High concentration liquid aqueous pyronaridine formulations

Abstract

The present invention relates to a liquid aqueous formulation of roxadustat for medical treatment, said liquid aqueous formulation comprising at least 70 wt.% water and at least 40 pmol / ml dissolved roxadustat, having a pH in the range of 2.5 to 5.8, and characterized in that the ratio of the osmolality of the formulation (in Osm / kg) to the concentration of roxadustat in the formulation (in mmol / mL) is less than 2.0. The present invention also relates to a sterile liquid aqueous formulation comprising at least 70 wt.% water and at least 40 pmol / ml dissolved roxadustat, having an osmolality in the range of 0.15 to 0.60 Osm / kg, a pH in the range of 2.5 to 5.8, and characterized in that the ratio of the osmolality of the formulation (in Osm / kg) to the concentration of roxadustat in the formulation (in mmol / mL) is less than 2.0. The concentrated aqueous roxadustat formulation of the present invention is well suited for intramuscular administration, but can also be appropriately administered subcutaneously, intranasally, orally, intraperitoneally or intravenously.

Description

Technical Field

[0001] This invention relates to a high-concentration liquid aqueous phenazine formulation, which is particularly suitable for intramuscular or intranasal administration, for example.

[0002] The present invention further relates to the use of the above-mentioned phenazine preparations in medical treatment (such as malaria treatment). Background Technology

[0003] Malaria is a serious and sometimes fatal disease caused by a parasite that typically infects humans, found in mosquitoes that bite. People with malaria usually experience severe illness, including high fever, chills, and flu-like symptoms. There are four types of Plasmodium parasites that infect humans: malignant Plasmodium , Plasmodium vivax , Plasmodium ovale and Plasmodium triflorum .also, Plasmodium norotri (A type of Plasmodium that naturally infects Southeast Asian macaques) can also infect humans, causing malaria that is transmitted from animals to humans (“zoonotic” malaria). Plasmodium falciparum It is the type of malaria most likely to cause severe infection and can lead to death if left untreated.

[0004] The World Health Organization estimates that in 2020, there were 241 million clinical cases of malaria globally, and 627,000 people died from the disease, the majority of whom were children in Africa. Because of the high incidence and mortality rates, malaria places a tremendous burden on the economies of many countries. Since many malaria-affected countries are already relatively poor, the disease perpetuates a vicious cycle of disease and poverty.

[0005] Phenylephrine is an antimalarial drug. It was first manufactured in 1970 and has been used clinically in China since the 1980s. In a small (n = 88) malaria study in Cameroon, phenylephrine showed a 100% cure rate, compared to 60% for chloroquine. It is also being investigated as a potential anticancer drug and a treatment for Ebola virus.

[0006] Artesunate / flunarizine (marketed under the brand name Pyramax®) is an oral fixed-dose combination drug used to treat malaria. Each Pyramax tablet contains 180 mg of flunarizine tetraphosphate and 60 mg of artesunate. Pyramax tablets should be taken orally in a single daily dose for three consecutive days (1–4 tablets per day, depending on body weight).

[0007] The combination of phenazine and artesunate is being investigated as a potential treatment for moderate to severe SARS-CoV-2.

[0008] Shao ( A review of the antimalarial drug pyronaridine.A report in *Chin Med J (Engl)* (1990, 103:428-434) stated that intramuscular injection of a 4% solution of 48 µmol / kg phenazine in rabbits caused a mild local irritation reaction, which completely disappeared within 2 weeks.

[0009] Adegoke et al ( Determination of the phyiscochemical properties of Pyronaridine - a new antimalarial drug [Physicochemical properties of the novel antimalarial drug pyronaridine determined] The physicochemical properties of phenazine were studied in Pak. J. Pharm. Sci., 2006, Vol. 19(1), 1-6. Pharazine tetraphosphate was found to be slightly soluble in water and very slightly soluble in other solvents. At the end of the article, the authors concluded that their research indicated a need to improve the properties of phenazine to enhance its usefulness, and they hypothesized that selecting other salt forms with better solubility, partition, and distribution properties might help improve the poor bioavailability of the drug that has been observed.

[0010] Croft et al. Review of pyronaridine anti-malarial properties and product Characteristics [Overview of the antimalarial properties and product characteristics of phenazine] The Malaria Journal (2012, 11:270) reported that cyclophosphamide is effective against... Plasmodium falciparum (Including chloroquine-resistant strains) It has high potency. The authors report that phenazine is administered as phenazine tetraphosphate (56.89% base) (a bitter, yellow, odorless powder), and that the drug is produced in China as oral tablets, an injection for intramuscular administration, and has also been administered intravenously. The authors further observed that the free base of phenazine is 'very slightly' soluble in water, while the tetraphosphate is 'slightly' soluble (1.46%) in water, which facilitates oral absorption.

[0011] Chu et al. Pyronaridine: a review of its clinical pharmacology in the Treatment of malaria: A clinical pharmacological review of cyclophosphamide in the treatment of malaria. A comprehensive review published in J Antimicrob Chemother 2023;78: 2406-2418 provides the clinical pharmacology of phenazine in the treatment of malaria. The paper notes that phenazine tetraphosphate, as a monotherapy, is available in the Chinese market under the brand name Malaridine, and is formulated as enteric-coated tablets (100 mg phenazine base) for oral administration and as an intramuscular drip (80 mg phenazine base / 2 mL).

[0012] Currently, there are no available phenazine formulations with sufficient concentration for intramuscular or intranasal administration to humans. Summary of the Invention

[0013] The inventors have developed a sufficiently concentrated and sufficiently low molality osmolality liquid aqueous phenazine formulation for intramuscular administration. The liquid aqueous phenazine formulation of the present invention comprises at least 40 µmol / ml of dissolved phenazine, has a pH in the range of 2.5 to 5.8, and a ratio of the molality osmolality of the formulation (in Osm / kg) to the concentration of phenazine in the formulation (in mmol / mL) less than 2.0.

[0014] Accordingly, one aspect of the present invention relates to a liquid aqueous formulation of phenazine for medical treatment, the liquid aqueous formulation comprising at least 70 wt.% water and at least 40 µmol / ml of dissolved phenazine, having a pH in the range of 2.5 to 5.8, and characterized in that the ratio of the weight molar osmolality concentration of the formulation (in Osm / kg) to the concentration of phenazine in the formulation (in mmol / mL) is less than 2.0.

[0015] The inventors have discovered that by dissolving a phenazine base in water with a strong acid, resulting in a formulation with a pH in the range of 2.5 to 5.8 and a weight-molar osmolality to phenazine concentration ratio of less than 2.0, a high-concentration aqueous phenazine liquid formulation suitable for safe intramuscular administration can be obtained. By using a phenazine base instead of, for example, phenazine tetraphosphate, the inventors have been able to prepare isotonic high-concentration phenazine solutions.

[0016] Therefore, another aspect of the present invention relates to a sterile liquid aqueous formulation comprising at least 70 wt.% water and at least 40 µmol / ml of dissolved phenazine, having a weight mol osmolality concentration in the range of 0.15 to 0.60 Osm / kg, a pH in the range of 2.5 to 5.8, and characterized in that the ratio of the weight mol osmolality concentration of the formulation (in Osm / kg) to the concentration of phenazine in the formulation (in mmol / mL) is less than 2.0.

[0017] The concentrated aqueous phenazine formulation of the present invention is ideally suited for intramuscular administration, but may also be appropriately administered intranasally, orally, or intravenously. Detailed Implementation

[0018] A first aspect of the invention relates to a liquid aqueous formulation of phenazine for medical treatment, the liquid aqueous formulation comprising at least 70 wt.% water and at least 40 µmol / ml of dissolved phenazine, having a pH in the range of 2.5 to 5.8, and characterized in that the ratio of the weight molar osmolality concentration of the formulation (in Osm / kg) to the concentration of phenazine in the formulation (in mmol / mL) is less than 2.0.

[0019] As used herein, the term "rhodinil" refers to the substance 4-[(7-chloro-2-methoxy-1,5-dihydrobenzo[b][1,5]naphthidin-10-yl)imino]-2,6-bis(pyrrolidine-1-ylmethyl)cyclohexyl-2,5-dien-1-one (CAS No.: 74847-35-1). The molar mass of rhodinil is 518.05 g / mol.

[0020] As used in this article, the term "malaria" refers to a disease selected from [the text is incomplete]. Plasmodium falciparum , Plasmodium vivax , Plasmodium ovale , Triple malaria protozoan and Plasmodium norotri Malaria infection.

[0021] The medical treatment of the present invention is preferably a medical treatment for humans.

[0022] The treatment of the present invention preferably includes administration of a liquid aqueous phenazine formulation to provide a dose of at least 1 µmol / kg body weight, more preferably a dose of 2 to 100 µmol / kg body weight, and most preferably a dose of 3 to 60 µmol / kg body weight.

[0023] Treatment may suitably include administration of aqueous phenazine formulations via various routes of administration, including intramuscular, subcutaneous, intravenous, intranasal, oral, or intraperitoneal administration. More preferably, treatment includes intramuscular, intranasal, or intravenous administration of aqueous phenazine formulations.

[0024] In a preferred embodiment of the invention, the treatment comprises intramuscular administration of an aqueous phenazine formulation. According to a particularly preferred embodiment, the aqueous phenazine formulation is administered intramuscularly to provide a dose of at least 1 µmol / kg body weight, more preferably 2 to 30 µmol / kg body weight, and most preferably 3 to 20 µmol / kg body weight of phenazine.

[0025] In another preferred embodiment of the invention, the treatment comprises intranasal administration of an aqueous phenazine formulation. According to a particularly preferred embodiment, the aqueous phenazine formulation is administered intranasally to provide a dose of at least 0.5 µmol / kg body weight / day, more preferably 1 to 30 µmol / kg body weight / day, and most preferably 2 to 20 µmol / kg body weight / day of phenazine.

[0026] The treatment of the present invention preferably includes intranasal administration of an aqueous phenazine preparation at least once a day for a period of 2-10 days, preferably for a period of 3-6 days.

[0027] In yet another preferred embodiment, the treatment comprises oral administration of an aqueous phenazine formulation. According to a particularly preferred embodiment, the aqueous phenazine formulation is administered orally to provide a dose of at least 2 µmol / kg body weight / day, more preferably 4 to 50 µmol / kg body weight / day, and most preferably 5 to 20 µmol / kg body weight / day of phenazine.

[0028] The treatment of the present invention preferably includes oral administration of an aqueous phenazine preparation at least once a day for a period of 2-10 days, preferably 3-6 days.

[0029] Preferred embodiments of the present invention relate to diseases caused by parasites or viruses, and more preferably to the treatment of diseases caused by parasites.

[0030] Examples of parasitic diseases treatable according to the present invention include malaria and diseases caused by metaparasites, such as schistosomiasis and cysticercosis. Preferably, the disease treated is malaria or is caused by parasites... Echinococcus granulosus This causes cystic echinococcosis.

[0031] Examples of viral diseases that can be treated according to the present invention include SARS-CoV-1, SARS-CoV-2, Ebola virus disease, and Marburg virus disease.

[0032] According to a particularly preferred embodiment, the medical treatment according to the invention includes the treatment of malaria, especially by... malignant Plasmodium Malaria caused by [the disease].

[0033] The treatment of this invention is particularly suitable for treating severe malaria. Severe malaria occurs when infection is complicated by severe organ failure or when the patient has blood or metabolic abnormalities. The manifestations of severe malaria include the following: • Cerebral malaria, accompanied by behavioral abnormalities, altered consciousness, seizures, coma, or other neurological abnormalities. Severe anemia caused by hemolysis (destruction of red blood cells). • Hemoglobinuria caused by hemolysis (urine contains hemoglobin) • Acute respiratory distress syndrome (ARDS), an inflammatory response in the lungs that inhibits oxygen exchange, can occur even after the number of parasites has decreased due to treatment. • Blood clotting abnormalities • Low blood pressure caused by cardiovascular failure Acute kidney injury • Hyperparasitemia, in which more than 5% of red blood cells are infected with Plasmodium parasites. • Metabolic acidosis (excessive acidity in the blood and tissue fluid), usually associated with hypoglycemia. Treatment of malaria according to the present invention may include the co-administration of one or more other drugs. According to a preferred embodiment, treatment includes the co-administration of artesunate. Combined treatment using artesunate is particularly suitable for treating severe malaria.

[0034] The aqueous phenazine formulation used for medical treatment according to the present invention is preferably a sterile liquid aqueous phenazine formulation specified below.

[0035] Another aspect of the invention relates to a sterile liquid aqueous formulation comprising at least 70 wt.% water and at least 40 µmol / ml of dissolved phenazine, having a weight mol osmolality concentration in the range of 0.15 to 0.60 Osm / kg, a pH in the range of 2.5 to 5.8, and characterized in that the ratio of the weight mol osmolality concentration (in Osm / kg) of the formulation to the concentration of phenazine in the formulation (in mmol / mL) is less than 2.0.

[0036] The liquid aqueous formulation of the present invention preferably contains 100-400 µmol / ml, more preferably 150-330 µmol / ml, and most preferably 180-300 µmol / ml of dissolved phenazine.

[0037] The amount of phenazine together with water is preferably at least 90 wt.%, more preferably at least 93 wt.%, and most preferably 95-98 wt.%.

[0038] The liquid aqueous formulation preferably does not contain undissolved phenazine.

[0039] The weight molar osmotic pressure concentration of the aqueous phenazine formulation is preferably in the range of 0.18 to 0.50 Osm / kg, and most preferably in the range of 0.20 to 0.40 Osm / kg.

[0040] The ratio of the weight molar osmolality (in Osm / kg) of the aqueous phenazine formulation to the phenazine concentration (in mmol / mL) in the same formulation is preferably in the range of 1.2 to 1.9, more preferably in the range of 1.3 to 1.8, and most preferably in the range of 1.4 to 1.7.

[0041] The pH of the aqueous phenazine formulation is preferably in the range of 3.0 to 5.7, more preferably in the range of 4.0 to 5.6, and most preferably in the range of 4.5 to 5.5.

[0042] The aqueous phenazine formulation preferably contains at least 72 wt.% water, more preferably 75-95 wt.% water, and most preferably 80-92 wt.% water.

[0043] Due to the buffering capacity of phenazine, it is advantageous to use a strong acid to lower the pH, otherwise it would be difficult (if not impossible) to achieve the desired near-isotonic osmolar concentration. Therefore, in preferred embodiments, the aqueous phenazine formulation contains at least 1 mmol of phenazine per mmol, more preferably 1.5-5 mmol, and most preferably 2-4 mmol of one or more strong acids selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, and sulfuric acid.

[0044] According to particularly preferred embodiments, the aqueous phenazine formulation contains at least 1 mmol of phenazine per mmol, more preferably 1.5-5 mmol, and most preferably 2-4 mmol of hydrochloric acid.

[0045] According to particularly preferred embodiments, the combination of water, phenazine, and a strong acid accounts for at least 92 wt.%, more preferably at least 96 wt.%, and most preferably at least 99 wt.% of the aqueous phenazine formulation.

[0046] The aqueous phenazine formulation of the present invention is preferably phosphate-free, or has a phosphate content of less than 2 mmol / mmol phenazine, more preferably less than 1 mmol / mmol phenazine, and even more preferably less than 0.3 mmol / mmol phenazine.

[0047] The liquid aqueous formulation of the present invention preferably contains 100-400 µmol / ml, more preferably 150-330 µmol / ml, and most preferably 180-300 µmol / ml of dissolved phenazine.

[0048] Preferably, the aqueous phenazine formulation contains 0-200 µmol / ml of phosphate, more preferably 0-100 µmol / ml of phosphate, and even more preferably 0-50 µmol / ml of phosphate. Most preferably, the aqueous phenazine formulation is phosphate-free.

[0049] The inventors have discovered that the stability of aqueous phenazine formulations can be improved by incorporating L-methionine. Accordingly, in preferred embodiments, the aqueous phenazine formulation contains 0.2-10 mg / mL of L-methionine, more preferably 0.4-6 mg / mL of L-methionine, and most preferably 0.8-4 mg / mL of L-methionine.

[0050] Another aspect of the present invention relates to a method for preparing the liquid aqueous formulation described above herein, the method comprising combining a pyrrolidine base, water and an acid.

[0051] The acid used in this method is preferably selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, and sulfuric acid. Most preferably, the acid used in this method is hydrochloric acid.

[0052] The invention is further illustrated by the following non-limiting examples.

[0053] Example Comparison Example A An aqueous solution of phenazine tetraphosphate (molar amount equal to 910.03 g) was prepared and the osmotic concentration of the solution was determined. The results are shown in Table 1.

[0054] Table 1

[0055] A 200 mg / mL solution of phenazine tetraphosphate has a pH of 2.60.

[0056] Comparison Example B Experiments were conducted to determine the physical appearance of phenazine base at a concentration of 10 mg / mL in different solvents. The results are summarized in Table 2.

[0057] Table 2

[0058] Example 1 Aqueous phenazine solutions according to the present invention were prepared based on the formulations shown in Table 3.

[0059] Table 3

[0060] The solution was prepared by weighing 1200 mg of free phenazine base into a glass vial and then neutralizing the base with the required amount of pre-diluted 1 M HCl. A set value of 5650 mg of 1 M HCl was used to achieve pH 5.0. Finally, all contents were diluted to 10.0 ml with MilliQ water.

[0061] The resulting phenazine solution is a deep orange to black color, free of undissolved substances, and has a pH of 5.0. The osmotic pressure of the solution is 0.342 Osm / kg.

[0062] The ratio of weight molar osmolality (in Osm / kg) to phenazine concentration (in mmol / mL) is equal to 0.342 / 0.232 = 1.48.

[0063] The solution of phenazine was sterilized by passing it through a sterile 0.22 µm polyethersulfone filter.

[0064] The sterile solution thus obtained can be appropriately administered intramuscularly (e.g., in the gluteal muscle) at a dose of 5 mL to deliver 600 mg of phenazine.

[0065] Example 2 Aqueous phenazine solutions according to the present invention were prepared based on the formulations shown in Table 4.

[0066] Table 4

[0067] The solution was prepared by weighing the free base of phenazine and (if applicable) L-methionine into a glass vial. Next, the required amount of pre-diluted 1 M HCl was added to neutralize the base. To achieve pH 5.0, 5,650 mg of 1 M HCl was used. Finally, all contents were diluted to 10.0 ml with MilliQ water.

[0068] The solutions thus obtained were filled into individual clear glass HPLC vials with snap-caps. The stability of the solutions was investigated by storing them at 40°C, 25°C, and 5°C for up to 6 months. The concentration of phenazine was monitored by analyzing the solutions after 2, 3, 6, and 14 months and comparing the concentrations with the initial phenazine concentrations. The results are shown in Table 5.

[0069] Table 5 .

Claims

1. A liquid aqueous formulation of phenazine for medical treatment, said liquid aqueous formulation comprising at least 70 wt.% water and at least 40 µmol / ml dissolved phenazine, having a pH in the range of 2.5 to 5.8, and characterized in that... The ratio of the weight molar osmolality (in Osm / kg) of the formulation to the concentration of phenazine (in mmol / mL) in the formulation is less than 2.

0.

2. The aqueous liquid formulation of phenazine for medical treatment according to claim 1, wherein, This liquid aqueous formulation has a weight molar osmolality concentration in the range of 0.15 to 0.60 Osm / kg.

3. The aqueous liquid formulation of phenazine for medical treatment according to claim 1 or 2, wherein, This liquid aqueous formulation contains 100-400 µmol / mL of dissolved pyrrolidine.

4. Aqueous liquid formulation of phenazine for medical treatment according to any one of the preceding claims, wherein, This liquid aqueous preparation is administered intramuscularly.

5. The aqueous liquid formulation of phenazine for medical treatment according to claim 4, wherein, This liquid aqueous formulation is administered intramuscularly to provide a dose of at least 1 µmol / kg / day of cyclophosphamide.

6. The aqueous liquid formulation of phenazine for medical treatment according to any one of claims 1-3, wherein, This liquid aqueous preparation is administered intranasally.

7. The aqueous liquid formulation of phenazine for medical treatment according to claim 6, wherein, This liquid aqueous formulation is administered intranasally to provide a dose of at least 0.5 µmol / kg / day of phenazine.

8. The aqueous liquid formulation of phenazine for medical treatment according to any one of claims 1-3, wherein, This liquid aqueous formulation is administered orally to provide a dose of at least 2 µmol / kg / day of cyclophosphamide.

9. Aqueous liquid formulation of phenazine for medical treatment according to any one of the preceding claims, wherein, This liquid aqueous formulation contains at least 1 mmol of hydrochloric acid per mmol of phenazine.

10. Aqueous liquid formulation of phenazine for medical treatment according to any one of the preceding claims, wherein, This liquid, water-based preparation is used to treat diseases caused by parasites or viruses.

11. A sterile liquid aqueous preparation comprising at least 70 wt.% water and at least 40 µmol / ml of dissolved phenazine, having a weight molality osmolarity in the range of 0.15 to 0.60 Osm / kg and a pH in the range of 2.5 to 5.8, and characterized in that... The ratio of the weight molar osmolality (in Osm / kg) of the formulation to the concentration of phenazine (in mmol / mL) in the formulation is less than 2.

0.

12. The liquid aqueous formulation according to claim 11, wherein, This liquid aqueous formulation contains 100-400 µmol / mL of dissolved pyrrolidine.

13. The liquid aqueous formulation according to claim 11 or 12, wherein, This liquid aqueous formulation has a weight molar osmolality concentration in the range of 0.20 to 0.40 Osm / kg.

14. The liquid aqueous formulation according to any one of claims 11-13, wherein, This liquid aqueous preparation is phosphate-free, or the phosphate content is less than 2 mmol per mmol of phenazine.

15. The liquid aqueous formulation according to any one of claims 11-14, wherein, This liquid aqueous formulation contains at least 1 mmol of hydrochloric acid per mmol of phenazine.

16. A method for preparing a liquid aqueous formulation according to any one of claims 11-15, the method comprising combining a phenazine base, water and an acid.

17. The method according to claim 16, wherein, The acid is selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, and sulfuric acid.

18. The method according to claim 17, wherein, The acid is hydrochloric acid.

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