Orellanine preparation

Abstract

Disclosed is an aqueous pharmaceutical preparation containing orelanine.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical preparation containing orellanine.

Background Art

[0002] Orellanine (see formula I below) is a selective nephrotoxin that is present in relatively large amounts in some fungal species of the Cortinarius family.

Chemical

Summary of the Invention

[0003] Accordingly, the present invention provides an aqueous pharmaceutical preparation containing 0.1 to 40 mg / ml of orellanine and a buffer solution and having a pH in the range of 7 to 8. The present inventors have found that such a preparation is stable and suitable for administration by intravenous injection.

Modes for Carrying Out the Invention

[0004] The pharmaceutical preparation is an aqueous preparation. Water for injection (WFI) can be used for an aqueous preparation to be administered by intravenous injection.​ The amount of olelanin in the formulation is 0.1 - 40 mg / ml, and may be 0.5 - 30 mg / ml, olelanin 1 - 20 mg / ml, olelanin 5 - 15 mg / ml, olelanin 8 - 12 mg / ml, olelanin 9 - 11 mg / ml, or olelanin about 10 mg / ml. In an embodiment, the formulation contains olelanin 10 mg / ml. The amount of olelanin in the formulation can be measured by conventional methods including high performance liquid chromatography (HPLC). The inventors have found that the solubility of olelanin is pH-dependent. Outside the preferred pH range, the solubility of olelanin is low and it is not possible to obtain a concentration of olelanin preferred for pharmaceutical formulations for administration by intravenous injection. The pH of the formulation is in the range of 7 - 8, and may be in the range of 7.1 - 7.6, or in the range of 7.2 - 7.5, or in the range of 7.3 - 7.4. In an embodiment, the pH is about 7.4 (7.39 - 7.41). The pH can be measured by conventional methods, such as according to Ph.Eur. 2.2.3

[0005] The inventors have found that the concentration of the buffer has an influence on the solubility of olelanin. The concentration of the buffer may be 10 - 200 mM, or 20 - 180 mM, or 30 - 120 mM, or 40 - 80 mM, or 40 - 60 mM. The buffer may be a phosphate buffer such as NaH2PO4 - Na2HPO4 or citric acid - Na2HPO4. The buffer ensures that the pH is within the selected range. Further adjustment of the pH can be carried out by adding a base or an acid, such as adding sodium hydroxide or hydrochloric acid, for example, using 0.2 M sodium hydroxide or hydrochloric acid. The inventors have found that the weight osmolality of the formulation affects the solubility of olelanine. The weight osmolality of the formulation may be greater than 200 mOsm, or may be in the range of 250 - 350 mOsm. The weight osmolality can be measured by conventional methods, such as according to Ph.Eur 2.2.35. In one embodiment of the present invention, the weight osmolality of the formulation is such that it is within the isotonic range, i.e., the formulation has an isotonicity similar to that of blood.

[0006] The ionic strength of the formulation can be increased by adding sodium chloride. The formulation can contain 0.05 - 2% by weight of sodium chloride (weight based on the weight of the formulation), or 0.1 - 1% by weight of sodium chloride, or 0.5 - 0.9% by weight of sodium chloride, or about 0.8% by weight of sodium chloride. The inventors have found that by adjusting the pH and weight osmolality, a stable formulation with an olelanine concentration suitable for intravenous administration can be prepared. In an embodiment, the aqueous pharmaceutical formulation contains 0.1 - 40 mg / ml of olelanine, a phosphate buffer with a buffer concentration of 10 - 200 mM, and 0.1 - 1% by weight of sodium chloride, the pH of the formulation is in the range of 7 - 8, and optionally the weight osmolality of the formulation is greater than 200 mOsm. In another embodiment, the aqueous pharmaceutical formulation contains 1 - 20 mg / ml of olelanine, a phosphate buffer with a buffer concentration of 10 - 200 mM, and 0.1 - 1% by weight of sodium chloride, the pH of the formulation is in the range of 7 - 8, and optionally the weight osmolality of the formulation is greater than 200 mOsm.

[0007] In another embodiment, the aqueous pharmaceutical formulation contains 5 - 15 mg / ml of olelanine, a phosphate buffer with a buffer concentration of 10 - 200 mM, and 0.1 - 1% by weight of sodium chloride, the pH of the formulation is in the range of 7 - 8, and optionally the weight osmolality of the formulation is greater than 200 mOsm. In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 0.1 to 40 mg / ml, a phosphate buffer selected from NaH2PO4-Na2HPO4 or citric acid-Na2HPO4 with a buffer concentration of 10 to 200 mM, and 0.1 to 1% by mass of sodium chloride. The pH of the preparation is in the range of 7 to 8, and optionally the weight osmolarity of the preparation is greater than 200 mOsm. In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 1 to 20 mg / ml, a phosphate buffer selected from NaH2PO4-Na2HPO4 or citric acid-Na2HPO4 with a buffer concentration of 10 to 200 mM, and 0.1 to 1% by mass of sodium chloride. The pH of the preparation is in the range of 7 to 8, and optionally the weight osmolarity of the preparation is greater than 200 mOsm. In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 5 to 15 mg / ml, a phosphate buffer selected from NaH2PO4-Na2HPO4 or citric acid-Na2HPO4 with a buffer concentration of 10 to 200 mM, and 0.1 to 1% by mass of sodium chloride. The pH of the preparation is in the range of 7 to 8, and optionally the weight osmolarity of the preparation is greater than 200 mOsm.

[0008] In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 0.1 to 40 mg / ml, a phosphate buffer with a buffer concentration of 40 to 60 mM, and 0.5 to 0.9% by mass of sodium chloride. The pH of the preparation is in the range of 7.2 to 7.5, and optionally the weight osmolarity of the preparation is in the range of 250 to 350 mOsm. In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 1 to 20 mg / ml, a phosphate buffer with a buffer concentration of 40 to 60 mM, and 0.5 to 0.9% by mass of sodium chloride. The pH of the preparation is in the range of 7.2 to 7.5, and optionally the weight osmolarity of the preparation is in the range of 250 to 350 mOsm. In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 5 to 15 mg / ml, a phosphate buffer with a buffer concentration of 40 to 60 mM, and 0.5 to 0.9% by mass of sodium chloride. The pH of the preparation is in the range of 7.2 to 7.5, and optionally the weight osmolarity of the preparation is in the range of 250 to 350 mOsm. In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 0.1 to 40 mg / ml, a phosphate buffer selected from NaH2PO4-Na2HPO4 or citric acid-Na2HPO4 with a buffer concentration of 40 to 60 mM, and 0.5 to 0.9% by mass of sodium chloride, the pH of the preparation is in the range of 7.2 to 7.5, and optionally the weight osmolarity of the preparation is in the range of 250 to 350 mOsm.

[0009] In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 1 to 20 mg / ml, a phosphate buffer selected from NaH2PO4-Na2HPO4 or citric acid-Na2HPO4 with a buffer concentration of 40 to 60 mM, and 0.5 to 0.9% by mass of sodium chloride, the pH of the preparation is in the range of 7.2 to 7.5, and optionally the weight osmolarity of the preparation is in the range of 250 to 350 mOsm. In another embodiment, the aqueous pharmaceutical preparation contains olelanine at 5 to 15 mg / ml, a phosphate buffer selected from NaH2PO4-Na2HPO4 or citric acid-Na2HPO4 with a buffer concentration of 40 to 60 mM, and 0.5 to 0.9% by mass of sodium chloride, the pH of the preparation is in the range of 7.2 to 7.5, and optionally the weight osmolarity of the preparation is in the range of 250 to 350 mOsm. The preparation a) adding a buffer to the aqueous olelanine solution; b) stirring; c) adjusting the pH by adding a further buffer, by adding a base (e.g., sodium hydroxide), and / or by adding an acid (e.g., hydrochloric acid) and can be prepared by a method comprising.

[0010] Alternatively, the preparation a) adding olelanine (which may be in aqueous solution) to the buffer; b) stirring; c) adjusting the pH by adding additional buffer, by adding a base (e.g., sodium hydroxide), and / or by adding an acid (e.g., hydrochloric acid); It can be prepared by a method comprising: The steps of stirring and adjusting the pH can be repeated several times to ensure that the final pH is within the selected range. The inventors have observed that the pH of the formulation may change upon stirring, and as a result, adjusting the pH is necessary to achieve a pH within the selected range and to obtain a stable formulation.

[0011] The buffer can be prepared by combining buffer components (e.g., as NaH2PO4 and Na2HPO4, or as sodium citrate and Na2HPO4), water for injection, and sodium chloride. The formulation may be filtered (e.g., through a 0.22 μm PVDF filter) before being packaged. The formulation can be packaged in conventional containers such as glass injection vials. The vials can be stoppered with flip-off caps and sealed. The formulation can be used in a method for treating cancer such as renal cancer. The formulation can be used to treat patients suffering from or prone to renal cell carcinoma. The treatment method can be used to treat tumors localized in one or both of the patient's kidneys. Alternatively, the renal cell carcinoma may have metastasized such that at least one renal cell carcinoma tumor is present in at least one non-renal tissue.

[0012] The formulation is suitable for administration by intravenous injection. The formulation can be administered as a single dose, as a series of daily doses, or in an intermittent dosing regimen (e.g., multiple doses or dose sequences administered at intervals of 1 to about 30 days, 1 to about 14 days, or 1 to about 7 days). When administered in a single dose, a dose of about 1 mg / kg to about 100 mg / kg of orelanine, or a dose of about 2 mg / kg to about 25 mg / kg of orelanine, or a dose of about 5 mg / kg to about 15 mg / kg of orelanine can be administered to a patient. The formulation can be administered in more than two doses. The doses can be administered daily or intermittently (e.g., having at least one non-administration day between successive doses). When administered in multiple doses, a dose of about 0.5 mg / kg to about 10 mg / kg of orelanine, or a dose of about 1 mg / kg to 5 mg / kg of orelanine, or a dose of about 2 mg / kg of orelanine can be administered to a patient. When successive doses are administered intermittently, the successive doses can be administered at intervals of 2 to 7 days. The formulation can be administered to a patient in 3, 4, 5 or more than 6 doses, and each dose is administered at an interval of 3 to 5 days. The formulation can be administered to a patient in 4, 5, or more than 6 doses at intervals of 3 to 4 days, and each dose contains about 1 mg / kg to about 20 mg / kg of orelanine, or about 2 mg / kg to about 10 mg / kg of orelanine, or about 5 mg / kg of orelanine.

[0013] The daily dose of the formulation can be administered for at least 2 days. A typical daily dose administered to a patient is 0.1 to 10 mg / kg, or 1 to 5 mg / kg, or about 2 mg / kg. The treatment protocol can include daily administration of the formulation for 5 to about 30 days, or preferably 10 to 20 days, or most preferably about 14 days. In some examples, it may be desirable to combine the multiple intermittent dosing protocols, daily dosing protocols, or combinations thereof as described above with rest periods and / or recovery periods. Thus, in some examples, it may be desirable to administer the formulation according to a daily dosing method or an intermittent dosing method, measure the tumor response to the treatment, and then, if necessary, remove the renal cancer tumor or further reduce the size of the renal cancer tumor by performing the next daily dosing treatment or intermittent dosing treatment. Such dosing strategies are well known to those skilled in the field of oncology. In one embodiment of the present invention, a patient suffering from renal cell carcinoma can be treated with a formulation by continuously injecting about 0.5 to 5 mg of olelanine / kg bw, most preferably about 2 mg of olelanine / kg bw, for about 7 to 21 days continuously, or for about 14 days continuously, daily. One to five hours after each daily injection of the formulation, most preferably about 2 hours after such an injection, all the olelanine that has not been taken up into the tumor tissue is removed, and in order to minimize any unwanted side effects that may occur in the extracellular space, the patient undergoes hemodialysis for 1 to 5 hours, most preferably about 2 hours.

[0014] The above preferred dosages and dosage regimens are based on a human patient with renal cell carcinoma having a tumor mass of about 1 kg and a body weight of 70 kg. However, as will be readily known to those skilled in the field of cancer medicine, such preferred dosages and dosage regimens are determined to a significant extent by patient characteristics such as age, gender, body weight, general condition, and especially the tumor mass of the individual patient and the response to treatment. As always, the ultimate responsibility for selecting the appropriate dosage and treatment strategy lies with the physician in charge of the patient. The present invention will now be described by reference to examples that are not intended to limit the invention.

Examples

[0015] (Solubility study using buffer (pH-dependent solubility test)) In order to analyze the effect of pH on the solubility of olelanine, phase solubility studies were performed on different buffers. The following four buffers were tested. · 50 mM acetate buffer pH 4.5 - Prepared from 1.81 g of sodium acetate and 14 ml of acetic acid (2N), and adjusted to 1 l with water for injection (WFI). · 50 mM phosphate buffer pH 5.8 - Prepared from 92 ml of NaH2PO4 and 8 ml of Na2HPO4, and adjusted to 200 ml with WFI. · 50 mM phosphate buffer pH 6.8 - Prepared from 51 ml of NaH2PO4 and 49 ml of Na2HPO4, and adjusted to 200 ml with WFI. · Prepared from 19 ml of NaH2PO4 and 81 ml of Na2HPO4 in 50 mM phosphate buffer pH 7.4 and adjusted to 200 ml with WFI. The pH of each buffer was adjusted with a pH meter and 0.2 M sodium hydroxide or hydrochloric acid. The solubility of oleanolic acid in each buffer was tested by weighing 40 mg of oleanolic acid into a vial, adding 2 ml of buffer, stirring the suspension for 1 hour, and then stirring at 400 rpm at room temperature for 24 hours. Each suspension was centrifuged at 5300 rpm for 15 minutes. The supernatant was filtered through a 0.45 μm nylon filter. Table 1 shows the results of the test.

[0016]

Table 1

[0017] The maximum concentration of oleanolic acid obtained was 1.7 mg / ml in 50 mM phosphate buffer pH 7.4. (Solubility study using different buffer concentrations at pH 7.4) The effect of buffer concentration was evaluated. The following three buffers were used. · 25 mM phosphate buffer pH 7.4 - Prepared from 0.95 ml of NaH2PO4 (0.1 M) and 4.05 ml of Na2HPO4 (0.1 M) and adjusted to 20 ml with WFI. · 50 mM phosphate buffer pH 7.4 - Prepared from 19 ml of NaH2PO4 (0.1 M) and 81 ml of Na2HPO4 (0.1 M) and adjusted to 200 ml with WFI. · 100 mM phosphate buffer pH 7.4 - Prepared from 38 ml of NaH2PO4 (0.1 M) and 162 ml of Na2HPO4 (0.1 M) and adjusted to 200 ml with WFI. The pH of each buffer was adjusted with a pH meter and 0.2 M sodium hydroxide or hydrochloric acid.

[0018] The solubility of olelanine in each buffer was tested by weighing 20 mg of olelanine into a 5 ml vial, adding 2 ml of buffer, stirring the suspension for 3 hours, and then adjusting the pH to 7.4 with 0.2 M sodium hydroxide or hydrochloric acid. The suspension was then stirred at 400 rpm for 24 hours at room temperature. Each suspension was filtered through a 0.2 μm PVDF syringe filter. Table 2 shows the results of the test.

[0019]

Table 2

[0020] For these formulations, the solubility was in the range of 3.7 - 4.9 mg / ml. From the perspective of solubility and purity, 50 mM seems to be the optimal concentration of the buffer. (Solubility study using different buffer salts at pH 7.4) The following four buffers at pH 7.4 and 50 mM were evaluated. · 50 mM phosphate buffer pH 7.4 - Prepared from 19 ml of NaH2PO4 (0.1 M) and 81 ml of Na2HPO4 (0.1 M), adjusted to 200 ml with WFI. · 50 mM phosphate buffer pH 7.4 - Prepared from 28.1 ml of KH2PO4 (0.2 M) and 21.9 ml of NaOH (0.2 M), adjusted to 200 ml with WFI. · 50 mM Tris - HCl buffer pH 7.4 - Prepared from 54.7 ml of tris(hydroxymethyl)aminomethane (0.1 M) and 45.3 ml of hydrochloric acid (0.1 M), adjusted to 200 ml with WFI. · 50 mM citrate - phosphate buffer pH 7.4 - Prepared from 2.4 ml of citric acid monohydrate (0.2 M) and 47.6 ml of Na2HPO4 (0.2 M), adjusted to 200 ml with WFI. The pH of each buffer was adjusted with a pH meter and 0.2 M sodium hydroxide or hydrochloric acid.

[0021] The solubility of oleanolic acid in each buffer was tested by weighing 20 mg of oleanolic acid into a 5 ml vial, adding 2 ml of the buffer, stirring the suspension for 3 hours, and then adjusting the pH to 7.4 with 0.2 M sodium hydroxide or hydrochloric acid. The suspension was then stirred at 400 rpm at room temperature for 24 hours. Each suspension was filtered through a 0.2 μm PVDF syringe filter. Table 3 shows the results of the test.

[0022] [Table 3]

[0023] The best solubility was provided by the NaH2PO4-Na2HPO4 buffer and the citric acid-Na2HPO4 buffer. (Solubility study using different ionic strengths at pH 7.4) The ionic strength of the NaH2PO4-Na2HPO4 buffer was adjusted by adding sodium chloride. The ionic strength of the 50 mM NaH2PO4-Na2HPO4 buffer was estimated to be approximately 130 mM. The proportions of sodium chloride added to the buffer were 0.41%, 0.98%, and 2.15%. The solubility of oleanolic acid in each buffer was tested by weighing 20 mg of oleanolic acid into a 5 ml vial, adding 2 ml of the buffer, stirring the suspension for 3 hours, and then adjusting the pH to 7.4 with 0.2 M sodium hydroxide or hydrochloric acid. The suspension was then stirred at 400 rpm at room temperature for 24 hours. Each suspension was filtered through a 0.2 μm PVDF syringe filter. Table 4 shows the results of the test.

[0024] [Table 4]

[0025] By increasing the ionic strength, the weight osmolarity of the solution increased and the solubility of oleanolic acid was enhanced. (Formulation adjustment) The following two buffers were selected: 50 mM NaH2PO4-Na2HPO4 pH 7.4 and 50 mM citric acid-Na2HPO4, each with 0.70% sodium chloride added. By adding 0.70% sodium chloride, a solution close to isotonicity is obtained. The formulation was prepared by weighing 40 mg of olelanine into a 5 ml vial, adding 2 ml of buffer, stirring the suspension for 1 hour, adjusting the pH to 7.4 with sodium hydroxide (0.2 M), stirring the suspension at 400 rpm at room temperature for 24 hours, and adjusting the pH to 7.4 with sodium hydroxide (0.2 M). Each suspension was filtered through a 0.2 μm PVDF syringe filter. Table 5 shows the characteristics of the formulation.

[0026]

Table 5

[0027] By adjusting the pH to 7.4 twice, a solubility of more than 10 mg / ml was obtained. That is, first after 1 hour of stirring and again after overnight stirring.

Claims

1. Orellanine 0.1-40 mg / ml, Buffer and A pharmaceutical aqueous solution preparation containing, A pharmaceutical preparation with a pH in the range of 7 to 8.

2. A pharmaceutical preparation according to claim 1, comprising 1 to 20 mg / ml of oreranine.

3. A pharmaceutical preparation according to claim 2, comprising 5 to 15 mg / ml of oreranine.

4. The pharmaceutical preparation according to claim 1, wherein the pH is in the range of 7.1 to 7.

6.

5. The pharmaceutical preparation according to claim 4, wherein the pH is in the range of 7.2 to 7.

5.

6. The pharmaceutical formulation according to claim 1, wherein the concentration of the buffer solution is 10 to 200 mM.

7. The pharmaceutical preparation according to claim 6, wherein the concentration of the buffer solution is 40 to 60 mM.

8. The pharmaceutical formulation according to claim 1, wherein the buffer is a phosphate buffer.

9. The buffer is NaH 2 PO 4 -Na 2 HPO 4 or sodium citrate 2 HPO 4 The pharmaceutical preparation according to claim 8.

10. The pharmaceutical preparation according to claim 1, wherein the weight osmolality is greater than 200 mOsm.

11. The pharmaceutical preparation according to claim 10, wherein the weight osmolality is in the range of 250 to 350 mOsm.

12. A pharmaceutical preparation according to claim 1, comprising sodium chloride.

13. A pharmaceutical preparation according to claim 12, comprising 0.1 to 1% by mass of sodium chloride.

14. A pharmaceutical preparation according to any one of claims 1 to 13, for the treatment of cancer.

15. The pharmaceutical preparation according to claim 14, wherein the cancer is renal cancer.

16. The pharmaceutical formulation according to claim 14, wherein the treatment comprises the step of providing the formulation to a patient in need thereof for at least two days, the daily dose being 0.1 to 10 mg / kg of body weight.

17. The pharmaceutical formulation according to claim 14, wherein the treatment comprises the step of intravenously administering the formulation to a patient in need thereof.

18. A method for preparing a pharmaceutical preparation according to any one of claims 1 to 13, a) The step of adding a buffer to an aqueous solution of olelanine, or the step of adding olelanine, which may be in aqueous solution form, to a buffer, b) The step of stirring, c) The step of adjusting the pH by adding further buffer, by adding a base (e.g., sodium hydroxide), and / or by adding an acid (e.g., hydrochloric acid). A method that includes this.

19. The method according to claim 18, wherein steps (b) and (c) are performed at least twice.

20. A kit for intravenous administration, comprising a pharmaceutical formulation according to any one of claims 1 to 13, packaged in a container, and further comprising information for providing the formulation to a patient in need thereof.