TWI229600B - Pharmaceutical composition for parenteral administration for the treatment of hepatocellular carcinoma - Google Patents

Abstract

The invention discloses a pharmaceutical composition for parenteral administration for the treatment of hepatocellular carcinoma, which comprises a therapeutically effective amount of arsenic trioxide and a pharmaceutically acceptable carrier.

Description

1229600 A7 B7 5. Description of the invention (1) Background of the invention Arsenic trioxide can be used as a herbicide and can also be used to kill rodents and insects. It can be used as a parasiticicide in medicine, and it is also a component of traditional Chinese medicine. Its main medical purpose is to treat anaemia and dyspepsia. It can also be used to treat tooth marrow diseases. Devitalizing agent; it also treats psoriasis, syphilis, and rheumatosis with the principle of "poisoning with poison". In the 1820's, trioxide was considered a potential co-carcinogen for human malignancies, especially bladder, skin, and lung cancers. Due to its carcinogenicity, today's medical uses are mostly only for the treatment of trypanosomiasis associated with the central nervous system. However, in recent years, research has found that arsenic trioxide is used as an extremely effective antileukemia agent (antileukaemic agent), which can effectively treat acute promyelocytic leukemia, and has been seen in many literatures. For example, Blood, Vol. 88, No. 3, 1996, ρ 1052-1061, European Journal of Cancer, Vol. 35, No. 8, 1999, ρ · 1258-1263, and The New England Journal of Medicine, Vol. 339, No. 19, 1998, p. 1341-1348. However, the aforementioned prior art does not describe the efficacy of arsenic trioxide in the treatment of tumors, especially in the treatment of liver tumor-related diseases. Furthermore, there is currently no clinically effective drug for the treatment of hepatocellular carcinoma. Most of the current hepatocellular carcinoma therapies for metastatic or local therapies are transcatheter -4-

O: \ 72 \ 72584.DOC \ WCK This paper size is applicable to Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1229600 A7 B7 V. Description of the invention (2) Transcatheter arterial embolization or alcohol Percutaneous ethanol injection, or systemic chemotherapeutics, such as the use of D 0χ 〇rubici η, Tamoxifen in high dose combined with Doxorubicin or EA-PFL, etc. Although the remission rate of these agents can reach i 5 to 30%, most patients with hepatocellular carcinoma are often accompanied by cirrhosis and its complications (such as decreased white blood cells, decreased platelets, or liver function incompensation), making it impossible to use the whole body. Sex chemotherapy. Chemotherapy is not effective in prolonging overall patient survival. SUMMARY OF THE INVENTION The present invention has unexpectedly discovered that arsenic trioxide is formulated as a medicinal medicinal composition for injection and can be used to treat diseases related to liver tumors. The object of the present invention is to provide a pharmaceutical composition for injection for treating liver tumors. Brief illustration of the figure Figure 1 shows the inhibitory effect of the compound (dioxygen trioxide) used in the present invention and the control compound (mitomycin) on the liver tumor cell line H c-4 Figure 2 shows the used in the present invention The inhibitory effect of the compound (dioxygen) and the control compound (mitomycin) on the liver tumor cell line Hep 3 β. Figure 3 shows the inhibitory effect of the compound (arsenic trioxide) used in the present invention and the control group compound (mitomycin) on the liver tumor cell line He p G 2

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Line 1229600 A7 B7

5. Description of the invention (3) Fruits. Figure 4 shows the inhibitory effect of the compound (diquinone trioxide) used in the present invention on liver tumor cell lines HC-4, Hep 3B and HepG2. Detailed description of the invention The arsenic trioxide (chemical formula As " 3) in the present invention is a white or transparent amorphous lump or crystal, which is not easily soluble in water and has a sweet taste. It is highly toxic and is generally used in the manufacture of glass, enamels and other substances. The pharmaceutical composition for injection of the present invention has excellent curative effect on diseases related to liver tumors. "Hepatic tumor" as referred to in this article generally includes two types of tumor cells: hepatocellular carcinoma and cholangiocarcinoma. It can also be divided into liver cell carcinoma (liver cell carcinoma) and liver fibrous layer. Fibrolamellar variant, intrahepatic bile duct carcinoma, mixed hepatocellular cholangiocarcinoma, and undifferentiated hepatocellular carcinoma. "Medically effective amount" mentioned herein refers to an amount sufficient to provide a therapeutic effect when administered to a mammal in need of such treatment. "Medically effective amount will vary with the individual and condition being treated, the weight and age of the individual, and condition. The severity of the drug, the mode of administration of the drug, and other factors, which can be determined by professionals in the pharmaceutical industry. A medically effective amount suitable for use in the pharmaceutical composition of the invention

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f Dioxin concentration, preferably 0.01 to 10 mg / g, and most preferably (U to) t g / g. The "medical composition of the present invention is adjusted to g Jicheng injection dosage form for administration, which can be made into any ^ Know injection dosage forms, such as powder injections; Dongjing injections, emulsification injections, oily injections, microlipid injections, etc. "Pharmaceutically acceptable carrier" in this context means any drug suitable for injection Known inert carriers, such as organic solvents, stabilizers, chelating agents, P square rotants, emulsifiers, suspending agents, diluents or gelling agents. The "medical composition" of the present invention may be familiar with this technology. A conventional method is used to prepare the above-mentioned injection form. For example, in a preferred embodiment of the present invention, an appropriate amount of arsenic trioxide, disodium ethylenediamine tetraacetate, buffer solution and water are uniformly mixed to make a general injection. The following examples are provided to further illustrate the feasibility of the present invention, so as to make the technical content of the present invention more specific, but not intended to limit the scope of the present invention. Others skilled in the art are based on the know-how All the changes and improvements of the invention which can be achieved through teaching can all belong to the scope of the present invention. Example 1: The arsenic trioxide propellant of the present invention 1 ^ g / set and its price and its preparation method Ingredients_ Content Arsenic trioxide 10 Mg disodium ethylenediamine tetraacetate 4 mg O: \ 72 \ 72584.DOC \ tsp

1229600 A7 B7 V. Description of the invention (5)

Buffer solution proper amount of water for injection (USP) to 10.0 ml First dissolve dioxin trioxide and disodium ethylenediamine tetraacetate in an appropriate amount of water for injection (WF I), then add water for injection to 90% of the target volume, and then Use a buffer solution (which can use acetic acid, hydrochloric acid, sodium hydroxide, sodium dicarbonate or phosphate buffer) to adjust the physiologically acceptable pH value of the human body. Finally, water for injection was added to adjust the total volume to 10 ml. Example H The composition of one arsenic trioxide 1 lyophilized injection of the present invention and its preparation method Ingredient content • Arsenic trioxide 10 亳 g mannitol 0.5 mg citric acid 0.8 mg buffer solution appropriate amount of water for injection (USP) to 10.0 ml Dissolve arsenic trioxide, mannitol and citric acid in an appropriate amount of water for injection (WFI), then add water for injection to a target volume of 90/0, and then adjust to an physiologically acceptable pH with an acid-base reagent or buffer. p Η value. Finally, water for injection was added to adjust the total volume to 10 liters, and the solution was placed in a freeze dryer to make a freeze-dried injection. Example 3: The antitumor effect of the arsenic trioxide injection of the present invention 1. Test compound and concentration O: \ 72 \ 72584.DOC \ tsp -8-This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm)

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Line 1229600 A7 _ B7 V. Description of the invention (6) The arsenic trioxide compound was dissolved in a 100% D M S 〇 solution and subjected to ultrasonic vibration for 30 minutes. Subsequently, it was diluted with distilled water to obtain a 40% D M S 0 test solution of dioxin trioxide compound solution with working concentrations of 10,000, 1,000, 1,000, 100, 1 and 1 μM, respectively. At the time of the test, the final analysis concentration obtained by using the 1000-fold dilution method was 1000′1, 1, 0, and 0.001 μM, and the test was performed in the culture medium. 2. Cell lines The liver tumor cell lines used for the medical efficacy test of the present invention are shown in Table 1. Table 1. Cell line source cell strain types HC-4. Hep 3B ATCC HB-8064 human liver tumor cells from Hokkaido University HepG2 ATCC HB-8065 Human liver tumor cells 3. Analytical method 100 pL of liver tumor cell suspension (cell line HC-4, Hep 3B or He pG2) (approximately 2.5 x 103 / well) in 96-well microdrops In the dish, the culture environment is 37 ° C and the concentration of carbon dioxide is 5%. After 24 hours, 100 pL of growth medium, 2 u 1 of dioxin trioxide test solution, mitomycin (mitomycin), or a vehicle (40% DMS 0) were added to the wells, respectively. Incubate for 72 hours, and the test concentrations of dikun trioxide are 100, 10, 1, 0.1, and 0.01 μM, respectively. At the end of the culture period, pour out the medium in the microtiter tray, and add 200 pL of fresh medium and 20 pL of -9- O: \ 72 \ 72584.DOC \ tsp to each well. A4 specification (210X297 mm) 1229600 A7 __B7 V. Description of the invention (7) An alamarBlue reaction reagent is reacted for 6 hours. Finally, the fluorescence intensity was analyzed at 530nm and 590nm with a Spectra flour Plus fluorescence reader. The results obtained are shown in Table 2 and Table 3 and Figures 1 to 4. 7F · Table 2 Test cell cell growth percentage (mean soil SEM, n = 2) Trioxide control empty starting point control carrier concentration (Time.) Culture group 100 10 1 0.1 0.01 HC-4 • 100 0 100 • 91 ± 5 6 ± 0 94 ± 6 96 ± 5 98 soil 4 Hep 3B • 100 0 100 • 86 ± 1 -1 ± 2 85 soil 1 100 ± 5 92 ± 6 HepG2 • 100 0 100 -64 ± 9 75 ± 8 109 ± 3 107 ± 2 100 ± 3 The growth percentage value of the above test is -1 0 0 in the control blank group and 1 in the body culture group 0 0, the starting point (T ime 〇) is 0. Table 3 Growth percentage of control test cell lines (mean ± SEM, n = 2) Trioxide control Null start point Control loading concentration (μM) Diarsenic white group (TimeG) culture group 10 1 0.1 0.01 0.001 HC-4 • 100 0 100 -91 ± 2 -29 ± 14 8 ± 0 50 ± 2 96 ± 4 Hep3B • 100 0 100 -65 ± 5 • 11 ± 5 22 ± 3 55 ± 8 95 ± 6 HepG2 • 100 0 100 • 98 ± 5 -36 ± 1 29 ± 0 71 ± 4 102 + 6 -10-

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The growth percentage value of the above test was "0" in the control blank group, 100 in the control carrier culture group, and 0 (TimmeG) as the starting point. Based on the above test results, the ICsG, TGI & LC "values of arsenic trioxide and mitomycin for each liver tumor cell line are further calculated, as shown in Table 4. Table 4 Test cell line ^ 50 (μΜ) TGI ( pM) LC5〇 (m «M) Arsenic trioxide HC-4 5.0 11 24 Mitomycin 0.0 17 0.16 1.4 Arsenic trioxide Hep 3 B 3.6 9.8 26 Mitomycin-0.025 0.3 7 5.2 Arsenic trioxide Hep G 2 18 37 7 6 Silk Mitomycin 0.047 0.25 1.3 IC 50 indicates the concentration that inhibits 50% of cell growth; T GI indicates the concentration that inhibits total growth; LC 50 indicates the concentration that causes 50% of cells to die. From Table 2 above The results to Table 4 show that the present invention uses diquinone to combat the growth of liver tumor cells and has a good inhibitory effect. -11-

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Claims (1)

^^ 25668 Patent Application Chinese Application for Patent Scope Replacement (October 1993), Patent Application Park A8 B8 C8 D8 \ Cj ^ (° n 2. 3. 4. 5. 6. 7 · One use An injectable pharmaceutical composition for treating liver tumors, which includes a medically effective amount of tritiated dioxin and a pharmaceutically acceptable carrier including a chelating agent at a concentration of 0.2 to 0.4 mg / ml. According to the scope of patent application No. 1 The pharmaceutical composition for injection, wherein the liver tumor is hepatocellular carcinoma and cholangiocarcinoma. The pharmaceutical composition for injection according to the first patent application scope, wherein the liver tumor is hepatocellular carcinoma ( liver cell carcinoma), fibrolamellar variant, intrahepatic bileduct carcinoma, mixed hepatocellular cholangiocarcinoma, and undifferentiated hepatocellular carcinoma. According to the scope of patent application, the first The pharmaceutical composition for injection according to any one of items 3 to 3, wherein the concentration of arsenic trioxide is 0.01 to 10 mg / g. A pharmaceutical composition for injection, wherein the concentration of dikun trioxide is 0.1 to 1 mg / g. The pharmaceutical composition for injection according to item 1 of the patent application scope, wherein the mixture is ethylenediamine tetraacetic acid disodium (EDTA) An injectable pharmaceutical composition for treating liver tumors, comprising a medically effective amount of dioxin and an emulsifier and a stabilizer (the ratio of which is 0.1 to 0.2 mg / ml) 1: 1 to 1: 2) pharmaceutically acceptable carriers. This paper size applies the ZTE National Standard (CNS) A4 specification (210X 297 mm), less ~-1229600-C8 ^ __ D8_ VI. Application scope of patent 8. The pharmaceutical composition for injection according to item 7 of the scope of patent application, of which Liver tumors are hepatocellular carcinoma and cholangiocarcinoma. 9. The pharmaceutical composition for injection according to item 7 of the scope of patent application, wherein the liver tumors are liver cell carcinoma, fibrolamellar variant, and intrahepatic bile duct carcinoma. ), Mixed hepatocellular cholangiocarcinoma and undifferentiated hepatocellular carcinoma. 10. The pharmaceutical composition for injection according to any one of claims 7 to 9 of the scope of the patent application, wherein the concentration of arsenic trioxide is from 0.01 to mg / g. 11. The pharmaceutical composition for injection according to item 10 of the scope of patent application, wherein the concentration of the di-emulsified mono-god is 0.1 to 1 mg / g. 12. The pharmaceutical composition for injection according to item .9 of the application, wherein the emulsifier is mannitol. 13. The pharmaceutical composition for injection according to item 9 of the application, wherein the stabilizer is citric acid or ascorbic acid. O: \ 72 \ 72584-931027.DOC \ su ^ ΐ Paper Ruler G.CNS) Specifications (21GX297 Gongcheng)