Freeze-dried Sovastatin-based Injectable Formulations And Methods Of Making

Abstract

A freeze-dried injectable formulation based on sovastatin comprising sovastatin in the range of from about 0.01 to about 0.02 weight percent, trisodium citrate dihydrate being present in the range of from about 20 to about 80 weight percent, and mannitol being present in the range of from about 20 to about 80 weight percent. The composition comprises sovastatin; trisodium citrate; the invention relates to a redissolved liquid composition of a freeze-dried pharmaceutical composition of sovatide, mannitol and water or 0.9% sodium chloride aqueous solution, and a method for preparing the freeze-dried pharmaceutical composition of sovatide, which comprises the following steps: dissolving sovatide, trisodium citrate dihydrate and mannitol in water for injection; enabling the solution to pass through a 0.2 mu membrane filter for filtering; filling each vial to a target filling volume; and the filled vials are freeze-dried.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This international application claims priority to U.S. Non-Provisional Patent Application No. 18 / 343,087, filed on June 28, 2023, and U.S. Non-Provisional Patent Application No. 18 / 478,528, filed on September 29, 2023. The contents of each of these applications are incorporated herein by reference. Technical Field

[0003] The present invention relates to a freeze-dried injectable pharmaceutical formulation based on thiabendazole for treating various neurological diseases and conditions (including but not limited to cerebral ischemic stroke and intracranial hemorrhage) and a method for preparing the same, by providing an intravenous administration route and avoiding the pain or irritation caused by conventional procedures. Background Art

[0004] There are a wide variety of diseases and conditions that cause discomfort and can be fatal, yet have very limited treatment options. A small number of these diseases and conditions include neurological disorders or diseases, including stroke, Alzheimer's disease, spinal cord injury, cognitive impairment, neurofibromatosis, Huntington's disease, Parkinson's disease, neonatal hypoxic-ischemic encephalopathy, and multi-infarct dementia. Although various drugs have been discovered and synthesized to address these diseases and conditions, concerns remain regarding their effectiveness and potential side effects.

[0005] Neurological conditions (such as ischemic stroke, intracranial hemorrhage and brain injury) usually cause cerebral edema. Stroke is one of the most common causes of death and is also an important cause of serious long-term disability. Ischemic stroke caused by arterial occlusion is the cause of most strokes. Various treatment options have been developed to treat stroke. Mannitol is a kind of such treatment option because it reduces brain volume by reducing total water content, reduces blood volume by vasoconstriction, and reduces CSF volume by reducing water content. Mannitol can also improve cerebral perfusion by reducing viscosity or changing erythrocyte rheology.

[0006] However, the administration of mannitol alone is insufficient to reduce morbidity and mortality in stroke patients. Furthermore, the need to monitor fluid and electrolyte levels, serum osmolality, and renal, cardiac, and pulmonary function during and after mannitol infusion makes the treatment process cumbersome.

[0007] Another treatment option available for stroke is the use of tissue plasminogen activator (tPA). However, the use of tPA is limited to a short time window of <4.5 hours after symptom onset and it also carries the risk of intracranial hemorrhage.

[0008] Sovateltide is one such drug with therapeutic potential for treating such diseases / conditions, preferably neurological conditions. Sovateltide, or succinyl-(glutamyl(9)-alanyl(11,15))-endothelin-1(8-21), is a compound with a 15-amino acid chain. Sovateltide mimics the properties of a natural peptide called endothelin-1 (ET-1). ET-1 is known to activate two types of receptors in the body, endothelin A (ETAR) and endothelin B (ETBR). Sovateltide is designed to specifically activate ETBR, making it a selective agonist for this receptor type. In fact, aqueous solutions of sovateltide tend to deteriorate during storage.

[0009] US8623823 Disclosed the use of ET B Methods of administering ET receptor agonists (e.g., IRL-1620) for the treatment of stroke or cerebrovascular accident. B Receptor agonists are used alone or in combination with a second agent that can be used to treat stroke or other cerebrovascular accidents.

[0010] US10561704 Disclosed are compositions and methods for treating neuropsychiatric disorders in vertebrates and humans. More specifically, the present invention provides the use of the endothelin-B receptor agonist IRL-1620 as a neuroprotective and neuroregenerative agent at appropriate doses. Thus, in one aspect, the present disclosure provides a method for treating a neuropsychiatric disorder comprising administering a therapeutically effective amount of an endothelin-B receptor agonist to a patient in need thereof to treat the neuropsychiatric disorder. In some embodiments, the endothelin-B receptor agonist is co-administered with an additional agent to treat the neuropsychiatric disorder. In some embodiments, the additional agent is selected from an antidepressant, an anti-inflammatory agent, a CNS stimulant, a neuroleptic, and an antiproliferative agent.

[0011] RU2739382C1 Disclosed is the technical field: medicine. Content: The present invention relates to the treatment of stroke, and in particular to a new method for treating stroke by administering imatinib. Imatinib is administered to the patient at a dose of 650-1600 mg / day on the first day, and is administered to the patient at a dose of 650-1200 mg / day for at least 2 consecutive days, preferably at least 3 consecutive days, and most preferably at least 4 consecutive days. Administration of imatinib for more than 3 days improves the neurological outcome of patients after stroke and improves the functional independence of patients. Effect: The present invention improves the treatment of acute ischemic or hemorrhagic stroke and broadens the therapeutic window of thrombolysis. RU'382 discloses that imatinib is administered intravenously as a first dose and then The use of subsequent doses of imatinib administered orally to treat stroke. The drug is administered via a larger diameter, so the reaction time is longer.

[0012] US8361459B2Disclosed are cells derived from postnatal tissues, such as the umbilical cord and placenta, and methods of using them to regenerate, repair, and improve neural tissue and improve behavior and neurological function in stroke patients. US'459 mentions that The use of postpartum derived cells to treat stroke and other acute neurodegenerative disorders. However, the methods mentioned in the prior art are They are very specific for stroke and acute neurodegenerative diseases and therefore have a narrow range of applications.

[0013] Conventionally, many compositions and methods capable of treating various neurological disorders have been developed. However, since such compositions and methods use only a single drug, the neurological prognosis of such compositions is insufficient.

[0014] However, such approaches either involve the use of multiple doses of drugs with a high response time or have a narrow spectrum of action.

[0015] In order to overcome the above-mentioned shortcomings, there is a need in the art to develop lyophilized sovastatin-based pharmaceutical formulations for the treatment of various neurological disorders (e.g., cerebral ischemic stroke, intracranial hemorrhage) that are in lyophilized form to prevent the formulation from deteriorating and that can be administered via the intravenous route to prevent any chance of irritation to the patient that typically occurs during traditional treatment.

[0016] Purpose of the Invention

[0017] The main object of the present invention is to overcome the disadvantages of the prior art.

[0018] One object of the present invention is to develop formulations containing lyophilized sovastatin for the treatment of various neurological diseases and conditions (including but not limited to cerebral ischemic stroke, intracranial hemorrhage) that exhibit greater inherent stability compared to sovastatin solutions.

[0019] Another object of the present invention is to develop a formulation that can be administered via the intravenous route or the intraarterial route.

[0020] Another object of the present invention is to develop formulations that are patient compliant.

[0021] Another object of the present invention is to develop a formulation containing very small amounts of sovastatin, thereby preventing any chance of side effects due to an overdose of sovastatin.

[0022] The foregoing and other objects, features and advantages of the present invention will become apparent upon further review of the following detailed description of the preferred embodiments, as illustrated in the accompanying drawings. SUMMARY OF THE INVENTION

[0024] The present invention relates to lyophilized injectable formulations based on sovastatin for use in the treatment of various neurological disorders and diseases for administration intravenously or via the intra-arterial route and methods for their preparation.

[0025] According to one embodiment of the present invention, a pharmaceutical formulation for treating stroke comprises: i) an endothelin-B receptor analog in a dosage range of 0.00001 mg / kg-1 mg / kg, and ii) a drug in a dosage range of 0.0015 g / kg-5 g / kg.

[0026] According to a second embodiment of the present invention, endothelin-B receptor analogs include but are not limited to sovastatin, BQ-3020, [Ala 1,3,11,15 ]-endothelin, sarafotoxin S6c, endothelin-3, and the drugs include but are not limited to mannitol and citicoline.

[0027] According to one embodiment of the present invention, a lyophilized sovastatin-based injectable formulation comprises: i) an active pharmaceutical ingredient in the range of 0.01-0.02 wt %, ii) at least two soluble excipients in the range of 20-80 wt %, and iii) water for injection in the range of 1-2 wt %.

[0028] According to a third embodiment of the present invention, a lyophilized solvatide-based injectable formulation comprises: i) solvatide present in a range of about 0.01 to about 0.02 weight %, trisodium citrate dihydrate present in a range of about 20 to about 80 weight %, and mannitol present in a range of about 20 to about 80 weight %.

[0029] According to a fourth embodiment of the present invention, a method for preparing a lyophilized sovastatin-based injectable formulation comprises the following steps: i) dissolving trisodium citrate dihydrate in water for injection to obtain a mixture, ii) adding the active pharmaceutical ingredient, sovastatin, to the mixture, and then adding mannitol to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, followed by sterile filtering the solution and aseptically filling the filtered solution into a vial, and iv) half-stoppering the vial and then loading the vial into a freeze dryer for freeze drying to obtain a lyophilized sovastatin-based injectable formulation.

[0030] While the invention has been described and illustrated with particular reference to preferred embodiments thereof, it will be apparent that variations therein may be made which fall within the scope of the invention.

[0031] BRIEF DESCRIPTION OF THE DRAWINGS

[0032] These and other features, aspects and advantages of the present invention will become better understood with reference to the following description, appended claims and accompanying drawings, in which:

[0033] Figure 1 Depicted is a graphical representation of the modified Rankin scale for patients treated with control and sovastatin together with mannitol;

[0034] Figure 2 Depicted is a graphical representation of the number of patients with cerebral ischemic stroke treated with control and sovastatin together with mannitol and having a modified Rankin scale of 0-2 at 90 days after randomization;

[0035] Figure 3 Depicts a graphical representation of the number of patients with cerebral ischemic stroke treated with control and sovastatin with mannitol and having an improvement of 2 or more from baseline on the modified Rankin scale at 90 days after randomization;

[0036] Figure 4 Depicted is a graphical representation of the modified Rankin scale for patients treated with control and sovastatin together with citicoline;

[0037] Figure 5 Depicts a graphical representation of the number of patients with cerebral ischemic stroke treated with control and sovastatin together with citicoline and having a modified Rankin scale of 0-2 at 90 days after randomization;

[0038] Figure 6 Depicts a graphical representation of the number of patients with cerebral ischemic stroke treated with control and sovastatin together with citicoline and having an improvement of 2 or more from baseline on the modified Rankin scale at 90 days after randomization;

[0039] Figure 7 Graph depicting the modified Rankin scale for patients treated with control and sovastatin together with cerebrolysin;

[0040] Figure 8 Depicts a graphical representation of the number of patients with cerebral ischemic stroke treated with control and sovastatin together with cerebrolysin and having a modified Rankin scale of 0-2 at 90 days after randomization;

[0041] Figure 9 Depicts a graphical representation of the number of patients with cerebral ischemic stroke treated with control and sovastatin together with cerebrolysin and having an improvement of 2 or more from baseline on the modified Rankin scale at 90 days after randomization;

[0042] Figure 10 Depicts a graphical representation of the modified Rankin scale for patients treated with control and sovastatin together with thrombolytics;

[0043] Figure 11 Depicted is a graphical representation of the number of patients with cerebral ischemic stroke treated with control and sovastatin together with thrombolytics and with a modified Rankin scale of 0-2 at 90 days after randomization;

[0044] Figure 12 Depicts a graphical representation of the number of patients with cerebral ischemic stroke who were treated with control and sovastatin together with thrombolytics and had a modified Rankin scale improvement of 2 or more from baseline at 90 days after randomization;

[0045] Figure 13 A flow chart depicting the process of developing a lyophilized sovastatin-based injectable formulation is shown. Detailed Description of the Invention

[0047] The following description includes a preferred best mode of one embodiment of the present invention. It will be apparent from the description of the present invention that the present invention is not limited to the embodiments described, but rather the present invention also includes various modifications and embodiments thereof. Therefore, this description should be regarded as illustrative rather than restrictive. Although the present invention is susceptible to various modifications and alternative constructions, it should be understood that the present invention is not intended to be limited to the specific form disclosed, but rather, the present invention covers all modifications, alternative constructions, and equivalents that fall within the spirit and scope of the present invention as defined in the claims.

[0048] In any embodiment described herein, the open-ended terms "comprising," "including," etc. (which are synonymous with "including," "having," and "characterized by") may be replaced by the corresponding partially enclosed phrases "consisting essentially of," etc. or the corresponding enclosed phrases "consisting of," etc.

[0049] As used herein, the singular forms "a," "an," and "the" and the like, without a quantifier, refer to both the singular and the plural, unless expressly stated to be directed to the singular only.

[0050] As used herein, the term "pharmaceutical dosage form" refers to the specific form or formulation in which a drug or drug substance is formulated and administered to a patient for therapeutic purposes. Such dosage forms can vary in their physical state (e.g., solid, liquid, or semisolid) and their route of administration (e.g., oral, topical, intravenous, or inhalation).

[0051] As used herein, the term "freeze-dried or lyophilized" powder or cake or formulation refers to any solid material that has undergone the freeze-drying or freeze-drying process of an aqueous solution. Ideally, a freeze-dried formulation is obtained by freeze-drying a solution consisting of an aqueous solvent.

[0052] As used herein, the term "stable pharmaceutical composition" refers to a formulation of a drug or active ingredient that retains its physical, chemical, and therapeutic properties for an extended period of time under appropriate storage conditions.

[0053] The present invention is a pharmaceutical formulation for treating neurological disorders, comprising an endothelin-B receptor analog and a drug, wherein the analog exhibits synergistic effects with the drug, resulting in enhanced therapeutic effects of the formulation against various neurological disorders (eg, stroke).

[0054] The endothelin-B receptor analogs disclosed herein are selected from the group consisting of: sovastatin, BQ-3020, [Ala 1 ,3,11,15 ]-endothelin, sarafotoxin S6c, endothelin-3. The drugs disclosed herein include but are not limited to mannitol and citicoline.

[0055] Sovastatin as an active ingredient includes a liquid injection with a solvent, which contains at least one soluble excipient, wherein the soluble excipient includes mannitol, trisodium citrate, anhydrous citric acid, disodium hydrogen citrate, disodium hydrogen phosphate, sodium chloride, and hydroxypropyl cyclodextrin, wherein the soluble excipient has multiple purposes of serving as an osmotic pressure regulator, a pH regulator, and a solubility enhancer.

[0056] The osmotic pressure regulator (buffer), pH regulator and solubility enhancer are selected from sodium chloride, trisodium citrate, anhydrous citric acid, disodium hydrogen citrate, disodium hydrogen phosphate, hydroxypropyl β-cyclodextrin and mannitol and mixtures thereof.

[0057] The freeze-dried filler is selected from the group consisting of sodium chloride, trisodium citrate, anhydrous citric acid, disodium hydrogen citrate, disodium hydrogen phosphate, hydroxypropyl beta-cyclodextrin and mannitol, and mixtures thereof.

[0058] In one aspect, the present invention relates to a lyophilized pharmaceutical composition comprising: a) solvatide; b) trisodium citrate; and c) mannitol. In one embodiment, a) solvatide is present in a range of about 0.01% to about 0.02% by weight; b) trisodium citrate dihydrate is present in a range of about 20% to about 80% by weight; and c) mannitol is present in a range of about 20% to about 80% by weight. In another embodiment, a) solvatide is present in a range of about 30 μg; b) trisodium citrate is present in a range of about 50 mg; and c) mannitol is present in a range of about 160 mg.

[0059] Specifically, the present invention describes a lyophilized solvatide-based injectable formulation comprising: i) solvatide present in a range of about 0.01 to about 0.02 weight percent, trisodium citrate dihydrate present in a range of about 20 to about 80 weight percent, and mannitol present in a range of about 20 to about 80 weight percent.

[0060] The formulations herein are prepared as lyophilized formulations and can be administered by intravenous or intraarterial routes.

[0061] The formulation is administered via a catheter, with or without mechanical assistance, or during an embolectomy or thrombectomy procedure.

[0062] Formulations are prepared in the form of nanoparticles, gels or hydrogels, nanoemulsions, microparticles, colloidal suspensions, sterile suspensions, solutions, aerosols, and powders.

[0063] The formulation comprises a biodegradable polymer, wherein the biodegradable polymer is poly(lactic-co-glycolic acid) (PLGA) or pegylated PLGA (PEG-PLGA).

[0064] The formulation includes additives including, but not limited to, polyvinyl alcohol (PVA) or other known nanoparticle stabilizers.

[0065] The lyophilized pharmaceutical composition comprises about 30 μg of sovastatin; about 50 mg of trisodium citrate dihydrate; and about 160 mg of mannitol.

[0066] The lyophilized pharmaceutical composition further comprises: no more than 2% of total impurities; or no more than 0.5% of any unspecified impurities; or no more than 1.0% of D-His-sovaccinium impurity.

[0067] The lyophilized pharmaceutical composition contains: no more than 0.7% total impurities; or no more than 0.15% any unspecified impurities; or no more than 0.22% D-His-sovaccinium impurity. The composition is reconstituted with 0.9% sodium chloride solution and water for injection.

[0068] A method of treating a neurological condition or disease comprising administering a lyophilized pharmaceutical composition to a patient in need thereof.

[0069] The neurological condition or disease is selected from the group consisting of stroke, Alzheimer's disease, spinal cord injury, cognitive impairment, neurofibromatosis, Huntington's disease, Parkinson's disease, neonatal hypoxic-ischemic encephalopathy, traumatic brain injury, and multi-infarct dementia.

[0070] A reconstituted liquid composition is prepared comprising sovastatin, trisodium citrate, mannitol, and water or a 0.9% sodium chloride aqueous solution. In one embodiment, the sovastatin, trisodium citrate, and mannitol in the reconstituted liquid composition are provided as lyophilized powders.

[0071] A liquid pharmaceutical composition wherein sovastatin is present in a range of about 0.01 to about 0.02 weight percent; trisodium citrate dihydrate is present in a range of about 20 to about 80 weight percent, and mannitol is present in a range of about 20 to about 80 weight percent.

[0072] Additionally, sovastatin is present at about 30 μg; trisodium citrate is present at about 50 mg; and mannitol is present at about 160 mg in the liquid composition.

[0073] In the liquid composition, sovastatin is present at a concentration of 6 μg / ml.

[0074] The liquid composition further comprises: no more than 2% total impurities; or no more than 0.5% of any unspecified impurities; or no more than 1.0% of the D-His-sovaceptin impurity.

[0075] The liquid composition further comprises: no more than 0.7% total impurities; or no more than 0.15% of any unspecified impurities; or no more than 0.22% of the D-His-sovaccinium impurity.

[0076] For the reconstituted liquid composition, the osmotic pressure is 240 to 310 mOsm / L, and preferably about 286 mOsm / L. The pH of the liquid composition is 7.0 to 8.5.

[0077] The present invention also includes a kit or co-package comprising: a vial of a lyophilized pharmaceutical composition containing sovastatin; and an ampoule / vial containing a 0.9% aqueous sodium chloride solution or water for injection. The kit or co-package of the present invention, wherein the vial of the lyophilized pharmaceutical composition containing sovastatin comprises: approximately 30 μg of sovastatin; approximately 50 mg of trisodium citrate dihydrate; and approximately 160 mg of mannitol.

[0078] The kit or co-pack includes a plurality of vials containing the lyophilized pharmaceutical composition of sovastatin and a plurality of ampoules / vials of 0.9% sodium chloride aqueous solution or water for injection.

[0079] In addition, the kit or co-package comprises 3 vials of the lyophilized pharmaceutical composition of sovastatin and 3 ampoules / vials of 0.9% sodium chloride aqueous solution or water for injection.

[0080] The present invention relates to a lyophilized sovastatin-based injectable formulation and a method for preparing the same, which are useful in treating various neurological disorders and diseases without causing any side effects to the human body due to the use of a relatively small amount of sovastatin in the formulation.

[0081] According to one embodiment of the present invention, a lyophilized sovastatin-based injectable formulation comprises: i) an active pharmaceutical ingredient in the range of 0.01-0.02 wt %, ii) at least two soluble excipients in the range of 20-80 wt %, and iii) water for injection in the range of 1-2 wt %.

[0082] In one aspect, the present invention relates to a method for preparing a lyophilized pharmaceutical composition of solvatide, comprising: i) dissolving solvatide, trisodium citrate dihydrate, and mannitol in water for injection; ii) filtering the solution through a 0.2μ membrane filter; iii) filling each vial to a target fill volume; and iv) lyophilizing the filled vials. In one embodiment, the solution in step i) is stirred at 300-350 rpm. In another embodiment, the pH of the solution in step i) is adjusted to a pH of approximately 7.5 to 8.5.

[0083] The method for preparing a lyophilized solvatide-based injectable formulation comprises the following steps: i) dissolving trisodium citrate dihydrate in water for injection to obtain a mixture, ii) adding the active pharmaceutical ingredient, solvatide, to the mixture, followed by adding mannitol to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, followed by sterile filtering the solution and aseptically filling the filtered solution into vials, and iv) half-stoppering the vials and then loading the vials into a freeze dryer for lyophilization to obtain a formulation (e.g., Figure 13 (depicted in ).

[0084] The process is carried out at about 20°C to 30°C.

[0085] In this method, the pH of the solution is adjusted to a pH of about 7.5 to 8.5.

[0086] In one embodiment, the present invention comprises sovastatin as an active ingredient, including a liquid injection with a solvent, which contains at least one soluble excipient, wherein the soluble excipient includes mannitol, trisodium citrate, anhydrous citric acid, disodium hydrogen citrate, disodium hydrogen phosphate, sodium chloride, hydroxypropyl cyclodextrin, wherein the soluble excipient has multiple purposes of serving as an osmotic pressure regulator, a pH regulator, and a solubility enhancer.

[0087] In another embodiment, the osmotic pressure regulator (buffer), pH regulator and solubility enhancer are selected from sodium chloride, trisodium citrate, anhydrous citric acid, disodium hydrogen citrate, disodium hydrogen phosphate, hydroxypropyl β-cyclodextrin and mannitol and mixtures thereof.

[0088] In another embodiment, the freeze-dried filler is selected from the group consisting of sodium chloride, trisodium citrate, anhydrous citric acid, disodium hydrogen citrate, disodium hydrogen phosphate, hydroxypropyl β-cyclodextrin, mannitol, and mixtures thereof.

[0089] The subject matter of the present disclosure is further illustrated by the following specific but non-limiting examples.

[0090] The pharmaceutical formulation for treating stroke comprises 0.0003 mg / kg of sovastatin and 0.75 to 1.25 g / kg of mannitol or 0.0003 mg / kg of sovastatin and 15 to 30 mg / kg of citicoline.

[0091] For the proposed administration of the formulation comprising mannitol and sovastatin, the mannitol infusion is followed by the sovastatin. Similarly, for the proposed administration of the formulation comprising citicoline and sovastatin, the citicoline infusion is followed by the sovastatin.

[0092] According to another embodiment of the present invention, sovastatin is administered before, after, or simultaneously with mannitol or citicoline.

[0093] According to another embodiment of the present invention, an example of treatment with mannitol and solvatide is as follows: treatment is started when a patient with neurological impairment is admitted to the hospital, and a mannitol solution is administered intravenously. The dosage can range from 0.1 g / kg to 5.0 g / kg per day. The average dose of mannitol during a 24-hour administration period is approximately 0.75 to 1.25 g / kg, and the duration of mannitol treatment can last for approximately 3 to 10 days, with most patients receiving mannitol for approximately 6 days. Another method of administering mannitol is to infuse 100 ml of 20% mannitol every four hours for five to six days. Solvatide is administered as an intravenous push injection at a dose of 0.3 μg / kg over a period of multiple intervals over one minute.

[0094] According to another embodiment of the present invention, an example of treatment with citicoline and solvatide is as follows: treatment is initiated within 24 hours of the onset of neurological deficit symptoms. Citicoline is administered intravenously at a dose ranging from 1.5 mg / kg to 75 mg / kg every 12 hours for the first three to six days, followed by oral administration at a dose of 1000 mg every 12 hours for six weeks. Solvatide is administered at a dose of 0.3 μg / kg as an intravenous bolus over one minute at intervals.

[0095] A prospective, multicenter, randomized, phase III study was conducted in patients aged 18 to 78 years with acute ischemic stroke. Patients with radiographically confirmed ischemic stroke were eligible for enrollment if they presented up to 24 hours after symptom onset and had a modified Rankin Scale (mRS) score of 3 to 4 or a NIHSS (National Institutes of Health Stroke Scale) score greater than 5. Patients with intracranial hemorrhage and those receiving endovascular treatment were excluded.

[0096] Patients were randomly assigned in a 1:1 ratio, with 80 patients assigned to the sorafenib group and 78 patients assigned to the control (saline) group. Each patient received standard of care (SOC) and was followed for 90 days. On days 1, 3, and 6, study drug (normal saline or sorafenib (0.3 μg / kg)) was administered as a 1-minute intravenous bolus, divided into three doses at 3 ± 1 hour intervals (total sorafenib dose of 0.9 μg / kg / day).

[0097] The primary objective was to determine neurological outcomes based on the modified Rankin Scale (mRS), National Institutes of Health Stroke Scale (NIHSS), and Barthel Index (BI) scores from day 1 to day 90. In addition, quality of life was measured using the EuroQol-EQ-5D (EQ-5D) and stroke-specific quality of life (SSQoL) scores at 60 and 90 days of treatment.

[0098] A total of 158 patients with acute ischemic stroke were enrolled in this trial, of whom 137 completed 90-day follow-up. Patients received either saline (n = 70; 45 men and 25 women) or sovastatin (n = 67; 45 men and 22 women) within 24 hours of stroke onset. Patients in the control and sovastatin cohorts received the trial drug (mannitol or citicoline) 16.85 ± 0.74 and 17.40 ± 0.67 hours after stroke onset, respectively (mean ± SEM, p = 0.583). Baseline characteristics and SOC were similar between the two cohorts. Mean Alberta Stroke Program Early Computed Tomography Score (ASPECTS) scores were similar in the control (7.44) and sovastatin (7.61) groups, indicating similar infarct size in the two groups.

[0099] At day 90 after randomization, the modified Rankin Scale was measured in 31 patients in the control group and 30 patients in the sorafenib group in patients with ischemic stroke treated with mannitol (N = 61). Figure 1 and Table 1 depict graphical and tabular representations of the modified Rankin Scale for patients treated with control and sovastatin together with mannitol, respectively. It was found that the modified Rankin Scale was significantly reduced in patients treated with mannitol and sovastatin compared to those patients who received mannitol.

[0100] Table 1: Modified Rankin Scale in patients treated with control and solvatide with mannitol

[0101]

[0102] refer to Figure 2and Table 2 depict graphical and tabular representations, respectively, of the number of patients with ischemic stroke who were treated with control and sovastatin together with mannitol and who had a modified Rankin scale of 0-2 at 90 days after randomization; significantly more patients who were treated with mannitol and the endothelin-B agonist sovastatin had a modified Rankin scale of 0-2 at 90 days after randomization than did patients who were treated with mannitol.

[0103] Table 2: Number of patients with ischemic stroke treated with control and sovastatin with mannitol and with a modified Rankin scale of 0-2 at 90 days after randomization

[0104]

[0105] refer to Figure 3 Table 3 and Table 3 depict graphical representations and tabular representations, respectively, of the number of patients with ischemic stroke who were treated with a control and sovastatin plus mannitol and who had an improvement of 2 or more on the modified Rankin scale compared to baseline at 90 days after randomization. The number of patients with ischemic stroke who had an improvement of 2 or more on the modified Rankin scale compared to baseline at 90 days after randomization was significantly greater in the group treated with mannitol and the endothelin-B agonist sovastatin than in the group treated with mannitol. Thus, sovastatin improved neurological outcomes compared to mannitol.

[0106] Table 3: Number of patients with ischemic stroke treated with control and sovastatin plus mannitol who had an improvement of 2 or more from baseline on the modified Rankin Scale at 90 days after randomization

[0107]

[0108] The modified Rankin Scale was measured in 23 patients in the control group and 24 patients in the sorafenib group at day 90 after randomization in patients with ischemic stroke treated with citicoline (N = 47). Figure 4 and Table 4, depict graphical and tabular representations of the modified Rankin Scale for patients treated with control and sovastatin together with citicoline, respectively. The modified Rankin Scale was found to be significantly lower in patients treated with citicoline and the endothelin-B agonist sovastatin compared to those patients receiving citicoline.

[0109] Table 4: Modified Rankin Scale for Patients Treated with Control and Sovastatin with Citicoline

[0110]

[0111]

[0112] refer to Figure 5 and Table 5, depict graphical and tabular representations, respectively, of the number of patients with ischemic stroke who were treated with control and sovastatin together with citicoline and had a modified Rankin scale of 0-2 at 90 days after randomization; significantly more patients with ischemic stroke who were treated with citicoline and the endothelin-B agonist sovastatin had a modified Rankin scale of 0-2 at 90 days after randomization than did patients treated with citicoline.

[0113] Table 5: Number of patients with ischemic stroke treated with control and sovastatin with citicoline and with a modified Rankin scale of 0-2 at 90 days after randomization

[0114]

[0115] refer to Figure 6 Table 6 and Table 6 depict graphical representations and tabular representations, respectively, of the number of patients with ischemic stroke who were treated with a control and with sovastatin in combination with citicoline and who had an improvement of 2 or more on the modified Rankin scale compared to baseline at 90 days after randomization; the number of patients with ischemic stroke who had an improvement of 2 or more on the modified Rankin scale compared to baseline at 90 days after randomization was significantly greater in the group treated with citicoline and the endothelin-B agonist sovastatin than in the group treated with citicoline. Thus, sovastatin improved neurological outcomes compared to citicoline.

[0116] Table 6: Number of patients with ischemic stroke treated with control and sovastatin with citicoline who had an improvement of 2 or more from baseline on the modified Rankin scale at 90 days after randomization

[0117]

[0118]

[0119] According to another embodiment of the present invention, a formulation comprising sovastatin and cerebrolysin was also prepared. On day 90 after randomization, the modified Rankin Scale was measured in 14 patients in the control group and 12 patients in the sovastatin group in patients with cerebral ischemic stroke treated with cerebrolysin (N=26).

[0120] refer to Figure 7and Table 7, depict graphical and tabular representations of the modified Rankin Scale for patients treated with control and sovastatin together with cerebrolysin, respectively. Patients treated with cerebrolysin and the endothelin-B agonist sovastatin were found to have similar neurological outcomes compared to those receiving cerebrolysin.

[0121] Table 7: Modified Rankin Scale for patients treated with control and sovastatin together with cerebrolysin

[0122]

[0123] refer to Figure 8 and Table 8 , depict graphical and tabular representations, respectively, of the number of patients with ischemic stroke who were treated with control and with sovastatin together with cerebrolysin and had a modified Rankin scale of 0-2 at 90 days after randomization; the number of patients with ischemic stroke who were treated with cerebrolysin and the endothelin-B agonist sovastatin at 90 days after randomization was also very similar compared to patients treated with cerebrolysin.

[0124] Table 8: Number of patients with ischemic stroke treated with control and sovastatin together with cerebrolysin and with a modified Rankin scale of 0-2 at 90 days after randomization

[0125]

[0126] refer to Figure 9 Table 9 depicts a graphical representation and a tabular representation, respectively, of the number of patients with ischemic stroke who were treated with a control and with sovastatin plus cerebrolysin and who had an improvement of 2 or more on the modified Rankin Scale compared to baseline at 90 days after randomization. The number of patients with ischemic stroke who had an improvement of 2 or more on the modified Rankin Scale compared to baseline at 90 days after randomization was similar in the group treated with cerebrolysin and the endothelin-B agonist sovastatin compared to the group treated with cerebrolysin. Therefore, sovastatin did not improve neurological outcomes compared to cerebrolysin.

[0127] Table 9: Number of patients with ischemic stroke treated with control and sovastatin plus cerebrolysin who had an improvement of 2 or more from baseline on the modified Rankin Scale at 90 days after randomization

[0128]

[0129] According to another embodiment of the present invention, a formulation comprising sovastatin and a thrombolytic agent was also prepared. On day 90 after randomization, the modified Rankin scale of the control group patients and sovastatin-treated cerebral ischemic stroke patients (N=29) treated with thrombolytic agents was measured in 20 patients in the control group and 9 patients in the sovastatin group.

[0130] refer to Figure 10 and Table 10, depict graphical and tabular representations of the modified Rankin Scale for patients treated with control and sovastatin together with thrombolytics, respectively. Patients treated with thrombolytics and the endothelin-B agonist sovastatin were found to have similar neurological outcomes compared to those who received thrombolytics.

[0131] Table 10: Modified Rankin Scale for patients treated with control and sovastatin with thrombolytics

[0132]

[0133]

[0134] refer to Figure 11 and Table 11 depict graphical and tabular representations, respectively, of the number of patients with ischemic stroke who were treated with control and sovastatin together with thrombolytics and had a modified Rankin scale of 0-2 at 90 days after randomization; the number of patients with ischemic stroke who were treated with thrombolytics and the endothelin-B agonist sovastatin at 90 days after randomization was also very similar compared to patients treated with thrombolytics.

[0135] Table 11: Number of patients with ischemic stroke treated with control and sovastatin with thrombolytics and with a modified Rankin scale of 0-2 at 90 days after randomization

[0136]

[0137] refer to Figure 12 Table 12 depicts a graphical representation and a tabular representation, respectively, of the number of patients with ischemic stroke who were treated with a control and with sovastatin plus thrombolytics and who had an improvement of 2 or more on the modified Rankin Scale compared to baseline at 90 days after randomization. The number of patients with ischemic stroke who had an improvement of 2 or more on the modified Rankin Scale compared to baseline at 90 days after randomization was similar in the group treated with thrombolytics and the endothelin-B agonist sovastatin compared to the group treated with thrombolytics. Therefore, sovastatin did not improve neurological outcomes compared to thrombolytics.

[0138] Table 12: Number of patients with ischemic stroke treated with control and sovastatin plus thrombolytics who had an improvement of 2 or more from baseline on the modified Rankin Scale at 90 days after randomization

[0139]

[0140] Therefore, the combination of sovastatin and mannitol or sovastatin and citicoline is effective in treating neurological disorders because sovastatin can enhance the efficacy of mannitol and citicoline, thereby improving neurological prognosis and can help treat stroke, intracranial hemorrhage, brain injury and other neurological disorders that cause brain edema.

[0141] More Examples

[0142] Example 1

[0143] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) dissolving 4.800 gm of mannitol in 75 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture, and then adding 1.500 gm of trisodium citrate dihydrate to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the stopper on the vial and then loading the vial into a freeze dryer for lyophilization to obtain a lyophilized sovastatin-based injectable formulation. Table 1 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0144] Table 1: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 1)

[0145] Element unit Amount used Sovastatin MG 0.990 Mannitol GM 4.800 Trisodium citrate GM 1.500 Water for injection ML QS to 75.0

[0146] Example 2

[0147] Method for preparing the formulation: i) adding 0.3 gm of mannitol to 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of solvatide to the mixture, and then adding 1% trisodium citrate dihydrate to obtain a solution, iii) checking the pH of the solution and adjusting it to 7.0 to obtain the volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the stopper on the vial, and then loading the vial into a freeze dryer for lyophilization to obtain the formulation. Table 2 shows the composition and ingredients used to prepare the lyophilized solvatide-based injectable formulation.

[0148] Table 2: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 2)

[0149]

[0150]

[0151] Example 3

[0152] A method for preparing a lyophilized sovastatin-based injectable formulation is described, comprising the steps of: i) adding 0.3 gm of mannitol to 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture, followed by 35 mg of 1% trisodium citrate dihydrate to obtain a solution, iii) checking the pH of the solution and adjusting it to pH 5.10 using a 1% citric acid solution to obtain a volume of the solution, followed by filtering the solution and filling the filtered solution into vials, and iv) half-stoppering the vials and then loading the vials into a freeze dryer for lyophilization to obtain a formulation. Table 3 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0153] Table 3: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 3)

[0154] Element unit Amount used Sovastatin MG 0.990 Trisodium citrate MG 35 Mannitol GM 0.3 Anhydrous citric acid GM 1% solution for pH adjustment Water for injection ML QS to 30.0

[0155] Example 4

[0156] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) adding 0.3 gm of mannitol to 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture and dissolving it with 0.1% sodium hydrogen phosphate solution to obtain a solution, iii) checking the pH of the solution and adjusting it to pH 7.51 to obtain a volume of the solution, followed by filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 4 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0157] Table 4: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 4)

[0158] Element unit Amount used Sovastatin MG 0.990 Disodium hydrogen citrate GM 1% solution for pH adjustment Mannitol GM 0.3 Water for injection ML QS to 30.0

[0159] Example 5

[0160] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) adding 0.3 gm of mannitol to 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture and dissolving it with 9.0 mg of sodium hydrogen phosphate solution to obtain a solution, iii) checking the pH of the solution and adjusting it to pH 6.47 using a 1% citric acid solution to obtain a volume of the solution, followed by filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 5 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0161] Table 5: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 5)

[0162] Element unit Amount used Sovastatin MG 0.990 Disodium hydrogen phosphate MG 9.0 Mannitol GM 0.3 Anhydrous citric acid GM 0.1% solution for pH adjustment Water for injection ML QS to 30.0

[0163] Example 6

[0164] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) adding 0.6 gm of mannitol and 60.0 mg of sodium chloride to 30 ml of water for injection to obtain a mixture, ii) adding 1.980 mg of sovastatin to the mixture and dissolving it with 25.0 mg of sodium sulfate to obtain a solution, iii) checking the pH of the solution and adjusting it to pH 7.32 using a 1% citric acid solution to obtain a volume of the solution, followed by filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 6 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0165] Table 6: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 6)

[0166] Element unit Amount used Sovastatin MG 1.980 Sodium chloride MG 60.0 Mannitol GM 0.6 sodium sulfate MG 25.0 Disodium hydrogen phosphate GM 1% solution for pH adjustment Water for injection ML QS to 30.0

[0167] Example 7

[0168] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) adding 0.150 gm of mannitol and 0.150 gm of hydroxypropyl β-cyclodextrin to 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture and dissolving it with 1% trisodium citrate to obtain a solution, iii) checking the pH of the solution and adjusting it to 7.10 with a 1% trisodium citrate dihydrate solution to obtain a volume of the solution, followed by filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 7 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0169] Table 7: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 7)

[0170] Element unit Amount used Sovastatin MG 0.990 Hydroxypropyl β-cyclodextrin GM 0.150 Mannitol GM 0.150 Trisodium citrate GM 1% solution for pH adjustment Water for injection ML QS to 30.0

[0171] Example 8

[0172] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) adding 0.150 gm of mannitol and 0.150 gm of hydroxypropyl β-cyclodextrin to 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture and dissolving it with 30 mg of trisodium citrate to obtain a solution, iii) checking the pH of the solution and adjusting it to pH 5.51 using a 1% citric acid solution to obtain a volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 8 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0173] Table 8: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 8)

[0174] Element unit Amount used Sovastatin MG 0.990 Hydroxypropyl β-cyclodextrin GM 0.150 Mannitol GM 0.150 Anhydrous citric acid GM 1% solution for pH adjustment Trisodium citrate MG 30.0 Water for injection ML QS to 30.0

[0175] Example 9

[0176] A method for preparing a lyophilized sovastatin-based injectable formulation is described, comprising the following steps: i) adding 1.500 gm of mannitol to 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture and dissolving it with a 1% solution of trisodium citrate dihydrate to obtain a solution, iii) checking the pH of the solution and adjusting it to pH 7.25 using a 1% solution of trisodium citrate dihydrate to obtain a volume of the solution, followed by filtering the solution and filling the filtered solution into vials, and iv) half-stoppering the vials and then loading the vials into a freeze dryer for lyophilization to obtain a formulation. Table 9 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0177] Table 9: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 9)

[0178] Element unit Amount used Sovastatin MG 0.990 Mannitol GM 1.500 Trisodium citrate GM 1% solution for pH adjustment Water for injection ML QS to 30.0

[0179] Example 10

[0180] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) dissolving 0.900 gm of sodium chloride in 30 ml of water for injection to obtain a mixture, ii) adding 0.900 mg of sovastatin to the mixture and dissolving it with a 1% solution of trisodium citrate dihydrate, followed by adding 0.300 mg of Kollid PF-12 and stirring to dissolve it to obtain a solution, iii) checking the pH of the solution and adjusting it to pH 6.80 with a 1% solution of trisodium citrate dihydrate to obtain a volume of the solution, followed by filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 10 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0181] Table 10: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 10)

[0182] Element unit Amount used Sovastatin MG 0.990 Sodium chloride GM 0.900 Kollidon PF-12 GM 0.300 Trisodium citrate GM 1% solution for pH adjustment Water for injection ML QS to 30.0

[0183] Example 11

[0184] A method for preparing a lyophilized sovaccin-based injectable formulation is described, comprising the steps of: i) dissolving 1.500 gm of mannitol in 30 ml of water for injection to obtain a mixture, ii) adding 0.300 gm of trisodium citrate dihydrate to the mixture and stirring to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, followed by filtering the solution and filling the filtered solution into vials, and iv) half-stoppering the vials and then loading the vials into a freeze dryer for lyophilization to obtain the formulation. Table 11 shows the composition and ingredients used to prepare the lyophilized sovaccin-based injectable formulation.

[0185] Table 11: Compositions and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 11)

[0186] Element unit Amount used Sovastatin - - Mannitol GM 1.500 Trisodium citrate GM 0.300 Water for injection ML QS to 30.0

[0187] Example 12

[0188] The method for preparing a lyophilized sovaccin-based injectable formulation includes the following steps: i) dissolving 1.500 gm of mannitol in 30 ml of water for injection to obtain a mixture, ii) adding 0.600 gm of trisodium citrate dihydrate to the mixture and stirring to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the stopper on the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 12 shows the composition and ingredients used to prepare the lyophilized sovaccin-based injectable formulation.

[0189] Table 12: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 12)

[0190] Element unit Amount used Sovastatin - - Mannitol GM 1.500 Trisodium citrate GM 0.600 Water for injection ML QS to 30.0

[0191] Example 13

[0192] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) dissolving 1.500 gm of mannitol in 30 ml of water for injection to obtain a mixture, ii) adding 0.990 mg of sovastatin to the mixture and dissolving it with 0.900 gm of trisodium citrate dihydrate to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the stopper on the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 13 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0193] Table 13: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 13)

[0194]

[0195]

[0196] Example 14

[0197] The method for preparing a lyophilized sovaccin-based injectable formulation includes the following steps: i) dissolving 7.500 gm of mannitol in 100 ml of water for injection to obtain a mixture, ii) adding 2.500 gm of trisodium citrate dihydrate to the mixture to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the stopper on the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 14 shows the composition and ingredients used to prepare the lyophilized sovaccin-based injectable formulation.

[0198] Table 14: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 14)

[0199] Element unit Amount used Sovastatin - - Mannitol GM 7.500 Trisodium citrate GM 2.500 Water for injection ML QS to 100.0

[0200] Example 15

[0201] The method for preparing a lyophilized sovaccin-based injectable formulation includes the following steps: i) dissolving 7.500 gm of mannitol in 100 ml of water for injection to obtain a mixture, ii) adding 2.500 gm of trisodium citrate dihydrate to the mixture and stirring to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the stopper on the vial and then loading the vial into a freeze dryer for lyophilization to obtain a formulation. Table 15 shows the composition and ingredients used to prepare the lyophilized sovaccin-based injectable formulation.

[0202] Table 15: Compositions and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 15)

[0203] Element unit Amount used Sovastatin - - Mannitol GM 7.500 Trisodium citrate GM 2.500 Water for injection ML QS to 100.0

[0204] Example 16

[0205] The method for preparing a lyophilized sovastatin-based injectable formulation includes the following steps: i) dissolving 1.500 gm of trisodium citrate dihydrate in 75 ml of water for injection to obtain a mixture, ii) adding 0.900 mg of sovastatin to the mixture, and then adding 4.800 gm of mannitol to obtain a solution, iii) checking the pH of the solution to obtain the volume of the solution, then filtering the solution and filling the filtered solution into a vial, and iv) half-stoppering the stopper on the vial and then loading the vial into a freeze dryer for lyophilization to obtain a lyophilized sovastatin-based injectable formulation. Table 16 shows the composition and ingredients used to prepare the lyophilized sovastatin-based injectable formulation.

[0206] Table 16: Composition and ingredients for preparing lyophilized sovastatin-based injectable formulations (Example 16)

[0207]

[0208] Results of different tests:

[0209] Referring to Table 16, a tabular representation of the results of various experiments conducted to study certain parameters is depicted.

[0210] Table 16: Test results

[0211]

[0212] The results of Example 1 were analyzed according to the description, clarity, osmolality and measured internal specifications and were found to meet these standards after one month of storage at a temperature of 25° C. Due to these findings, a pilot batch was produced for further testing and investigation.

[0213] refer to Figure 1 , depicting a flow chart of the process of developing an injectable formulation involving several steps. In the flow chart, the production process of 30 μg of sovastatin injection is explained.

[0214] In the process specification:

[0215] Referring to Table 17, a tabular representation of an overview of the specific parameters and requirements that must be met during formulation manufacturing is depicted. During the manufacturing process, various parameters are closely monitored to ensure that the final product meets the required standards. In particular, the parameters are product description, solution pH, assay, sterility, bacterial endotoxin testing, and bioburden. Following analysis, it is determined that the prognosis or results obtained are within the predefined or specified ranges or limits.

[0216] Table 17: Overview of specific parameters and requirements that must be met during formulation manufacture

[0217]

[0218] Determination of freeze drying cycle:

[0219] The vials filled with the filtered solution are then lyophilized by placing them into a freeze dryer. This process involves three main stages: i) freezing, ii) primary drying, and iii) secondary drying. i) The primary drying stage is particularly critical and involves sublimating ice from the frozen product. In this case, the freeze drying cycle for lyophilizing the formulation is selected as follows. First, the freezing stage is set to -45°C. This extremely low temperature ensures that the substance freezes quickly and efficiently. This step is crucial to prevent the formation of large ice crystals that could damage the product's structure and composition.

[0220] After the freezing stage, ii) the primary drying stage begins. The temperature is set to -20°C for a specified time. During this time, ice crystals in the substance begin to sublime, changing from a solid to a gaseous state. This process removes most of the water from the product. iii) The next stage, primary drying, is set to 0°C for a specified time. This slower process ensures that all remaining water is removed from the product. Finally, in the final stage of primary drying, the temperature is set to +10°C for a specified time. At this temperature, any remaining ice in the product sublimates, and the drying process is complete. The final product is a freeze-dried formulation that can be stored long-term at 2°C-8°C or 22°C-28°C.

[0221] The use of freeze drying yields a number of benefits which can be summarized as follows:

[0222] a) The present invention relates to an injectable pharmaceutical product with good physical appearance and rapid re-dissolution properties. The incorporation of mannitol in appropriate proportions into the formulation enables the development of simple and uncomplicated formulations with excellent flexibility for producing injectable solvatide. The addition of mannitol facilitates freezing of the solution, and the resulting dried, frozen product exhibits enhanced solubility and clarity upon reconstitution. Furthermore, impurity levels are low, and product stability can be effectively monitored, ensuring well-regulated quality.

[0223] b) The product exhibits long-term stability and excellent quality. One aspect of this embodiment relates to a lyophilized formulation of sovastatin. In addition, the lyophilized formulation is packaged in vials or another container suitable for pharmaceutical use.

[0224] Therefore, it can be concluded that the product derived from the freeze-drying process has a significantly low moisture content, with a limit of less than 7.0%, and can be stored at low temperatures or room temperature. The product exhibits no impurity content and retains its original color as a freeze-dried injectable powder / cake. Upon redissolution, the resulting solution maintains its clarity and does not exhibit significant changes. This generally demonstrates a high level of product stability. Bacterial endotoxin test results meet regulatory requirements and were performed using the gel-gel technique Limulus Amebocyte Lysate (LAL) reagent.

[0225] Finished product specifications:

[0226] Refer to Table 18, which summarizes the various parameters established to ensure the quality and consistency of the finished product. Lyophilized solvastatin injection was analyzed for specific parameters, including its description, identification, reconstitution time, and appearance after reconstitution. The analytical results showed that all of these parameters met the required specifications, indicating that the product passed the necessary quality standards. The reconstituted solution was found to be within the specified pH range of 7.0-8.5, with a pH of 8.32. The moisture content, determined using the Karl Fischer method, was 2.61% by weight, well within the permitted limit of no more than 7%. Bacterial endotoxins were found to meet regulatory requirements, and the particle content of particles ≥10 μm and ≥25 μm was 81% and 0.2% per container, respectively, both within limits. No specific impurities were detected, and unidentified impurities and total impurities were 0.39% and 0.87%, respectively. An assay value of 95.65% and an osmolality of 275 mOsm / L were observed. These results from the batch analysis confirm the successful development of the manufacturing process. Lyophilized sovastatin injection 30 μg should be stored between 2 and 8°C, protected from light.

[0227] Table 18: Summarizes the various parameters established to ensure the quality and consistency of the finished product (i.e. disclosed in Example 1)

[0228]

[0229]

[0230] Packaging Configuration:

[0231] The formulation in the vial is further packaged. Its configuration is mentioned below:

[0232] ● Glass Vial - 5 mL, USP Type 1, amber, tubular glass vial.

[0233] ●Rubber Stopper-13mm Slotted Grey Bromobutyl

[0234] butyl) "RFU" rubber stopper.

[0235] ●Aluminum Seal - 13mm aluminum flip-top seal.

[0236] See Tables 19 and 20 for a tabular representation of the container and closure system specifications and results, respectively.

[0237] Table 19: Specifications for 13mm aluminum flip-top tear-off seals

[0238] Sr.No. describe Specifications (mm) 1 Inner diameter of aluminum seal 13.00-13.60 2 Outer diameter of aluminum seal 13.50-14.10 3 Diameter of plastic disc 14.50-15.00 4 Height of plastic tray 2.90m-3.30 5 Internal height of aluminum seal 6.10-6.40 6 Assembly height 7.62-8.38 7 Thickness of aluminum cover 0 18-0 20 8 Number of bridge pieces 06 9 Approved drawing number FO / VS / DE-13

[0239] As shown in Table 19, very precise measurements were taken using the packaging assembly aluminum flip-top tear-off seal (13 mm). The seal was designed to fit securely on containers with an inner diameter of 13.32 to 13.40 mm, while its outer diameter ranged from 13.78 to 13.90 mm. The diameter of the plastic disc (which is an integral part of the seal and is used to cover the top of the vial) was 14.75 to 14.83 mm, and its height was 3.02 to 3.10 mm. The internal height of the aluminum seal (i.e., the distance between the bottom of the seal and the top of the plastic disc) ranged from 6.25 mm to 6.32 mm. The assembled height of the seal (i.e., the distance between the bottom of the seal and the top of the plastic disc) ranged from 7.81 mm to 8.02 mm. The thickness of the aluminum lid was 0.18 mm, which ensured a secure seal. In addition, the seal conformed to the approved drawings, which ensured that it met all necessary requirements.

[0240] Table 20: Specifications of 13mm grey bromobutyl “RFU” rubber stoppers

[0241]

[0242]

[0243] As shown in Table 20, the 13 mm slotted gray bromobutyl "RFU" rubber stopper met the required physical parameters. The disc diameter was 12.68 mm, while the collar or flange thickness was 2.21 mm, with a maximum height of 10.47 mm. The rubber stopper had a diameter of 7.87 mm and exhibited optimal fit to the vial. The sample's infrared spectrum was consistent with that of the reference standard, enabling accurate identification of the stopper. The appearance of solution A was consistent with acceptable opalescence and color standards, with the solution's opalescence no more opalescent than that of the opalescence standard, OS3, and no darker than that of the reference solution, BYS6. The stopper exhibited no significant acidity or alkalinity (0.19 mL) and was within the specified range, while light absorption (0.0028) and reducing substances (1.9 mL) were within acceptable limits. The heavy metal concentration was less than 20 ppm, and the residue upon evaporation was minimal, at 2.1 mg, demonstrating compliance with the required parameters. The rubber stopper passed sterilization testing, demonstrating that it did not soften or become sticky, and there were no visual changes in the closure. Furthermore, the liquid particle test results showed 17 particles ≥10 μm and 1 particle >25 μm, meeting the established specifications. Sterility testing indicated no growth was observed, and endotoxin testing revealed an endotoxin concentration of less than 0.25 EU / ml, demonstrating compliance with the required quality standards. Therefore, the 13 mm slotted gray bromobutyl "RFU" rubber stopper successfully met the necessary physical and chemical specifications, demonstrating its suitability for use.

[0244] Referring to Table 21, a tabular representation of the specifications of the amber vials is depicted. The results presented show that a detailed study of the physical parameters of the 5 ml amber vials was conducted. The height of the vials was measured and found to range from 48.00 mm to 48.73 mm, with an average height of 48.36 mm. Additionally, the rim height of the vials was observed to vary between 3.60 mm and 3.91 mm, with an average height of 3.75 mm. The diameter of the vial body was found to range from 16.16 mm to 16.37 mm, with an average diameter of 16.27 mm. Furthermore, the outer diameter of the mouth of the vials was determined to be 12.94 mm to 13.02 mm, with an average diameter of 12.98 mm. Furthermore, the inner diameter of the mouth of the vials was found to vary between 6.93 mm and 7.18 mm, with an average diameter of 7.05 mm. Furthermore, both the T1 body wall thickness and the T2 bottom wall thickness of the vials were determined to meet the specified standards. Detailed data collected from the analysis ultimately demonstrated that the 5ml amber vial was well within the specified parameters and met the required standards. The vial underwent rigorous visual testing and was found to meet industry standards. The vial was constructed from borosilicate amber USP-type glass that met structural requirements. Furthermore, the product was free of dust, dirt, or any foreign particles that could affect its quality or functionality. Beyond that, the vial was evaluated for chips or bubbles that could compromise its integrity. Based on the results, it met the established standards, indicating that it was free of any visible defects (Table 21). Furthermore, the vial was subjected to various tests, including a glass particle test, a surface glass test, and an arsenic test, with the results indicating compliance with the standards (Table 21).

[0245] Table 21: Specifications of amber vials

[0246]

[0247]

[0248] Stability studies:

[0249] The stability of 30 μg of sovastatin for injection was studied in USPI type glass vials stored at 25°C ± 2°C and 60% RH ± 5% RH for no less than 12 months, with each vial containing 30 μg of sovastatin. The pH of the initial lyophilized injection was 7.91, and after 12 months the pH was 7.67, which was within the specified limits. The reconstitution time, clarity of the reconstituted solution, moisture content, sterility, bacterial endotoxin limit, particulate matter, and related substances (%) all met regulatory requirements and were within the specified limits. Initially, the assay percentage of the lyophilized sovastatin injection was 100.41%, and after storage at 25°C ± 2°C and 60% RH ± 5% RH for no less than 12 months, the assay value was 98.81%. The results confirmed that the formulation remained stable for no less than 12 months at a storage temperature of 25°C ± 2°C.

[0250] Similarly, a developed lyophilized injectable formulation of sovastatin stored in the temperature range of 2°C-8°C was stable for storage for not less than 36 months, and the results met the specified reconstitution time, clarity of the reconstituted solution, moisture content, sterility, bacterial endotoxin limit, particulate matter, and related substances (%). Initially, the pH of the lyophilized injection stored at 5°C ± 3°C was 8.18, and after 36 months, the pH was 7.77. Initially, the assay percentage of the lyophilized sovastatin injection stored at 5±3°C was 106.00%, and after storage of the formulation for not less than 36 months, the assay value was 95.58%. The results indicate that the formulation remains stable for not less than 36 months at a storage temperature of 5°C ± 3°C.

[0251] Forced degradation studies:

[0252] Referring to Table 22, a tabular representation of forced degradation studies of Sovastatin Injection is depicted. Different conditions were set for forced degradation (chemical degradation), including acid degradation, alkali degradation; and peroxide degradation, respectively; 1 mL-5N HCl / 3 hours RT; 1 mL-5N NaOH / 3 hours RT; 1 mL-30% H2O2 / 3 hours RT. Different conditions were set for forced degradation (physical degradation), including thermal degradation and humidity degradation, respectively; 3 hours / 105°C; 95% RH / 24 hours. Different forced degradation conditions were set to observe photolytic degradation and samples covered with aluminum foil (dark conditions); final packaging; primary packaging (labeled); primary packaging (unlabeled) and 1.2 million Lux-Hours and 200 Watt-Hour / m 2 (Open conditions). The study results show that 30 μg of lyophilized sovaccinium injection exhibits sensitivity to degradation by acids, bases, peroxides, and photolysis. Chromatographic and peak purity analysis of the degraded samples demonstrated uniform analyte peaks with no co-eluting peaks from the degradation process. Sovaccinium peak purity was maintained in all chromatograms under various stress conditions.

[0253] Table 22: Forced degradation study of Sovaccin Injection

[0254]

[0255]

[0256] Validation of the assay method for Sovastatin Injection:

[0257] Chromatographic parameters for method validation of the assay for Sovastatin Injection: For the experiments, an analytical system containing an Xtimate C18 (250 x 4.6 mm), 5 μm or Chromcore 120 C18 (250 x 4.6 mm), 5 μm or equivalent analytical column was used. The flow rate was maintained at 0.8 mL / min, and the injection volume was 80 μL. The detection wavelength was set to 215 nm, and the total run time was 30 minutes. The autosampler temperature was 5° C., and the column oven temperature was 40° C. The measured retention time was 13 ± 2 minutes.

[0258] Mobile phase for determination:

[0259] Mobile phase-A, (10 mM KH2PO4 & 10 mM NaCl, pH-7.5)

[0260] Mobile phase-B (methanol)

[0261] Referring to Table 23, a tabular representation of the method validation summary for the determination of sovastatin injection is depicted. The method was validated for the determination of sovastatin in 30 μg of sovastatin for injection by HPLC. After data evaluation, the method was found to be precise, linear, accurate, robust, durable, and specific. Since the results of all validation parameters were within the acceptance criteria, it was concluded that the analytical method is suitable for the determination of sovastatin by HPLC. Based on the solution stability study, the sovastatin standard solution was stable at 5°C for up to 24 hours, the sovastatin injection sample solution was stable at 5°C for up to 26 hours, and the sample solution of the in-process bulk solution was stable at 5°C for up to 25 hours (Table 23).

[0262] Table 23: Validation Summary of the Sovastatin Injection Assay

[0263]

[0264]

[0265]

[0266]

[0267]

[0268] Summary of validation of related substance methods:

[0269] Chromatographic parameters for method validation of related substances of sovastatin injection (Method 1): Method-1 utilizes chromatographic conditions to separate and quantify specific compounds of interest. The analytical columns used are Xtimate C18 columns or ChromCore120C18 columns, both with dimensions of 4.6X 250mm and 5m. A Ghost Buster column with a size of 4.6X 50mm is used to detect any unexpected peaks, commonly known as ghost peaks, that may interfere with the results. The flow rate is set at 0.5mL / min and the injection volume is 90μL. The detection wavelength is 215nm and the autosampler temperature is maintained at 5°C. The column oven temperature is set to 50°C and the run time of the method is 120min. Under these chromatographic conditions, the retention time of the compound of interest is approximately 66.0 minutes. These parameters ensure the separation and accurate quantification of the compounds in the sample.

[0270] Mobile phase for unspecified impurities (Method 1):

[0271] Mobile phase A (KH2PO4 + sodium chloride in HPLC water).

[0272] Mobile phase-B (methanol).

[0273] With reference to Table 24, a tabular representation of the method validation summary for related substances in Sovaccinium Injection (Method 1) is depicted. After data evaluation, the method was found to be precise, linear, accurate, robust, durable, and specific. Since the results for all validation parameters were within the acceptance criteria, it was concluded that the analytical method was suitable for measuring organic impurities (unspecified) in lyophilized Sovaccinium by HPLC. Based on solution stability studies, at 5°C, the standard solution was stable for up to 60 hours, and the sample solution was stable for up to 12 hours (Table 24).

[0274] Table 24: Summary of method validation for related substances in Sovaccinium Injection (Method 1)

[0275]

[0276]

[0277]

[0278]

[0279]

[0280] Chromatographic parameters for method validation of related substances of sovastatin injection (method 2): The given information describes the conditions and parameters used in the analytical column for chromatographic analysis. An Ultisil XB-C30 column or a ChromCore C30 column with a size of 4.6X 250mm and a particle size of 5μm was used as the analytical column. A Ghost-Buster column with a size of 4.6X 50mm was used for removal. The flow rate was maintained at 0.5mL / min, and an injection volume of 80μL was used. The detection wavelength was set to UV at 215nm, and the autosampler temperature was maintained at 5°C and the column oven temperature was set at 50°C. The standard run time was 30 minutes, while the sample solution was 80 minutes. The retention time used for analysis was between 6 and 12 minutes, and the relative retention time relative to sovastatin was approximately 0.2. These parameters are critical for obtaining accurate and reproducible results in chromatographic analysis.

[0281] Mobile phase for indicated impurities (Method 2): Mobile phase A (0.05 M 1-octanesulfonic acid sodium salt, pH 7.5);

[0282] Mobile phase-B (methanol).

[0283] With reference to Table 25, a tabular representation of the method validation summary for related substances in solvatide injection (Method 2) is depicted. After data evaluation, the method was found to be precise, linear, accurate, robust, durable, and specific. Since the results for all validation parameters were within the acceptance criteria, it was concluded that the analytical method was suitable for the measurement of organic impurities (designated -D-His-solvatide) in lyophilized solvatide by HPLC. Based on solution stability studies, at 5°C, the standard solution was stable for up to 48 hours, the unspiked sample solution was stable for up to 40 hours, and the control sample solution was stable for up to 39 hours (Table 25).

[0284] Table 25: Summary of validation of related substances in Sovastatin injection (Method 2)

[0285]

[0286]

[0287]

[0288]

[0289]

[0290] Although the field of the present invention has been described herein with reference to specific embodiments, this description is not intended to be interpreted in a limiting sense. Various modifications of the disclosed embodiments and alternative embodiments of the present invention will become apparent to those skilled in the art upon reference to the description of the present invention.

Claims

1. A freeze-dried pharmaceutical composition comprising: a) Sovastatin; b) trisodium citrate; and c) Mannitol.

2. The lyophilized pharmaceutical composition of claim 1, wherein the composition is a lyophilized powder.

3. The lyophilized pharmaceutical composition of claim 1, wherein the sovastatin is present in the range of about 0.01 to about 0.02 weight %.

4. The lyophilized pharmaceutical composition of claim 1, wherein the trisodium citrate dihydrate is present in the range of about 20 to about 80 weight percent.

5. The lyophilized pharmaceutical composition of claim 1, wherein the mannitol is present in the range of about 20 to about 80 weight %.

6. The lyophilized pharmaceutical composition of claim 1, wherein the sovastatin is present at about 30 μg; trisodium citrate is present at about 50 mg; and mannitol is present at about 160 mg.

7. The lyophilized pharmaceutical composition of claim 1, wherein the composition further comprises sodium chloride.

8. The lyophilized pharmaceutical composition of claim 1, wherein the composition further comprises water.

9. A freeze-dried pharmaceutical composition comprising: a) about 30 μg of sovastatin; b) about 50 mg of trisodium citrate dihydrate; and c) about 160 mg of mannitol.

10. The lyophilized pharmaceutical composition according to any one of the preceding claims, wherein the composition comprises: i) not more than 2% of total impurities; or ii) not more than 0.5% of any unspecified impurity; or iii) not more than 1.0% of D-His-Sovacrine impurity.

11. The lyophilized pharmaceutical composition according to any one of claims 10, wherein the composition comprises: i) not more than 0.7% of total impurities; or ii) not more than 0.15% of any unspecified impurity; or iii) not more than 0.22% of D-His-Sovacidin impurity.

12. The lyophilized pharmaceutical composition according to any one of the preceding claims, which is reconstituted with 0.9% sodium chloride solution.

13. The lyophilized pharmaceutical composition according to any one of the preceding claims, which is reconstituted with water for injection.

14. A method of treating a neurological condition or disease comprising administering to a patient in need thereof the lyophilized pharmaceutical composition of any one of the preceding claims.

15. The method of claim 14, wherein the neurological disorder or disease is selected from the group consisting of stroke, Alzheimer's disease, spinal cord injury, cognitive impairment, neurofibromatosis, Huntington's disease, Parkinson's disease, neonatal hypoxic-ischemic encephalopathy, traumatic brain injury, and multi-infarct dementia.

16. The method of claims 14-15, wherein the neurological disorder or disease is stroke.

17. The method of claims 14-16, wherein the lyophilized pharmaceutical composition according to any one of the preceding claims is administered intravenously.

18. A reconstituted liquid composition comprising: a) Sovastatin; b) trisodium citrate; c) mannitol; and d) Water or 0.9% sodium chloride aqueous solution.

19. The liquid composition of claim 18, wherein the sovastatin, trisodium citrate and mannitol are provided as a lyophilized powder.

20. The liquid composition of claim 18, which is prepared by reconstituting lyophilized powders of sovastatin, trisodium citrate and mannitol in water or a 0.9% sodium chloride aqueous solution.

21. The liquid composition of claim 18, wherein the sovastatin is present in the range of about 0.01 to about 0.02 weight %.

22. The liquid composition of claim 18, wherein the trisodium citrate dihydrate is present in the range of about 20 to about 80 weight percent.

23. The liquid composition of claim 18, wherein the mannitol is present in the range of about 20 to about 80 weight percent.

24. The liquid composition of claim 18, wherein solvatide is present at about 30 mg; trisodium citrate is present at about 50 mg; and mannitol is present at about 160 mg.

25. The liquid composition of claims 18-24, wherein the sovastatin is present in the liquid composition at a concentration as described above.

26. The liquid composition of claims 18-25, wherein the composition comprises: i) not more than 2% of total impurities; or ii) not more than 0.5% of any unspecified impurity; or iii) not more than 1.0% of D-His-Sovacrine impurity.

27. The liquid composition of claim 26, wherein the composition comprises: i) not more than 0.7% of total impurities; or ii) not more than 0.15% of any unspecified impurity; or iii) not more than 0.22% of D-His-Sovacidin impurity.

28. The liquid composition of claims 18-27, wherein the reconstituted liquid composition has an osmotic pressure between 240 and 310 mOsm / L.

29. The liquid composition of claim 28, wherein the reconstituted liquid composition has an osmotic pressure of about 286 mOsm / L.

30. The liquid composition of claims 18-29, wherein the reconstituted liquid composition has a pH of about 7.0 to 8.

5.

31. The liquid composition of claims 18-30, which is administered intravenously.

32. A method of treating a neurological condition or disease comprising administering to a patient in need thereof a liquid composition according to any one of the preceding claims 18-31.

33. The method of claim 32, wherein the neurological disorder or disease is selected from the group consisting of stroke, Alzheimer's disease, spinal cord injury, cognitive impairment, neurofibromatosis, Huntington's disease, Parkinson's disease, neonatal hypoxic-ischemic encephalopathy, traumatic brain injury, and multi-infarct dementia.

34. The method of claim 33, wherein the neurological disorder or disease is stroke.

35. A kit or co-package comprising: i) a vial containing a lyophilized pharmaceutical composition of sovastatin; and ii) An ampoule containing 0.9% sodium chloride aqueous solution or water for injection.

36. The kit or co-package of claim 35, wherein the vial containing the lyophilized pharmaceutical composition of sovastatin comprises: a) about 30 μg of sovastatin; b) about 50 mg of trisodium citrate dihydrate; and c) about 160 mg of mannitol.

37. The kit or co-package of claim 35, wherein the kit or co-package comprises a plurality of vials containing the lyophilized pharmaceutical composition of sovastatin and a plurality of ampoules of 0.9% sodium chloride aqueous solution or water for injection.

38. The kit or co-package according to claim 37, wherein the kit or co-package comprises three vials of the lyophilized pharmaceutical composition of sovastatin and three ampoules of 0.9% aqueous sodium chloride solution or water for injection.

39. The kit or co-package of claims 37-38, wherein the vial of lyophilized pharmaceutical composition containing sovastatin comprises: a) about 30 μg of sovastatin; b) about 50 mg of trisodium citrate dihydrate; and c) about 160 mg of mannitol.

40. A method for preparing a lyophilized pharmaceutical composition of sovastatin, comprising: i) dissolving sovastatin, trisodium citrate dihydrate and mannitol in water for injection; ii) filtering the solution through a 0.2μ membrane filter; iii) filling each vial to the target fill volume; and iv) Lyophilize the filled vials.

41. The method of claim 40, wherein the stopper on the vial is half-stoppered, and then the vial is loaded into a freeze dryer for freeze drying to obtain a freeze-dried sovastatin-based injectable formulation.

42. The method of claim 40, wherein the solution in step i) is stirred at 300-350 rpm.

43. The method of claim 40, wherein the pH of the solution in step i) is adjusted to a pH of about 7.5 to 8.

5.

44. A lyophilized sovastatin-based injectable formulation comprising: active pharmaceutical ingredient in the range of 0.01-0.02% by weight; at least two soluble excipients in the range of 20-80 wt. %; and Water for injection in the range of 1-2 wt%.

45. The formulation of claim 1, wherein the active pharmaceutical ingredient is sovastatin.

46. ​​The formulation of claim 1, wherein the soluble excipients include but are not limited to mannitol, trisodium citrate dihydrate, anhydrous citric acid, disodium hydrogen citrate, disodium hydrogen phosphate, sodium chloride, hydroxypropyl beta-cyclodextrin.

47. The formulation of claim 1 or 3, wherein the soluble excipient is desirably mannitol as a lyophilization bulking agent and trisodium citrate dihydrate as a buffering agent.

48. A method for preparing the lyophilized sovastatin-based injectable formulation according to claim 1, comprising the steps of: dissolving the trisodium citrate dihydrate in the water for injection to obtain a mixture; adding the active pharmaceutical ingredient to the mixture, and then adding the mannitol to obtain a solution; checking the pH of the solution to replenish the volume of the solution, followed by sterile filtering the solution and filling the sterile filtered solution into vials; and The stopper on the vial was half-stoppered and the vial was then loaded into a freeze dryer for lyophilization to obtain the formulation.

49. The method of claim 5, wherein the vials filled with the filtered solution are lyophilized by freezing at -45°C, then drying at -20°C, drying at 0°C, and then drying at +10°C.

50. The method of claim 5, wherein the vial is removed from the freeze dryer and sealed to obtain the lyophilized sovastatin-based injectable formulation.

51. The method of claim 5, wherein the formulation is administered intravenously.

52. The method of claim 5, wherein the vials are washed and depyrogenated prior to filling the sterile filtered solution into the vials.

53. The method of claim 5, wherein the lyophilized sovastatin-based injectable formulation is stored at 2-8 degrees Celsius or 22 degrees to 28 degrees Celsius.

Images