WO2000053212A1 - Pharmaceutical composition containing cyclosporin - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing cyclosporin, the active ingredient of which is cyclosporin, which characterized in that choosing a pharmaceutical organic acid such as medium or long chain saturated or unsatured fatty acid, substituted carboxylic acid etc., or an admixture of one or more acids or fish oil as a hydrophobic ingredient, depending on the need of various dosage form, not adding water or adding appropriate amount water to made a hydrophilic medium, the formulation of the composition is suitable in preparation of dosage forms such as soft capsule, paste, eye dops, oral liquid and injection etc.

Description

 FIELD OF THE INVENTION

 The present invention relates to a cyclosporine-containing pharmaceutical composition, the active component of which is cyclosporine; ethanol or propylene glycol or a mixture thereof as a solvent or a surfactant auxiliary; an HLB (hydrophilic-lipophilic balance) value of 10 to 19 The hydrophilic surfactant is a solubilizer; it is characterized by the selection of medium or long chain saturated or unsaturated fatty acids, substituted carboxylic acids and other pharmaceutically acceptable organic acids or a mixture of one or more of these acids or fish oil as the lipophilic component ; According to the different requirements of each dosage form, the hydrophilic matrix can be made without adding water or adding appropriate water. The composition can be applied to the preparation of dosage forms such as soft gum liniments, ointments, eye drops, oral solutions and injections.

 More specifically, the present invention relates to a composition of poorly soluble drugs such as cyclosporine, and the formulation of the composition can be applied to the preparation of dosage forms such as soft gel tinctures, ointments, eye drops, oral solutions, and injections.

Background technique

Cyclosporine, also known as Cyclosporin A (Cyclosporin Δ or cyclossporine, referred to as CsA), is a cyclic polypeptide compound consisting of 11 amino acid residues. It is mainly used in organ transplantation and bone marrow transplantation for clinical resistance. Exclusion therapy and treatment of autoimmune diseases. It acts on the early stage of lymphocyte proliferation, and its repressive effect on cells is reversible. It does not affect the hematopoietic function of the bone marrow, does not cause bone marrow suppression, leukocytes and thrombocytopenia. It is a new and highly effective immunosuppressant. Cyclosporine is a white crystalline powder with a molecular weight of 1202.64. It is a highly lipophilic and hydrophobic substance and is almost insoluble in water. It is easily soluble in organic solvents such as methanol, ethanol, acetone, ether, and chloroform. Due to the above reasons, cyclosporine is rarely absorbed by the human body after oral administration, and its bioavailability is poor. In the early 1980s, Sandoz scientists -After dissolving the spores in ethanol, it is mixed with a surfactant Labraf il M 1944 CS with an HLB value of 3 and a vegetable oil to prepare an oily preparation. This formulation becomes an emulsion after adding water, but the emulsion is very unstable, and it is usually temporarily diluted with milk or fruit juice before taking. The oral bioavailability of this preparation is largely affected by the body condition of the patient, which varies greatly from individual to individual, with a relative bioavailability of 4 to 60%. This preparation is difficult to maintain an effective therapeutic concentration. In addition, the main toxic and side effects of cyclosporine are liver and kidney toxicity. Increasing the dosage will also increase the toxic and side effects. Because this preparation uses ethanol as the solvent, the ethanol content will change with time during the processing and storage of soft capsules, which will cause the crystallization of cyclosporine, thereby reducing the bioavailability of cyclosporine, so the stability Worse.

 In order to improve the bioavailability of cyclosporine, reduce the disparity in bioavailability between individuals and the inconvenience caused by diluting the cyclosporine liquid composition with fruit juice or milk before oral administration, many researchers have tried to develop a This enables cyclosporine to be distributed in water in a molecular state, so that the absorption of cyclosporine in the human body is not affected by the amount of bile secreted and the fat content of the food used, thereby reducing the individual differences of cyclosporine and improving A new cyclosporine formulation for its bioavailability. To this end, Sandoz Pharmaceuticals has developed a new cyclosporine liquid composition and made a new oral solution and a soft gel tincture. Hanmi Pharmaceutical Industries has also developed a new prescription. Cyclosporine soft capsules, both capsules and the new oral solution of "San Taoist" are currently available on the market under the trade names "Sandimmun Neoral" and "Implanta" ) ".

Object of the invention

 An object of the present invention is to find and develop a cyclosporine liquid composition capable of making cyclosporine more stable.

 The present invention is based on the research on the solubilization of poorly soluble drugs.

-1- The combination of surface activity, surfactant, solubilizer, and oily components has been studied for a long time, and a new cyclosporine liquid composition has not been discovered in the prior art. This composition can make cyclosporine more stable, and it is not affected by the migration of surfactant ingredients during processing and storage. It can maintain the specific ratio even in the presence of water or the packaging is not sealed. Better stability of existing cyclosporine formulations. The proportion of the active ingredient cyclosporine in the composition can be from 0.5 to 15% (weight ratio). Within this range, the emulsification of the composition in water is excellent. The bioavailability of the new combined cyclosporine oral preparation is greatly improved compared to the original cyclosporine preparation, and it has consistent bioequivalence with commercially available Xinshan Diming. Brief description of the invention

 The invention relates to a cyclosporine-containing pharmaceutical composition, which includes:

 (1) cyclosporine as an active component;

 (2) ethanol or propylene glycol or a mixture thereof as a solvent or a surfactant auxiliary;

 (3) a hydrophilic surfactant having a HLB value of 10 to 19 as a solubilizer;

 (4) pharmaceutically acceptable organic acids such as medium or long-chain saturated or unsaturated fatty acids, substituted carboxylic acids, or a mixture of one or more of the acids or fish oil as lipophilic components;

 (5) According to the different requirements of each dosage form, the hydrophilic matrix can be made without adding water or adding appropriate water;

 The composition can be prepared into dosage forms such as soft gel liniments, ointments, eye drops, oral solutions and injections.

 Detailed description of the invention

According to the present invention, the present invention relates to a cyclosporine-containing pharmaceutical composition, which includes: (1) cyclosporine as an active component;

 (2) ethanol or propylene glycol or a mixture thereof as a solvent or a surfactant auxiliary;

 (3) a hydrophilic surfactant having a HLB value of 10 to 19 as a solubilizer;

 (4) Medium-long chain saturated or unsaturated fatty acids, substituted carboxylic acids, or other pharmaceutically acceptable organic acids or mixtures of one or more of these acids or fish oils as lipophilic components;

 (5) According to the different requirements of each dosage form, the hydrophilic matrix can be made without adding water or adding appropriate water;

 The composition can be prepared into dosage forms such as soft gel liniments, ointments, eye drops, oral solutions and injections.

 According to the present invention, the solvent or surfactant auxiliary as the active ingredient escapes from ethanol or propylene glycol or a mixed solution of ethanol and propylene glycol, which have good solubility for fat-soluble drugs, wherein the ratio of ethanol to propylene glycol is 1: 0.1 to 10 (Weight ratio), preferably the ratio of ethanol and propylene glycol mixed with the co-solvent is 1: 0.5-5 (weight ratio), and the optimal ratio is 1: 1 to 3 (weight ratio).

 According to the present invention, the solubilizer of the fat-soluble drug of the present invention is selected from pharmaceutically acceptable hydrophilic surfactants having an HLB value of 10 to 19 to promote the equilibrium between the hydrophilic and lipophilic portions of the composition to form a stable emulsion. Such surfactants with an HLB value of 10 to 19 are polyoxyethylene castor oil derivatives, such as Cremophor EL, Cremophor RH40, Cremophor 60, or Tweens, such as Tween 80, Tween 65, Tween 20, or Meze. , Such as Myrj 52. Particularly preferred are polyoxyethylene castor oil derivatives.

According to the present invention, the pharmaceutical composition of the present invention is characterized by: Of pharmaceutically acceptable organic acids such as saturated or unsaturated fatty acids, substituted carboxylic acids, or a mixture of one or more of these acids, or fish oil as the lipophilic component. The application of this oily ingredient makes the composition more stable and simpler than other compositions of the original invention. In the present invention, medicinal organic acids such as medium-long-chain saturated or unsaturated fatty acids, substituted carboxylic acids, or a mixture of one or more of the acids or carboxylic acids in fish oil may be in an ester state with the alcohols in the composition. Exist, or it can exist in a free state. The medium and long-chain saturated fatty acids as the oily component are C _{8 -28} carboxylic acids containing 8 to 28 carbon atoms, and the medium and long chain unsaturated fatty acids as the oily component are C containing 1 to 24 carbon atoms. _{1 () -24} mono-, di-, tri-enoic acid, substituted carboxylic acid lactic acid, 70% DHA fish oil, etc. Preferably the oil component is a fatty acid containing 14-- (I, II, triene) acid or mixture of saturated and unsaturated ₂₂ to 22 carbon atoms.

 According to the present invention, another feature of the pharmaceutical composition of the present invention is that according to different uses of the composition, water may not be added, and a certain proportion of water may be added to form a hydrophilic matrix, wherein the ratio of the active component to water is 1: 0 to 1000 (weight ratio). If a certain proportion of water is added to the cyclosporine oral solution, the temperature at which the composition solidifies or flocculates can be reduced, so that it can still remain turbid at relatively low temperatures. This feature can also be used in the preparation of hydrophilic ointments and eye drops.

 According to the present invention, according to different clinical needs, the formulation formula containing cyclosporin or other fat-soluble drugs, the composition can be suitable for encapsulation of gelatin into soft capsules, and can also be made into ointments, eye drops, Oral solutions and injections.

 When the composition of the present invention is made into various dosage forms, as needed, auxiliary materials or additives required for the specific dosage form may be added, such as antioxidants, flavoring agents, penetration enhancers, pH regulators, preservatives, etc. , Not restricted by the content listed. The preparation of different dosage forms can be performed according to the conventional requirements of the dosage form.

The following examples will explain the present invention in more detail. However, it should It is understood that these examples are not intended to limit the present invention.

 Example 1. Preparation of cyclosporine oral solution

Component Dosage (g) Cyclosporine 100 ethanol and propylene glycol mixed solution 230 polyoxyethylene castor oil 400 oleic acid 220 vitamin E 2 distilled water balance 1000 ml

Example 2. Preparation of Cyclosporin Capsules

 Components Amount (g) Cyclosporine 50 Mixed liquid of ethanol and propylene glycol 100 Polyoxyethylene castor oil 200 Refined fish oil 130 1000 capsules capsules Example 3. Preparation of cyclosporine eye drops

Component dosage (g) Cyclosporine 20 ethanol and propylene glycol mixed solution 50 5 1 Polyoxyethylene castor oil 90 Stearic acid 60 Vitamin E 1

 Saline

 1000 liters of the oral solution prepared in the above Example 1 (that is, the neocyclosporine oral solution hereinafter-a tube called neospin) and the original prescription cyclosporine capsule (referred to as sispin capsule) The relative bioavailability of the human body was compared with the gelatine tincture of Johor Bahru. The results are as follows:

Twelve male healthy volunteers were orally administered cyclosporine capsules, which were developed and provided by Hangzhou Zhongmei Huadong Pharmaceutical Co., Ltd.-Cyspin capsules (called Cyspin capsules, sold in the market) and Xinhuan Pharmacokinetic parameters of sporesin oral liquid-a-Xinsisiping (tube called Xinsisiping) and commercially available imported microemulsified cyclosporine capsules——Xinshandi gelatin capsule Compare. Whole blood concentration was measured by HPLC method, using 3P87 and NDST programs, pharmacokinetic analysis and paired t test were performed according to statistical moments. The results show that the AUC (area under the plasma concentration-time curve, the main parameter for determining bioavailability) after oral administration of cysteinpine peptone, cynothepine, and Xinshandi gelatin glutamate were 11.43 ± 2. 48, 16 77 ± 2. 49 and 16. 39 ± 3. 54mg / l * h; Cmax (concentration when blood concentration reaches peak value) 1. 56 ± 0. 25, 2. 38 ± 0. 38 and 2. 47 ± 0. 42mg / l; Tmax (time when blood concentration reaches its peak) 2. 04 ± 0.54, 2.00 ± 0.56 and 1.62 ± 0. 38h. The AUC, Cmax, and Tmax in the main pharmacokinetic parameters of the three are significantly different from that of Sispin and Xinsiping and Xinshan gelatin; but there is no significant difference between Xinsping and Xinshanji Sexual difference. Relative production of Sethping gelatine, New Sethping gelatine and commercially available Xinshan gelatine The bioavailability was 73.4 ± 25.2% and 105.0 ± 17.9%, respectively. The results showed that Xinsaispin and Xinshandi gelatin tincture were bioequivalent in humans.

 Twelve adult male healthy volunteers were selected for the experiment. After a clinical laboratory examination of blood, urine routine, liver and kidney function, electrocardiogram and related immunological indicators were normal, after fully understanding the drug's pharmacology, efficacy and adverse reactions, sign the informed consent form of the volunteer. No medication was taken within two weeks before and during the test. During the test period, no drugs were allowed, smoking, alcohol and tea were not allowed, and they did not participate in heavy exercise activities.

 Cyclosporine reference, provided by Hangzhou Zhongmei Huadong Pharmaceutical Co., Ltd. (9201C, Zhejiang Pharmaceutical Inspection, purity 0. 976 mg I mg)

 The internal standard, cyclosporine D (referred to as CsD, is another component in cyclosporin as an internal standard), with a purity of 98.4%, is provided by the Sichuan Institute of Antibiotic Industry and formulated into 1.2 mg / l of Solution standby

 The acetonitrile used in the experiment was pure domestic chromatographic grade I and was produced by Zhejiang Huangyan Chemical Experiment Factory. The remaining reagents and organic solvents are domestic analytical grades. The methanol used to prepare the purified mobile phase needs to be re-distilled. The water used is ultrapure water.

 The high-performance liquid chromatograph is Shimadzu LC-6A, including SPD ~ 6AV visible ultraviolet detector; CT0-6A column oven; SCL-6A system controller; FCN-2AH high-pressure switching valve; CR-4A data processor. Domestic LD4-2 centrifuge and TGL-16 high-speed centrifuge. P-133 microcomputer.

 On the eve of the test, fasting was performed after dinner to 4 hours after administration. Twelve people were randomly divided into three groups of four. Among them, 4 people in the first group took 500 mg (20 capsules) of cyspin capsules, 4 people in the second group took 500 mg (5 ml) of neospin drugs, and 4 people in the third group took 500 mg (20 mg) Grains), the three groups were swallowed (washed) with 300 ml Guozhen liquid. Low-fat meals were taken at a uniform standard 4 hours after administration.

Subjects received 3 cross-dose (Sespin capsules, Neospin, Xinshandiming Capsules), one at a time, each test interval is 7 days. During the experiment, there were rescue measures and doctor supervision.

 Venous blood was collected at 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 8.0, 12 and 24 hours after oral administration. Blood samples were collected in heparin anticoagulation test tubes and stored at -40 ° C for testing after vigorous shaking.

Direct column injection of whole blood samples was used for determination. Take 1ml of whole blood sample into a 5ml plastic centrifuge tube with stopper, add 1.0ml of CsD working solution, 1.0ml of methanol and 1.0ml of n-hexane precisely, mix by vortexing for lmin, centrifuge for 10min (16000r / min), discard the n-hexane layer Remove the supernatant in a 1.5ml centrifuge tube, centrifuge for 10min (16000r / min), and take 1.0ml for analysis. The purification column was an RP2 column (30 X 4.6mm, 25-40μιη), which was self-packed by the homogenization method. The purified mobile phase was methanol: water (65:35), with a flow rate of 1 ml / min. The purification time is 10min, and the tangent time is lmin. The separation was performed using a Shim-pack CLC-0DS column (150 x 6 allowance, 5μιη), and a pre-column (0DS, 10 X 4.6 mm ΙΟμπι) was added. The column temperature was 70 ", and the mobile phase was analyzed. For acetonitrile: 0.02511101 / 1 ammonium phosphate (82:18, ρΗ2.5), flow rate is 1ml / min, detection wavelength is 210nm, instrument sensitivity is 0.02AUFS. The retention times of CsA and CsD measured by this method are 9.2min and 11.2 min. Quantified by internal standard peak height ratio, linear relationship in the range of 0.025 ~ 3mg / l, cadaver 1.04 * 10- 3x- 4.06 * 10-3, R = 0.9999, based on the signal-to-noise ratio of 3, the lowest total The blood test concentration is 0.01mg / l. Within this range, take 1 ml of high, medium and low concentrations of CsA standard solution, add 1 ml of blank whole blood, 1 ml of CsD working solution and 1 ml of n-hexane, and mix by vortexing for 1 min. After the measurement according to the above method, the CsA peak height (H) was obtained. Then, 1 ml of the CsA standard solution of the same concentration was added, 1 ml of water was added, 1 ml of the CsD working solution, and the mixture was vortexed for 1 min. The peak height (H0) of CsA is obtained. The purification recovery rate is H / H0. The average purification recovery rate is 98.5% (Table 1). The average method recovery rate is 99.3% (Table ² ). It was also measured that the intra-day RSD was 2.3%, and the intra-day RSD was 2.6% (Table

3).

 Its average blood concentration-one-time curve is shown in Figure 1. The abscissa indicates the time (hour) after taking the medicine, and the ordinate indicates the blood drug concentration (mg / 1). The triangle points in the figure represent the average blood concentration time curve after taking Xinshan Diming soft capsule, the square points represent the average blood concentration time curve after taking Sispin soft capsule, and the diamond points represent new The mean blood concentration-time curve after cysteinpine oral solution.

 Pharmacokinetic parameters and bioavailability F value

(1) The practical pharmacokinetic calculation program 3P87 and the new drug statistical program NDST formulated by the Chinese Pharmacological Society were used for processing on the P-133 microcomputer. According to the changes in blood drug concentration, one- and two-compartment model curve fitting was performed, and the results showed that the pharmacokinetics of oral cyclosporine conformed to the one-compartment model. In the actual calculation, parameters such as AUC and MRT were calculated according to the statistical moment. Cma and Tmax were read out based on the actual blood concentration time data, and T _{1/2 was} calculated according to the one-compartment model. Paired t test was used for statistical processing.

 Among them, the AUC of Sespin capsule, Xin Sespin and imported Xinshandiming were 11. 43 ± 2. 48, 16. 77 ± 2. 49 and 16. 39 ± 3. 54mg / l * h;

1.56 ± 0. 25, 2. 38 ± 0. 38 and 2. 47 + 0. 42mg / l; Tmax 2. 04 ± 0. 54,

2. 00 ± 0. 56 and 1. 62 ± 0. 38h.

(2) Calculation of bioavailability (F value):

 Sethping capsule or New Sething AUC

F = X 100% Johor Bahru Gelatin 嚢 AUC Relative Bioavailability of Sethping Grub, New Sethpin and Johor Bahru Gelatin

(F value) of 7 ³ .4 ± 25.2% and 105.0 ± 17.9%, respectively, calculated according to the mean of 69.7% and 102.3%, respectively. Purification recovery table for determination of CsA content in whole blood by HPLC column switching method (N = 5)

CSA concentration H H0 Purified recovery RSD (%) Average

 (MF / L) Rate (%)

 0.101 729 740 98.51 2.01

 0.504 3566 3641 97.95 1.83 98.51

2.018 14456 14593 99.06 1.05 Determination of CsA content in whole blood by HPLC column switching method Method recovery table (N = 5) Adding amount Measured amount Recovery average SD

 (MF / L) (MG / L) (%) (%)

0.101 0.101 99.81

0.504 0.496 98.41 99.31 0.64

2.018 2.012 99.70

Precision table for determination of CsA content in whole blood by HPLC column switching method Concentration (MF / L) RSD N (time) during the day RSD N (time) during the day

0.102 1.76 5 1.88 3

0.505 1.48 5 2.34 3

2.070 2.27 5 2.57 3

Claims

Rights request 1. A cyclosporine-containing pharmaceutical composition comprising the following main ingredients:  (1) cyclosporine as an active component;  (2) ethanol or propylene glycol or a mixture thereof as a solvent or a surfactant auxiliary;  (3) a hydrophilic surfactant having a HLB value of 10 to 19 as a solubilizer; (4) medium-long-chain saturated or unsaturated fatty acids as a lipophilic component, pharmaceutically acceptable organic acids such as substituted carboxylic acids, or a mixture of one or more of these acids, or fish oil; (5) According to the different requirements of each dosage form, the hydrophilic matrix can be made without adding water or adding appropriate water;  The composition can be prepared into dosage forms such as soft gel liniments, ointments, eye drops, oral solutions, and injections.  2. The composition according to claim 1, wherein as a solvent or a surfactant auxiliary is ethanol, propylene glycol or a mixture of both.  3. The composition according to claim 1, wherein the ratio of ethanol to propylene glycol as the mixed co-solvent is 1: 0.1 to 10 (weight ratio).  4. The composition according to claim 1, wherein the ratio of ethanol to propylene glycol as a mixed co-solvent is 1: 0.5 to 5 (weight ratio).  5. The pharmaceutical composition according to claim 1, wherein the surfactant having an HLB value of 10 to 19 is a polyoxyethylene castor oil derivative or a Tween type or a Meze type. 6. A pharmaceutical composition according to claim 1, wherein the saturated fatty acid as the oil component the long chain carboxylic acid is C ₈ _ _28. 7. A pharmaceutical composition according to claim 1, wherein the oil component as long-chain unsaturated fatty acid is a C, di-, tri- acid _{1 ()} _ _24. 8. The pharmaceutical composition of claim 7, wherein as the oil component c polyunsaturated fatty acid is mono-, di-, tri- acid ₁₄ _ _22.  The pharmaceutical composition according to claim 1, wherein the substituted carboxylic acid as the oily component is lactic acid.  The pharmaceutical composition according to claim 1, wherein the fish oil as the oily component is fish oil having a DHA content of 70% by weight.  The pharmaceutical composition according to claim 1, wherein a medium-long-chain saturated or unsaturated fatty acid, a substituted carboxylic acid, or a pharmaceutically acceptable organic acid such as an oily component or a mixture of one or more of them is used.  The pharmaceutical composition according to claim 1, wherein the ratio of the active ingredient to water is 1: 0 to 1000 (weight ratio).  The pharmaceutical composition according to claim 1, wherein the composition can be prepared into gelatin-encapsulated soft capsules, ointments, eye drops, oral solutions, injections and the like.