CLIAMS:We claim,
1. A freeze dried composition of atracurium besylate comprising atracurium besylate; an acid to adjust the pH and polyvinyl Pyrolidone.
2. The formulation of claim 1, wherein the polyvinylpyrrolidone comprises Povidone K12.
3. The freeze dried composition according to claim 1, wherein, the acid is benzene sulphonic acid.
4. The freeze dried composition according to claim 1, wherein the composition comprises 2 to 5% of polyvinylpyrrolidone.
5. The freeze dried composition according to claim 1, wherein the composition comprises 3% of polyvinylpyrrolidone.
6. The freeze dried composition according to claim 1, wherein the benzene sulphonic acid is used to adjust the pH between 3.0-3.5.
7. A process for preparation of freeze dried composition of atracurium besylate which comprises:
a. Dissolving polyvinyl pyrrolidone in a part of cold water for injection, followed by addition of Atracurium Besylate to obtain solution;
b. Adjusting the pH of the solution upto 3.0-3.5;
c. Making up the required volume with cold water for injection followed by aseptical filtration; and
d. Filling the solution in 5 ml Amber Tubular vial and Lyophilised.
8. The process as claimed in claim 7 wherein the solution contains 2% w/v to 5% w/v of polyvinylpyrrolidone.
9. The process as claimed in claim 7 wherein the solution contains 3% w/v of polyvinylpyrrolidone.
10. The process as claimed in claim 7 wherein the solution is chilled to 2°C -8°C.
11. The process as claimed in claim 7 wherein the pH is adjusted with the solution of benzenesulphonic acid.
12. The process as claimed in claim 7 wherein the solution pH is between 3.0 and 3.5.
13. A freeze dried lyophilized form of Atracurium Besylate injection according to any one of the preceding claims comprises 2% to 5% polyvinylpyrrolid0one to increase the purity and stability of Atracurium Besylate.
14. A freeze dried lyophilized form of Atracurium Besylate injection according to claim 13, wherein 2 to 5% polyvinylpyrrolidone is dissolved in aqueous solvent.

Dated this 14th day of May, 2014

Dr. P. Aruna Sree
(Regn.No.: IN/PA 998)
Agent for the Applicant
Gopakumar Nair Associates
,TagSPECI:

FORM 2
THE PATENTS ACT 1970
(39 of 1970)
AND
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rule13)
1. TITLE OF THE INVENTION:

“A STABLE LYOPHILIZED INJECTION OF ATRACURIUM BESYLATE”

2. APPLICANT:

(a) NAME: NEON LABORATORIES LTD.

(b) NATIONALITY: Indian Company incorporated under the
Companies Act, 1956

(c) ADDRESS: 140, Damji Shamji Industrial Complex, Mahakali Caves
Road, Andheri (East), Mumbai - 400093, Maharashtra, India.

3.PREAMBLE TO THE DESCRIPTION:
The following specification describes the nature of this invention and the manner in which it is to be performed:

Technical filed:

The present invention provides highly stabilized freeze dried composition of Atracurium Besylate. More particularly the invention provides a freeze dried composition of atracurium besylate comprising Atracurium Besylate; a stabilizer; an acid to adjust the pH with water as principle solvent. The invention further relates to process of preparation of stable freeze dried composition of atracurium besylate.

Background and prior art:

Atracurium besylate is classified as an intermediate-duration non-depolarizing neuromuscular-blocking agent, used adjunctively in anesthesia to facilitate endotracheal intubation and to provide skeletal muscle relaxation during surgery or mechanical ventilation.

Currently available (Marketed) formulation of Atracurium Besylate is a sterile solution of Atracurium Besylate in water for injection which shows high degradation and there by development of Impurities on storage. Majority of impurities specified for Atracurium Besylate are the isomeric form of Atracurium Besylate. Such isomeric impurities are majorly generated when the API is in solution form.

The administration of Atracurium besylate through the vascular pathway, such as intravenous injection has an advantage of an immediate response and the response may also be easily controlled.

Atracurium injection involves stability problems mainly caused by chemical and physical factors such as light, heat and pH-sensitivity. The stability of the solution is dependant on physical and chemical factors, thus limiting the shelf life of the solution, results in degradation products.

Also, the atracurium besylate injection needs to be stored at temperature between 2.0°C & 8.0°C, and at a pH between 3.0 & 4.0. However, with extended storage time, the stability of atracurium besylate is decreased and related impurities increased thus affects the efficacy of the drug.

In the above context, there are few prior arts which have tried to overcome the problems involved in the injectable atracurium besylate. One such solution is disclosed in CN10053173.

CN10053173 discloses freeze dried composition of atracurium. The composition adopts atracurium or its pharmaceutically acceptable salt or optical isomer as active ingredient, which comprises atracurium, sugar and an organic acid. The invention also discloses a method for preparing the composition.

GB2445746 discloses Carbonated water may be used as a solvent for freeze drying. A crude organic compound may be purified by dissolving it in carbonated water to form a solution and subsequently freeze drying the solution to yield the compound with a reduced level of residual solvent. The carbonated water preferably has a pH less than 6 prior to dissolution and is optionally further carbonated during or after dissolution to restore a pH of less than 8. One such organic compound is atracurium.

In view of the above, it is evident that there is long standing need in the art to provide atracurium besylate in stable lyophilized injectable form with low impurity profile.

Therefore, the object of the invention is to provide a stable lyophilized injectable atracurium besylate with low impurity profile and thus to have long therapeutic stability.

Detailed description of the invention:

In accordance with the objective, the invention provides stable lyophilized injectable atracurium besylate with low impurity profile by minimizing the degradation.

The rational of current invention is to minimize the degradation of atracurium besylate throughout its shelf life and to provide the resultant dosage with maximum stability and with low impurity profile. Therefore, the invention provides a lyophilized product that allows the API in anhydrous form and there by reduces the degradation rate and provides maximum stability with reduction in impurities generation.

The purpose of providing the freeze dried atracurium besylate is to reduce the generation of impurities.

Accordingly, the instant invention provides a freeze dried composition of atracurium besylate comprising atracurium besylate; Povidone K12, an organic acid to adjust the pH with water as a principle solvent.

In a preferred embodiment, the instant freeze dried composition comprises atracurium besylate; Benzene Sulphonic Acid to adjust the pH and Povidone K12 and water as a principle solvent.

The freeze dried composition according to the invention contains 2% w/v to 5% w/v of polyvinylpyrrolidone.

In an embodiment, the freeze dried composition may preferably contain 3% w/v of polyvinylpyrrolidone.

According to the preferred embodiment, the pH of the freeze dried composition is adjusted with benzenesulphonic acid. The pH of freeze dried composition is preferably maintained between 3.0 and 3.5.

In another preferred embodiment, the invention provides a process for preparation of freeze dried composition which comprises:
a) Dissolving Povidone K12 in a part of cold water for injection, followed by addition of atracurium besylate;
b) Adjusting the pH of the solution between 3.0 to 3.5;
c) Making up the required volume with cold water for injection followed by aseptical filtration;
d) Filling the solution in 5 ml amber vial and Lyophilised.
The solution according to the invention contains 2% w/v to 5% w/v of polyvinylpyrrolidone.

In an embodiment, the solution may preferably contain 3% w/v of polyvinylpyrrolidone.
In an embodiment, the solution is chilled to 2°C -8°C.

According to the process the pH of the solution is adjusted with solution of benzenesulphonic acid. The pH is preferably adjusted between 3.0 and 3.5.

A freeze dried lyophilized form of Atracurium Besylate injection according to invention comprises 2% to 5% polyvinylpyrrolidone to increase the purity and stability of Atracurium Besylate.

A freeze dried lyophilized form of Atracurium Besylate injection according to the invention, wherein, the 2 to 5% polyvinylpyrrolidone is dissolved in aqueous solvent.

In yet another preferred embodiment, the invention provides stability studies of the instant composition, wherein, the Assay of the drug is in the range of N.L.T. 90% -N.M.T. 115% with total impurities – N.M.T. 10.0% and Laudanosine impurity N.M.T 3.0% at a temperature of Below 25°C.

Several different trials were conducted & tested for the stability. Some of these trials are discussed below in brief.

The following examples, which include preferred embodiments, will serve to illustrate the practice of this invention, it being understood that the particulars shown are by way of example and for purpose of illustrative discussion of preferred embodiments of the invention.

Example 1:
Ingredients Quantity/ ml
Atracurium Besylate 12.5 mg
Benzene Sulphonic acid Solution q.s to pH 3.0 to 3.5
WFI Qs 1 mL

Procedure:
a) Dissolving Atracurium Besylate in a part of cold water for injection, followed by adjusting the pH of the solution 3.0 to 3.5 with solution of Benzene sulphonic acid;
b) Making up the required volume with cold water for injection followed by aseptical filtration and
c) Filling the solution in 5 mL Amber Tubular vial and Lyophilised.
The results are described in Table 1 herein below:
Stage

Assay
pH Water Chromatography Impurity
Acidic Comp. Laud-
anosine Cis-Isomer Hydro. Comp. Trans-Isomer Hydro. Comp Cis-Isomer Monohydrate Trans-Isomer Monohydrate Total Impurity
N.M.T. 6.0% N.M.T. 3.0% N.M.T. 6.0% N.M.T. 6.0% N.M.T. 3.0% N.M.T.
3.0% N.M.T. 10.0%
Initial 100.0 3.22 0.89 4.97 0.054 1.04 3.39 0.051 0.077 9.94

Example 2:
Ingredients Quantity/ ml
Atracurium Besylate 12.5 mg
TBA 0.75 ml
Benzene Suphonic acid Solution q.s to pH 3.0 to 3.5
WFI Qs 1 mL

Procedure:
a) Dissolving Atracurium Besylate in a part of water for injection, followed by addition and mixing of co-solvent TBA;
b) adjusting the pH of the solution to 3.2 ± 0.1; Making up the required volume with cold water for injection followed by aseptical filtering ; and
c) Filling the solution in 5 mL Amber Tubular vial, and Lyophilised.
The results are described in Table 2 herein below:

Stage

Assay
pH Water Chromatography Impurity
Acidic Comp. Laud-
anosine Cis-Isomer Hydro. Comp. Trans-Isomer Hydro. Comp Cis-Isomer Monohydrate Trans-Isomer Monohydrate Total Impurity
N.M.T. 6.0% N.M.T. 3.0% N.M.T. 6.0% N.M.T. 6.0% N.M.T. 3.0% N.M.T.
3.0% N.M.T. 10.0%
Initial 103.84% 3.20 0.54 1.23% 0.050% 0.30% 0.92% 0.055% 0.081% 2.92%
25°C/
1 Month 95.9% 3.24 0.58 2.29% 4.07% 2.36% 2.47% 1.02% 1.04% 14.25%

Example 3:

Ingredients Quantity/ ml
Atracurium Besylate 12.5 mg
Povidone K12 50 mg
Benzene Sulphonic acid Solution q.s to pH 3.0 to 3.5
WFI Qs 1 mL

Procedure:
a) Dissolving Stabilizer Povidone K12 in a part of cold water for injection, followed by addition of Atracurium Besylate;
b) Adjusting the pH of the solution between 3.0 and 3.5 with solution of Benzene sulphonic acid; making up the required volume with cold water for injection followed by aseptical filtering; and
c) Filling the solution in 5 mL Amber Tubular vial and Lyophilised.
The results are described in Table 3 herein below:

Stage

Assay
pH Water Chromatography Impurity
Acidic Comp. Laud-
anosine Cis-Isomer Hydro. Comp. Trans-Isomer Hydro. Comp Cis-Isomer Monohydrate Trans-Isomer Monohydrate Total Impurity
N.M.T. 6.0% N.M.T. 3.0% N.M.T. 6.0% N.M.T. 6.0% N.M.T. 3.0% N.M.T.
3.0% N.M.T. 10.0%
Initial 108.75 3.30 0.88 0.03 0.25 Nil 0.03 0.12 0.35 1.07
1M/2-8°C 107.66 3.37 0.87 0.03 0.39 Nil 0.02 0.17 0.53 1.25
2M/2-8°C 103.95 3.31 0.89 0.02 0.36 0.01 0.03 0.18 0.52 1.55
3M/2-8°C 102.77 3.35 0.85 0.01 0.36 0.01 0.02 0.16 0.50 1.26
6M/2-8°C 103.83 3.23 0.87 0.03 0.42 0.01 0.03 0.18 0.57 1.68
1M/25°C 106.44 3.32 0.89 0.03 0.81 Nil 0.02 0.32 1.01 2.33
2M/25°C 108.37 3.29 0.85 0.03 1.34 0.01 0.03 0.51 1.67 3.98
3M/25°C 108.88 3.28 0.86 0.02 1.58 0.01 0.04 0.56 1.89 4.41
6M/25°C 102.19 3.34 0.85 0.04 3.48 0.01 0.03 1.10 3.87 8.90

Example 4:
Ingredients Quantity/ ml
Atracurium Besylate 12.5 mg
Povidone K12 50 mg
TBA 0.15 ml
Benzene Sulphonic acid Solution q.s to pH 3.0 to 3.5
WFI Qs 1 mL

Procedure:
a) Dissolving Povidone K12 in a part of cold water for injection, followed by addition of TBA and Atracurium Besylate;
b) Adjusting the pH of the solution between 3.0 and 3.5 using solution of Benzene sulphonic acid; Making up the required volume with cold water for injection followed by aseptical filtration.
c) Filling the solution in 5 mL Amber Tubular vial and Lyophilised.
The results are described in Table 4 herein below:
Stage

Assay
pH Water Chromatography Impurity
Acidic Comp. Laud-
anosine Cis-Isomer Hydro. Comp. Trans-Isomer Hydro. Comp Cis-Isomer Monohydrate Trans-Isomer Monohydrate Total Impurity
N.M.T. 6.0% N.M.T. 3.0% N.M.T.
6.0% N.M.T.
6.0% N.M.T.
3.0% N.M.T.
3.0% N.M.T. 10.0%
Initial 109.28 3.26 0.84 0.025 0.238 Nil 0.030 0.12 0.338 1.038
25°C/
1Month 105.78 3.31 0.91 0.040 0.89 Nil 0.029 0.32 1.04 2.55

Example 5
Ingredients Quantity/ ml
Atracurium Besylate 12.5 mg
Povidone K 12 30 mg
Benzene Sulphonic Acid Solution q.s to pH 3.0 to 3.5
WFI Qs 1 mL

Dissolving Kollidon 12 PF in a part of cold water for injection, followed by addition of Atracurium Besylate. Adjust the pH of the solution between 3.0 and 3.5 with 0.2 % solution of Benzene sulphonic acid. Make up the required volume with cold water for injection followed by aseptically filtration. Filling the solution in 5 mL Amber Tubular vial and Lyophilised.
The results are described in Table 6 herein below:
Stage

Assay
pH Water Chromatography Impurity
Acidic Comp. Laud-
anosine Cis-Isomer Hydro. Comp. Trans-Isomer Hydro. Comp Cis-Isomer Monohydrate Trans-Isomer Monohydrate Total Impurity
N.M.T. 6.0% N.M.T. 3.0% N.M.T.
6.0% N.M.T.
6.0% N.M.T.
3.0% N.M.T.
3.0% N.M.T. 10.0%
Initial 107.93 3.22 1.59% 0.0605% 0.0641% 0.024% 0.0608% 0.024% 0.0976% 0.439%
1M/2-8°C 107.08 3.27 1.67 0.03 0.08 Nil 0.027 Nil 0.23 0.48
2M/2-8°C 106.15 3.28 1.89 0.036 0.056 0.013 0.059 0.047 0.090 0.658
3M/2-8°C 107.04 3.25 2.07 0.028 0.060 0.012 0.038 0.037 0.092 0.495
6M/2-8°C 107.66 3.27 2.53 0.049 0.055 0.014 0.047 0.034 0.084 0.724
1M/25°C 107.27 3.29 2.03 0.09 0.07 0.25 0.024 Nil 0.086 0.46
2M/25°C 106.30 3.25 2.47 0.159 0.073 0.042 0.153 0.055 0.109 0.906
3M/25°C 106.88 3.29 3.08 0.303 0.103 0.078 0.248 0.052 0.141 1.218
6M/25°C 105.75 3.26 3.75 0.509 0.116 0.121 0.392 0.162 0.162 1.731