weight dextran and
100 mg of glucose in 20 mL of the mobile phase.
[0087] Test sample solutions of iron sucrose complex for analysis were prepared by transferring about 250 mg of
45 each iron sucrose complex to a 10-mL volumetric flask, diluting to the line with mobile phase, and mixing. Test samples
which were reaction aliquots were prepared by diluting 1 mL of the reaction mixture to 10 mL with mobile phase.
[0088] About 25 mL of each standard solution and test sample solution was injected (Shimadzu auto injector SIL10A-
VP) onto the column. Chromatograms were recorded and the retention times and peak areas of all components above
the detectability threshold were measured. The analyte retention times were as follows:
50
Iron sucrose complex peak : About 27 minutes
Sucrose (free) peak : About 38 minutes
[0089] The retention times of the standard solutions and their molecular weights were plotted to generate a third order
55 (cubic) calibration curve. The correlation coefficient obtained was not less than 0.98. The molecular weight of the complex
was calculated using the calibration curve. The molecular weight distribution curve of the each sample was sliced into
fractions. The weight-average molecular weight (Mw) was calculated according to the formula:
9
EP 1 733 058 B1
5 wherein AT is the area of each fraction of the sample distribution; and MT is the corresponding mean molecular weight
of each fraction as determined from its retention time on the calibration curve. The molecular weight di stribution curve
obtained for the Injection conformed to the following parameters:
Mw = 34,000-60,000 Da,
10 Mn = not less than 24,000 Da, and
Mw/Mn = not more than 1.7.
15 [0090] About 50 mg of iron sucrose complex was finely crushed using an agate pestle and mortar. The crushed sample
was placed over the trough plate of the Horizontal Attenuated Total Reflectance (HATR) assembly of a Perkin Elmer
Spectrum 1 FTIR Spectrometer. The spectrum was recorded (4 scans, 4000 to 800 cm-1) and corrected for background
signal.
[0091] About 5 mg of iron sucrose complex was dissolved in 1.5 mL of D2O and transferred into an NMR sample tube.
The proton NMR was recorded (-5 to 20 ppm, Varian 400 MHz NMR spectrometer) using standard parameters with 3-
trimethylsilylpropionic acid sodium salt (TSP) as an internal standard.
25
Determination of Iron content by Atomic Absorption Spectroscopy (AAS)
[0092] An iron content calibration curve was prepared by plotting absorbances at the iron emission line at 248.3 nm
versus concentration (mg per mL) for a series of standard iron solutions. The absorbances were measured with a Perkin
30 Elmer 5000 atomic absorption spectrophotometer equipped with an iron hollow-cathode lamp and air-acetylene flame,
and using a calcium chloride solution as a blank. Reaction aliquots from reactions performed in the preparation of the
iron sucrose complexes of the invention were dissolved in water, and the absorbance at 248.3 nm were recorded. The
content of iron in the samples prepared from reaction aliquots was determined according to the prepared calibration curve.
35 Elemental Analyses
[0093] Elemental analyses (carbon and hydrogen) were performed by Atlantic MicroLabs, Norcross, GA.
Determination of Degree of Crystallinity of Iron Sucrose Complexes by X-ray Powder Diffraction Analyses
40
[0094] X-ray powder diffraction analyses were performed on the dried purified products of the reactions performed
in the preparation of the iron sucrose complexes of the invention. Each sample of iron sucrose complex of the invention
was analyzed as a fine powder. The dried purified reaction products required no additional processing before X-ray
diffraction analysis. The powder sample to be analyzed was placed onto a zero background holder and inserted into a
45 Philips PW1800 XR diffractometer. The X-ray analysis comprised Cu radiation over the angular range (theta) of 5˚ to
60˚ with a step size of 0.03˚. The analyses at each step required from about 5 to about 30 seconds depending on the
degree of crystallinity of the sample.
[0095] Ferric chloride hexahydrate (5 g, 18.5 mmol) was dissolved in deionized water (20 mL) at a temperature of
about 20˚ C. To the stirred ferric chloride solution was added sodium carbonate 30% w/v aqueous solution dropwise
55 with the pH of the ferric chloride solution monitored during the addition (first base addition). The addition was stopped
when the pH reached 2.2. The pH of the mixture was monitored using a pH meter. The temperature of the mixture was
maintained at about 20˚ C. Following the addition of sodium carbonate, the resulting mixture was dark brown to reddish
brown in color. The mixture was allowed to stand for about 30 min., during which time the pH of the reaction mixture
10
EP 1 733 058 B1
was observed to drop to 1.7. Additional sodium carbonate solution (second base addition) was added as the pH of the
mixture was monitored. A gelatinous precipitate appeared at a pH of about 3.0. The addition of the sodium carbonate
solution was continued until the pH of the mixture reached 4.0, yielding a suspension of ferric hydroxide. Following the
addition of sodium carbonate solution, the reaction mixture was allowed to stand for about 10 minutes to allow the
5 precipitate to settle. The precipitated ferric hydroxide was then collected by filtration and washed with water (25 mL).
The wet filter cake (about 15 g) was made into a slurry in water (about 20 mL).
10 [0096] To a 100 mL three necked round bottom flask, fitted with reflux condenser and stirrer assembly, was added
water (15 mL) and sucrose (30 g). The resulting mixture was heated in an oil bath maintained at 120 ˚C for about 10
min. The temperature of the mixture reached about 100-105˚C. Sodium hydroxide (about 2 mL, 20 % w/v) was added
to the heated reaction mixture. Then the slurry of ferric hydroxide prepared in Step 1 was added to the sucrose mixture
over about 15 min. The reaction mixture formed a clear dark brown solution after the addition of the suspension of ferric
15 hydroxide was completed. Following the addition of the ferric hydroxide slurry, the temperature of the reaction mixture
was maintained at about 100 to 105˚C for about 2 hrs. The reaction mixture was subsequently cooled to ambient
temperature (20 to 25˚C). An aliquot (1 mL) of the reaction mixture was removed for GPC analysis to confirm the weight
average molecular weight of the product iron sucrose complex (about 55,000 Daltons).
[0097] A water-miscible organic solvent (ethanol, about 350 mL) was added to the reaction mixture formed in Step 2,
at about 25˚C with stirring. A dark brown precipitate formed and was collected by filtration. The collected product was
further purified by dissolving it in water (10 mL) and subsequently adding ethanol (50 mL) to the dissolved product to
25 precipitate a purified product. The precipitate was collected by filtration, washed with ethanol, and dried under vacuum
at about 50˚C. The purified product was identified as iron sucrose complex and was analyzed by GPC. The GPC analysis
showed that the complex corresponding to the principle peak was present in greater than 95% purity and that the weight
average molecular weight remained at about 55,000 Daltons. This peak corresponds to the iron sucrose peak obtained
on analysis of the marketed iron sucrose in sucrose (VENOFER®), shown in Figure 3. The molecular weight and weight
30 average molecular weight values were determined using calibration curves and third order fitting. The procedure for
molecular weight determination of iron sucrose was the same as that which is described in USP 26, page 1016. The
content of Fe, C and H were determined for the precipitated Iron sucrose complex: Fe, 47% ; C, 25%; H, 7%;
[0100] The procedure of Example 1 was followed, except that the second base addition was continued until the pH
45 reached 7.0.
[0101] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 65,000 Daltons.
[0102] The procedure of Example 1 was followed, except that the second base addition was continued until the pH
reached 8.3.
[0103] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
55 corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 90,000 Daltons (Fig. 4).
11
EP 1 733 058 B1
[0104] The procedure of Example 1 was followed, except that in step 2, the 20% sodium hydroxide solution (2 mL)
was added after the completion of the addition of the slurry of ferric hydroxide, and in Step 3, the water miscible solvent
5 was methanol.
[0105] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 120,000 Daltons.
[0106] The procedure of Example 1 was followed, except that the first base addition comprised addition of the base
(sodium carbonate 30% w/v aqueous solution) as a single portion of 3.3 mL (about 1 equivalent based on the amount
of ferric salt).
15 [0107] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 50,000 Daltons. The GPC analysis of the purified product is shown in Figure 1. This peak corresponds to the
iron sucrose peak obtained on analysis of the marketed iron sucrose in sucrose (VENOFER®), shown in Figure 3.
[0108] Powder diffraction analysis was obtained for a sample of the dried purified product. The powder diffraction
20 analysis is reproduced in Fig. 5. The powder diffraction analysis indicates the presence of an amorphous product which
is substantially free of crystalline sucrose .
25 [0109] The procedure of Example 1 was followed, except that the first base addition comprised batchwise addition of
the base (sodium carbonate 30% w/v aqueous solution) in a single portion of 6.6 mL (about 2 equivalents based on the
amount of ferric salt).
[0110] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
30 was about 45,000 Daltons.
[0115] The procedure of Example 7 was followed, except that the second base addition was continued until the pH
reached 7.0.
12
EP 1 733 058 B1
[0116] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 60,000 Daltons.
5 Example 10: Preparation of Iron sucrose Complex and Analysis of Variation in Weight Average Molecular
Weight over Time
[0117] The procedure of Example 9 was followed, except for the following changes in Steps 1 and 2.
[0118] Step 1: The amount of the first base addition was 3.3 mL of 30% wt/Vol, aqueous sodium carbonate.
10 [0119] Step 2: One aliquot of the reaction mixture was removed immediately following completion of the addition of
the ferric hydroxide slurry. The reaction mixture was then maintained at 100-105˚C for about two hours, and then cooled
to ambient temperature (20 to 25˚C). A second aliquot of the reaction mixture was removed. GPC analysis was performed
on both the first and second aliquots. The weight average molecular weight determined for the first aliquot was about
85,000 Daltons. The weight average molecular weight determined for the second aliquot was about 50,000 Daltons.
15 [0120] The GPC analyses of the final purified and dried product indicated a weight average molecular weight of about
50,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of the marketed iron sucrose in
sucrose (VENOFER®), shown in Figure 3.
[0125] The procedure of example 9 was followed except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt./Vol, aqueous sodium carbonate, the water-miscible organic solvent used to precipitate the complex in Step
2 was isopropanol (350 mL), and the water-miscible organic solvent used to precipitate the purified complex in Step 2
35 was isopropanol (50 mL).
[0126] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 50,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of the marketed iron
sucrose in sucrose (VENOFER®), shown in Figure 3.
40
Example 13: Preparation of Iron sucrose Complex
[0127] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt./Vol, aqueous sodium carbonate, the water-miscible organic solvent used to precipitate the complex in Step
45 2 was acetone (350 mL), and the water-miscible organic solvent used to precipitate the purified complex in Step 2 was
acetone (50 mL).
[0128] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 50,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of the marketed iron
50 sucrose in sucrose (VENOFER®), shown in Figure 3.
[0129] The procedure of Example 9, except the amount of the first base addition in Step 1 was 3.3 mL of 30% wt/Vol,
55 aqueous sodium carbonate, the water-miscible organic solvent used to precipitate the complex in Step 2 was acetonitrile
(350 mL), and the water-miscible organic solvent used to precipitate the purified complex in Step 2 was acetonitrile (50
mL).
[0130] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
13
EP 1 733 058 B1
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 50,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of
the marketed iron sucrose in sucrose (VENOFER®), shown in Figure 3.
[0131] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt/Vol, aqueous sodium carbonate, and the purified product was dried under vacuum at a temperature of 120˚C.
Also, the water-miscible organic solvent used to precipitate the complex and to precipitate the purified complex in Step
10 2 was acetonitrile (350 mL and 50 mL, respectively).
[0132] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 50,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of
the marketed iron sucrose in sucrose (VENOFER®), shown in Figure 3.
15
Example 16: Preparation of Iron sucrose Complex
[0133] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt/Vol, aqueous sodium carbonate, and the amount of sucrose used in step 2 was 15 g. Also, the water-miscible
20 organic solvent used to precipitate the complex and to precipitate the purified complex in Step 2 was acetonitrile (350
mL and 50 mL, respectively).
[0134] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 75,000 Daltons.
25
Example 17: Preparation of Iron sucrose Complex
[0135] The procedure of Example 9 was followed, except the first base addition in Step 1 was the addition of 1 g of
sodium carbonate added as a solid. Also, the water-miscible organic solvent used to precipitate the complex and to
30 precipitate the purified complex in Step 2 was acetonitrile (350 mL and 50 mL, respectively).
[0136] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 55,000 Daltons.
[0137] The procedure of Example 9 was followed, except the first base addition in Step 1 was solid sodium bicarbonate
(first addition was 1.6g added all at once). Also, the water-miscible organic solvent used to precipitate the complex and
to precipitate the purified complex in Step 2 was acetonitrile (350 mL and 50 mL, respectively).
40 [0138] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 55,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of
the marketed iron sucrose in sucrose (VENOFER®), shown in Figure 3.
[0139] The procedure of Example 9 was followed, except the first added base in Step 1 was solid sodium bicarbonate
(first addition was 1.6g added all at once), and the second added base was sodium hydroxide (20% aqueous solution).
Also, the water-miscible organic solvent used to precipitate the complex and to precipitate the purified complex in Step
50 2 was acetonitrile (350 mL and 50 mL, respectively).
[0140] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 80,000 Daltons.
[0141] The procedure of Example 9 was followed, except that the first added base in Step 1 was tris-hydroxyethylami-
nomethane (first addition is 2.2 g added all at once), and the second added base was sodium hydroxide (20% aqueous
14
EP 1 733 058 B1
solution). Also, the water-miscible organic solvent used to precipitate the complex and to precipitate the purified complex
in Step 2 was acetonitrile (350 mL and 50 mL, respectively).
[0142] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
5 molecular weight was about 160,000 Daltons.
[0143] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
10 of 30% wt/Vol, aqueous sodium carbonate, and in Step 2 the heating temperature was 80˚C and the heating time was
about 4 hours.
[0144] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 80,000 Daltons.
15
Example 22: Preparation of Iron sucrose Complex
[0145] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt/Vol, aqueous sodium carbonate. Also, in Step 2, the solvent was 5 mL of water and 5 mL of ethanol, the
20 reaction temperature was about 80˚C, and the heating time was about 4 hours.
[0146] The GPC analyses of the reaction aliquot of Step 2 and of the final purified and dried product indicated that
the complex corresponding to the principle peak was present in greater than 95% purity and that the weight average
molecular weight was about 90,000 Daltons.
[0147] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt/Vol, aqueous sodium carbonate.
[0148] In Step 2, the GPC analysis of the aliquot showed a weight average molecular weight of the product was about
30 50,000 Daltons.
[0149] Also in Step 3, the reaction mixture was concentrated by vacuum distillation at 35˚C, prior to the addition of
ethanol (50 mL) which served to precipitate the iron sucrose complex. In addition, the purification of Step 3 was carried
out by dissolving the product in a 40% wt/Vol. sucrose solution in water (10 mL) and subsequently adding ethanol (50
mL) to the dissolved product to precipitate a purified product. The precipitate was collected by filtration, washed with
35 ethanol, and dried under vacuum at about 50˚C.
[0150] The purified product was identified as iron sucrose complex co-precipitated with sucrose and was analyzed
by GPC. The GPC analysis, shown in Figure 2, showed that the weight average molecular weight of the product was
about 50,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of the marketed iron sucrose
in sucrose (VENOFER®), shown in Figure 3.
40
Example 24: Preparation of Iron sucrose Complex
[0151] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt/Vol, aqueous sodium carbonate.
45 [0152] In Step 2, the GPC analysis of the aliquot showed a weight average molecular weight of the product was about
50,000 Daltons.
[0153] Also in Step 3, the reaction mixture was concentrated by vacuum distillation at 35˚C prior to the addition of
ethanol (50 mL) which served to precipitate the iron sucrose complex. In addition, the purification of Step 3 was carried
out by dissolving the product in a 20% wt/Vol. sodium chloride solution in water (10 mL) and subsequently adding ethanol
50 (50 mL) to the dissolved product to precipitate a purified product. The precipitate was collected by filtration, washed with
ethanol, and dried under vacuum at about 50˚C.
[0154] The purified product was identified as iron sucrose complex co-precipitated with sodium chloride confirmed by
the test for chloride ions using silver nitrate. The product was also analyzed by GPC.
[0155] The procedure of Example 9 was followed, except the amount of the first base addition in Step 1 was 3.3 mL
of 30% wt/Vol, aqueous sodium carbonate, and the ferric salt employed in Step 1 consisted of a combination of ferric
15
EP 1 733 058 B1
chloride hexahydrate (2.5 g, 9.25 mmol) and ferric nitrate nonahydrate (3.75 g, 9.25 mmol).
[0156] Also in Step 3, the reaction mixture was concentrated by vacuum distillation at 35˚C prior to the addition of
ethanol (50 mL) which served to precipitate the iron sucrose complex.
[0157] The GPC analyses of the reaction aliquot and of the final purified and dried product indicated that the complex
5 corresponding to the principle peak was present in greater than 95% purity and that the weight average molecular weight
was about 50,000 Daltons. This peak corresponds to the iron sucrose peak obtained on analysis of the marketed iron
sucrose in sucrose (VENOFER®), shown in Figure 3.
Example 26: Isolation of Iron sucrose Complex via Concentration of the Reaction Mixture According to Step
10 2 of Example 6
[0158] The procedure of Example 6 was followed, except the reaction mixture of Step 2 was subjected to vacuum
distillation at 50 to 60˚C to reduce the volume of the reaction mixture to about 30% of its original volume. The temperature
of the resulting mixture was adjusted to 25˚C, and ethanol (50 mL) was added with stirring. A dark brown precipitate
15 formed. The precipitate was collected by filtration. The collected product was purified by dissolution in water (10 mL).
Addition of ethanol (50 mL) to the dissolved product served to form a precipitate. This precipitate was collected by
filtration, washed with ethanol and dried under vacuum at about 50˚C. The product was identified as iron sucrose complex
co-precipitated with sucrose. Analysis (GPC) of the purified product, and of the aliquot removed in Step 2, yielded a
weight average molecular weight for the peak corresponding to the iron sucrose complex of about 70000 Daltons.
20
Example 27 Preparation (18 mmol scale) of Iron sucrose Complex without Isolating Ferric Hydroxide by Filtra-
tion
[0160] To the above-prepared suspension of ferric hydroxide at a pH of 1.7, was added sucrose (30 g). The resulting
35 mixture was heated to a temperature of about 80˚C. Sodium carbonate was added to adjust the pH to 7.0. The resulting
mixture was heated to a temperature of 100˚C and stirred at 100˚C for about 15 minutes. Sodium hydroxide (about 3
mL, 20 % w/v) was added to the heated reaction mixture. The reaction mixture formed a clear dark brown solution after
the addition of the suspension of ferric hydroxide was completed. Following the addition of the sodium hydroxide solution,
the temperature of the reaction mixture was maintained at about 100 to 105˚C for about 3 hrs. The reaction mixture was
40 subsequently cooled to ambient temperature (20 to 25˚C). An aliquot (1 mL) of the reaction mixture was removed for
GPC analysis to confirm the weight average molecular weight of the product iron sucrose complex (about 80000 Daltons).
45 [0161] The reaction mixture was concentrated by vacuum distillation at 50 to 60˚ C to about 30% of its original volume.
Ethanol (about 50 mL) was added to the concentrated reaction mixture at 25 ˚C with stirring. A dark brown precipitate
formed and was collected by filtration. The collected product was purified further by dissolving it in water (10 mL) and
subsequently adding ethanol (50 mL) to the dissolved product to precipitate a purified product. The precipitate was
collected by filtration, washed with ethanol, and dried under vacuum at about 50˚C. The purified product was identified
50 as iron sucrose complex and was analyzed by GPC.
16
EP 1 733 058 B1
the addition of sodium carbonate solution, the resulting mixture was dark brown to reddish brown in color. The mixture
was allowed to stand for about 30 min., during which time the pH of the reaction mixture was observed to drop to 1.7.
Additional sodium carbonate solution was added as the pH of the mixture was monitored. A gelatinous precipitate
appeared at a pH of about 3.0. Additional sodium carbonate solution was added until the pH of the mixture reached 4.0,
5 yielding a suspension of ferric hydroxide. Following the addition of sodium carbonate solution, the reaction mixture was
allowed to stand for about 10 minutes to allow the precipitate to settle. The precipitated ferric hydroxide was then collected
by filtration and washed with water (250 mL). The wet filter cake (about 150 g) was made into a slurry in water (about
200 mL).
[0163] To a 1000 mL three necked round bottom flask, fitted with reflux condenser and stirrer assembly, was added
water (50 mL) and sucrose (300 g). The resulting mixture was heated in an oil bath maintained at about 120 ˚C for about
20 min. The temperature of the mixture reached about 100-105˚C. Sodium hydroxide (about 20 mL, 20 % w/v) was
15 added to the heated reaction mixture. Then the slurry of ferric hydroxide prepared in Step 1 was added over about 25
min. The reaction mixture formed a clear, dark brown solution within one minute after the addition of the suspension of
ferric hydroxide was completed. Following the addition, the temperature of the reaction mixture was maintained at about
100 to 105˚C for about 2 hrs. The reaction mixture was subsequently cooled to ambient temperature (20 to 25˚C). An
aliquot (1 mL) of the reaction mixture was removed for GPC analysis to confirm the weight average molecular weight
20 of the product iron sucrose complex (about 50,000 Daltons). This peak corresponds to the iron sucrose peak obtained
on analysis of the marketed iron sucrose in sucrose (VENOFER®), shown in Figure 3.
25 [0164] Ethanol (about 3500 mL) was added into the reaction mixture formed in Step 2, at 25˚C with stirring. A dark
brown precipitate formed and was collected by filtration. The collected product was purified further by dissolving it in
water (100 mL) and subsequently adding ethanol (500 mL) to the dissolved product to precipitate a purified product.
The precipitate was collected by filtration, washed with ethanol, and dried under vacuum at about 50˚C. The purified
product was identified as iron sucrose complex and was analyzed by GPC, which yielded a weight average molecular
30 weight of about 50,000 Daltons for the principal peak, which was present at greater than 95% purity.
50 [0166] To a 100 mL three necked round bottom flask, fitted with reflux condenser and stirrer assembly, was added
water for injection (5 mL) and sucrose (15.2 g). The resulting mixture was heated in an oil bath maintained at 120 ˚C for
about 10 min. The temperature of the mixture reached about 100-105˚C. Sodium hydroxide (about 2 mL, 20 % w/v,
made with water for injection) was added to the heated reaction mixture. Then the slurry of ferric hydroxide prepared in
Step 1 was added to the sucrose mixture over about 15 min. The reaction mixture formed a clear dark brown solution
55 after the addition of the suspension of ferric hydroxide was completed. Following the addition of the ferric hydroxide
slurry, the temperature of the reaction mixture was maintained at about 100 to 105˚C for about 2 hrs. The reaction
mixture was subsequently cooled to ambient temperature (20 to 25˚C). An aliquot (1 mL) of the reaction mixture was
removed for GPC analysis to confirm the weight average molecular weight of the product iron sucrose complex (about
17
EP 1 733 058 B1
50,000 Daltons). The reaction mixture was concentrated by vacuum distillation at 35˚C. The resulting concentrate was
diluted with water to yield a solution containing 20 mg/mL of Ferric iron suitable for injection.
Example: 30: Preparation of Iron Sucrose Complex and Isolation by Freeze Drying
5
[0167] Iron sucrose complex was prepared by the process described in Steps 1 and 2 of Example 1.
[0168] After completion of the preparation of iron sucrose complex as in Step 2 of Example 1, the reaction mixture
was transferred to a round bottom flask and cooled to about -75˚C to freeze the reaction mixture. The frozen reaction
mixture was then freeze dried using a VIRTIS, model 12 EL freeze drying apparatus at a pressure of about 25 millitorr
10 for a time interval of about 12 hours.
Example: 31: Preparation (1.8 mol scale) of Iron Sucrose Complex and Isolation by Centrifugation
30 [0170] To a 10 liter three-necked round bottom flask, fitted with reflux condenser and stirrer assembly, was added
water (500 mL) and sucrose (3000 g). The resulting mixture was heated in an oil bath maintained at about 120˚C for
about 20 min. The temperature of the mixture reached about 100-105˚C. Sodium hydroxide (about 200 mL, 20 % w/v)
was added to the heated reaction mixture. Then, the slurry of ferric hydroxide prepared in Step 1 was added to the
reaction mixture over a time interval of about 25 min. The reaction mixture formed a clear, dark brown solution within
35 one minute after the addition of the suspension of ferric hydroxide was completed. Following the addition, the temperature
of the reaction mixture was maintained at about 100 to 105˚C for about 2 hrs. The reaction mixture was subsequently
cooled to ambient temperature (20 to 25˚C). An aliquot (1 mL) of the reaction mixture was removed for GPC analysis
to confirm the weight average molecular weight of the product iron sucrose complex (about 49,000 Daltons). This peak
corresponds to the iron sucrose peak obtained on analysis of the marketed iron sucrose product (VENOFER®), shown
40 in Figure 3.
[0171] Ethanol (about 5 000 mL) was added to the reaction mixture formed in Step 2, at 25˚C with stirring. A dark
45 brown precipitate formed. The precipitate was collected by centrifugation at 1500 rpm using a Rousselet-Robatel model
RC-30 centrifuge. The mixture containing the precipitated iron sucrose complex was fed to the centrifuge over about 30
min, and the centrifugation was continued for about 30 minutes after the feed was complete.
[0172] The collected precipitate was purified further by dissolving it in water (1000 mL) and subsequently adding
ethanol (5000 mL) to the dissolved product to precipitate a purified product. The precipitate was collected by centrifugation
50 as described above. The collected purified product was washed with ethanol, and dried under vacuum, at about 50˚C.
The purified product was identified as iron sucrose complex. GPC analysis yielded a weight average molecular weight
of about 49,000 Daltons for the principal peak, which was present at greater than 95% purity.
[0173] The present invention may be embodied in other specific forms without departing from the essential attributes
thereof and, accordingly, reference should be made to the appended claims, rather than to the foregoing specification,
55 as indication the scope of the invention.
18
EP 1 733 058 B1
Claims
1. A process of preparing iron sucrose complex containing no more than 15 % (wt./wt.) of excipients, comprising the
steps of:
5
(a) preparing ferric hydroxide by a process comprising the steps of:
(i) providing a reaction mixture comprising a ferric salt dissolved in an aqueous medium;
(ii) adding to the reaction mixture from 1 to 2 equivalents of a first base based on the amount of ferric salt
10 in the reaction mixture;
(iii) allowing the reaction mixture to equilibrate for a time interval greater than 10 minutes;
(iv) adding a second base to the equilibrated reaction mixture, in an amount sufficient to adjust the pH of
the reaction mixture to a selected pH; and
(v) collecting the ferric hydroxide from the reaction mixture;
15
(b) reacting the ferric hydroxide and sucrose in an aqueous reaction mixture comprising sodium ions, at a molar
ratio of sucrose to ferric hydroxide of from 2:1 to 50:1, for a selected time interval, at a selected temperature,
and at a pH in the range from 6.5 to 13; and
(c) isolating the iron sucrose complex from the aqueous reaction mixture.
20
2. A process according to claim 1, wherein the selected temperature of step (b) is in the range from 75˚C to 120˚C.
3. A process according to claim 1, wherein the selected time interval of step (b) is in the range from 2 minutes to 40 hours.
25 4. A process according to any of claims 1 to 3 wherein the isolated iron sucrose complex comprises crystalline iron
sucrose complex.
5. A process according to any of claims 1 to 3 wherein the isolated iron sucrose complex comprises amorphous iron
sucrose complex free of crystalline sucrose as determinable by X-ray powder diffraction.
30
6. A process according to any preceding claim wherein the isolated iron sucrose complex contains from 1 to 60% ferric
iron by weight.
7. A process according to claim 1, wherein the reaction mixture of step (b) comprises ferric iron in an amount from 0.2
35 % w/w to 8% w/w based on the weight of the reaction mixture.
8. A process according to any preceding claim, wherein the ferric salt is ferric chloride, ferric nitrate, or a mixture thereof.
9. A process according to claim 1, wherein the first and second bases are independently selected from the group
40 consisting of alkali metal carbonates, alkali metal bicarbonates, alkali metal hydroxides, and water-soluble amines.
10. A process according to claim 9, wherein the first and second base are independently selected from the group
consisting of sodium carbonate, sodium bicarbonate, sodium hydroxide, and tris-hydroxymethylaminoethane.
45 11. A process according to claim 1 wherein one or two equivalents of the first base are added to the reaction mixture
in step (a).
12. A process according to claim 1, wherein the first base is added to the reaction mixture in step (a) continuously at a
constant addition rate of from 0.02 to 0.2 equivalents of the base per minute, based on the amount of ferric salt,
50 until the pH of the resulting mixture is from 2.0 to 2.5.
13. A process according to claim 1, wherein the selected pH in step (a)(iv) is in the range from 3.5 to 9.
14. A process according to claim 13, wherein the selected pH is 4.0, 7.0, or 8.3.
55
15. A process according to claim 1 wherein the ferric hydroxide is collected from the reaction mixture in step (a)(v) by
filtration.
19
EP 1 733 058 B1
16. A process according to claim 15, further comprising the step of forming a slurry of the collected ferric hydroxide in
an aqueous solvent.
17. A process according to claim 16, wherein the aqueous solvent comprises water, or a mixture of water and a water-
5 miscible organic solvent.
18. A process according to any preceding claim, wherein the step of isolating the iron sucrose complex from the reaction
mixture comprises
10 (a) forming a mixture by adding to the temperature-adjusted reaction mixture of claim 1 at least one water-
miscible organic solvent in an amount sufficient to precipitate iron sucrose complex; and
(b) collecting the precipitated iron sucrose complex, from the mixture formed in step (a).
19. A process according to any preceding claim, wherein the step of isolating the iron sucrose complex from the aqueous
15 reaction mixture comprises freeze drying the aqueous reaction mixture; centrifugation of the aqueous reaction
mixture; or concentrating the reaction mixture to form a residue comprising iron sucrose complex.
20. The process according to any preceding claim, further comprising purifying the isolated iron sucrose complex, said
purifying comprising the steps of:
20
(a) dissolving the isolated iron sucrose complex in an aqueous solvent;
(b) forming a mixture by adding to the solution formed in step (a) at least one water-miscible organic solvent in
an amount sufficient to precipitate iron sucrose complex from the solution; and
(c) collecting the purified iron sucrose complex from the mixture formed in step (b).
25
21. A process of preparing a co-precipitate comprising iron sucrose complex and sucrose, comprising the steps of:
(a) preparing an isolated iron sucrose complex according to the process of claim 1;
(b) dissolving the isolated iron sucrose complex in an aqueous solvent to form a solution;
30 (c) forming a mixture by adding to the solution formed in step (b) at least one water-miscible organic solvent in
an amount sufficient to precipitate iron sucrose complex from the solution;
(d) collecting the purified iron sucrose complex from the mixture formed in step (c);
(e) dissolving the purified iron sucrose complex product formed in step (d) in an aqueous sucrose solution;
(f) forming a mixture by adding to the iron sucrose complex solution formed in step (e) at least one water-
35 miscible organic solvent in an amount sufficient to co-precipitate iron sucrose complex and sucrose; and
(g) collecting the co-precipitate formed in step (f).
22. A process according to claim 21, wherein the ratio of purified iron sucrose complex to aqueous sucrose solution is
in the range from 1:0.5 to 1:10 by weight.
40
23. A process according to claim 21, wherein the aqueous sucrose solution has a concentration in the range from 10%
to 50% weight/volume.
24. A process for preparing an aqueous solution of sucrose and iron sucrose complex, comprising the steps of:
45
(a) preparing an isolated iron sucrose complex according to the process of claim 1;
(b) dissolving the isolated iron sucrose complex in an aqueous solvent to form a solution;
(c) forming a mixture by adding to the solution formed in step (b) at least one water-miscible organic solvent in
an amount sufficient to precipitate iron sucrose complex from the solution;
50 (d) collecting the purified iron sucrose complex from the mixture formed in step (c); and
(e) dissolving the purified iron sucrose complex produced according to step (d) in a solution of sucrose in water.
25. A process of preparing an aqueous solution of sucrose and iron sucrose complex, comprising the steps of:
55 (a) preparing an isolated iron sucrose complex according to the process of claim 1;
(b) dissolving the isolated iron sucrose complex in an aqueous solvent to form a solution;
(c) forming a mixture by adding to the solution formed in step (b) at least one water-miscible organic solvent in
an amount sufficient to precipitate iron sucrose complex from the solution;
20
EP 1 733 058 B1
(d) collecting the purified iron sucrose complex from the mixture formed in step (c);
(e) dissolving the purified iron sucrose complex product formed in step (d) in an aqueous sucrose solution;
(f) forming a mixture by adding to the iron sucrose complex solution formed in step (e) at least one water-
miscible organic solvent in an amount sufficient to co-precipitate iron sucrose complex and sucrose;
5 (g) collecting the co-precipitate formed in step (f); and
(h) dissolving the collected co-precipitate in water.
Patentansprüche
10
1. Ein Verfahren zum Zubereiten eines Eisensaccharosekomplexes, der nicht mehr als 15 % (Gewicht/Gewicht) an
Hilfsstoffen enthält, das die folgenden Schritte beinhaltet:
(a) Zubereiten von Eisenhydroxid mittels eines Verfahrens, das die folgenden Schritte beinhaltet:
15
(i) Bereitstellen eines Reaktionsgemisches, das ein in einem wässrigen Medium aufgelöstes Eisensalz
beinhaltet;
(ii) Hinzufügen zu dem Reaktionsgemisch von von 1 bis 2 Äquivalenten einer ersten Base, basierend auf
der Menge an Eisensalz in dem Reaktionsgemisch;
20 (iii) Äquilibrierenlassen des Reaktionsgemisches für ein Zeitintervall von mehr als 10 Minuten;
(iv) Hinzufügen einer zweiten Base zu dem äquilibrierten Reaktionsgemisch in einer Menge, die ausreichend
ist, um den pH des Reaktionsgemisches auf einen ausgewählten pH einzustellen; und
(v) Entnehmen des Eisenhydroxids aus dem Reaktionsgemisch;
25 (b) Reagierenlassen des Eisenhydroxids und der Saccharose in einem wässrigen Reaktionsgemisch, das Na-
triumionen beinhaltet, bei einem Molverhältnis von Saccharose zu Eisenhydroxid von von 2:1 bis 50:1 für ein
ausgewähltes Zeitintervall bei einer ausgewählten Temperatur und einem pH im Bereich von 6,5 bis 13; und
(c) Isolieren des Eisensaccharosekomplexes aus dem wässrigen Reaktionsgemisch.
30 2. Verfahren gemäß Anspruch 1, wobei die ausgewählte Temperatur aus Schritt (b) im Bereich von 75 ˚C bis 120 ˚C liegt.
3. Verfahren gemäß Anspruch 1, wobei das ausgewählte Zeitintervall aus Schritt (b) im Bereich von 2 Minuten bis 40
Stunden liegt.
35 4. Verfahren gemäß einem der Ansprüche 1 bis 3, wobei der isolierte Eisensaccharosekomplex einen kristallinen
Eisensaccharosekomplex beinhaltet.
5. Verfahren gemäß einem der Ansprüche 1 bis 3, wobei der isolierte Eisensaccharosekomplex einen amorphen
Eisensaccharosekomplex beinhaltet, der frei von kristalliner Saccharose ist, wie mittels Pulverröntgenaufnahme
40 bestimmbar ist.
6. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei der isolierte Eisensaccharosekomplex von 1 bis
60 Gew.-% dreiwertiges Eisen enthält.
45 7. Verfahren gemäß Anspruch 1, wobei das Reaktionsgemisch aus Schritt (b) dreiwertiges Eisen in einer Menge von
0,2 Gew.-% bis 8 Gew.-% beinhaltet, basierend auf dem Gewicht des Reaktionsgemisches.
8. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei das Eisensalz Eisenchlorid, Eisennitrat oder ein
Gemisch davon ist.
50
9. Verfahren gemäß Anspruch 1, wobei die erste und zweite Base unabhängig aus der Gruppe, bestehend aus Alka-
limetallcarbonaten, Alkalimetallbicarbonaten, Alkalimetallhydroxiden und wasserlöslichen Aminen, ausgewählt sind.
10. Verfahren gemäß Anspruch 9, wobei die erste und zweite Base unabhängig aus der Gruppe, bestehend aus Na-
55 triumcarbonat, Natriumbicarbonat, Natriumhydroxid und Tris-hydroxymethylaminoethan, ausgewählt sind.
11. Verfahren gemäß Anspruch 1, wobei in Schritt (a) ein oder zwei Äquivalente der ersten Base zu dem Reaktions-
gemisch hinzugefügt werden.
21
EP 1 733 058 B1
12. Verfahren gemäß Anspruch 1, wobei die erste Base in Schritt (a) zu dem Reaktionsgemisch bei einer konstanten
Hinzufügungsrate von von 0,02 bis 0,2 Äquivalenten der Base pro Minute hinzugefügt wird, basierend auf der Menge
an Eisensalz, bis der pH des sich ergebenden Gemisches von 2,0 bis 2,5 beträgt.
5 13. Verfahren gemäß Anspruch 1, wobei der ausgewählte pH in Schritt (a)(iv) im Bereich von 3,5 bis 9 liegt.
14. Verfahren gemäß Anspruch 13, wobei der ausgewählte pH 4,0, 7,0 oder 8,3 beträgt.
15. Verfahren gemäß Anspruch 1, wobei das Eisenhydroxid aus dem Reaktionsgemisch in Schritt (a)(v) mittels Filtration
10 entnommen wird.
16. Verfahren gemäß Anspruch 15, das ferner den Schritt des Bildens einer Schlämme des entnommenen Eisenhy-
droxids in einem wässrigen Lösungsmittel beinhaltet.
15 17. Verfahren gemäß Anspruch 16, wobei das wässrige Lösungsmittel Wasser oder ein Gemisch aus Wasser und
einem wassermischbaren organischen Lösungsmittel beinhaltet.
18. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei der Schritt des Isolierens des Eisensaccharose-
komplexes aus dem Reaktionsgemisch Folgendes beinhaltet:
20
(a) Bilden eines Gemisches durch Hinzufügen mindestens eines wassermischbaren organischen Lösungsmit-
tels zu dem temperatureingestellten Reaktionsgemisch aus Anspruch 1 in einer Menge, die ausreichend ist,
um den Eisensaccharosekomplex zu präzipitieren; und
(b) Entnehmen des präzipitierten Eisensaccharosekomplexes aus dem in Schritt (a) gebildeten Gemisch.
25
19. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei der Schritt des Isolierens des Eisensaccharose-
komplexes aus dem wässrigen Reaktionsgemisch das Gefriertrocknen des wässrigen Reaktionsgemisches; das
Zentrifugieren des wässrigen Reaktionsgemisches; oder das Konzentrieren des Reaktionsgemisches zum Bilden
eines Rests, welcher den Eisensaccharosekomplex beinhaltet, beinhaltet.
30
20. Verfahren gemäß einem der vorhergehenden Ansprüche, das ferner das Reinigen des isolierten Eisensaccharo-
sekomplexes beinhaltet, wobei das Reinigen die folgenden Schritte beinhaltet:
40 21. Verfahren zum Zubereiten eines Co-Präzipitats, das einen Eisensaccharosekomplex und Saccharose beinhaltet,
das die folgenden Schritte beinhaltet:
(a) Zubereiten eines isolierten Eisensaccharosekomplexes gemäß dem Verfahren aus Anspruch 1;
(b) Auflösen des isolierten Eisensaccharosekomplexes in einem wässrigen Lösungsmittel zum Bilden einer
45 Lösung;
(c) Bilden eines Gemisches durch Hinzufügen mindestens eines wassermischbaren organischen Lösungsmit-
tels zu der in Schritt (b) gebildeten Lösung in einer Menge, die ausreichend ist, um den Eisensaccharosekomplex
aus der Lösung zu präzipitieren;
(d) Entnehmen des gereinigten Eisensaccharosekomplexes aus dem in Schritt (c) gebildeten Gemisch;
50 (e) Auflösen des in Schritt (d) gebildeten gereinigten Eisensaccharosekomplexprodukts in einer wässrigen
Saccharoselösung;
(f) Bilden eines Gemisches durch Hinzufügen mindestens eines wassermischbaren organischen Lösungsmittels
zu der in Schritt (e) gebildeten Eisensaccharosekomplexlösung in einer Menge, die ausreichend ist, um den
Eisensaccharosekomplex und Saccharose zu co-präzipitieren; und
55 (g) Entnehmen des in Schritt (f) gebildeten Co-Präzipitats.
22. Verfahren gemäß Anspruch 21, wobei das Gewichtsverhältnis von gereinigtem Eisensaccharosekomplex zu wäss-
riger Saccharoselösung im Bereich von 1:0,5 bis 1:10 liegt.
22
EP 1 733 058 B1
23. Verfahren gemäß Anspruch 21, wobei die wässrige Saccharoselösung eine Konzentration im Bereich von 10 % bis
50 % (Gewicht/Volumen) aufweist.
24. Verfahren zum Zubereiten einer wässrigen Lösung aus Saccharose und Eisensaccharosekomplex, das die folgen-
5 den Schritte beinhaltet:
(a) Zubereiten eines isolierten Eisensaccharosekomplexes gemäß dem Verfahren aus Anspruch 1;
(b) Auflösen des isolierten Eisensaccharosekomplexes in einem wässrigen Lösungsmittel zum Bilden einer
Lösung;
10 (c) Bilden eines Gemisches durch Hinzufügen mindestens eines wassermischbaren organischen Lösungsmit-
tels zu der in Schritt (b) gebildeten Lösung in einer Menge, die ausreichend ist, um den Eisensaccharosekomplex
aus der Lösung zu präzipitieren;
(d) Entnehmen des gereinigten Eisensaccharosekomplexes aus dem in Schritt (c) gebildeten Gemisch; und
(e) Auflösen des gemäß Schritt (d) hergestellten gereinigten Eisensaccharosekomplexes in einer Lösung aus
15 Saccharose in Wasser.
25. Verfahren zum Zubereiten einer wässrigen Lösung aus Saccharose und Eisensaccharosekomplex, das die folgen-
den Schritte beinhaltet:
20 (a) Zubereiten eines isolierten Eisensaccharosekomplexes gemäß dem Verfahren aus Anspruch 1;
(b) Auflösen des isolierten Eisensaccharosekomplexes in einem wässrigen Lösungsmittel zum Bilden einer
Lösung;
(c) Bilden eines Gemisches durch Hinzufügen mindestens eines wassermischbaren organischen Lösungsmit-
tels zu der in Schritt (b) gebildeten Lösung in einer Menge, die ausreichend ist, um den Eisensaccharosekomplex
25 aus der Lösung zu präzipitieren;
(d) Entnehmen des gereinigten Eisensaccharosekomplexes aus dem in Schritt (c) gebildeten Gemisch;
(e) Auflösen des in Schritt (d) gebildeten gereinigten Eisensaccharosekomplexprodukts in einer wässrigen
Saccharoselösung;
(f) Bilden eines Gemisches durch Hinzufügen mindestens eines wassermischbaren organischen Lösungsmittels
30 zu der in Schritt (e) gebildeten Eisensaccharosekomplexlösung in einer Menge, die ausreichend ist, um den
Eisensaccharosekomplex und Saccharose zu co-präzipitieren;
(g) Entnehmen des in Schritt (f) gebildeten Co-Präzipitats; und
(h) Auflösen des entnommenen Co-Präzipitats in Wasser.
35
Revendications
1. Un procédé pour préparer un complexe fer - saccharose ne contenant pas plus de 15 % (en poids/poids) d’excipients,
comprenant les étapes consistant à :
40
(a) préparer de l’hydroxyde de fer ferrique par un procédé comprenant les étapes consistant à :
(i) fournir un mélange réactionnel comprenant un sel de fer ferrique dissous dans un milieu aqueux ;
(ii) ajouter au mélange réactionnel de 1 à 2 équivalents d’une première base rapporté à la quantité de sel
45 de fer ferrique dans le mélange réactionnel ;
(iii) laisser le mélange réactionnel s’équilibrer pendant un intervalle de temps supérieur à 10 minutes ;
(iv) ajouter une deuxième base au mélange réactionnel équilibré, dans une quantité suffisante pour ajuster
le pH du mélange réactionnel à un pH sélectionné ; et
(v) recueillir l’hydroxyde de fer ferrique dans le mélange réactionnel ;
50
(b) faire réagir l’hydroxyde de fer ferrique et le saccharose dans un mélange réactionnel aqueux comprenant
des ions sodium, à un rapport molaire du saccharose à l’hydroxyde de fer ferrique allant de 2 : 1 à 50 : 1,
pendant un intervalle de temps sélectionné, à une température sélectionnée, et à un pH compris dans la gamme
allant de 6,5 à 13 ; et
55 (c) isoler le complexe fer - saccharose du mélange réactionnel aqueux.
2. Un procédé selon la revendication 1, dans lequel la température sélectionnée de l’étape (b) est comprise dans la
gamme allant de 75 ˚C à 120 ˚C.
23
EP 1 733 058 B1
3. Un procédé selon la revendication 1, dans lequel l’intervalle de temps sélectionné de l’étape (b) est compris dans
la gamme allant de 2 minutes à 40 heures.
4. Un procédé selon n’importe lesquelles des revendications 1 à 3 dans lequel le complexe fer - saccharose isolé
5 comprend un complexe fer - saccharose cristallin.
5. Un procédé selon n’importe lesquelles des revendications 1 à 3 dans lequel le complexe fer - saccharose isolé
comprend un complexe fer - saccharose amorphe dépourvu de saccharose cristallin comme pouvant être déterminé
par diffraction des rayons X sur poudres.
10
6. Un procédé selon n’importe quelle revendication précédente dans lequel le complexe fer - saccharose isolé contient
de 1 à 60 % de fer ferrique en poids.
7. Un procédé selon la revendication 1, dans lequel le mélange réactionnel de l’étape (b) comprend du fer ferrique
15 dans une quantité allant de 0,2 % en poids/poids à 8 % en poids rapporté au poids du mélange réactionnel.
8. Un procédé selon n’importe quelle revendication précédente, dans lequel le sel de fer ferrique est du chlorure de
fer ferrique, du nitrate de fer ferrique, ou un mélange de ceux-ci.
20 9. Un procédé selon la revendication 1, dans lequel les première et deuxième bases sont indépendamment sélection-
nées dans le groupe consistant en carbonates de métal alcalin, bicarbonates de métal alcalin, hydroxydes de métal
alcalin, et amines hydrosolubles.
10. Un procédé selon la revendication 9, dans lequel la première et la deuxième base sont indépendamment sélection-
25 nées dans le groupe consistant en carbonate de sodium, bicarbonate de sodium, hydroxyde de sodium, et trishy-
droxyméthylaminoéthane.
11. Un procédé selon la revendication 1 dans lequel un ou deux équivalents de la première base sont ajoutés au
mélange réactionnel dans l’étape (a).
30
12. Un procédé selon la revendication 1, dans lequel la première base est ajoutée au mélange réactionnel à l’étape (a)
de façon continue à une vitesse d’ajout constante allant de 0,02 à 0,2 équivalent de la base par minute, rapporté
à la quantité de sel de fer ferrique, jusqu’à ce que le pH du mélange résultant soit de 2,0 à 2,5.
35 13. Un procédé selon la revendication 1, dans lequel le pH sélectionné dans l’étape (a)(iv) est compris dans la gamme
allant de 3,5 à 9.
14. Un procédé selon la revendication 13, dans lequel le pH sélectionné est 4,0, 7,0, ou 8,3.
40 15. Un procédé selon la revendication 1 dans lequel l’hydroxyde de fer ferrique est recueilli dans le mélange réactionnel
à l’étape (a)(v) par filtration.
16. Un procédé selon la revendication 15, comprenant en outre l’étape consistant à former une boue de l’hydroxyde
de fer ferrique recueilli dans un solvant aqueux.
45
17. Un procédé selon la revendication 16, dans lequel le solvant aqueux comprend de l’eau, ou un mélange d’eau et
d’un solvant organique miscible à l’eau.
18. Un procédé selon n’importe quelle revendication précédente, dans lequel l’étape consistant à isoler le complexe
50 fer - saccharose du mélange réactionnel comprend
(a) former un mélange en ajoutant au mélange réactionnel à température ajustée de la revendication 1 au moins
un solvant organique miscible à l’eau dans une quantité suffisante pour précipiter le complexe fer - saccharose ;
et
55 (b) recueillir le complexe fer - saccharose précipité dans le mélange formé à l’étape (a).
19. Un procédé selon n’importe quelle revendication précédente, dans lequel l’étape consistant à isoler le complexe
fer - saccharose du mélange réactionnel aqueux comprend la lyophilisation du mélange réactionnel aqueux ; la
24
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centrifugation du mélange réactionnel aqueux ; ou la concentration du mélange réactionnel pour former un résidu
comprenant le complexe fer - saccharose.
20. Le procédé, selon n’importe quelle revendication précédente, comprenant en outre la purification du complexe fer
5 - saccharose isolé, ladite purification comprenant les étapes consistant à :
21. Un procédé pour préparer un coprécipité comprenant un complexe fer - saccharose et du saccharose, comprenant
les étapes consistant à :
22. Un procédé selon la revendication 21, dans lequel le rapport du complexe fer - saccharose purifié à la solution de
saccharose aqueuse est compris dans la gamme allant de 1 : 0,5 à 1 : 10 en poids.
30 23. Un procédé selon la revendication 21, dans lequel la solution de saccharose aqueuse a une concentration comprise
dans la gamme allant de 10 % à 50 % en poids/volume.
24. Un procédé pour préparer une solution aqueuse de saccharose et de complexe fer - saccharose, comprenant les
étapes consistant à :
35
(a) préparer un complexe fer - saccharose isolé selon le procédé de la revendication 1 ;
(b) dissoudre le complexe fer - saccharose isolé dans un solvant aqueux pour former une solution ;
(c) former un mélange en ajoutant à la solution formée à l’étape (b) au moins un solvant organique miscible à
l’eau dans une quantité suffisante pour précipiter le complexe fer - saccharose à partir de la solution ; et
40 (d) recueillir le complexe fer - saccharose purifié dans le mélange formé à l’étape (b) ; et
(e) dissoudre le complexe fer - saccharose purifié produit selon l’étape (d) dans une solution de saccharose
dans de l’eau.
25. Un procédé pour préparer une solution aqueuse de saccharose et de complexe fer - saccharose, comprenant les
45 étapes consistant à :
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26
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27
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28
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29
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This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European
patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be
excluded and the EPO disclaims all liability in this regard.
• GUPTA et al. Kidney Int., May 1999, vol. 55 (5), • Raising the Bar for Quality Drugs. Chemical and En-
1891-8 [0003] gineering News. American Chemical Society, 19
• CHARYTAN et al. Am J Kidney Dis., February 2001, March 2001, 26-31 [0008]
vol. 37 (2), 300-7 [0004] • FISHBANE et al. Semin Dial., November 2000, vol.
13 (6), 381-4 [0008]
32