Preparation of stable ferric derisomaltose

Abstract

The present invention relates to the preparation of stable Ferric derisomaltose with high purity. The present invention further relates to the preparation of stable Ferric derisomaltose with % dimer content above 2.9% by weight, based on the total weight of the hydrogenated oligosaccharide.

Description

[0001] “PREPARATION OF STABLE FERRIC DERISOMALTOSE”

[0002] CROSS REFERENCE TO THE RELATED APPLICATIONS

[0003] This application claims priority benefits of the earlier Indian provisional application no IN 202441029535 filed on Apr 11, 2024.

[0004] FIELD OF THE INVENTION:

[0005] The present invention relates to the preparation of stable Ferric derisomaltose with high purity.

[0006] The present invention further relates to the preparation of stable Ferric derisomaltose having the content of dimer saccharide in said hydrogenated oligosaccharide is 2.9% by weight or more, based on the total weight of the hydrogenated oligosaccharide, preferably 3% by weight or more, and more preferably 3.2% by weight or more.

[0007] BACKGROUND OF THE INVENTION:

[0008] Ferric derisomaltose is an iron carbohydrate complex with a matrix structure composed of interchanging layers of ferric hydroxide and the carbohydrate derisomaltose. Derisomaltose consists of linear, hydrogenated isomaltooligosaccharides with an average molecular weight of 1000 Da and a narrow molecular weight distribution that is almost devoid of mono- and disaccharides. Ferric derisomaltose has an average molecular weight of 155,000 Da.

[0009] Ferric derisomaltose is approved under the brand name MONOFERRIC® on Jan 16, 2020 for treatment of iron deficiency anemia (IDA) in adult patients. Ferric Derisomaltose is disclosed as product in Pharmacosmos US8815301B2.

[0010] US9439969B2 discloses the preparation of Ferric derisomaltose wherein the dimer content is less than 2.9% by weight or less, based on the total weight of the hydrogenated oligosaccharide. This patent discloses that the dimer content should be less than 2.9% by weight for a stable iron oligosaccharide compound. The desired dimer content is obtained by membrane filtration. US11851504B2 discloses iron oligosaccharide comprising hydrogenated oligosaccharide in stable association with ferric oxyhydroxide, wherein the hydrogenated oligosaccharide is having average molecular weight of less than 3,000 Daltons and iron oligosaccharide has an in-vitro half-life of 18.3 hours or more. The patent discloses relation of dimer content with molecular weight, half-life, and stability of the iron oligosaccharide.

[0011] The inventors of the present invention has surprisingly found that the increase in dimer content to 3.0%-5.0% by weight did not have significant change in in-vitro half-life and stability of the product. Hence, the inventors of the present invention has developed a process for the preparation of Ferric derisomaltose which is stable with high purity with dimer content in the said hydrogenated oligosaccharide is greater than 2.9% by weight.

[0012] SUMMARY OF THE INVENTION:

[0013] One aspect of the present invention relates to the process for the preparation of stable Ferric derisomaltose with high purity.

[0014] One aspect of the present invention relates to preparation of stable Ferric derisomaltose having the content of dimer saccharide in said hydrogenated oligosaccharide is 2.9% by weight or more, based on the total weight of the hydrogenated oligosaccharide, preferably 3% by weight or more and more preferably 3.2% by weight or more.

[0015] Another aspect of the present invention relates to preparation of stable Ferric derisomaltose having the content of dimer saccharide in said hydrogenated oligosaccharide is 2.9% by weight or more, based on the total weight of the hydrogenated oligosaccharide, wherein the iron oligosaccharide compound has an in in-vitro half-life of 18.3 hours or less, preferably in-vitro half-life of 18.1 hours or less and more preferably in-vitro half-life of 18.0 hours or less.

[0016] Another aspect of the present invention relates to the process for the preparation of stable Ferric derisomaltose, which comprises a) Purification of dextran 1 ; b) Reduction of dextran 1 to dextran; c) Reaction of dextran with ferric oxyhydroxide to obtain Ferric derisomaltose; and d) Purification of Ferric derisomaltose.

[0017] Another aspect of the present invention relates to the process for the purification of Dextran 1 (Formula III), which comprises a) dissolving dextran in water; b) slow addition of dextran 1 solution to a suitable solvent; c) stirring the reaction mass to 25-35 °C; d) filtering the reaction mass; and e) dried to obtain pure Dextran 1.

[0018] BRIEF DESCRIPTION OF THE DRAWINGS:

[0019] FIG. 1 shows a plot of the in-vitro t* vs. the % dimer content of the iron oligosaccharide product (Table 1)

[0020] DETAILED DESCRIPTION OF THE INVENTION:

[0021] The present application will now be described more fully hereinafter with reference to the accompanying examples and experiments, in which illustrative embodiments of the application are shown. This application may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the application to those skilled in the art.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.

[0023] One embodiment of the present invention relates to the process for the preparation of stable Ferric derisomaltose, having high purity and % dimer content 2.9% by weight or more, based on the total weight of the hydrogenated oligosaccharide, which is represented by the scheme 1

[0024] Scheme 1

[0025] Another embodiment of the present invention relates to the process for the purification of Dextran 1 (Formula III), which comprises a) dissolving dextran 1 in water; b) slow addition of dextran 1 solution to a suitable solvent; c) stirring the reaction mass to 25-35 °C; d) filtering the reaction mass; and e) dried to obtain pure Dextran 1.

[0026] In another embodiment of the present invention, step b) involves reduction of dextran 1 in the presence of reducing agent and stirred at 25-35 °C, followed by pH adjustment pH 6.0-6.5 using HC1 solution. The reaction mass is filtered and dried to obtain reduced dextran (Formula II).

[0027] In another embodiment of the present invention, reducing agent is selected from but not limited to sodium borohydride, lithium aluminium hydride, diborane, ferrous sulfate, oxalic acid, formic acid, ascorbic acid, potassium iodide preferably sodium borohydride and ferrous sulfate.

[0028] In another embodiment of the present invention step c) involves reaction of dextran with ferric oxyhydroxide followed by acid hydrolysis in presence of mineral acid. pH of the reaction mass was adjusted to pH 11-14 using sodium hydroxide, stirred and filtered. pH of the filtrate was adjusted using HC1 to pH 5-6 and dissolved in suitable solvent, stirred, filtered and dried to obtain Ferric derisomaltose.

[0029] In another embodiment of the present invention, step d) involves purification of ferric derisomaltose by dissolving crude Ferric derisomaltose in water, stirring followed by pH adjustment between 5-7 using HCl / NaOH solution. Filtering the reaction mass using micron filtration and dissolving the filtrate in Retarded Ethanol, stirred, filtered and dried to obtain pure Ferric Derisomaltose.

[0030] In another embodiment of the present invention, ferric oxyhydroxide is prepared using Ferric chloride hexahydrate, by dissolving in water and in presence of sodium carbonate, followed by pH adjustment pH 6-8 using HC1, stirred, filtered and dried to obtain Ferric oxyhydroxide.

[0031] In another embodiment of the present invention, suitable solvents used is selected from ethanol, methanol, propanol, isopropanol, butanol, pentanol, hexanol and cetyl alcohol, preferably methanol and ethanol. In another embodiment of the present invention, mineral acid selected from but not limited to hydrochloric acid, sulphuric acid, nitric acid, phosphoric acid, hydrobromic acid and hydrofluoric acid, preferably hydrochloric acid.

[0032] In another embodiment of the present invention, the Ferric derisomaltose was stable for atleast 30 days even when the % dimer content was in the range of 3.0%- 4.5% by weight, more preferably in the range of 3.0%-5.0% by weight, based on the total weight of the hydrogenated oligosaccharide.

[0033] In another embodiment of the present invention relates to pharmaceutical formulation consisting of Ferric derisomaltose which is stable for atleast 12 months even when the % dimer content was in the range of 3.0%-4.5% by weight, more preferably in the range of 3.0%-5.0% by weight, based on the total weight of the hydrogenated oligosaccharide.

[0034] In another embodiment of the invention, the employed hydrogenated oligosaccharide is hydrogenated dextran having a weight average molecular weight (Mw) between 500 and 3000 Daltons, a number average molecular weight (Mn) above 500 Daltons, wherein 90% by weight of said dextran has molecular weights less than 3500 Daltons, and the Mw of the 10% by weight fraction of the dextran having the highest molecular weights is below 4500 Daltons.

[0035] In yet another embodiment of the present invention relates to Kinetic degradation studies of Ferric Derisomaltose.

[0036] A range of iron oligosaccharide compounds with differing contents of disaccharide were analyzed with respect to their rate of hydrolysis and apparent molecular weight, both of which are indicative of the quality of the respective compounds. Besides, it was found that the thermostability of iron oligosaccharides is a function of their apparent molecular weight (MP). Thus, such compounds are unstable to an unsatisfactory degree, if the apparent molecular weight markedly exceeds a value of 160,000 Daltons upon storage at an elevated temperature for three months as test solutions. Analysis of the stability and quality of Ferric Derisomaltose:

[0037] To analyse indicative quality of the compound, Ferric derisomaltose (Iron isomaltoside 1000) was analysed with respect rate of hydrolysis (Reductive Kinetic Degradation study / Acid soluble FeOOH by Ultraviolet-Visible Spectroscopy).

[0038] The acidic hydrolysis of the FeOOH in the compound of FeOOH and oligosaccharide was analyzed by quantifying the decreasing FeOOH concentration with UV- spectroscopy. The rate of hydrolysis as expressed by the half - life ( ti / 2 ) is an important indicative parameter of the compound. The UV- Visible Spectrophotometer used was a UV 2080N (Analytical Technologies Limited) with UV Professional Analysis Software. The hydrolysis of Fe3+was measured by optical absorbance at 287.3 nm using time interval 1 minute, in a Quartz Glass Cuvette of path length 1cm (10 mm).

[0039] Material used:

[0040] 1) Water (Process OR Milli-Q)

[0041] 2) Hydrochloric acid (Grade-AR, make: SDFCL / s d fine-chem limited)

[0042] 3) Sodium Chloride (Grade-AR, make: SDFCL / s d fine-chem limited)

[0043] The absorbance of Ferric derisomaltose (10 mg Fe / 1) in 0.9% NaCl / 0.24 M HC1 was measured at 287.3 nm from t = 0 min to t = 48 h, unless otherwise specifically mentioned. The half-life (t * ) is defined as the time at which the absorbance is the half of the value compared the absorbance at t = 0. Initial absorbance after dilution of the iron preparation at t = 0 min was set to 1 according to 100% undissolved FeOOH and all other measurements were normalized for this. Ln(normalized absorbance) on Y-Axis was plotted against time on X-Axis time in hours and fitted with a second degree polynomial equation (f(x) = ax2 + bx + c) (R2 > 0.99). From the constant a, b, and c of second degree polynomial equation, Half-life 11 / 2 was calculated from f at2+bt+c (t0.5) = ln(0.5). The results are shown in Table 1 and Table 2. Table 1: Batch I

[0044] Table 2: Batch II As shown in Table 2, the apparent molecular weight is in a desirable range, viz., about 130 kD to about 150 kD, when the amount of dimer in the oligosaccharide is more than 2.9%. The same holds true for the rate of hydrolysis as expressed by the half-life. The Half-life (ti / 2) is defined as the time at which the absorbance is the half of the value compared the absorbance at t=0.

[0045] The molecular weight of the Ferric derisomaltose of the present invention is in the desirable range of viz. about 130 kD to about 160 kD even when the amount of dimer in the oligosaccharide is 3.2% to 4.5%, based on the total weight of the hydrogenated oligosaccharide. The In-vitro half-life of the iron oligosaccharide (Ferric derisomaltose) of the present invention did not show any significant change and found to be little lower than 18 hrs., even when the % dimer content was above 2.9%, based on the total weight of the hydrogenated oligosaccharide, further results are depicted in Table 1 and Table 2 and shown in Figure 1.

[0046] In another embodiment of the present invention relates to molecular weight determination by Gel permeation technology (GPC) using refractive index high performance liquid chromatograph (RI-HPLC), the method of analysis is given below:

[0047] Method of Analysis:

[0048] Instrumentation: High Performance Liquid Chromatograph (HPLC) equipped with gradient capability

[0049] Data handling system: Empower-3 chromatographic software equipped with GPC software with RID Detector.

[0050] Column:

[0051] Make : Shodex

[0052] Name : OHpak SB-804 HQ (Product code - F6429103)

[0053] Column Size : 8.0mmID x 300mmL

[0054] Particle size : 10 pm

[0055] Pore size : 2000 A Guard Column:

[0056] Make : Shodex (F6709430)

[0057] Name : OHpak SB-G 6B

[0058] Column Size : 6.0mmID x 50mmL

[0059] Particle size : 10 pm

[0060] Reagents:

[0061] 1. Water (Mili-Q or HPLC grade is preferable)

[0062] 2. Di-Sodium hydrogen orthophosphate dihydrate (AR grade)

[0063] 3. Sodium dihydrogen orthophosphate Anhydrous (AR grade)

[0064] Preparation of Sample solution:

[0065] Weigh about 850 mg of sample (appx. Iron content 24%) into autoclave vial and add 2 ml of diluent, sonicate it to dissolve and autoclave it at 120°C for 20 minutes. Cool the solution to room temperature. (Appx. 100000 ppm of Iron)

[0066] Procedure:

[0067] Separately inject the Diluent, Calibration standard solutions, system suitability solution and sample solution into the chromatograph using above chromatographic parameters.

[0068] In another embodiment of the present invention relates to Ferric Derisomaltose is used, wherein the iron content is 5-50% by weight.

[0069] In another embodiment of the present invention relates to Ferric Derisomaltose of the present invention, wherein the polydispersity index is not more than (NMT) 2.0 (Mw / Mn), preferably NMT 1.8 (Mw / Mn).

[0070] In another embodiment of the present invention relation to a pharmaceutical composition consisting of Ferric Derisomaltose in powder form dissolved in aqueous medium, pH adjuster and stabilizer.

[0071] In another embodiment, the present invention relates to injectable formulations of Ferric Derisomaltose, packaged into container systems such as ampoules, vials. In another embodiment of the present invention relates to stability Ferric deriso maltose which is illustrated in the table 3 and table 4 below:

[0072] Table 3:

[0073] 5 Table 4: Process for the preparation of Ferric derisomaltose as disclosed herein are given in the examples below. The following examples are for illustration only and are not intended in any way to limit the scope of the present application.

[0074] EXAMPLES

[0075] Example 1: Preparation of pure Dextran 1 (Formula III)

[0076] 50g of Dextran 1 is dissolved in 100ml of water at 25-35 °C. Stir the reaction mass. The reaction mass was slowly added to methanol at 25-35 °C followed by rinsing with water and stirred for 40min. The reaction mass was filtered and suck dried the material under nitrogen atmosphere. The reaction mass is dried under vacuum at below 50 °C to obtain pure dextran.

[0077] Yield: 40-50%

[0078] Example 2: Preparation of reduced dextran (Formula II)

[0079] 50gm of purified dextran is dissolved in 100ml of water and stirred the reaction mass until a clear solution is obtained at 25-35 °C. The reaction mass was cooled to 10-15 °C. To the reaction mass 50% solution of sodium hydroxide (1.12gm) and sodium borohydride (4gm) was added and stirred at 24-30 °C for 16hr. pH of the reaction mass was adjusted between 6.0-6.5 using HC1 solution. The reaction mass was added dropwise to methanol taken in another RBF at below 40 °C for 45-60 min. The reaction mass filtered and suck dried under vacuum upto 70- 80min. The reaction mass was dried under vacuum at below 50 °C to obtain reduced dextran.

[0080] Yield: 85-90%

[0081] Example 3: Preparation of Ferric Derisomaltose (Formula la)

[0082] 50gm of Ferric chloride hexahydrate is dissolved in water and stirred until a clear solution is obtained at 25-35 °C. The pH of the solution was adjusted to 6.3 to 6.8 using sodium carbonate solution and if the pH was above 6.8 the pH was adjusted using HC1 solution. Stir the reaction mass at 25-35 °C. Filter the reaction mass and suck dry the material under vacuum. The reaction mass was rinsed with water. The reaction mass was added to the reactor and charged with water and then with reduced dextran filtrate, prepared by dissolving reduced dextran in water, stirring and filtering. The reaction mass was stirred for 5-10min at 20-30 °C. Citric acid monohydrate was added to the reaction mass and stirred. pH of the reaction mass was adjusted between pH 12.8-13.4 using sodium hydroxide solution. The reaction mass was heated to 70-90 °C until a clear solution is obtained. The reaction temperature was increased to 90-100 °C and stirred. The reaction mass was cooled to 25-35 °C and filtered through micron filter. The pH of the filtrate was adjusted to pH 5.2-5.5 using HC1 and stirred at 25-35 °C. Methanol (2L) was added to the filtrate and stirred at below 35 °C. The reaction mass was filtered and suck dried under nitrogen atmosphere and dried under vacuum at below 60 °C to obtain Ferric Derisomaltose.

[0083] Yield: 50-60%.

[0084] Example 4: Purification of Ferric Derisomaltose (Formula I)

[0085] 50gm of Ferric derisomaltose is dissolved in 200ml water and heated to a reaction temperature of 90- 110 °C. The reaction mass was cooled to 50-60 °C and 7g of sodium chloride was added to the reaction mass. The reaction mass was stirred for l-2hr. The reaction mass was cooled to 20-30 °C. The pH of the reaction mass was adjusted to 5.8-6.3 using 5% hydrochloric acid or 5% Sodium hydroxide solution. The reaction mass was filtered through 1 micron filter and then through 0.2 micron filter. The filtrate was added to the retarder ethanol at 20-30 °C. The reaction mass was stirred and filtered and washed with fine filter Retarder Ethanol. The solid obtained was dried in hot air oven at 50-70 °C to obtain pure Ferric derisomaltose.

[0086] Yield: 90%; Chloride content: not more than 2.5%.

[0087] The process described above is used to prepare Batch I and Batch II materials, with slight modifications in the reaction time and cooling temperature.

[0088] Without wishing to be bound to a theory, the process described in the present invention is believed to be stable process for the preparation of Ferric Derisomaltose which is commercially scalable, economical, stable and with improved yield along with high purity.

[0089] While the illustrative embodiments of the invention have been described with particularity, it will be understood that various other modifications will be apparent to and can be readily made by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is not intended that the scope of the claims appended hereto be limited to the examples and descriptions set forth hereinabove but rather that the claims be construed as encompassing all the features of patentable novelty which reside in the present invention, including all features which would be treated as equivalents thereof by those skilled in the art to which the invention pertains.

Claims

We Claim:1) A process for the preparing of stable Ferric derisomaltose, wherein the Ferric derisomaltose obtained has an in-vitro half-life of 22.1 hours or lower, preferably in-vitro half-life of 18.3 hours or lower, wherein the Ferric derisomaltose is stable for at least 12 months with % dimer content in the range of 3.0%-5.0% by weight, based on the total weight of the hydrogenated oligosaccharide.2) A process for the preparing of stable Ferric derisomaltose, wherein an iron oligosaccharide compound comprising a hydrogenated oligosaccharide in stable association with ferric oxyhydroxide, the hydrogenated oligosaccharide having a weight average molecular weight (Mw) between 500 and 3,000 Daltons, wherein the content of dimer saccharide in said hydrogenated oligosaccharide is 3.0% by weight or above, based on the total weight of the hydrogenated oligosaccharide, wherein the Ferric derisomaltose obtained is stable for atleast 12 months and has an in-vitro half-life of 22.1 hours or lower, preferably in-vitro half-life of 18.3 hours or lower.3) A pharmaceutical formulation consisting of Ferric derisomaltose, which is stable for at least 12 months with % dimer content in the range of 3.0%-5.0% by weight, based on the total weight of the hydrogenated oligosaccharide.4) A process for the preparation of stable Ferric derisomaltose as claimed in claims 2, which comprises a) purification of dextran 1 ; b) reduction of dextran 1 to dextran; c) reaction of dextran with ferric oxyhydroxide to obtain Ferric derisomaltose; and purification of Ferric derisomaltose.5) The process for the preparation of Ferric derisomaltose as claimed in claim 1 and claim 2, wherein the purification is carried out using a suitable solvent selected from ethanol, methanol, propanol, isopropanol, butanol, pentanol, hexanol, and cetyl alcohol, preferably methanol and ethanol.6) The process for the preparation of Ferric derisomaltose as claimed in claim 1 and claim 2, wherein step b) is carried out in present of reducing agent selected from but not limited to sodium borohydride, lithium aluminium hydride, diborane, ferrous sulfate, oxalic acid, formic acid, ascorbic acid, potassium iodide preferably sodium borohydride and ferrous sulfate.7) The process for the preparation of Ferric derisomaltose as claimed in claim 1 and claim 2, wherein step c) is carried out in the presence of mineral acid selected from but not limited to hydrochloric acid, sulphuric acid, nitric acid, phosphoric acid, hydrobromic acid, and hydrofluoric acid, preferably hydrochloric acid.

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