Innovative preparation and crystallization of iosimenol

Abstract

The present invention generally relates to a process of preparing iosimenol and a process of preparing a crystal of iosimenol, as well as a crystal of iosimenol prepared by these processes.

Claims

1. A crystal of iosimenol characterized by a powder x-ray diffraction pattern having four or more 20.2 peaks and selected from about 8.1, 9.6, 9.9, 10.0, 10.7, 15.4, 16.9, 18.0, 18.6, 18.9, 20.1, 20.4, 21.9, 22.2, 22.5, 24.8, 26.1, 26.8, 27.5, 28.9, 29.4, 29.7, 30.5, 34.1 and 34.6, wherein measurement of said crystal is at a temperature of about 293 K.

2. The crystal of iosimenol of claim 1 characterized by a powder x-ray diffraction pattern having four or more 20.2 peaks and selected from about 8.1, 20.4, 21.9, 22.2, 22.5, 26.8 and 30.5, wherein measurement of said crystal is at a temperature of about 293 K.

3. A crystal of iosimenol characterized by unit cell parameters at T=293K substantially equal to the following: a=21.8919(16) , b=9.8210(9) , c=20.0233(12) , =90, =94.955(1), =90, volume 4289(6) .sup.3 and a monoclinic P21/a space group.

4. A process for preparing iosimenol of formula: ##STR00011## comprising reacting CVI of formula: ##STR00012## with an alkylating agent introducing 2,3-dihydroxypropyl group in the presence of an inorganic base in a solvent comprising 2-methoxyethanol.

5. The process of claim 4, wherein the alkylating agent introducing 2,3-dihydroxypropyl group is selected from the group consisting of 3-halo-1,2-propanediol and glycidol.

6. The process of claim 4, wherein the alkylating agent introducing 2,3-dihydroxypropyl group is 3-halo-1,2-propanediol.

7. The process of claim 4, wherein the inorganic base is selected from the group consisting of an alkali metal hydroxide and an alkaline earth metal hydroxide.

8. The process of claim 4, wherein the inorganic base is lithium hydroxide, calcium hydroxide, sodium hydroxide, potassium hydroxide or mixture thereof.

9. The process of claim 4, wherein the reaction to prepare iosimenol is done in the presence of a metal halide besides an inorganic base.

10. The process of claim 9, wherein the metal halide is selected from the group consisting of CaCl.sub.2, ZnCl.sub.2 and MgCl.sub.2.

11. A process for preparation of the crystalline iosimenol of claim 1 from a saturated or supersaturated solution of iosimenol, comprising: Step 1: suspending deionized iosimenol in a solvent mixture comprising water and one or more organic solvents selected from the group consisting of C.sub.1-C.sub.6 linear or branched alkanols, alkoxyalkanols, C.sub.2-C.sub.8 aliphatic ethers, and C.sub.4-C.sub.6 cyclic ethers, Step 2: subjecting the mixture to heat and/or ultrasonic to completely dissolve the iosimenol in the mixture, Step 3: continuing to subject the solution to the same or different heat and/or ultrasonic to deposit the crystalline ioseminol of claim 1, and Step 4: collecting the resulting crystalline ioseminol of claim 1 on a filter.

12. The process of claim 11, wherein the heating in Step 2 and/or Step 3 is done with microwave.

13. The process of claim 11, wherein the organic solvent in Step 1 is selected from the group consisting of methanol, ethanol, n-propanol, 2-propanol, n-butanol, butanol, sec-butanol, tert-butanol, pentanols including isoamylalcohols, hexanols and 2-methoxyethanol, 2-ethoxyethanol, 1-methoxy-2-propanol and 2-isopropoxyethanol.

14. The process of claim 11, wherein the solvent mixture in Step 1 contains up to 20% water.

15. The process of claim 11, wherein the crystallization process in Step 3 may be initiated by adding a seed of iosimenol crystal while or after the temperature is raised.

16. The process of claim 11, wherein trometamol is used to buffer pH during crystallization process.

17. The process of claim 11, wherein Steps 2 and 3 are done at 70 C.-140 C.

18. The process of claim 11, wherein the concentration of iosimenol as the starting material in Step 1 is 10 w/v %-60 w/v %.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 shows comparison of the powder patterns of samples JM 070415A to JM 070415D, JM-090415E, and JM-050315B.

(2) FIG. 2 shows crystalline iosimenol at ultrasound, typical agglomerates consisting of Sample I-1.

(3) FIG. 3 shows typical spray-dried particles of amorphous iosimenol, Sample I-2.

(4) FIG. 4 shows spray-dried amorphous iosimenol, Sample I-2.

(5) FIG. 5 shows Sample I-3, crystalline iosimenol. Image shows a wide particle size distribution and particle shape diversity.

(6) FIG. 6 shows Sample I-3, crystalline iosimenol.

DESCRIPTION OF EMBODIMENTS

(7) Preparation of Crude Iosimenol

(8) The concentration of CVI in the reaction mixture is 5 to 20 w/w %, preferably 10 to 15 w/w %.

(9) The amount of 3-halo-1,2-propanediol or glycidol is 2.0 to 4.0 moles per one mole of CVI, preferably 3.6 to 3.8 moles.

(10) The inorganic base used in the reaction that iosimenol is prepared from CVI and 3-halo-1,2-propanediol or glycidol includes an alkali metal hydroxide and an alkaline earth metal hydroxide, for example, lithium hydroxide, calcium hydroxide, sodium hydroxide, potassium hydroxide, or mixture thereof.

(11) The amount of the inorganic base is 3.0 to 5.0 moles per one mole of CVI, preferably 4.0 moles.

(12) The metal halide used in the reaction that iosimenol is prepared from CVI and 3-halo-1,2-propanediol or glycidol includes CaCl.sub.2, ZnCl.sub.2 and MgCl.sub.2, preferably CaCl.sub.2. The CaCl.sub.2 includes CaCl.sub.2 (H.sub.2O).sub.x wherein x=0, 2, 4, and 6, the ZnCl.sub.2 includes ZnCl.sub.2(H.sub.2O).sub.y wherein y=0, 1, 1.5, 2.5, 3 and 4, and the MgCl.sub.2 includes MgCl.sub.2(H.sub.2O).sub.z wherein z=0, 2, 4 and 6.

(13) The amount of the metal is 3.0 to 5.0 moles per one mole of CVI, preferably 4.0 moles.

(14) The solvent used in the reaction that iosimenol is prepared from CVI and 3-halo-1,2-propanediol or glycidol is 2-methoxyethanol or a solvent comprising 2-methoxyethanol. The solvent comprised besides 2-methoxyethanol includes, for example, glycerin, the amount of glycerin is 0.2 to 0.3 grams per one gram of CVI, preferably 0.25 grams. The 2-methoxyethanol is added in an amount of 2.5 to 6 mL per gram of CVI, preferably 3.0 to 3.5 mL.

(15) The reaction temperature when iosimenol is prepared from CVI and 3-halo-1,2-propanediol or glycidol is 20 C. to 80 C., preferably 30 C. to 70 C., more preferably 40 C. to 50 C.

(16) The reaction time when iosimenol is prepared from CVI and 3-halo-1,2-propanediol or glycidol is 10 hours to 70 hours, preferably 16 hours to 48 hours, more preferably 18 hours to 24 hours.

(17) The impurities produced when crude iosimenol is prepared are mainly IMP1, IMP2, IMP3, and IMP4 which are over-alkylated compounds. The structures thereof are presumed as shown below.

(18) ##STR00008## ##STR00009##

(19) Deionization of Crude Iosimenol

(20) Deionized crude iosimenol as the starting material in the crystallizing process can be prepared from crude iosimenol as mentioned below. Obtained crude iosimenol contains inorganic salts and other ionic organic impurities. Aqueous solution of crude iosimenol can be deionized by nanofiltration or by ion-exchange resins.

(21) (1) Deionized crude iosimenol can be solidified by proper conventional methods, e.g. by precipitation of concentrated solution in alcohol or by spray drying. Concentration of the aqueous solution can be made by heat evaporation under reduced pressure or by nanofiltration combined with reverse osmosis.

(22) (2) Also another possibility is a direct use of deionized crude iosimenol as aqueous solution and concentrated via azeotropic distillation with 2-methoxyethanol. Water content is monitored and after water removal by the distillation the ratio of crystallization solvent is adjusted.

(23) The quality of the starting deionized crude iosimenol can affect the purity of crystallized iosimenol, i.e., achieved purity of crystallized material depends on the purity of starting crude deionized iosimenol. Lower quality material (below 92%) requires double crystallization to achieve purity above 98%.

(24) Starting material for crystallization must be deionized and reasonably pure. At the current stage of development, we are able to achieve final purity of iosimenol crystals around 97%, if we apply this method on deionized crude iosimenol API with starting purity 86-90% of HPLC area. To achieve purity over 98%, it can be presumed that the purity of starting material must be over 94%. We assume that this starting purity 94% is achievable, if the synthesis of intermediates according to Patent Literature 1 will be well optimized.

(25) Crystalline Iosimenol

(26) The crystals of iosimenol can be characterized via crystallography techniques, such as (but not limited to) X-ray diffraction, neutron diffraction, electron diffraction, and/or the like. In some embodiments, the iosimenol crystals can be characterized by x-ray diffraction patterns, or by one or more lattice parameters, or combinations thereof, for example as described herein.

(27) The crystal of iosimenol can be characterized by a powder x-ray diffraction pattern having four or more 20.2 peaks and selected from about 8.1, 9.6, 9.9, 10.0, 10.7, 15.4, 16.9, 18.0, 18.6, 18.9, 20.1, 20.4, 21.9, 22.2, 22.5, 24.8, 26.1, 26.8, 27.5, 28.9, 29.4, 29.7, 30.5, 34.1 and 34.6, wherein measurement of said crystal is at a temperature of about 293 K.

(28) Character of the crystal of iosimenol can be measured with the measurement instruments used in the working examples described below, but should not be limited thereto.

(29) General Procedure of Crystallization

(30) Some conditions of crystallizing procedure are shown below, but should not be limited thereto.

(31) (1. Composition of Solvent Mixture)

(32) Composition of the crystallization solvent mixture is a fundamental parameter of the crystallization. The solvents used in the purification are mainly composed of a mixture of 2-methoxyethanol and water, whose ratio in volume is selected from the range of 98:2 to 80:20. A stoichiometric amount of water is needed in the solvent mixture, but more than 10% water can reduce the yield in the purification. Besides 2-methoxyethanol, other solvents may be used such as one or more C.sub.1-C.sub.6 linear or branched alkanols, alkoxyalkanols, and C.sub.2-C.sub.8 aliphatic or C.sub.4-C.sub.6 cyclic ethers. The alkanols include methanol, ethanol, n-propanol, 2-propanol, n-butanol, i-butanol, sec-butanol, tert-butanol, pentanols including isoamylalcohols, and hexanols; the alkoxyalkanols include 2-methoxyethanol, 2-ethoxyethanol, 1-methoxy-2-propanol, and 2-isopropoxyethanol; and the ethers include cyclopentyl methyl ether, di-t-butyl ether, diethyl ether, diglyme, diisopropyl ether, dimethoxyethane, dimethoxymethane, 1,4-dioxane, ethyl t-butyl ether, methoxyethane, methyl t-butyl ether, 2-methyltetrahydrofuran, morpholine, tetrahydrofuran, and tetrahydropyran.

(33) (2. Providing Energy)

(34) Supply of energy such as heat and ultrasonic, specifically heat, to the crystallization mixture is useful to promote the crystallization in high purity. It is also possible to heat the mixture with microwave energy. Temperature during crystallization process should be above 50 C., preferably 70-140 C., more preferably 80-110 C., ideally 90-100 C. Higher temperature, ultrasonic and microwaves can promote the kinetics of the crystallization process. The most convenient temperature for crystallization is at the boiling point of the used solvent mixture. The pressure applied is atmospheric, or elevated, if required temperature exceeds the boiling point of the solvent mixture at atmospheric pressure. Heat and mass transfer is achieved by stirring or by ultrasound. Seeding of crystallization mixture may be necessary.

(35) (3. Time of Crystallization)

(36) Time of crystallization is not limited as long as a sufficient amount of the crystal is deposited. In general, the time is about 10 to 200 hours, preferably about 20 to 150 hours. Short time thereof can lead to low yield, whereas long time thereof can increase decomposition products. In order to shorten the time of crystallization, microwave or ultrasonic can assist the crystallization with heat.

(37) (4. Concentration of Iosimenol in Crystallization Solvent)

(38) Concentration of the starting material iosimenol in the solvent mixture for crystallization is 10 to 60 w/v %, preferably 20 to 50 w/v %, more preferably 25 to 40 w/v %.

(39) (5. Seeding)

(40) Seeding of crystallization mixture with small amount of crystalline iosimenol increases yield and purity and dramatically reduces (about 50%) crystallization time. The amount of crystalline iosimenol added for seeding is not limited. To get a rough idea, 0.1 to 10 w/w % per crude iosimenol can be used.

(41) Specific Condition of Innovative Crystallization of Iosimenol

(42) Main parameters having influence on crystallization of iosimenol can be: composition and pH of crystallization solution temperature of crystallization process time of crystallization presence of intentionally added crystallization centers (seeding) mixing of solution during crystallization process concentration of iosimenol in crystallization solution presence of ionic materials in crude iosimenol purity of starting crude iosimenol
but not all the parameters need to be explicitly defined.

(43) Although contrast media are relatively stable compounds, usually after a long-term exposure to heat, certain degradation may occur. The most vulnerable are amide groups and covalently bound iodine. In case of iosimenol, the acidic hydrogens on aliphatic carbon of malonyl group may cause cleavage of the bridge at higher temperature and can lead to form monomeric impurities. These all mentioned degradations are more intensive under strong acidic or strong basic conditions. Using our experience in formulation of iosimenol, we used trometamol as a buffer to control pH between 6-7 from the viewpoint of influencing of color. Ideal conditions for iosimenol crystallization are when the crystallization is performed at shortest process time at lowest possible temperature under suitable and controlled pH between 6-7 by buffer from the viewpoint of influencing of color.

(44) Through visual observation (change of color solution to slightly yellow-brown), we found that, between 80-110 C., the change of color is not intensive and no new impurities or increased level of known impurities are observed. Therefore, ideal conditions for crystallization are temperatures between 80-110 C., pH 6-7 and as short as possible time for crystallization.

(45) Reduction of time necessary for completion of crystallization was achieved by microwave assisted crystallization or by ultrasound. Both methods significantly reduce time of crystallization.

(46) The concentration of iosimenol in crystallization solvent mixture has a significant influence on yield and purity.

(47) We found that this phenomenon is driven mainly by the ratio between iosimenol and 2-methoxyethanol rather than other cosolvents (water, n-butanol). The optimal ratio is iosimenol/2-methoxyethanol about 40/100. Any significant deviation from this ratio lowers the yield and purity of obtained crystals.

EXAMPLES

(48) Hereinafter, the present invention is illustrated by the following examples, but should not be construed to be limited thereto, and it is possible to vary each condition unless the variation is beyond the range of the present invention.

Example 1. Preparation of Iosimenol (Laboratory Scale)

(49) ##STR00010##

(50) 250 mL three neck round-bottomed flask equipped with a magnetic stirrer was charged with 5,5-[(1,3-dioxo-1,3-propanediyl)diimino]bis[N-(2,3-dihydroxypropyl)-2,4,6-triiodo-1,3-benzenedicarboxamide] (CVI) (36.0 g, 0.027 mol), methoxyethanol (108 mL), lithium hydroxide monohydrate (4.54 g, 0.110 mol), glycerin (9.0 g) and anhydrous calcium chloride (12.0 g, 0.108 mol). The mixture was heated to 50-55 C. Then, 3-chloro-1,2-propanediol (11.35 g, 0.103 mol) was added thereto, maintaining the internal temperature at 40-45 C. The reaction mixture was heated for 21 hours at 40-45 C. After this time, the reaction was considered complete; the reaction mixture was precipitated by ethanol (220 mL) at 55-60 C. Cooled suspension (at 10-12 C.) was filtered and washed with methanol (150 mL). The crude product with salts contained 36.8 g of iosimenol (92% of theory).

(51) The crude product containing iosimenol and salts was dissolved in water (500 mL) at pH=1.9-2.0 and at temperature of 40-45 C. Acidity of the solution was adjusted with hydrochloric acid (1:2). The solution was after 10 minutes of stirring cooled to 10-15 C. Conductivity of the solution was adjusted by stirring the solution with anion-exchanger resins (Purolite A-400, 100 g) and cation-exchanger resins (Purolite C-100, 48 g). The mixture was stirred for 2 hours at 20-30 C. with adding of 500 mL of water. The mixture with resins (conductivity of 0.8 S/cm) was filtered and washed with water (100 mL). The filtrate was concentrated on rotary evaporator and dried to solid form. Then solid was transferred to glass drying tray and dried in a vacuum oven at 55-60 C. under nitrogen atmosphere. The obtained crude iosimenol is a white-off deionized powder (27.28 g, 68.2% of theory) with HPLC purity 93.22 area %. Overalkyls: imp. IMP1+IMP2: 2.55%, imp. IMP3: 0.29% and imp. IMP4: 0.56%.

(52) Following the working examples disclosed in Patent Literature 1 (U.S. Pat. No. 5,698,739 B) and Patent Literature 2 (WO 2009/091758), each crude iosimenol was prepared and their characteristics were analyzed. The results are shown in Table 1 along with the result of Example 1.

(53) TABLE-US-00001 TABLE 1 Yield Impurity profile (area %) Process (% th.) Iosimenol IMP1 + IMP2 IMP3. IMP4. U.S. Pat. No. 81.3 73.68 3.11 3.24 2.46 5,698,739 A U.S. Pat. No. 68.8 79.75 3.55 1.83 3.18 8,680,334 B2 Example 1 92.0 93.22 2.55 0.29 0.56 note: Yield is directed to crude product containing crude iosimenol and salts.

Example 2. Preparation of Iosimenol (Commercial Scale)

(54) 250 L glass-lined reactor equipped with a stirrer was charged with CVI (36.0 kg, 27.07 mol), methoxyethanol (108 L), lithium hydroxide monohydrate (4.54 kg, 108.2 mol), glycerin (9.0 kg) and anhydrous calcium chloride (12.0 kg, 108.1 mol). The mixture was heated to 55-60 C. Then, 3-chloro-1,2-propanediol (11.35 kg, 102.7 mol) was added thereto, maintaining the internal temperature at 50-55 C. The reaction mixture was heated for 16 hours at 55-60 C. After this time, the reaction mixture was precipitated by ethanol (220 L) at 50-55 C. Cooled suspension (at 10-15 C.) was centrifuged and washed with ethanol (60 L). The crude product with salts contained 36 kg of iosimenol (90% of theory).

(55) The crude product containing iosimenol and salts was dissolved in water (140 L) at pH=1.5-2.0 and at temperature of 48-52 C. Acidity of the solution was adjusted with hydrochloric acid (1:2). After 10 minutes of stirring, the solution was cooled to 15-20 C. pH of the solution was adjusted with a solution of sodium hydrogen carbonate to pH of 5.2-5.8. Main portion of the salts was removed by using of nanofiltration unit to achieve the conductivity between 0.2-0.7 mS/cm. The target conductivity of the solution was adjusted by stirring the solution with anion-exchanger resin (Purolite A-400, 8-10 kg) and cation-exchanger resin (Purolite C-100, 2-3 kg). The mixture with resins was stirred for 2 hours at 20-30 C. When conductivity 0.3-1.0 S/cm was achieved, the resins were filtered off and washed with water (20 L). The filtrate was charcoaled two times (20.86 kg) for 1 hour at 55-60 C. The cooled filtrate (15-20 C.) was concentrated on reverse osmosis unit to a density 1.40-1.41 g/ml (about 50 w/w %). The concentrated solution of iosimenol was charged into ethanol (300 L) during 1 hour at 55-60 C. The suspension of iosimenol was cooled and the precipitated iosimenol was centrifuged and washed with ethanol. The solid was dried at max. 50 C. for 12 hours. The dried iosimenol was dissolved into 20-30 w/w % aqueous solution and solidified by spray drying.

(56) Yield of iosimenol is 26 kg (65% of theory) with HPLC purity 91.61%. Overalkyls: imp. IMP1+IMP2: 2.75%, imp. IMP3: 0.44% and imp. IMP4: 0.41%.

(57) Preparation of Seed for Crystallization

(58) First crystals of iosimenol were obtained during the initial experiments on crystallization. Amorphous crude iosimenol (75 g) was firstly purified by column chromatography (achieved purity 98.1%) and then crystallized in mixture of 2-methoxyethanol (50 mL) and methanol (7 mL). The crystallization mixture was refluxed for 24 hours. Then the mixture was diluted with a mixture of water and methanol (8 mL and 10 mL, respectively). Then, fraction of solvent (10 mL) was distilled-off during 1 hour. The remaining mixture was maintained for next 24 hours under reflux and then cooled to 65 C. during 5 hours. Then 2-methoxyethanol was added (70 mL) and the obtained mixture was heated to reflux. The first traces of solid appeared after 5 days and prolonging of reflux for further 15 days caused substantial crystallizing of the material. The crystals were separated and used as seeding in future experiments.

(59) Preparation of Crystal of Iosimenol

(60) Three typical processes of crystallization of iosimenol are shown below in three examples.

(1) Crystallization of Iosimenol Using Conventional HeatingExample 3

(61) Nanofiltered, deionized and charcoaled aqueous solution of crude iosimenol (purity 96-97% HPLC area) was concentrated under reduced pressure to dryness. 40 grams (calculated on anhydrous base) of the iosimenol was suspended in solvent mixture containing 2-methoxyethanol (100 mL) water (2.5-7 mL) and n-butanol (5-10 mL) in a 250 mL three-necked flask equipped with a condenser. The suspension was stirred and heated up to 85 C. until clear solution is obtained, then heated to reflux and seeds of iosimenol crystals (0.5-1.0 g) were added. The mixture was maintained under reflux. The first crystals appeared after 6-12 hours. The course of crystallization was checked by monitoring of remaining dissolved iosimenol in sample of liquid phase and when remaining iosimenol stayed unchanged in two consecutive testing, the suspension was filtered at 80-90 C. The obtained solid material was washed with ethanol (100 mL) at 60-70 C. The HPLC purity of the crystals was 98.5-99.2% and the crystallization yield was 50-53%. Total time of the crystallization was 72 hours.

(2) Crystallization of Iosimenol Using UltrasoundExample 4

(62) Iosimenol and solvent mixture, in ratio and amount as described in previous Example 3, were conventionally heated to reflux and ultrasonicated (20 kHz, pulse mode). Seeds of iosimenol crystals in proportion to deionized crude iosimenol corresponding to Example 3 were added. The mixture was maintained under reflux and ultrasonicated. The course of crystallization was checked in the same way as in Example 3. The suspension of crystalline iosimenol was filtered at 80-90 C. and obtained solid material washed with ethanol at 60-70 C. The HPLC purity of the crystals was 99.1% and the crystallization yield was 45%. Total time of the crystallization was 24 hours.

(3) Crystallization of Iosimenol Using Microwave IrradiationExample 5

(63) Iosimenol and solvent mixture were prepared and dissolved as described in previous Example 3. The obtained solution was transferred into a flask with crimp top, magnetic stir bar added, seeded with iosimenol crystals in the ratio to crude iosimenol corresponding to Example 3, and tightly closed. Then the stirred solution was irradiated by microwave in Biotage equipment with a preset target temperature 90 C. The obtained suspension of crystalline iosimenol was filtered at 80-90 C. and the solids were washed with ethanol at 60-70 C. The HPLC purity of the crystals was 99.0% and the crystallization yield was 40%. Total time of the crystallization was 16 hours.

(64) Powder Diffraction Characterization of Iosimenol Obtained by Innovative Crystallization

(65) (1) Source of Sample Used

(66) Seven batches of powdered iosimenol were provided, JM-070415A to JM-070415D, JM-090415E, JM-160415 and JM-050315B (see, Table 2). Small amounts of the provided samples were picked for X-ray powder diffraction (XRPD) analysis. Samples JM-070415A to JM-070415D and JM-090415E have got almost the same powder diffraction patterns and are identical with the diffractogram JM-050315B (reference sample). Powder patterns of the samples JM-070415A to JM-070415D and JM-090415E have got the same number of peaks with respect to iosimenol JM-050315B, so they do not contain any other crystalline phase (see, FIG. 1).

(67) TABLE-US-00002 TABLE 2 Samples used for XRPD analysis Experiment: JM-050315B* JM-070415A JM-070415B JM-070415C JM-070415D JM-090415E JM-160415** Main purpose of the Preparation crystals for crystallographic analysis experiment: Starting Iosimenol, 40 g, 98.75% 20 g, 98.10% 20 g, 98.10% 40 g, 98.10% 40 g, 98.10% 40 g, 98.10% 40 g 98.93% purity: 2-Methoxyethanol: 100.0 mL 50.0 mL 50.0 mL 100.0 mL 100.0 mL 100.0 mL 100.0 mL n-Butanol: 5.0 mL 0.0 mL 2.5 mL 0.0 mL 5.0 mL 5.0 mL 10.0 mL Water: 2.65 mL 0.0 mL 0.0 mL 2.65 mL 2.65 mL 2.65 mL 5.0 mL Trometamol: no no no no no no no Tested temperature range: 93-97 C. 94-99 C. 94-100 C. 94-97 C. 92-97 C. 95-97 C. 95-97 C. Seeding: 0.05 g no no no no 2 g 0.05 g Crystallization observed: yes yes yes yes yes After After 38 min. 30 min. Time of the crystallization: 84 hours 168 hours 168 hours 168 hours 168 hours 96 hours 72 hours Water content in crystals 0.8% 1.1% 0.9% 1.0% 0.9% 1.0% n.a. by KF: Purity of the obtained 99.03% 94.31% 98.64% 96.93% 97.77% 98.47% 99.13% crystal (% area by HPLC): Yield: 65.7% 12.8% 36.0% 36.7% 45.3% 54.3% 1.0% Purify of the mother liquor 96.26% 84.51% 96.19% 88.04% 90.17% 90.41% n.d. (% area by HPLC) *Second crystallization (starting material was a crystalline form), **only this crystallization was without mixing, n.d. = not detected, n.a. = not applicable

(68) (2) Sample Preparation

(69) The samples were examined with an optical microscope and it was found that the size of the crystallite is below 10 micrometers. Unfortunately no suitable single-crystals were observed.

(70) (3) Data Collection

(71) X-ray powder diffraction data were collected at room temperature with an X'Pert PRO 8-8 powder diffractometer with parafocusing Bragg-Brentano geometry using CuK radiation (=1.5418 , U=40 kV, I=30 mA). Data were scanned with an ultrafast detector X'Celerator over the angular range 5-80 (2) with a step size of 0.0167 (2) and an accumulative counting time 20.32 s.Math.step.sup.1. Indexing procedure was performed on more precise data scanned with an accumulative counting time 162.88 s.Math.step.sup.1. Data evaluations were performed using the software package HighScore Plus.

(72) (4) References to Programs Used HighScore Plus, Full Powder Pattern Analysis Software, V3.0e, PANALYTICAL, Almelo, Holland. Boultif, A. and Lur, D. (2004). Powder pattern indexing with the dichotomy method, J. Appl. Crystallogr. 37, 724-731.

(73) (5) Results of the Comparison of the Measured Samples

(74) All seven samples have got almost the same X-ray powder patterns. The number of the peaks for all the seven samples in the 2 range (5-20, reasonably separated peaks) is the same and the positions of the peaks are the same within the experimental errors. The relative intensities of the peaks are as well almost the same. Therefore we can state that these seven powdered samples have got almost randomly oriented crystallites. See the following Table 3 and diffractograms in FIG. 1.

(75) TABLE-US-00003 TABLE 3 PXRD Peak Positions (degrees 2 0.2) JM-070415A JM-070415B JM-070415C JM-070415D JM-090415E JM-160415 JM-050315B 8.2 8.1 8.2 8.1 8.1 8.1 8.1 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.9 9.9 9.9 9.9 9.9 9.9 9.9 10.0 10.0 10.1 10.0 10.0 10.0 10.0 10.7 10.6 10.7 10.7 10.7 10.7 10.7 15.5 15.4 15.5 15.4 15.4 15.5 15.4 16.9 16.8 16.9 16.9 16.9 16.9 16.9 18.0 18.0 18.1 18.0 18.0 18.1 18.0 18.6 18.6 18.6 18.5 18.5 18.6 18.5 19.0 18.9 19.0 19.0 18.9 18.9 18.9 20.1 20.1 20.1 20.1 20.0 20.0 20.0 20.4 20.3 20.4 20.4 20.4 20.4 20.4 22.0 21.9 22.0 21.9 21.9 22.0 21.9 22.2 22.1 22.2 22.2 22.2 22.2 22.1 22.5 22.4 22.5 22.4 22.4 22.5 22.5 24.8 24.8 24.9 24.8 24.8 24.8 24.8 26.2 26.1 26.2 26.1 26.1 26.2 26.1 26.9 26.8 26.9 26.8 26.8 26.9 26.8 27.6 27.5 27.6 27.5 27.5 27.6 27.5 29.0 28.9 29.0 28.9 28.9 29.0 28.9 29.4 29.3 29.4 29.4 29.4 29.4 29.4 29.7 29.7 29.8 29.7 29.7 29.7 29.7 30.6 30.5 30.6 30.5 30.5 30.5 30.5 34.1 34.0 34.1 34.1 34.0 34.1 34.0 34.6 34.6 34.6 34.5 34.5 34.6 34.6

(76) (6) Results of the Unit Cell Parameters for the Sample Iosimenol JM-050315B

(77) The automatic indexing of results obtained using evaluation software DICVOL04 (reference Boultif, A. and Lur, D. (2004). Powder pattern indexing with the dichotomy method, J. Appl. Crystallogr. 37, 724-731) has shown, that the compound C.sub.31H.sub.36I.sub.6N.sub.6O.sub.14 is monoclinic with space group P2.sub.1/a, and the unit-cell parameters were least-square refined to the values:

(78) TABLE-US-00004 a = 21.8919(16) , = 90 b = 9.8210(9) , = 94.955(1) c = 20.0233(12) , = 90 Volume 4289(6) .sup.3

(79) Crystal Class: monoclinic P2.sub.1/a

(80) Cell determined from 138 reflections

(81) Cell 2 range=5-80

(82) Temperature 293K

(83) Evaluation of Particle Morphology, Skeletal and Apparent Density of Crystalline and Sprayed (Amorphous) Iosimenol

(84) The objective was to evaluate a solid iosimenol, which was obtained from its solution as a powder form by crystallization, ultrasound assisted crystallization and spray drying. Samples identified as crystallized iosimenol were our main object of interest, while spray-dried iosimenol were studied in order to find the characteristic differences. Each type of iosimenol was represented by one sample.

(85) Particle morphology was evaluated by Scanning Electron Microscopy (SEM) combined with ion microscope FIB-SEM Tescan Lyra3GMU equipped with a number of detectors EDS, EBSD, STEM, EBIC and TOF-SIMS. Due to electric non-conductivity of iosimenol, its molecules accumulate electric charge leading to deterioration of SEM images. Therefore, the samples were coated with a thin layer of platinum for effective removal of electric charges to obtain high quality SEM images.

(86) Skeletal (true) density of the material was measured using helium pycnometer Micomeritics Multivolume Pycnometer 1305 and using mercury porosimeter Micromeritics AutoPore IV.

(87) Apparent (gravity) density, i.e. density covering all open or closed cavities or pores was also measured.

(88) Three samples of iosimenol were analyzed. Results of testing of individual samples obtained from the skeletal density (helium pycnometry and mercury intrusion porosimetry) and apparent density are shown in the Table 4.

(89) TABLE-US-00005 TABLE 4 Comparison of skeletal and apparent (true) density of crystalline and spray-dried iosimenol API Sample .sub.He [g cm.sup.3] .sub.Hg [g cm.sup.3] .sub.a [g cm.sup.3] I-1 JM-280415; 2.3700 2.3032 1.057 ultrasound, crystallization I-2 PSD 420131115/165; 2.1974 1.9837 1.193 spray-dried I-3 JM-210415TKR; 2.4022 2.3624 1.135 crystallization .sub.He - skeletal density by helium pycnometry, .sub.Hg - skeletal density by mercury porosimetry, .sub.a - apparent density

(90) Slightly lower skeletal density by mercury porosimetry was observed in comparison with skeletal densities by helium pycnometer. These differences are caused by physical properties of the probe used. Smaller and more agile helium is able to explore also cavities that are accessible only through very narrow necks. Based on small difference between densities, we can assume that number of narrow necks (inaccessible for mercury) is very low.

(91) Values of skeletal density indicate significant impairment of spray-dried sample I-2, which can be interpreted as the existence of closed porosity, which occurs quite frequently in spray-dried samples. Closed porosity is also observed in the electron microscope, see in Drawings.

(92) Mercury intrusion curve for sample I-1 shows the majority of intrusion volume of mercury in pressure, which can be interpreted as the characteristic size of narrow necks with values of 200 to 300 nm. Such values are consistent with the observed objects in the image of sample I-1 (see, FIG. 2) where hexagonal blocks are clearly visible constituting the agglomerate sample I-1 of approximately 500 to 2,000 nm. Since these numbers are in good agreement with values from the mercury intrusion curve, we can assume that the surface of sample I-1 is formed only by the geometrical shape of non-porous particles.

(93) Images of sample I-2 (see, FIG. 3 and FIG. 4) reveals two types of spherical particles formed by spray drying. Some are almost smooth, some are slightly deformed. Sample I-2 indicates the presence of a significant quantity of pores with size in the interval of 7-20 nm.

(94) Mercury intrusion curve for sample I-3 shows a gradual filling under of corresponding cylindrical pore with diameter of 100 nm, which can be characterized as narrow necks formed during agglomeration of particles of a sample I-3 (see, FIG. 5, FIG. 6).

(95) Summary of Crystal

(96) XRPD, Scanning Electron Microscopy (SEM) combined with ion microscope FIB-SEM, skeletal density were performed to identify the material obtained by innovative crystallization of deionized crude iosimenol. The obtained results confirmed crystalline structure of iosimenol, uniformity of obtained crystalline phases, i.e. crystals contain only one polymorph.

(97) Crystalline samples of iosimenol, which were prepared by crystallization under various conditions, show identical X-ray powder diffraction. The unit cell of crystals was characterized as monoclinic space group P2.sub.1/a, with the following parameters:

(98) TABLE-US-00006 a = 21.8919(16) , = 90 b = 9.8210(9) , = 94.955(1) c = 20.0233(12) , = 90

(99) Scanning Electron Microscopy (SEM) combined with ion microscope FIB-SEM provide images of iosimenol solids and those prepared via innovative crystallization shows geometry typical for crystalline phases. Crystals are organized in tetragonal or hexagonal blocks constituting the agglomerates.

INDUSTRIAL APPLICABILITY

(100) The role of invented crystallization is either to fully replace the current HPLC purification technology which is very expensive or to be used in combination with substantially simplified HLPC purification.