TW200946140A - Stable-type Cucurbitacin medicinal liquid compositions - Google Patents

Abstract

The present invention provides a stable-type Cucurbitacin medicinal liquid compositions, which comprises: Cucurbitacin, liquid-state fat, and vitamin E, wherein the weight ratio between Cucurbitacin and the liquid-state fat is 1:100 ~ 1:5000, and the weight of vitamin E is 1% ~ 50% of that of the liquid-state fat. The liquid-state fat is the fat which is in the liquid state at room temperature, which can be selected from MCT (Medium-Chain Triglyceride), LCT (Long-Chain Triglyceride), STG (structured-triglycerides), soybean oil, peanut oil, olive oil, or coconut oil, or the mixture of two or more than two of them. The liquid-state fats such as MCT, etc. can dissolve Cucurbitacin well to enhance the anti-tumor effect of Cucurbitacin, and to reduce the individual difference. Vitamin E can increase the stability of Cucurbitacin. The molecular-state stable-type Cucurbitacin liquid compositions of the present invention can be further made into soft capsule and liquid-type hard capsule, and can be used for oral administration or external use. By adding emulsifier to make it into emulsion, the present invention can be applied by injection, oral administration or external use.

Description

200946140 IX. Description of the Invention [Technical Fields of the Invention] The present invention belongs to the field of medical technology and is a stable pharmaceutical liquid composition relating to the known substance Cucurbitacins. [Previous technique] Cucurbitacins belong to a class of tetracyclic triterpenes compounds in which 19-methyl groups are present at the C-9 position, mainly distributed in Cucurbitaceae plants, in cruciferous and scrophulariaceae. It has also been found in higher plants such asaceae, Begonia, Du Yingke, and Sijimu. Qiu Minghua, a researcher at the Kunming Institute of Botany, Chinese Academy of Sciences, led a team of scientists to discover 229 kinds of almost all cucurbitacin compounds based on a series of new cucurbitacin found, and proposed a new structural classification model from the previous five categories. The structure type is reclassified into 12 categories and is recognized by the international academic community. At present, the cucurbitacin approved by China is also known as cucurbitacin BE, which is an extract of φ Chinese traditional medicine melon (alias: bitter clove), which mainly contains cucurbitacin B and E. The content of cucurbitacin B in the existing extract is More than 60%, its quality standards are included in China's ministerial standards. Cucurbitacin tablets are used to treat chronic hepatitis ("cucurbita tablets", Pharmacopoeia, 1986, 21 (6): 357), and found in 13 hospitals in Shanghai, Beijing, Chongqing and other places, the effective rate is 75.2. %, the apparent efficiency is 44.6%. It has been observed clinically that cucurbitacin tablets can comprehensively improve the common symptoms and main signs of chronic hepatitis, and have obvious effects of S-GPT, turbidity (TTT, ΖηΤΤ) and biliary redness. Does not cause S-GPT rebound, has a significant corrective effect on protein inversion and high-global egg 200946140 white blood, can also improve the non-specific cellular immunity of patients with chronic hepatitis, no obvious side effects. In addition, cucurbitacin tablets can also be used to treat primary liver cancer. After clinical observation in six hospitals in Shanghai, Beijing, Guangxi, etc., 169 cases were counted, with an effective rate of 9% and a remarkable efficiency of 39%. Clinical observations show that compared with 5-fluorouracil, the former is superior to the control group in improving symptoms, eliminating liver pain, reducing tumors, prolonging survival, and restoring physical strength, and has no side effects of general chemotherapy drugs ( 0 <Treatment of hepatitis, liver cancer new drug cucurbitacin tablets>, Chinese herbal medicine 1987, 18 (10): 21; &lt; Clinical observation of 50 cases of primary liver cancer treated with cucurbitacin&gt;, new drugs and clinical 1984, 3(2): 21- 2 2; "Pharmacological and clinical application of cucurbitacin", Chinese herbal medicine, 1 992, 23 (1 1 ): 605~60 8). Recently, the authors have combined cucurbitacin tablets with chemotherapeutic drugs to treat advanced liver cancer. The median survival time was increased from 6.1±7.12 months to 12.5±7.54 months for single chemotherapeutic drugs. Clinical efficacy of liver cancer>, Cancer, 1 989, 8 (6): 434-436); another literature found that cucurbitacin b, E φ has a strong killing effect on nasopharyngeal carcinoma cells; Long-term medication (6 weeks) can significantly inhibit liver fibrosis and prevent the formation and development of hepatic steatosis and cirrhosis. The Chinese patents that have been filed and published include: “Nano-cucurbitacin preparation drugs and preparation methods” (Patent No.: 0 1 1 03 65 8.3); “cucurbitacin cyclodextrin inclusion compound and its preparation” (patent Case No.: 02 1 5 3 64 7.3 ); "cucurbitacin liposome group and its preparation" (patent number: 02 1 4463 3.4); "cucurbitacin drip nine and its preparation method" (patent number: 2003) 1 0 1 00943.3 ); "cucurbitacin drip nine and its preparation process" (patent number: 200310113362.3) cucurbitacin drug drop nine and its preparation method 20090026 (patent number: 03 1 34669.3); "cucurbitacin liquid type group Fang and its preparation" (Patent No.: 2004 1 002 1 53 6.8); "Prescription of cucurbitacin Nine Formulation and its preparation method" (Patent No.: ZL 20041 00650 85.8); "New cucurbitacin soft capsule and its Preparation process" (Patent No.: 2005 1 0063 660.5); "Chinese medicine cucurbitacin soft capsule and its preparation method" (Patent No.: 200510097038.6); "A cucurbitacin drip nine and its preparation method" (Patent No.: 200510013633.7 ). φ Although there are such a large number of patents and literatures, there is no report on the use of MCT (medium-chain triglyceride, MCT) or other liquid fats to dissolve cucurbitacin, and no vitamin E is used. A report on improving the stability of cucurbitacin. SUMMARY OF THE INVENTION An object of the present invention is to provide a stable cucurbitacin pharmaceutical liquid composition in which a solvent is capable of dissolving cucurbitacin well, and φ should be capable of improving the stability of cucurbitacin. When it is used for the preparation of a medicine, the anti-tumor effect of the medicine can be improved; the individual difference can be reduced; and the soft capsule and the liquid type hard capsule for oral or external use can be further prepared; the emulsifier and the like can be added, and the compound can be compounded according to a conventional method. The molecular state cucurbitacin-stabilized liquid composition prepared by dissolving cucurbitacin in a solvent is dispersed to form an emulsion, which can be injected, orally or externally. It can be used for the treatment of chronic hepatitis, improve the non-specific cellular immunity of patients with chronic hepatitis, increase the concentration of white blood cells, inhibit the proliferation of liver fibrosis, prevent the formation and development of hepatic steatosis and cirrhosis; Liver cancer and nasopharyngeal cancer. 200946140 The technical solution for achieving the above object of the invention is a stable cucurbitacin pharmaceutical liquid composition, characterized in that the composition comprises the following components: cucurbitacin, liquid fat and vitamin E, wherein the weight of cucurbitacin and liquid fat The ratio is 1: 1〇〇~1: 5000; vitamin E is 1%~50% of the weight of liquid fat. [Embodiment] The liquid fat described in the above scheme refers to a fat mash which is liquid at normal temperature, and may be selected from the group consisting of medium chain triglyceride (MCT), long chain triglyceride (LCT), structural oil, soybean oil. , peanut oil, olive oil, or coconut oil, or a mixture of two or more of them. The composition of the above two liquid fats (for example, a mixture of medium chain triglycerides and soybean oil) is such that the weight ratio of the two liquid fats is 1 • 0 〜1 _·10. The present invention recommends that the liquid fat be a medium chain triglyceride, or a mixture of medium chain glycerol triester and soybean oil, or a structural oil, or a mixture of medium chain triglycerides and a structural oil.

The term "structural oil" as used herein, also known as structured-triglycerides (STG), is a triterpene glycerol molecule formed by lactonization of MCT and LCT. For related information, see, for example, (1) Nordenstrom J, Thorne A, Olivercrona A. "Metabolic effects of infusion of a structured-triglyceride emulsion in healthy subjects [J]. Nutrition, 1 995, 1 1 (3 ) :2 6 9-2 74 ; and (2) S ands trp R, H y 11 and er A, Ko rner U, et 200946140 al . Structured triglycerides to postoperative patients · a safety and tolerance study [J]. J Parenter Enteral Nutr, 1993, 17(2): 153-157. The preferred composition ratio of the above medicinal liquid composition is: cucurbitacin / liquid fat = 1: 500~1 : 3000 (weight ratio); vitamin E is the weight of liquid fat 1 %~20%, the better ratio is: Vitamin E is 5%~20% of the weight of liquid fat. The invention recommends: when vitamin E is 1%~9% of the weight of liquid fat, the vitamin E is resistant. Tocopherol with higher oxidizing power. A better combination of the above medicinal liquid compositions is: medium fat triglyceride, cucurbitacin / medium chain triglyceride (MCT) = 1: 800~1 : 1200 (weight) Ratio), while vitamin E is medium chain triglyceride (MCT) 10% by weight: citricin 1 part by weight, medium chain triglyceride (MCT) 800-1200 parts by weight, vitamin E 80-120 parts by weight. Stable cucurbitacin medicinal liquid composition of the present invention The cucurbitacin component is one or more compounds selected from the group consisting of cucurbitacin, which can be extracted from Cucurbitaceae, Cruciferae, Scrophulariaceae, Begonia, Du Yingke, Quercus and some large fungi; It can be said that cucurbitacin can be obtained from the cucurbit of the Cucurbitaceae L. (Cucumismelo L.), which can also be obtained by synthesis or purchase. The better cucurbitacin is cucurbitacin. BE (Received by the Ministry of Health of the People's Republic of China, the standard for the preparation of traditional Chinese medicine preparations in the 19th volume), the main component of cucurbitacin BE is cucurbitacin B, the content of leucovorin B is more than 60%, and strontium may include bismuth e 2009200946140 cucurbitacin B, dihydrocucurin B and other components. In the present invention, cucurbitacin can also be a single compound of high purity, such as HPLC area normalization method to determine the purity of cucurbitacin B greater than 99%, purity More than 95% Lo heterologous cyclin E or cucurbitacin B. The medium chain triglyceride (MCT) is a compound or mixture of a triglyceride of a fatty acid having 8 to 12 linear carbon atoms, more preferably 8 to 10 linear carbon atoms, such as Miglyol 812, Miglyol 810. , Miglyol 818, can also use Miglyol 829 (

Caprylic/Capric/Triglyceride or Miglyol 840 (Proylene Glycol Dicaprylate/Dicaprate, propylene glycol diester of C8~C10 saturated vegetable fatty acids). The MCT recommended for use in the present invention is caprylic/capric glycerin, such as Miglyol 812, Miglyol 810 (see the Medium Chain triglycerides of the European Pharmacopoeia). The vitamin E is a tocopherol, a tocotrienol, a tocopherol or a derivative of a tocopherol (e.g., an ester such as tocopherol acetate), or a mixture thereof. Among them, tocopherol acetate or tocopherol or a mixture thereof is preferred, and a more preferable solution is to use the α-form tocopheryl acetate or tocopherol or a mixture thereof, the stable cucurbitacin medicinal liquid composition for injection, oral or In the preparation of the external emulsion, the composition is directly emulsified or a conventional pharmaceutical excipient is added. For the preparation of the oral or topical preparation, the preparation involved includes a soft capsule or a liquid type hard capsule. -10- 200946140 The liquid fat such as MCT in the present invention can dissolve cucurbitacin well. When used in the preparation of a drug, the anti-tumor effect of the drug can be improved; the individual difference can be reduced; and the use of vitamin E can greatly improve the stability of cucurbitacin. We combine liquid fat (such as MCT) with vitamin E to form a composite solvent, and use the composite solvent to dissolve cucurbitacin to obtain a stable liquid composition of molecular cucurbitacin. The composition can be further prepared into soft capsules and The liquid type hard capsule can be used for oral or external use; the emulsifier and the like can be added, and the molecular state cucurbitacin-stabilized liquid prepared by dissolving cucurbitacin in a complex solvent can be dispersed to form an emulsion according to a conventional method. This emulsion can be injected, alpha or topical. It is used to treat chronic hepatitis, improve non-specific cellular immunity in patients with chronic hepatitis, increase white blood cell concentration, inhibit liver fibrosis, prevent liver steatosis and cirrhosis. It can also be used to treat primary disease. Liver cancer and nasopharyngeal cancer. The formulation of the stable liquid composition is such that the total amount of cucurbitin per unit dosage form is 0.1 mg to 1 mg, and the dose for human use is 0.3 mg ❹ ~3 mg/day, which is administered once to four times. In order to further illustrate the contents of the present invention and its efficacy, the following examples are provided. The examples are provided for illustrative purposes only and are not intended to limit the scope of the invention. EXAMPLES The sources of the materials, equipment and animals used in the examples are described below. Ingredients: Tocopherol acetate (purchased in BSF, Chinese Pharmacopoeia -11 - 200946140 2nd edition of the second edition), tocopherol (purchased in BSF); cucurbitacin BE (purchased in Tianjin Pharmaceutical Research Institute Industry limited liability company); MCT (purchased in Shanghai Dongsheng Industrial Co., Ltd. or Tieling North Asia Pharmaceutical Oil Co., Ltd.), soybean oil (purchased from Tieling North Asia Pharmaceutical Oil Co., Ltd.) ° Reagents: methanol, acetonitrile for chromatographic purity 'Other reagents such as phosphoric acid are analytically pure. ❹ HPLC conditions: column: Diamonsil C18 (20〇x4.6mm, 5μιη, Dima); mobile phase: acetonitrile_water monophosphate (volume ratio 45:55:0.1); UV detection wavelength: 228nm; column temperature: 30 ° C; flow rate: lmL.miiT1; injection amount: 20μΙν. Animals: All animals are from the Animal Department of Shenyang Pharmaceutical University or the Animal Department of China Medical University.

Example 1: Solubility of cucurbitacin BE in soybean oil or MCT Q According to the solubility measurement method, cucurbitacin BE (cucurbitacin B content greater than 60%, commercially available) in soybean oil and MCT (medium chain triglyceride) was determined. The solubility in the results is: about 1.88 mg of cucurbitacin per g of soybean oil, about 3.36 mg of cucurbitacin per g of medium chain triglyceride (MCT), so the solubility of MCT on cucurbitacin BE is about soybean oil. 2 times. Example 2: Antitumor effect of cucurbitacin BE soybean oil or MCT oil solution Using H22 as a model (see Example 20 for establishment and method of animal model), cucurbitacin BE (commercially available, after purification, cucurbitacin B) Content -12- 200946140 Controlled in 62%~65%, the other components are mainly cucurbitacin E, isoquercetin b, dihydrocucurin B) soybean oil solution and MCT oil solution anti-tumor effect (dose according to total cucurbitacin Calcium, 0.2 mg/kg body weight, intragastric administration, continuous administration for 8 days), the tumor inhibition rates were 32.6% and 41.2%, respectively. From this, it can be seen that MCT is used as a solvent to increase the tumor inhibition rate of cucurbitacin. Example 3: Antioxidant action Since cucurbitacin B contains a double bond, oxidative damage is likely to occur, and an antioxidant should be added. The antioxidant effects of adding 5% tocopheryl acetate or 5% sputum as antioxidants in MCT were investigated. The lmg cucurbitacin/gMCT oil solution was prepared and accelerated at 80 °C for 3 days. As a result, the cucurbitacin content decreased by 35% without antioxidants, and the cucurbitacin content decreased when 5% tocopherol or 5% tocopherol was added. About 17%. See Figures 1 to 4. Example 4: Concentration screening of antioxidant vitamin E Considering that vitamin E (this embodiment is tocopherol acetate) is contained in the pharmacopoeia of the People's Republic of China and has pharmaceutical specifications, we conduct in-depth research on vitamin E to MCT. The antioxidant concentrations of 0%, 1%, 5%, 10%, 20%, 30% and 50% were calculated at different concentrations (calculated as (vitamin E/MCT) X 100%). The sample solution containing the above different concentrations of vitamin E was prepared at a ratio of 1 mg of cucurbitacin per gram of MCT. After the samples were destroyed at 8 ° C for 4 days, the reduction of cucurbitacin in each sample solution was determined by HPLC. . Results The cucurbitacin decreased by about 41%, 35%, 28% ' 16%, -13- 200946140, and the antioxidants were 12%, 6% and 2%, respectively. When the vitamin E content was more than 5%, the effect was more obvious. The cucurbitacin content is basically not more than 30%. Example 5 = Preparation Cucurbitin BE solution (the ratio of cucurbitacin to MCT is 1: 1) (vitamin E is 50% of MCT) 〇

Cucurbitacin BE MCT 1 g l〇〇g Vitamin E (tocopherol acetate) 50g Mix MCT with vitamin E evenly, add cucurbitacin, and dissolve in heat to obtain a clear and transparent solution. The solution can be further prepared by the method of preparing soft capsules or liquid type hard capsules, each containing 151 mg | , which is equivalent to 1 mg of cucurbitacin per capsule. Emulsification can be carried out by adding a φ emulsifier to the solution to prepare an emulsion (10% MCT 20% MCT concentration emulsion) for injection, oral administration or external administration. Example 6: The cucurbitacin solution (with a ratio of cucurbitacin to MCT of 1:500) is 40% of MCT) Cucurbitacin BE lg MCT 500g Vitamin E (tocopherol acetate) 200g 8 0°C Conventional solution Or (Wei-14- 200946140 Mix MCT with vitamin E evenly, add cucurbitacin, dissolve it in heat to obtain a clear and transparent solution. This solution can continue to prepare soft capsules or liquid hard capsules according to the method, 425 per capsule Mg ΐ , equivalent to cucurbitin 0.6 mg per capsule. Emulsify with emulsifier of J in this solution to prepare emulsion (5% MCT 10% MCT emulsion) for injection, oral or topical administration 7 0. Conventional solution is appropriate or 〇 Example 7: The cucurbitacin solution (with a ratio of cucurbitacin to MCT of 1:500) is 30% of MCT) Cucurbitacin BE lg MCT 50〇g Vitamin E ( Tocopherol acetate) i5〇g (dimension MCT and vitamin E are mixed evenly 'add cucurbitacin' to dissolve in heat to obtain a clear and transparent solution. This solution can continue to prepare soft capsules or liquid hard capsules according to the method of sputum. 200 mg rich, equivalent to each grain There is cucurbitacin · 3 mg. Add 7 emulsifier to the solution to emulsifie 'can be prepared as an emulsion (5% MCT 10% MCT concentration emulsion)' for injection, oral or topical administration. 70 °C routine The solution is appropriate or Example 8: The formulation of the sucrose solution (the ratio of leucovorin to MCT is 5 〇〇) (V--15-200946140 Biotin E is 20% of MCT) Cucurbitacin BE lg MCT 500g Vitamin E (Acetic acid Tocopherol) i〇〇g Mix MCT with vitamin E evenly, add cucurbitacin, and dissolve it at 70 ° C to obtain a clear and transparent solution. This solution can continue to prepare soft capsules or liquid hard capsules according to the conventional method. Packing 3 60 mg of this solution, equivalent to 0.6 mg of cucurbitacin per capsule. Emulsifying the solution with an appropriate emulsifier to prepare an emulsion (5% MCT or 10% MCT emulsion) for injection Oral, topical administration. Example 9: Preparation of cucurbitacin solution (with a ratio of cucurbitacin to MCT of 1:1000) (vitamin E is 10% of MCT) _ Cucurbitacin BE lg MCT 1 000g Vitamin E (Tocopherol acetate) L〇〇g Mix MCT with vitamin E evenly, add 葫The solution is heated and dissolved at 60 ° C to obtain a clear and transparent solution. The solution can continue to prepare soft capsules or liquid hard capsules according to a conventional method, each of which contains 333 mg of the solution, which is equivalent to cucurbitacin·3 in each capsule. The emulsion can be prepared by adding an appropriate emulsifier to the solution to prepare an emulsion (10% MCT or -16 - 200946140 20% MCT concentration emulsion) for injection or oral administration. Example 1 Preparation: Preparation of cucurbitacin solution (with a ratio of cucurbitacin to MCT of 1:200) (vitamin oxime is 10% of MCT) Cucurbitacin BE 1 g MCT 200〇g 维生素 Vitamin E (tocopherol acetate) 200g MCT and vitamin E are evenly mixed, and cucurbitacin is added and dissolved at 60 ° C to obtain a clear and transparent solution. The solution can be further prepared by a conventional method of preparing soft capsules or liquid type hard capsules, each containing 440 mg of this solution, which corresponds to cucurbitacin 2 mg per pellet. Emulsification can be carried out by adding an appropriate emulsifier to the solution to prepare an emulsion (10% MCT or 20% M C T concentration emulsion) for injection or oral administration.实施 Example 11:

Preparation of cucurbitacin solution (the ratio of cucurbitacin to MCT is 10% of MCT) 1 : 3000 ) (Vegeurin be 1 g MCT 3000g Vitamin E (tocopherol acetate) 3 00g Mix MCT with vitamin E evenly ' Add cucurbitacin, dissolve at 60 ° C -17- 200946140 to obtain a clear and transparent solution. This solution can continue to prepare soft capsules or liquid hard capsules according to the conventional method, each 330 mg of this solution 'equivalent to each It contains cucurbitacin 0.1 mg. Emulsification can be prepared by adding an appropriate emulsifier to the solution to prepare an emulsion (30% MCT concentration emulsion) for injection or oral administration. Example 1 2 : Mesoruthenium solution ( Preparation of cucurbitacin and MCT ratio of 1:5000) (vitamin £ is 5% of MCT) Cucurbitacin BE lg MCT 5000g Vitamin E (tocopherol acetate) 250g Mix MCT with vitamin E and add cucurbitacin at 5〇 °C heating to dissolve to obtain a clear and transparent solution. The solution can continue to prepare soft capsules or liquid type hard capsules according to the conventional φ method. Each capsule contains 520 mg of this solution, which is equivalent to containing O.lmg in each capsule. The solution The medium is forced into an appropriate emulsifier for emulsification, and an emulsion (5% MCT or 10% MCT emulsion) can be prepared for injection or oral administration. Example 1 3: Cucurbitacin solution (cucurbitacin to MCT ratio) Formulation for 1:5000) (vitamin E is 1% of MCT) Cucurbitacin BE ^ -18- 200946140 5000g 50g

MCT Tocopherol ❹ Mix MCT with tocopherol, add cucurbitacin, and dissolve at 40 °C to obtain a clear and transparent solution. The solution can be further prepared by a conventional method of preparing soft capsules or liquid type hard capsules, each containing 510 mg of the solution' equivalent to 0.1 mg of cucurbitacin per capsule. Emulsification can be carried out by adding an appropriate emulsifier to the solution to prepare an emulsion (10% MCT or 25% M C T concentration emulsion) for injection or oral administration. Example 1 4: Preparation of a solution of cucurbitacin B (purity greater than 99% by HPLC area normalization) (the ratio of cucurbitacin to MCT is 1:1,000) (vitamin Ε is 10% of MC )) B lg ❹

MCT Vitamin E (tocopherol acetate) 1 000g 100g MCT and vitamin E are mixed evenly, and cucurbitacin is added and dissolved at 60 ° C to obtain a clear and transparent solution. The solution can be further prepared according to a conventional method of preparing a soft capsule or a liquid type hard capsule, each containing 220 mg of the solution, which corresponds to 0.2 mg of cucurbitacin per capsule. Emulsification can be carried out by adding an appropriate emulsifier to the solution to prepare an emulsion (10% MCT or 20% MCT emulsion) for injection, oral administration or external administration. -19- 200946140 Example 1 5: Preparation of cucurbitacin E (purity greater than 95% by HPLC area normalization) solution (with a ratio of cucurbitacin E to MCT of 1:1,000) (vitamin E is MCT 10) % ) Glucosamine E 1 g MCT 1 000g Vitamin E (tocopherol acetate) l〇〇g ❹ Mix MCT with vitamin E evenly, add cucurbitacin E, and dissolve at 60 °C to obtain a clear and transparent solution. The solution can be continued to prepare soft capsules or liquid type hard capsules according to a conventional method, each containing 33 0 mg of the solution, which corresponds to cucurbitacin E 0 • 3 mg per capsule. Emulsification can be carried out by adding an appropriate emulsifier to the solution to prepare an emulsion (10% MCT~20% MCT concentration milk emulsion! I), which can be administered by injection, oral or topical administration.实施 Example 1 6: Formulation stability test The cucurbitacin oil solution obtained by the above several different examples (the solution before the preparation of the emulsion without adding an emulsifier) is at 4 (TC force # speed # @, the results are shown in the table). 1. -20- 200946140 Table 1 Drug Stability Test Results Time (Month) Example 5 Example 7 Example 8 Example 9 Example 10 Example 11 0 100.0% 100.0% 100.0% 100.0% 100.0% 100.0% 1 99.8 % 99.1% 98.5% 99.5% 98.6% 98.5% 2 98.8% 98.6% 99.0% 99.2% 99.7% 97.9% 3 99.5% 99.3% 98.7% 98.9% 99.3% 99.6% It can be seen that the prepared cucurbitacin oil solution is very stable. Example 1 7: Preparation of cucurbitacin BE (manufactured by cucurbitacin B content greater than 60%, commercially available) Cucurbitin be 1 g MCT 1 000 g Vitamin E (tocopherol acetate) 50 g Phospholipid 120 g Glycerol 220 g Water added to 10000 ml Prepared according to conventional procedures Emulsion method The emulsion is prepared, the concentration of cucurbitin is 〇. 1 mg/ml; the prepared emulsion can be injected, orally or externally. Example 1 8 : Cucurbitacin B (purity greater than 99% by HPLC area normalization method) Preparation of Emulsions-21 - 200946140 Meng Lusu B 1 g MCT 1 〇〇g tocopherol 50g o-lip 1 0 0 g HS 15 (polyethylene glycol stearate 15) l〇g glycerol 220g hydraulic pressure to 1 0 0 0 m 1

The emulsion is prepared according to a conventional method for preparing an emulsion, and the concentration of cucurbitacin is 1 mg/ml; the prepared emulsion can be injected, orally or externally. Example 1 9 : Cucurbitacin E (purity greater than 95% as determined by HPLC area normalization) Preparation of emulsion Cucurbitacin E 5 g MCT 500 g Tocopherol 50 g Phospholipid 60 g HS 15 1 OOg Poloxamer F 6 8 2 0 0 g Glycerin 200g Water is added to 1 0 0 0 m 1 The emulsion is prepared according to a conventional method for preparing an emulsion, and the concentration of cucurbitacin is -22-200946140 0 · 5 mg/ml; the prepared emulsion can be injected, orally or externally used. Poloxamer is a polyoxyethylene block copolymer sold under the trade name Pluronic by BSF, Germany. Example 20: Antitumor pharmacodynamics (repeated three times) Establishment of a (-) tumor-bearing mouse model In vitro resuscitation of liver cancer H22 tumor cell, in an index set of cells, centrifuged at a speed of l〇〇〇r/min, The supernatant was washed twice with PBS, diluted with sterile physiological saline, and adjusted to 2 x 107 cells/ml of tumor cells. Healthy mice were selected, each of which was intraperitoneally injected with the above 0.2 m1. Note that one week after the ascites of the inoculated mice was observed, the abdomen of the inoculated mice was significantly enlarged, and the bulging was taken as the ascites milk white, diluted with sterile physiological saline in a sterile test tube. The tumor fineness φ was 1χ107/ml, and each was subcutaneously inoculated with 0.2 ml' to establish a H22 right subcutaneous inoculation model. (II) Experimental grouping and administration The mice with the established model were randomly divided into groups of 10 and 20 mice each, and numbered. The model control group (administered CMC-Na solution): cucurbitacin commercially available tablet group (硏 fine' in suspension, gavage, dose according to total cucurbitacin as 〇.2 group (administered dose according to total cucurbitacin) Calculated as 〇·2 mg kgd). Polyene oxypropylene. The product is harvested in the long-term, centrifuged to the concentration of the cell suspension, about ascites is the cell concentration of mouse liver cancer (control group volume of CMC-Na) MCT self-modeling -23- 200946140 The second day after the start of the administration, continuous administration ίο天, once a day. (3) Weigh the tumor and calculate the tumor inhibition rate for 24 hours, the cervical spine dislocation method to kill the animal, said Body weight, dissection of the tumor block, weighing the tumor with an electronic balance. Calculate the tumor inhibition rate according to the following formula: tumor inhibition rate = (1 - the average tumor weight of the drug-administered group / the average tumor weight of the control group 1%)値Use standard deviation to express (i±SD), result Table 2, mouse transplanted liver cancer H22 tumor inhibition result (X Shi SD) group animal body weight (g) tumor weight (g) tumor inhibition rate (%) (start / Finally) start the last (7 ± sd) model control group 20/20 19.66 23.18 2.56 ± 0.51 tablet group (1) 10/10 19.13 22.65 1.61±0.38 37.1 Tablet group (2) 10/10 18.96 23.31 1.76±0.43 31.2 Tablet group (3) 10/10 20.13 22.77 1.78±0.53 30.5 MCT group (1) 10/10 19.51 23.02 1.51±0.25 41.0 MCT Group (2) 10/10 19.62 22.88 1.57±0.31 38.7 MCT group (3) 10/10 19.73 23.22 1.45±0.28 43.3 Note: The MCT group was tested with the cucurbitacin oil solution of Example 9. The test results show that the tablet group The difference in tumor weight was large (s D 0.38' 0.43> 0.53), and the tumor inhibition rate was low (3 7.1 %, 3 1 · 2 % ' 3 0.5%, respectively): the difference in tumor weight was smaller in the MCT group. (SD is 〇·25 -24- 200946140, 0.31, 0.28), the tumor inhibition rate is higher (41.0%, 38.7°/., 43.3%, respectively), indicating that cucurbitacin is absorbed after cucurbitacin is dissolved by MCT. Improvement, individual difference was reduced, and curative effect was improved. Example 21 was substantially the same as Example 13, but the tocopherol was 9% (450 g) of MCT. Example 22 was substantially the same as Example 5, but MCT was changed therein. Using a mixture of MCT and soybean oil, the weight ratio of MCT to soybean oil is 1 ▲ : 1 〇; vitamin E is tocopherol acetate, vitamin E Medium chain triglycerides with 1% by weight of a mixture of soybean oil Gan large square; wherein cucurbitacin melon plants using gourd Cucumismelo L. stalk, stem end, Guadi extraction refining cucurbitacin obtained. Example 23 is substantially the same as Example 5, but wherein MCT is changed to a mixture of MCT and soybean oil, the weight ratio of MCT to soybean oil is 1:9; and the vitamin E is α-type tocotrienol and The dosage is 3% by weight of the mixture of medium chain triacetin and soybean oil; the cucurbitacin is cucurbit Φ. The content of cucurbitacin in the cucurbitacin BE is greater than 60%. Example 24' is substantially the same as Example 5, but wherein the MCT is changed to a mixture of MCT and soybean oil, the weight ratio of MCT to soybean oil is 1:8; the vitamin E is an ester derivative of tocopherol and The dosage is 5% by weight of the mixture of MCT and soybean oil; the cucurbitacin also contains cucurbitacin E, isoliquirin B, and dihydrocucurin. Example 25' is substantially the same as Example 5, but in which the MCT is changed to a mixture of MCT and soybean oil, the weight ratio of MCT to soybean oil is! The vitamin E described in '7' is an ester derivative of the second tocopherol, which is go -25- 200946140 g 〇 Example 26, which is basically the same as Example 5, but the MCT thereof is changed to a mixture of MCT and soybean oil. The weight ratio of MCT to soybean oil is 1:6; the vitamin E is a mixture of tocopherol acetate and tocotrienol. Example 27 is substantially the same as Example 5, but wherein the MCT is changed to a mixture of MCT and soybean oil, and the weight ratio of MCT to soybean oil is 1:5 ° 实施 Example 28, which is substantially the same as Example 5, but wherein The MCT was changed to a mixture of MCT and soybean oil. The weight ratio of MCT to soybean oil was 1:4 〇 Example 29, which was basically the same as Example 5, but the MCT was changed to a mixture of MCT and soybean oil, MCT and large. The weight ratio of soybean oil was 1:3 = Example 30, which was basically the same as Example 5, but the MCT was changed to a mixture of MCT and soybean oil, and the weight ratio of MCT to soybean oil was 1 〇: 2 °. 31, substantially the same as Example 5, but wherein the MCT is changed to a mixture of MCT and soybean oil, and the weight ratio of MCT to soybean oil is 1:1 ° Example 32, which is basically the same as Example 5, but the MCT therein is changed. Use Miglyol 829. Example 33 is substantially the same as Example 5, but the MCT therein is changed to Miglyol 840. -26- 200946140 Example 34, which is substantially the same as Example 5, but in which the vitamin E is changed to a mixture of tocopherol acetate and tocopherol 1:1. Example 35' is substantially the same as Example 5 except that the weight ratio of cucurbitacin/medium chain triglyceride is 1:800; vitamin E is 10 g. Example 36' is substantially the same as Example 5 except that the weight ratio of cucurbitacin/medium chain triglyceride is 1:1200; vitamin e is 1〇〇g. Example 3 7 is basically the same as Example 5 except that the liquid fat therein is a long-chain triglyceride (LCT).实施 Example 3 8 is basically the same as Example 5, but the liquid fat therein is a structural oil. Example 3 9 ' is basically the same as Example 5, but the liquid fat therein is soybean oil. Example 40' is substantially the same as Example 5 except that the liquid fat therein is peanut oil. Example 4 1 ' is basically the same as Example 5, but the liquid fat 其中 therein is olive oil. Example 42 was substantially the same as Example 5 except that the liquid fat therein was coconut oil. Example 4 3 is substantially the same as Example 5 except that the liquid fat therein is a mixture of a structural oil and a medium chain triglyceride 1. Example 44 is substantially the same as Example 5 except that the liquid fat therein is a mixture of soybean oil, structural oil and medium chain triglyceride 1: 1:1. Example 4 5 ' is substantially the same as Example 5' but the liquid fat therein is a mixture of structural oil, medium chain triglyceride and long chain triglyceride (LCT) 1: 1 -27- 200946140: 〇. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a HPLC diagram of 0 at the time when cucurbitin was not thermally destroyed in Example 3. Fig. 2 is an HPLC after rupture of a sample of cucurbitacin without vitamin bismuth in Example 3 at 80 °C for 3 days. In the cucurbitacin sample solution, 1 mg of cucurbitin per gram of MCT is contained; FIG. 3 is an HPLC diagram of the cucurbitacin sample solution of 5% acetic acid tocopherol added in Example 3 after being destroyed at 80 ° C for 3 days, wherein cucurbitacin 1 mg of cucurbitacin per gram of MCT in the sample solution; and FIG. 4 is an HPLC diagram of the cucurbitacin sample solution added with 5% tocopherol in Example 3 after being destroyed at 80 ° C for 3 days, wherein each gram of MCT in the cucurbitacin sample solution Contains 1 mg cucurbitacin. -28-

Claims (1)

200946140 X. Patent application garden 1. A stable cucurbitacin pharmaceutical liquid composition, characterized in that the composition comprises the following components: cucurbitacin, liquid fat and vitamin E, wherein the weight ratio of cucurbitacin to liquid fat is I : 100 ~ 1 : 5000 ; Vitamin E is 1% to 50% by weight of liquid fat. 2. The stable cucurbitacin pharmaceutical liquid composition according to claim 1, wherein the liquid fat is a fat which is liquid at room temperature, and is selected from the group consisting of medium chain triglycerides, long chain triglycerides, Structural oil, soybean oil, 〇 peanut oil, olive oil, or coconut oil, or a mixture of two or more of them. 3. The stable cucurbitacin pharmaceutical liquid composition according to claim 2, wherein the liquid fat is a mixture of two liquid fats and the weight ratio of the two liquid fats is 1: 0 ~ 1 : 1 0 . 4. The stable cucurbitacin pharmaceutical liquid composition according to claim 2, wherein the liquid fat is medium chain triglyceride, or a mixture of medium chain glycerol and soybean oil, or medium chain triglyceride. The cucurbitacin medicinal liquid composition according to any one of claims 1 to 4, wherein the pharmaceutically acceptable liquid composition has a component weight ratio of cucurbitacin/liquid Fat = 1: 500 ~ 1: 3000; Vitamin E is 1 ° / weight of liquid fat. ~20%. 6. The stable cucurbitacin pharmaceutical liquid composition of claim 5, wherein the vitamin E is 5% to 20% by weight of the liquid fat. 7. For example, the stable cucurbitacin medicinal liquid group -29-200946140 of the scope of patent application 5 wherein the liquid fat uses medium chain triglyceride; the weight ratio of cucurbitacin to medium chain triglyceride is 1: 800~1: 1200; Vitamin E is 10% by weight of medium chain triglyceride. 8. The stable cucurbitacin pharmaceutical liquid composition according to claim 4, wherein the pharmaceutical liquid composition has a component ratio of: cucurbitacin 1 part by weight, medium chain triglyceride 800 to 1 200 parts by weight. , vitamin E 80 ~ 120 parts by weight. 9. The stable cucurbitacin pharmaceutical liquid composition according to claim 8, wherein the vitamin E is a tocopherol, a tocotrienol, a tocopherol or a tocotrienol derivative, or a mixture thereof. 10. The stable cucurbitacin pharmaceutical liquid composition according to claim 9, wherein the vitamin E is tocopherol acetate or tocopherol or a mixture thereof. 1 1. A stable cucurbitacin pharmaceutical liquid composition according to claim 10, wherein when vitamin E is from 1% to 9% by weight of the liquid fat, ^ the vitamin E is derived from tocopherol. The stable cucurbitacin pharmaceutical liquid composition according to any one of claims 1, 9, 10, and 11, wherein the vitamin E is α-form acetate tocopherol or α-type tocopherol. 13. The stable cucurbitacin pharmaceutical liquid composition according to claim 8, wherein the medium chain triglyceride is a compound or a mixture of triglycerides of a fatty acid having 8 to 12 linear carbon atoms. 14. The stable cucurbitacin pharmaceutical liquid composition according to claim 13, wherein the medium chain triglyceride is selected from the group consisting of Miglyol 8 12, -30-200946140 Miglyol 810, Miglyol 818, and Miglyol 829 °. 15. The stable cucurbitacin pharmaceutical liquid composition according to claim 14, wherein the medium chain triglyceride is Miglyol 812 or Miglyol 810 〇16. The stable cucurbitacin medicinal liquid according to claim 1 A composition wherein the liquid fat is Miglyol 840. 1 7. The stable cucurbitacin medicinal liquid composition according to claim 1, wherein the cucurbitacin is a cucurbitaceae plant, a cruciferous family, a scrophulariaceae, a begonia family, a eucalyptus, a quaternary plant. And tetracyclic triterpenoids extracted from large fungi. 18. The stable cucurbitacin medicinal liquid composition according to claim 17, wherein the cucurbitacin is obtained by extracting from the stalk, stalk, and cucurbit of the Cucurbitaceae plant Cucumismelo L. 19. The stable cucurbitacin pharmaceutical liquid composition according to claim 17, wherein the cucurbitacin is cucurbitacin BE, high-purity cucurbitacin B or cucurbitacin E, or other kinds of cucurbitacin compounds, The content of cucurbitacin B in cucurbitacin BE is greater than 60%. 20. The stable cucurbitacin pharmaceutical liquid composition according to claim 19, wherein the high purity cucurbitacin B has a cucurbitacin B content greater than 95%; or the high purity cucurbitacin E has a greater content of cucurbitacin E 95%. 21. The stable cucurbitacin pharmaceutical liquid composition according to any one of claims 17 to 20, wherein the cucurbitacin further comprises cucurbitacin E, isoliquirin B, dihydrocucurin B, and the like Cucurbitacin. 22. The composition of the composition of the stabilized cucurbitacin medicinal liquid -31 - 200946140 as claimed in claim i, the form of the composition. 2 3. As claimed in the patent application, the composition is a type of agent. 24. As claimed in the patent application composition, the composition further comprises 1 HS 1 5, poloxamer F 6 8 , hydrazine containing an emulsifier, such that the composition is a stable cucurbitacin medicinal liquid according to item 1 of the emulsion.软 soft capsules, liquid hard capsules, or liquid pharmaceutically acceptable liquids of stable cucurbitacin according to item 1. The conventional pharmaceutical excipients are selected from phospholipids, and/or glycerin. -32-