Sterile injectable formulation based on rocuronium

Abstract

The present invention relates to an injectable formulation based on Rocuronium bromide and the relative preparation and sterilization process.

Description

[0001] STERILE INJECTABLE FORMULATION BASED ON ROCURONIUM

[0002] The present invention relates to an injectable formulation based on Rocuronium bromide and the relative preparation and sterilization process.

[0003] Rocuronium bromide is a muscle-relaxant drug used as an adjuvant for general anesthesia to provide relaxation of the skeletal muscles during surgical interventions and to facilitate tracheal intubation in patients of all ages (from full-term newborns to adolescents and adults. In adults, it is also indicated as an adjuvant in intensive care units, for example to facilitate intubation for short-term use.

[0004] All sterile injectable solutions based on Rocuronium currently on the market (such as, for example, Rocuronium 10 mg / ml Tamarang; Rocuronium 10 mg / ml ICMA; Rocuronium 10 mg / ml Fresenius; Esmeron) are sterilized by means of sterilizing filtration.

[0005] It is known that the sterilizing filtration process does not provide the same guarantees of sterilization as a sterilization process carried out using steam (see EMA / CHMP / CVMP / QWP / 850374 / 2015 guidelines) .

[0006] Furthermore, Rocuronium is unstable in aqueous solution. During the steam sterilization phase, Rocuronium degrades forming a high content of impurity C having the following formula: The formation of impurity C due to the hydrolysis of the acetyl group of Rocuronium can also reach concentration levels higher than 2%.

[0007] In order to avoid degradation of the active ingredient, the product is conserved and stored in a refrigerator at 2-8°C and its transportation consequently requires the cold chain.

[0008] EP3162370 Al describes an injectable formulation comprising rocuronium bromide at a concentration of 10 mg / ml, an isotonicizing agent (NaCl) at a concentration of 9 mg / ml, a pH adjusting agent (NaOH) at a concentration of 0.2 mg / ml, a chelating agent (citric acid) at a concentration of 6.8 mg / ml. The content of impurity C due to the hydrolysis of rocuronium in this formulation after sterilization is around 0.40%.

[0009] The authors of the present invention have now developed a new injectable formulation that slows down the hydrolysis and oxidation process of Rocuronium bromide minimizing the content of impurity C (at TO < 0.20%) . The injectable formulation based on Rocuronium bromide can be advantageously sterilized in an autoclave and transported and stored at a temperature not exceeding 25°C.

[0010] The present invention therefore relates to a sterile injectable formulation comprising:

[0011] Rocuronium bromide as an active ingredient at a concentration of 9.5 mg / ml - 10.5 mg / ml, preferably 10 mg / ml ;

[0012] - an isotonicizing agent at a concentration ranging from 5 mg / ml to 7 mg / ml; one or more pH adjusting agents present at a concentration ranging from 0.01 to 6 mg / ml; characterized in that it further comprises one or more ion-chelating agents at a concentration ranging from 0.01 to 1 mg / ml and nitrogen as inerting agent.

[0013] Said isotonicizing agent is selected from the group consisting of sodium chloride, dihydrate, mannitol and glucose monohydrate or a combination thereof. According to a preferred embodiment of the present invention said isotonicizing agent is sodium chloride (NaCl) . Said isotonicizing agent is preferably present in a concentration of 6 mg / ml.

[0014] According to a further preferred embodiment of the present invention said one or more pH adjusting agents are selected from the group consisting of sodium acetate trihydrate, glacial acetic acid and hydrochloric acid 34-37%. According to a particularly preferred embodiment of the invention all three pH adjusting agents sodium acetate trihydrate, glacial acetic acid and hydrochloric acid at 34-37% are simultaneously present in the formulation. The pH adjusting agents sodium acetate trihydrate and glacial acetic acid are each present at a concentration of 2 mg / ml. The hydrochloric acid at 34- 37% is present at a concentration ranging from 0.01 to 6 mg / ml, preferably 3 mg / ml.

[0015] The pH of the injectable formulation based on Rocuronium bromide according to the invention is preferably kept at between 3.5 - 3.6.

[0016] According to a further preferred embodiment of the invention, said one or more ion-chelating agents are present at a concentration ranging from 0.01 mg / ml to 1 mg / ml in the formulation. In a preferred embodiment, said ion-chelating agents are selected from EDTA acid disodium salt and citric acid monohydrate. In an even more preferred embodiment , both agents EDTA acid disodium salt and citric acid monohydrate are each present in the formulation at a concentration of 0 . 1 mg / ml .

[0017] In the case of the formulation according to the invention, the presence of citric acid in a reduced concentration enhances the chelating activity exerted by sodium edetate (EDTA) .

[0018] The experiments provided in the present patent application show the synergism between citrate - edetate at acid pH with blockage of the degradation reactions of rocuronium .

[0019] It is therefore the synergistic activity between the two excipients EDTA and citric acid present in small quantities that acts in a positive sense by counteracting the degradation of rocuronium and the formation of impurity C .

[0020] In a preferred embodiment , the formulation according to the invention has the following composition :

[0021] The inj ectable formulation based on Rocuronium bromide of the invention comprises nitrogen as an inerting agent. Nitrogen is added to the formulation to eliminate residual oxygen until a level of oxygen in the bulk has reached < 0.3 ppm.

[0022] According to a particularly preferred embodiment of the invention, the injectable formulation based on Rocuronium bromide is made sterile by autoclave sterilization. The autoclave sterilization is preferably effected using a non-standard cycle as provided for by the EMA / CHMP / CVMP / QWP / 850374 guidelines: T°C >110°C F0> 8 (lethality) .

[0023] This non-standard autoclave sterilization cycle allows the finished product to be sterilized without degrading the active ingredient, reducing the content of impurity C in the finished product.

[0024] In a preferred embodiment, the injectable formulation according to the invention has an osmolarity within the range of 290-310 mOsm / kg, preferably 300 mOsm / kg .

[0025] The present invention also relates to the sterile injectable formulation based on Rocuronium bromide as defined above for use in the medical field as a musclerelaxant adjuvant for general anesthesia. The sterile injectable formulation based on Rocuronium bromide according to the invention is preferably packaged in vials .

[0026] The present invention also relates to a vial comprising the injectable formulation based on Rocuronium bromide described above together with nitrogen as an inerting agent. Nitrogen is added to the formulation to eliminate residual oxygen until reaching an oxygen level in the bulk of < 0.3 ppm. The vial comprising the injectable formulation based on Rocuronium bromide according to the invention can be advantageously sterilized in an autoclave and not by filtration sterilization, significantly reducing the presence of impurity C in the finished product (at time 0 < 0.20%) .

[0027] The above vial can be white or transparent and contain a volume of 5 ml or 10 ml of the sterile injectable formulation based on Rocuronium bromide according to the invention.

[0028] Finally, the invention provides for a process for the preparation of the sterile injectable formulation based on Rocuronium bromide described above, comprising the following steps: a) cold dissolution at a temperature within the range of 2-15°C of the components sodium chloride, hydrochloric acid at 34-37%, sodium acetate trihydrate, EDTA acid disodium salt, citric acid monohydrate, glacial acetic acid in a volume of water for injectable preparations equal to 80% of the final volume; b) creating a vacuum, bubbling nitrogen; c) adding the active ingredient Rocuronium bromide and stirring until its complete dissolution; d) adjusting the pH to 3.50-3.55 with hydrochloric acid at 34%-37%, bubbling nitrogen and creating a vacuum; e) bringing to volume; f) bubbling nitrogen and creating a vacuum; g) transferring and distributing the solution into the vials under a nitrogen flow; h) autoclave sterilization.

[0029] The autoclave sterilization using a non-standard cycle was carried out as required by the EMA / CHMP / CVMP / QWP / 850374 guideline: T°C >110°C F0>8

[0030] (lethality) .

[0031] The whole production process from the preparation step to the vial filling step must be carried out under vacuum conditions and in the presence of nitrogen (oxygen content in the bulk < 0.3 ppm) .

[0032] The present invention will now be described for illustrative but non-limiting purposes, according to preferred embodiments with particular reference to the attached figures, in which:

[0033] - Figure 1 shows the chromatograms obtained by HPLC analysis relating to rocuronium bromide, impurity C and impurity B present in the formulation of the prior art (rocuronium HBr 10 mg / ml; citric acid 6.8 mg / ml; sodium hydroxide 0.2 mg / ml; sodium chloride 9 mg / ml) .

[0034] The following illustrative, but non-limiting, examples are provided for a better illustration of the invention .

[0035] EXAMPLES

[0036] Example 1: Development of an Injectable formulation based on Rocuronium bromide according to the invention

[0037] Six different formulations were prepared having the following composition:

[0038] 1) Formula 1: standard formulation (corresponding to that present on the Esmeron market) ) Formula 2 modified ) Formula 3 modified ) Formula 4 modified

[0039] 5) Formula 5 modified

[0040] 6) Formula 6 modified (corresponding to Formula 4 without nitrogen)

[0041] Modified Rocuronium bromide injectable formulations were produced using the following manufacturing process: a) cold dissolution at a temperature within the range of 2-15°C of the components provided for in the formulations indicated above in a volume of water for injectable preparations equal to 80% of the final volume; b) creating a vacuum, bubbling nitrogen; c) adding the active ingredient Rocuronium bromide and stirring until its complete dissolution (10 minutes) ; d) adjusting the pH to 3.50-3.55 with hydrochloric acid at 37%, bubbling nitrogen and creating a vacuum; e) bringing to volume up to 100%; f) bubbling nitrogen and creating a vacuum (oxygen < 0.3 ppm) ; g) dividing the vials under nitrogen by transferring the solution under a nitrogen flow; h) autoclave sterilization with a standard and non-standard cycle as provided by the EMA / CHMP / CVMP / QWP / 850374 guidelines (current edition) .

[0042] The standard sterilization cycle was carried out under the following conditions >121°C F0> 15: whereas the non-standard sterilization cycle was carried out under the following conditions (T°C >110°C F0> 8) .

[0043] At this point, stability tests were started under different conditions 2-8°C, 25°C and 40°C for all formulations with verification time points TO (sterile product just produced) , T1 (after 1 month) and T3 (after 3 months) .

[0044] The results obtained are indicated in the following Tables :

[0045] Based on the results of the development tests, the most stable formulation was identified. The most stable formulations after the sterilization and aging step at TO and T3, at 25°C and 40°C were Formulations 4 and 5 especially with reference to the content of impurity C (impurity C level: at time 0 < 0.20% calculated with the HPLC-UV analytical method) .

[0046] In light of the results obtained, the following sterile injectable formulation (SALF Formulation) was developed and validated in shelf life:

[0047] Table 1

[0048] The following Table 2 shows the results of the stability study after 6 months relating to 6 batches of the SALF formulation indicated in the previous Table 1 with reference to the content of impurity C at different temperatures . Table 2

[0049] The data confirm that the injectable formulation based on Rocuronium bromide conceived allows the product to remain stable at T°C < 25°C for at least 18 months with an impurity level C <2%. The data confirm that the injectable formulation based on Rocuronium bromide according to the invention is sterilizable in an autoclave and if stored in the refrigerator maintains an impurity level C of < 0.20 for at least 3 years. Example 2: Comparison study with Rocuronium bromide formulations available on the market

[0050] The injectable formulations based on Rocuronium bromide available on the market that were analyzed and compared with the formulation according to the invention are the following:

[0051] - Rocuronium 10 mg / ml (Tamarang) ;

[0052] - Rocuronium 10 mg / ml (ICMA) ;

[0053] - Rocuronium 10 mg / ml (Fresenius) .

[0054] The chemical composition of the above formulations is as follows: Table 3

[0055] The reference products taken from the market were analyzed without autoclave sterilization and after autoclave sterilization at 121°C F0>8 and compared with the injectable formulation sterilized according to the invention (SALF Formulation - Table 2) . The results obtained for the products on the market sterilized or non-sterilized are summarized in the following Table 4.

[0056] The results obtained for the sterilized injectable

[0057] SALF formulation are indicated in the following Table 5:

[0058] Table 5 Example 3: Comparative study with the injectable formulation based on rocuronium bromide of EP3162370 Al The injectable formulation based on rocuronium bromide described in patent application EP3162370 Al (rocuronium HBr 10 mg / ml; citric acid 6.8 mg / ml; sodium hydroxide tablets 0.2 mg / ml; sodium chloride 9 mg / ml) was analyzed by HPLC in order to verify the content of impurity C in the final product before and after sterilization.

[0059] The pH of the sterile formulation was 2.91 and the osmolarity was 352 mOSm / Kg. HPLC analysis

[0060] The HPLC analysis was carried out under the following conditions :

[0061] Table 6

[0062] The retention times of the different substances analyzed in the formulation are indicated in the following Table:

[0063] Table 7

[0064]

[0065] The results of the HPLC analysis of the formulation before and after sterilization are indicated in the following Table:

[0066] Table 8

[0067] From the HPLC analysis of the formulation of rocuronium bromide of the prior art, it is evident that the presence of sodium citrate in high quantities does not stabilize the product as the content of impurity C after sterilization reaches 0.40%.

[0068] Furthermore, impurity B is also formed at a titer of 0.10% (see chromatograms in Figure 1) .

[0069] This can be explained by the fact that the formulation of the prior art is characterized by a high concentration of citric acid (6.8 mg / ml) which exerts the function of acidifying the solution and as a preservative, but it does not allow storage at T°C < 25°C for the entire shelf-life of the formulation nor a reduction in the content of impurity C of < 0 . 2 % .

[0070] In the case of the formulation according to the invention, the presence of citric acid is reduced by approximately 70 times compared to the formulation described in the prior art .

[0071] In these concentrations , citric acid exerts the function of enhancing the chelating activity of sodium edetate (EDTA) .

[0072] Sodium edetate , in fact , has the capacity of complexing the ions that are present in solution that would increase the degradation kinetics of rocuronium bromide with the formation of impurity C .

[0073] Under these conditions , rocuronium in solution dissociates into rocuronium + HBr and, due to the presence of high concentrations of citrates , stable but pharmacologically inactive rocuronium-citrate molecules could be formed .

[0074] The rocuronium solution in the inj ectable formulation according to claim 1 , on the other hand, is perfectly isotonic with blood ( 301 mOsm / Kg) .

[0075] The rocuronium solution claimed in fact contains a reduced content of citric acid ( 0 . 1 mg / ml ) which enhances the chelating activity of sodium edetate and cannot react with the active ingredient rocuronium HBr, thus leaving it available for interaction with cellular receptors .

Claims

CLAIMS1. A sterile injectable formulation comprising:Rocuronium bromide as the active ingredient at a concentration of 9.5-10.5 mg / ml, preferably 10 mg / ml;- an isotonicizing agent at a concentration ranging from 5 mg / ml to 7 mg / ml;- one or more pH adjusting agents at a concentration within the range of 0.01-6 mg / ml; characterized in that it further comprises one or more ion-chelating agents at a concentration ranging from 0.01 to 1 mg / ml and nitrogen as inerting agent.

2. The sterile injectable formulation according to claim 1, wherein said isotonicizing agent is selected from the group consisting of sodium chloride, calcium chloride dihydrate and glucose monohydrate or a combination thereof .

3. The sterile injectable formulation according to any of claims 1-2, wherein said isotonicizing agent is present at a concentration of 6 mg / ml.

4. The sterile injectable formulation according to any of claims 1-3, wherein said one or more pH adjusting agents are selected from the group consisting of sodium acetate trihydrate, glacial acetic acid and hydrochloric acid at 34%-37% or a combination thereof.

5. The sterile injectable formulation according to any of claims 1-4, wherein sodium acetate trihydrate and glacial acetic acid are present at a concentration of 2 mg / ml and hydrochloric acid at 34%-37% is present at a concentration of 3 mg / ml.

6. The sterile injectable formulation according to any of claims 1-5, wherein said one or more ion-chelatingagents are selected from EDTA acid disodium salt and citric acid monohydrate or a combination thereof.

7. The sterile injectable formulation according to any of claims 1-6, wherein the EDTA acid disodium salt and citric acid monohydrate are each present at a concentration of 0.1 mg / ml.

8. The sterile injectable formulation according to any of claims 1-7, comprising:Rocuronium bromide 10 mg / mlNaCl 6 mg / mlEDTA acid disodium salt 0,1 mg / mlCitric acid monohydrate 0,1 mg / mlSodium acetate trihydrate 2 mg / mlGlacial acetic acid 2 mg / mlHydrochloric acid at 34-37% 3 mg / mlWater for injectable solutions as necessaryNitrogen until an oxygen content of < 0.3 ppm is obtained .

9. The sterile injectable formulation according to any of claims 1-9, wherein the sterilization is effected in an autoclave.

10. The sterile injectable formulation according to claim 10, wherein the autoclave sterilization is effected using a non-standard sterilization cycle T°C >110°C F0> 8.

11. The sterile injectable formulation according to each of claims 1-11, for use in the medical field as a musclerelaxant adjuvant for general anesthesia.

12. A vial containing the sterile injectable formulation according to each of claims 1-11.

13. A process for the preparation of the sterile injectable formulation according to each of claims 1-11,comprising the following steps: a) cold dissolution at a temperature within the range of 2-15°C of the components sodium chloride, hydrochloric acid at 34-37%, sodium acetate trihydrate, EDTA acid disodium salt, citric acid monohydrate, glacial acetic acid in a volume of water for injectable preparations equal to 80% of the final volume; b) creating a vacuum, bubbling nitrogen; c) adding the active ingredient Rocuronium bromide and stirring until its complete dissolution; d) adjusting the pH to 3.50-3.55 with hydrochloric acid at 34%-37%, bubbling nitrogen and creating a vacuum; e) bringing to volume; f) bubbling nitrogen and creating a vacuum; g) transferring and distributing the solution into the vials under a nitrogen flow; h) autoclave sterilization.

14. The process for the preparation of the sterile injectable formulation according to claim 14, wherein the autoclave sterilization step h) is effected using a non-standard sterilization cycle T°C >110°C F0> 8.

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