Process for the preparation of bortezomib mannitol ester

Abstract

A novel and improved process for preparation of bortezomib mannitol ester is derived, which process avoids excessive use of solvents, involves convenient, industrially feasible and economical techniques, and provides improvements in purity over processes known in the art.

Claims

1. A process for the preparation of bortezomib-mannitol ester (compound B), ##STR00009## comprising: (a) dissolving bortezomib (compound A) ##STR00010## in a first solvent to form a first solution wherein the first solvent comprises methylene dichloride, ethyl acetate or any combination thereof and wherein the first solution contains less than 1% water; (b) adding mannitol to the first solution; (c) removing the first solvent from the first solution to form a residue comprising bortezomib-mannitol ester; (d) adding a second solvent to the residue to form a suspension of bortezomib-mannitol ester in the second solvent wherein the second solvent comprises n-heptane, hexane, toluene, cyclohexane, diisopropyl ether, diethyl ether, or any combination thereof; and (e) isolating the bortezomib-mannitol ester from the second solvent.

2. The process according to claim 1 wherein the step (b) is carried out at a temperature of less than 35 C.

3. The process according to claim 1 wherein, after the mannitol is added to the first solution, the first solution is mixed for less than 30 minutes.

4. The process according to claim 1 wherein the mannitol is D-mannitol, and wherein the bortezomib-mannitol ester is bortezomib D-mannitol ester having the following structure ##STR00011##

5. The process according to claim 1 wherein the first solution is free of water.

6. The process according to claim 1 wherein the removing of the first solvent in step (c) comprises evaporating the first solvent from the first solution.

7. The process according to claim 6 wherein the step of evaporating comprises subjecting the first solution to a temperature less than 30 C.

8. The process according to claim 6 wherein the step of evaporating comprises subjecting the first solution to a pressure less than 1 atmosphere.

9. The process according to claim 1 wherein the first solvent is not frozen prior to the step of removing the first solvent from the first solution.

10. The process according to claim 1 wherein the step of isolating comprises filtering the bortezomib-mannitol ester from the second solvent.

11. The process according to claim 1, further comprising sterilizing the first solution by filtration.

12. The process according to claim 11 wherein the first solution is filtered after step (a) and before step (b), and wherein said mannitol is sterile.

13. The process according to claim 11 wherein the filtration is carried out by passing the first solution through a sub-micron filter.

14. The process according to claim 11 wherein the filtration is carried out by passing the first solution through a first filter having a pore size less than 0.50 microns.

15. The process according to claim 11 wherein the filtration is carried out by passing the first solution through a second filter having a pore size less than 0.25 microns.

16. The process according to claim 1 wherein the step (b) is carried out at a temperature of less than 30 C.

17. The process according to claim 1 wherein, after the mannitol is added to the first solution, the first solution is mixed for less than 20 minutes.

18. The process according to claim 6 wherein the step of evaporating comprises subjecting the first solution to a temperature less than 25 C.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIGS. 1 and 2 each show 1D .sup.1H-nuclear magnetic resonance (NMR) spectra of two samples of bortezomib-mannitol ester prepared in accordance with the present invention. Details regarding the NMR acquisition parameters are provided in example 7.

(2) FIG. 3 shows the .sup.13C NMR spectrum of a sample of bortezomib-mannitol ester prepared in accordance with the present invention, between 0 to 110 ppm.

(3) FIG. 4 shows the .sup.13C NMR spectrum of a sample of bortezomib-mannitol ester commercially available as Bortenat, between 0 to 110 ppm.

DETAILED DESCRIPTION OF THE INVENTION

(4) The inventors of the present invention have developed a novel and improved process for preparation of bortezomib-mannitol ester (hereinafter referred to as compound B) from bortezomib (hereinafter referred to as compound A).

(5) The process of the invention results in the formation of compound B substantially free of degradation impurity X. As used herein, the term substantially free of degradation impurity X refers to bortezomib-mannitol ester having about 0.3% by weight of impurity X or less, preferably about 0.2% by weight of impurity X or less, more preferably about 0.1% by weight of impurity X or less. The impurity X related to bortezomib-mannitol ester as determined by high performance liquid chromatography (HPLC).

(6) According to an aspect of the present invention, there is provided a process for the preparation of compound B that comprises;

(7) (a) dissolving bortezomib (compound A) into a first solvent to form a solution;

(8) and adding mannitol to the solution;

(9) (b) removing the solvent by distillation;

(10) (c) adding a second solvent to the residue;

(11) (d) stirring, filtering and drying the bortezomib-mannitol ester.

(12) Compound A used as a starting material may be prepared by the processes known in the prior art, for example as per the process described in the IN patent application No. 2638/MUM/2012, and in WO 14/041324, which are both incorporated herein by reference in their entirety.

(13) The process for preparation of compound B may comprise reacting compound A with a reduced sugar moiety, like mannitol, wherein mannitol is preferably of D-configuration.

(14) In accordance with the present invention, the process may comprise of adding compound A in a first solvent selected from methanol, isopropyl alcohol, ethanol, methylene dichloride, ethyl acetate and tetrahydrofuran, or any mixture thereof, that can be used in place of TBA (tertiary butyl alcohol) used by the prior art, to obtain a chemically stable compound B. A large amount of TBA is required to dissolve bortezomib, which is undesirable from both a cost perspective and disposal concerns. Use of TBA also involves a higher temperature in order to dissolve the bortezomib, as well as the use of water, which are thought to favour the formation of impurities such as impurity X. The solvents used in accordance with the present invention solves these problems associated with the TBA.

(15) An example of the novel process of preparing compound B is as depicted in scheme I:

(16) ##STR00007##

(17) In an aspect of the present invention, the process for preparing compound B comprises treating compound A with mannitol in a first solvent, without involving water, at a preferably lower temperature of 272 C., that thereby obtain the formation of compound B substantially free of the degradation impurity X.

(18) ##STR00008##

(19) The temperature for reaction is preferably 272 C., but it will be appreciated that other temperatures may be used, including less than 30 C., less than 31 C., less than 32 C., less than 33 C., less than 34 C., less than 35 C., less than 36 C. or less than 37 C. The temperature is preferably higher than 10 C.

(20) The decreased levels of impurity X provided by the present invention is confirmed by the following comparative HPLC data, as depicted in the Table 1.

(21) TABLE-US-00001 TABLE 1 Sl. Degradation No. Process Purity Impurity (X) 1 As per the process described in 98.40% 0.41% the patent application WO2002059130 2 As per the process described in 99.56% 0.05% this present invention

(22) The reaction time is preferably 15 minutes, but it will be appreciated that longer reaction times may be used, including less than 17 minutes, less than 19 minutes, less than 19 minutes, less than 20 minutes, less than 21 minutes, less than 23 minutes, less than 25 minutes, less than 27 minutes, less than 29 minutes, or less than 30 minutes. The reaction time is preferably longer than 5 minutes.

(23) The process of the present invention is particularly advantageous as it avoids the much expensive and time consuming freeze-drying or alcohol lyophilisation of compound B; instead may use a simpler, convenient, industrially feasible and economical vacuum drying process.

(24) A solid form of compound B is produced when compound A is treated with mannitol in a first solvent. The first solvent is preferably polar. Suitable polar solvents include methanol, isopropyl alcohol, ethanol, methylene dichloride, ethyl acetate, tetrahydrofuran, and mixtures thereof.

(25) The first solvent is preferably substantially free of water. The term substantially free of water as used herein means less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, less than 0.1%, less than 0.05%, or 0% [all percentages in (v/v)]. It will be appreciated that very small amounts of water may be present in the first solvent; however, such small amounts should be avoided where possible, and where a small amount is present, it is incidental, and its presence unintentional.

(26) The process further comprises adding a second solvent to the residue to form a suspension of bortezomib-mannitol ester in the second solvent, and isolating the bortezomib-mannitol ester from the second solvent.

(27) The second solvent is preferably water immiscible. More preferably, the second solvent used is selected from the group consisting of: n-heptane, hexane, toluene, cyclohexane, diisopropyl ether, diethyl ether, and any combination thereof. The second solvent is preferably substantially free of water.

(28) The process further comprises removing the first solvent from the first solution to form a residue comprising bortezomib-mannitol ester. Removal of the first solvent is preferably carried out by evaporating the first solvent from the first solution, for instance by heating or distilling the first solvent, and/or subjecting the first solution to a reduced pressure (such as a pressure less than 1 atmosphere).

(29) The reduced pressure is pressure is less than 1 atmosphere. The pressure may be less than 0.8 atmospheres, 0.6 atmospheres, 0.4 atmospheres, 0.3 atmospheres, 0.2 atmospheres, 0.1 atmospheres, 0.05 atmospheres, or 0.02 atmospheres. The pressure is preferably higher than 0.01 atmospheres.

(30) The temperature at which the first solvent is evaporated is preferably lower than ambient temperature, i.e. 30. More preferably the temperature is less than 28 C., 26 C., 25 C., 24 C., 23 C., or 22 C. A temperature higher than 10 C. is preferred.

(31) In an aspect, isolation of compound B may be carried out by filtering the solid. The filtrate may be washed with an amount of the second solvent. The solid is preferably dried under vacuum at 432 C., but other temperatures may be used, including than 47 C., less than 49 C., less than 51 C., less than 53 C., less than 55 C. or less than 57 C. The temperature is preferably higher than 20 C.

(32) Compound B (bortezomib-mannitol ester) obtained in accordance with the present invention advantageously shows better water solubility i.e. soluble in around 5 volumes water, than the poorly water soluble drug bortezomib.

(33) A further advantage of the present invention is to provide compound B with low levels of free boronic acid. In particular, the present invention provides a product where the ratio of bortezomib-mannitol ester to bortezomib is greater than 1:0.09, preferably greater than 1:0.08, and most preferably greater than 1:0.07.

(34) The process of the invention results in the formation of compound C substantially free of degradation impurity X. As used herein, the term substantially free refers to bortezomib-mannitol ester having about 0.3% by weight of impurity X or less, preferably about 0.2% by weight of impurity X or less, more preferably about 0.1% by weight of impurity X or less.

(35) The impurity X related to bortezomib mannitol ester may be determined by high performance liquid chromatography (HPLC).

(36) In another aspect the present invention provides sterile non-lyophilised bortezomib-mannitol ester.

(37) According to an aspect of the present invention, there is provided a process for the preparation of sterile non-lyophilised bortezomib-mannitol ester that comprises; (a) dissolving bortezomib (compound A) into a first solvent to form a solution; (b) filtering the solution through 0.45 micron filter followed by 0.22 micron filter in a sterile area; (c) adding sterile mannitol to the solution; (d) removing the solvent by distillation; (e) adding a second solvent to the residue; (f) stirring, filtering and drying the sterile non-lyophilised bortezomib-mannitol ester.

(38) The first solution may be sterilised by filtration. This can be achieved by passing the first solution through a sub-micron filter, such as a 0.22 micron filter. The first solution may optionally be passed through a 0.45 micron filter, which may assist in removing larger insoluble matter from the first solution, and thus decreasing the chance of blocking the 0.22 micron filter. Filters with even smaller pore sizes may be used, such as those with 50 nm, or 20 nm, if desired.

(39) The present invention also relates to an advantageous process of preparing bortezomib-mannitol ester as depicted in scheme 1, which being a chemically induced process may increase the scope of introducing variables or specific functional groups at certain structural positions of the drug ester, which can further open a new route of investigation of structural modifications of bortezomib-mannitol esters by subsequently monitoring their activities.

(40) According to the present invention, there is provided a pharmaceutical composition comprising the bortezomib-mannitol ester of the present invention, with one or more pharmaceutically acceptable excipients.

(41) According to the present invention, there is also provided a process of preparing the pharmaceutical composition comprising bortezomib-mannitol ester of the present invention along with pharmaceutically acceptable excipients.

EXAMPLES

Example 1: Preparation of Compound B

(42) In a round bottomed flask, 1.0 gram of bortezomib was added to 20 ml of methanol. The mixture was stirred until dissolution of the solid bortezomib. To this solution 5.0 grams of D-mannitol was added and the reaction mixture was stirred continuously for 15 minutes at a temperature of 272 C. This was followed by distillation process by which the solvent methanol was completely distilled under vacuum at below 25 C. 50 ml of n-heptane was then charged and continuous stirring was carried out for 30 minutes. The product obtained was then filtered and washed with n-heptane. The solid was then exposed to drying process under vacuum at 432 C., to get the final compound B.

(43) Dry Weight: 5.5 grams

(44) Purity: 99.36%

(45) LOD: 0.65%

Example 2: Preparation of Compound B

(46) In a round bottomed flask, 1.0 gram of bortezomib was added to 20 ml of methanol. The mixture was stirred until dissolution of the solid bortezomib. To this solution 8.0 grams of D-mannitol was added and the reaction mixture was stirred continuously for 15 minutes at a temperature of 272 C. This was followed by distillation process by which the solvent methanol was completely distilled under vacuum at below 25 C. 50 ml of n-heptane was then charged and continuous stirring was carried out for 30 minutes. The product obtained was then filtered and washed with n-heptane. The solid was then exposed to drying process under vacuum at 432 C., to get the final compound B.

(47) Dry Weight: 8.5 grams

(48) Purity: 99.33%

(49) LOD: 0.32%

Example 3: Preparation of Compound B

(50) In a round bottomed flask, 1.0 gram of bortezomib was added to 20 ml of methanol. The mixture was stirred until dissolution of the solid bortezomib. To this solution 2.0 grams of D-mannitol was added and the reaction mixture was stirred continuously for 15 minutes at a temperature of 272 C. This was followed by distillation process by which the solvent methanol was completely distilled under vacuum at below 25 C. 50 ml of n-heptane was then charged and continuous stirring was carried out for 30 minutes. The product obtained was then filtered and washed with n-heptane. The solid was then exposed to drying process under vacuum at 432 C., to get the final compound B.

(51) Dry Weight: 2.8 grams

(52) Purity: 99.46%

(53) LOD: 0.96%

Example 4: Preparation of Compound B

(54) In a round bottomed flask, 1.0 gram of bortezomib was added to 100 ml of ethanol. The mixture was stirred until dissolution of the solid bortezomib. To this solution 5.0 grams of D-mannitol was added and the reaction mixture was stirred continuously for 15 minutes at a temperature of 272 C. This was followed by distillation process by which the solvent ethanol was completely distilled under vacuum at below 25 C. 50 ml of n-heptane was then charged and continuous stirring was carried out for 30 minutes. The product obtained was then filtered and washed with n-heptane. The solid was then exposed to drying process under vacuum at 432 C., to get the final compound B.

(55) Dry Weight: 5.3 grams

Example 5: Preparation of Compound B

(56) In a round bottomed flask, 1.0 gram of bortezomib was added to 10 ml of methylene chloride The mixture was stirred until dissolution of the solid bortezomib. To this solution 5.0 grams of D-mannitol was added and the reaction mixture was stirred continuously for 15 minutes at a temperature of 272 C. This was followed by distillation process by which the solvent methylene chloride was completely distilled under vacuum at below 25 C. 50 ml of n-heptane was then charged and continuous stirring was carried out for 30 minutes. The product obtained was then filtered and washed with n-heptane. The solid was then exposed to drying process under vacuum at 432 C., to get the final compound B.

(57) Dry Weight: 5.5 grams

Example 6A: Preparation of Compound B

(58) In a round bottomed flask, 1.0 gram of bortezomib was added to 20 ml of isopropyl alcohol. The mixture was stirred until dissolution of the solid bortezomib. To this solution 5.0 grams of D-mannitol was added and the reaction mixture was stirred continuously for 15 minutes at a temperature of 272 C. This was followed by distillation process by which the solvent methanol was completely distilled under vacuum at below 25 C. 50 ml of n-heptane was then charged and continuous stirring was carried out for 30 minutes. The product obtained was then filtered and washed with n-heptane. The solid was then exposed to drying process under vacuum at 432 C., to get the final compound B.

(59) Dry Weight: 5.3 grams

Example 6B: Preparation of Sterile Non-Lyophilized Bortezomib-Mannitol Ester

(60) The following operation was carried in a sterile area in its entirety. In round bottom flask, 1.0 gram of bortezomib was added to 20 ml of methanol and the mixture was stirred to clear solution. The clear solution was filtered through a 0.45 micron filter followed by a 0.22 micron filter in the sterile area. To this clear filtrate was added 10.0 gram of sterile D-mannitol and the contents were stirred for 15 min at 272 C. Methanol was distilled completely under vacuum below 25 C. 50 ml of filtered n-heptane was added and stirred for 30 min. The product obtained was filtered and washed with filtered n-heptane. The solid was then exposed to drying process under vacuum at 432 C. and packed in sterile bags.

Example 7: NMR Analysis of Compound B

(61) Compound B formed in accordance with the invention was analysed using NMR.

(62) 1D .sup.1H-nuclear magnetic resonance (NMR) analysis was performed by dissolving compound B in approximately 0.7 mL solution of 0.9% NaCl (w/v) in 10% D.sub.2O/90% H.sub.2O (v/v) on a Varian 500 MHz NMR spectrometer using following parameters to determine a boronic acid to boronic ester ratio.

(63) Parameter Details:

(64) 1. Spectral width: 2 ppm to 18 ppm

(65) 2. Acquisition time: 3 s

(66) 3. Number of scans: 64

(67) 4. Pulse width: 7.4 s

(68) (PRESAT Mode used to suppress the water peak resonate at 4.8 ppm)

(69) 1D .sup.1H & .sup.13C nuclear magnetic resonance (NMR) analysis was also performed by dissolving compound B in DMSO-d6 solvent to confirm the ester formation between bortezomib and D-mannitol.

(70) The .sup.1H NMR spectrum recorded in duplicate revealed a boronic acid (i.e. bortezomib) to boronic ester (i.e. bortezomib-mannitol ester) ratio of approximately 0.06:1 and 0.07:1, as depicted in the FIG. 1 and FIG. 2, respectively. The region from 2.35 to 2.85 ppm represents the integrated signal from the protons of free bortezomib and ester wherein the peak at 2.73 ppm is assigned to the free bortezomib and the peak at 2.62 ppm is assigned to the bortezomib-mannitol ester.

(71) Following are the results of free bortezomib to bortezomib-mannitol ester ratio reported in the art for Bortenat and VELCADE samples and the one for compound B.

(72) TABLE-US-00002 free bortezomib:bortezomib- Sample Details mannitol ester ratio Bortenat 2 mg 0.27:1 Bortenat 3.5 mg 0.13:1 VELCADE 0.10:1 Compound B 0.07:1

(73) Further, comparison of the .sup.13C NMR spectra of the bortezomib-mannitol ester formed in accordance with the invention and generic Bortenat sample (in particular, comparison with FIGS. 3 and 4), shows that the .sup.13C NMR spectrum of compound B matches with .sup.13C NMR spectrum of lyophilized material of Bortenat sample. In particular, the .sup.13C NMR Spectrum recorded in DMSO-d6 solvent shows that the characteristic peaks for bortezomib-mannitol ester, which include 63.1 ppm, 73.7 ppm and 78.1 ppm, are observed in both the bortezomib mannitol ester samples of compound B and the generic Bortenat sample. Tabulated below are the comparison results of bortezomib mannitol ester samples of compound B and generic Bortenat sample for ester confirmation.

(74) TABLE-US-00003 Bortenat Injection Source Compound B sample .sup.13C Chemical shift CH.sub.2 63.09 62.96 (ppm) of three Extra CH 73.71 73.62 peaks due to ester CH 78.10 78.00 formation (CH.sub.2, CH, CH respectively).

(75) The .sup.13C NMR spectrum of compound B therefore matches with .sup.13C NMR spectrum of lyophilized material of Bortenat Injection sample.

(76) The NMR data shows that that boronic acid: boronic ester ratio of compound B (bortezomib-mannitol ester) is on lower side (0.07:1) as compared to the values of both commercially available VELCADE and Bortenat samples, thus confirming that the processes of the present invention provide bortezomib-mannitol ester which is superior over the prior art.

Example 8: Preparation of Bortezomib Mannitol Ester for Injection (3.5 mg Bortezomib/3.5 ml Vial)

(77) Formula A

(78) Compound B in 1:6 Ratio

(79) TABLE-US-00004 Ingredients mg/ml mg/vial Mixture of bortezomib mannitol ester and 7.0 24.5 mannitol, with 1:6 weight ratio of bortezomib to mannitol Mannitol 4.0 14.0 Water for injection q.s. to 1.0 ml q.s. to 3.5 ml

(80) Formula B

(81) Compound B in 1:5 Ratio

(82) TABLE-US-00005 Ingredients mg/ml mg/vial Mixture of bortezomib mannitol ester and 6.0 21.0 mannitol, with 1:5 weight ratio of bortezomib to mannitol Mannitol 5.0 17.5 Water for injection q.s. to 1.0 ml q.s. to 3.5 ml

(83) Manufacturing Process

(84) 1. In a stainless compounding vessel, compound B was added to water and stirred at room temperature to dissolve.

(85) 2. To this clear filtrate was added mannitol under stirring and stirred.

(86) 3. Made up the volume with water for injection.

(87) 4. Filtered through 0.22 m sterilizing grade filter.

(88) 5. Vials filled and partially stoppered. Placed filled vials in lyophiliser for lyophilisation.

(89) 6. After completion of lyophilisation, vials stoppered completely and sealed.

(90) While the present invention has been described in terms of its specific embodiments and examples, they are not to be construed as limiting. It will be appreciated that the invention may be modified within the scope of the appended claims.