LONG-ACTING ROPIVACAINE PHARMACEUTICAL COMPOSITION, PREPARATION METHOD THEREFOR AND USE THEREOF
20230080811 · 2023-03-16
Inventors
Cpc classification
A61K9/0019
HUMAN NECESSITIES
A61P29/00
HUMAN NECESSITIES
A61K47/24
HUMAN NECESSITIES
International classification
A61K47/24
HUMAN NECESSITIES
A61K47/44
HUMAN NECESSITIES
Abstract
Provided are a long-acting ropivacaine pharmaceutical composition, a preparation method therefor and use thereof. The pharmaceutical composition comprises: ropivacaine; a pharmaceutical solvent; a pharmaceutical phospholipid; a pharmaceutical oil; and an efficacy enhancer, an optional antioxidant and an optional acid-base regulator. The pharmaceutical composition has a controllable release behavior and a sustained release effect, can significantly reduce the peak plasma concentration of the drug, maintain a stable plasma concentration in the body, prolong the effective treatment time, reduce the effective therapeutic dose, improve the utilization of the drug, and reduce the risk of neurotoxicity. The pharmaceutical composition has a long-acting analgesic effect and can be used for pain treatment.
Claims
1. A long-acting ropivacaine pharmaceutical composition comprising ropivacaine; a pharmaceutical solvent; a pharmaceutical phospholipid; a pharmaceutical oil; an efficacy enhancer; an optional antioxidant and an optional acid-base regulator, wherein the efficacy enhancer is one or a combination of two or more selected from the group consisting of ω-3 fatty acids and metabolites thereof, and substances rich in ω-3 fatty acids or metabolites thereof.
2. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the pharmaceutical composition reduces Ma peak plasma concentration of a drug by 30% or more compared with ropivacaine hydrochloride injection at the same dose; and the pharmaceutical composition has a duration of action of 12 h or more.
3. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises, based on the total weight of the composition, in weight percentage, 0.5% to 10% of ropivacaine; 2% to 25% of the pharmaceutical solvent; 8% to 55% of the pharmaceutical phospholipid; 15% to 89% of the pharmaceutical oil; 0.5% to 10% of the efficacy enhancer; about 0% to 1% of the antioxidant; and about 0% to 8% of the acid-base regulator.
4. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the ω-3 fatty acids and metabolites thereof include ω-3 polyunsaturated fatty acids and metabolites thereof, and wherein the substances rich in ω-3 fatty acids or metabolites thereof include substances rich in α-linolenic acid, substances rich in α-linolenic acid metabolites eicosapentaenoic acid and docosahexaenoic acid, or combinations thereof.
5. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the ropivacaine includes ropivacaine free base and salts thereof, wherein the pharmaceutical solvent is one or more selected from the group consisting of benzyl alcohol, ethanol, propylene glycol, glycerin, isopropanol, N-methylpyrrolidone, dimethyl sulfoxide, liquid polyethylene glycol, dimethylacetamide, glycerol monoacetate, polyethylene glycol monomethyl ether, diethylene glycol monoethyl ether, ethyl lactate, ethyl acetate, propylene glycol diethyl ester, diethyl malonate, tetrahydrofuran polyethylene glycol ether, and benzyl benzoate, wherein the antioxidant is one or more selected from the group consisting of cysteine, α-tocopherol, α-tocopherol acetate, N-acetyl-L-cysteine, butylated hydroxyanisole, dibutylated hydroxytoluene, propyl gallate, tert-butyl hydroquinone, lipoic acid, tea polyphenol, L-ascorbyl palmitate, and glutathione, wherein the acid-base regulator is one or more selected from the group consisting of arginine, lysine, histidine, glycine, tromethamine, diethanolamine, ethylenediamine, meglumine, hydrochloric acid, acetic acid, anhydrous citric acid, ascorbic acid, lactic acid, tartaric acid, methanesulfonic acid, methionine, sodium hydroxide, and triethanolamine, wherein the pharmaceutical oil is one or more selected from the group consisting of natural vegetable oils, semi-natural oils artificially modified from natural vegetable oils, purified oils, and corresponding derivatives, and artificially synthesized oils, and wherein the pharmaceutical phospholipid is one or more selected from the group consisting of natural phospholipids, semi-synthetic phospholipids, and synthetic phospholipids.
6. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the pharmaceutical solvent is one or more selected from the group consisting of benzyl alcohol, propylene glycol, and ethanol, wherein the antioxidant is one or more selected from the group consisting of α-tocopherol, L-ascorbyl palmitate, and cysteine, wherein the natural vegetable oils include castor oil, sesame oil, soybean oil, sunflower oil, peanut oil, corn oil, rapeseed oil, olive oil, and cottonseed oil, wherein the semi-natural oils artificially modified from natural vegetable oils include hydrogenated castor oil, wherein the artificially synthesized oils include medium chain triglycerides, long chain triglycerides, triacetin, and ethyl oleate, wherein the natural phospholipid is selected from the group consisting of egg yolk lecithin, soybean phospholipid and combinations thereof, wherein the semi-synthetic phospholipid is selected from hydrogenated egg yolk lecithin, hydrogenated soybean phospholipid, and combinations thereof, and wherein the synthetic phospholipid is one or more selected from the group consisting of dipalmitoyl phosphatidylethanolamine, dipalmitoyl phosphatidic acid, dipalmitoylphosphatidylglycerol, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, distearoylphosphatidyl-choline, and dimyristoylphosphatidylcholine.
7. (canceled)
8. A method of pain treatment comprising administering a pain relief medicament including the long-acting ropivacaine pharmaceutical composition according to claim 1.
9. The method according to claim 8, wherein the pain relief medicament is administered by local injection.
10. The method according to claim 8, wherein the expected total daily dose of the long-acting ropivacaine pharmaceutical composition is 5-1000 mg of the active drug.
11. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the pharmaceutical composition has a duration of action of 24 h or more.
12. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the pharmaceutical composition has a duration of action of 48 h or more.
13. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises, based on the total weight of the composition, in weight percentage, 1% to 8% of ropivacaine; 3% to 23% of the pharmaceutical solvent; 10% to 50% of the pharmaceutical phospholipid; 20% to 76% of the pharmaceutical oil; 1% to 9% of the efficacy enhancer; about 0% to 0.5% of the antioxidant; and about 0% to 5% of the acid-base regulator.
14. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises, based on the total weight of the composition, in weight percentage, 1% to 5% of ropivacaine; 5% to 20% of the pharmaceutical solvent; 15% to 45% of the pharmaceutical phospholipid; 22% to 60% of the pharmaceutical oil; 2% to 8% of the efficacy enhancer; about 0% to 0.3% of the antioxidant; and about 0% to 3% of the acid-base regulator.
15. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the ω-3 fatty acids and metabolites thereof include α-linolenic acid and its metabolites eicosapentaenoic acid and docosahexaenoic acid, and combinations thereof, wherein the substances rich in α-linolenic acid include linseed oil, perilla seed oil, walnut oil, argan oil, and combinations thereof, and wherein the substances rich in α-linolenic acid metabolites eicosapentaenoic acid and docosahexaenoic acid include fish oil, laver oil and algal oil, and combinations thereof.
16. The long-acting ropivacaine pharmaceutical composition according to claim 1, wherein the efficacy enhancer is one or more selected from substances rich in ω-3 fatty acids or metabolites thereof.
17. The long-acting ropivacaine pharmaceutical composition according to claim 16, wherein the substances rich in ω-3 fatty acids or metabolites thereof are substances having an eicosapentaenoic acid content of no less than 15% and a docosahexaenoic acid content of no less than 10%.
18. The long-acting ropivacaine pharmaceutical composition according to claim 16, wherein the substances rich in ω-3 fatty acids or metabolites thereof are substances having an eicosapentaenoic acid content of no less than 20% and a docosahexaenoic acid content of no less than 10%.
19. The long-acting ropivacaine pharmaceutical composition according to claim 17, wherein the efficacy enhancer is one or more selected from fish oil and laver oil.
20. The method according to claim 8, wherein a pain treated is postoperative pain.
21. The method according to claim 9, wherein the local injection includes subcutaneous or intramuscular injection, direct instillation at the incision, incision infiltration, administration at the nerve plexus, and intra-articular injection.
Description
DRAWINGS
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MODE FOR CARRYING OUT THE INVENTION
[0102] The composition, preparation method and use of the present invention are further illustrated by providing the following examples and experimental examples, but the present invention is not limited thereto. The present invention is further described in detail below with reference to the examples, but a person skilled in the art should understand that the present invention is not limited to these examples and the used preparation methods. Moreover, a person skilled in the art can make equivalent substitutions, combinations, improvements or modifications to the present invention based on the description of the present invention, and all of them fall within the scope of the present invention.
EXAMPLE
Comparative Examples 1 and 2
[0103] According to the formulation listed in Table 1, the pharmaceutical oil and the pharmaceutical solvent were mixed uniformly, and ropivacaine was added. The resulting mixture was vortexed, and ultrasonicated to obtain a turbid and opaque liquid;
[0104] The drug ropivacaine was tried to be first dissolved in the pharmaceutical solvent, and the dissolution process was relatively rapid at this time. Then the prescribed amount of pharmaceutical oil was added. The resultant was vortexed and a phenomenon of white turbidity immediately appeared.
TABLE-US-00001 TABLE 1 Formulation of Pharmaceutical Composition Formulation (w/w)
Drug ropivacaine 2% 2% Ropi- hydro- vacaine chloride Pharma- castor oil 88% / ceutical medium / 88% oil chain tri- glyceride Pharma- benzyl 10% 10% ceutical alcohol solvent Phenomenon turbid and turbid and opaque opaque
Comparative Example 3: Ropivacaine Hydrochloride Injection
[0105] Ropivacaine hydrochloride as the active pharmaceutical ingredient and sodium chloride were weighed in the prescribed amounts, and placed into a vial. Purified water was added. The resultant was vortexed and ultrasonicated to be dissolved to be clear to obtain a uniform solution. The solution was adjusted to pH value=4.0-6.0 to obtain the ropivacaine hydrochloride injection.
TABLE-US-00002 TABLE 2 Formulation of Comparative Example 3 Formulation Ratio ropivacaine 10 mg/ml hydrochloride sodium chloride 7.1 mg/ml purified water appropriate amount
Comparative Example 4: Pro-Liposome Preparation of CN104427977B (Conventional Sustained-Release Preparation)
[0106] Ropivacaine hydrochloride monohydrate, lecithin PL-90G, castor oil, and cysteine hydrochloride in the prescribed amounts were added to a pre-weighed round bottom flask, and weighed. An excess of anhydrous ethanol was added. The flask was placed in an ultrasonic wave water bath and subjected to ultrosonication to completely disperse the phospholipid and completely dissolve each component, while ensuring that the amount of anhydrous ethanol exceeded the required final amount. At this time, the round bottom flask was connected to a suitable rotary evaporator, and subjected to evaporation under reduced pressure until the weight change of the round bottom flask indicated that the amount of anhydrous ethanol was less than or equal to 6%. The flask was allowed to be cooled to room temperature. If necessary, anhydrous ethanol was again added until it reached 6%. The resultant was mixed well, and the content was transferred to a glass vial, and stored at room temperature.
TABLE-US-00003 TABLE 3 Formulation of Comparative Example 4 Formulation Ratio (w/w) ropivacaine hydrochloride 4.78% monohydrate Lecithin PL-90G 53.91% castor oil 35.21% anhydrous ethanol 6.0% cysteine hydrochloride 0.1%
Comparative Example 5: Oil Solution Preparation of CN103142458B
[0107] Ropivacaine free base was weighed in the prescribed amount, and placed into a vial. Benzyl alcohol and benzyl benzoate were added. The resultant was stirred to dissolve the drug completely. Then soybean oil was added. Then the resultant was stirred evenly to obtain a clear oily solution.
TABLE-US-00004 TABLE 4 Formulation of Comparative Example 5 Formulation Ratio Ropivacaine free base 30 mg/ml Soybean oil 75% (v/v) Benzyl benzoate 15% (v/v) Benzyl alcohol 10% (v/v)
Comparative Example 6: Preparation Provided by CN109316602A
[0108] Ropivacaine free base and parecoxib were weighed in the prescribed amounts, and benzyl alcohol, benzyl benzoate and ethanol in the prescribed amounts were added in turn. The resultant was shaken and stirred until the drugs were completely dissolved. Then the lecithin E80 was added in the prescribed amount, and the resultant was shaken and stirred until it was completely dissolved. An appropriate amount of castor oil was added and shaken well. The resultant was placed for 5 h at 2-8° C. and at room temperature, respectively. The obvious needle-like precipitation was observed under both conditions.
TABLE-US-00005 TABLE 5 Formulation of Comparative Example 6 Formulation Ratio (w/w) Ropivacaine free base 8% Parecoxib 3% Lecithin E80 26% Benzyl benzoate 15% Benzyl alcohol 10% Ethanol 10% Castor oil 28%
Comparative Example 7: Preparation Provided by CN109316602A
[0109] Ropivacaine free base and parecoxib were weighed in the prescribed amounts, and benzyl alcohol, benzyl benzoate and ethanol in the prescribed amounts were added in turn. A plug was covered. The resultant was stirred until the drugs were completely dissolved. The egg yolk lecithin PC-98T was then added, and the resultant was shaken and stirred until it was completely dissolved. An appropriate amount of soybean oil was added to the prescribed amount. The resultant was mixed well.
TABLE-US-00006 TABLE 6 Formulation of Comparative Example 7 Formulation Amount Ropivacaine free base 3% w/v Parecoxib 1.5% w/v Ethanol 10% v/v Benzyl benzoate 20% v/v Benzyl alcohol 10% v/v egg yolk lecithin PC-98T 20% w/v Soybean oil added to 100% v/v
Comparative Examples 8 and 9
[0110] Ropivacaine hydrochloride, propylene glycol, soybean lecithin S100, castor oil, and meglumine in the prescribed amounts were added to a pre-weighed round bottom flask, and weighed. An excess of anhydrous ethanol was added. The flask was placed in an ultrasonic wave water bath, and subjected to ultrasonication to dissolve each component completely. At this time, the round bottom flask was connected to a suitable rotary evaporator, and subjected to evaporation under reduced pressure until the weight change of the round bottom flask indicated that the anhydrous ethanol had been completely removed. The flask was allowed to be cooled to room temperature. The fish oil was added in the prescribed amount. The resultant was mixed well, and the content was transferred to a glass vial, and stored at room temperature.
TABLE-US-00007 TABLE 7 Formulation Information of Comparative Examples 8 and 9 Ratio of Ratio of Comparative Comparative Example 8 Example 9 Formulation (w/w) (w/w) Ropivacaine hydrochloride 4.5% 4.5% Soya bean lecithin S100 42% 42% Castor oil 38.2% 23.2%.sup. Propylene glycol 15% 15% Fish oil 3322 / 15% Meglumine 0.3% 0.3%
Example 1
[0111] Preparation of Long-Acting Pharmaceutical Compositions
[0112] The preparation process of the compositions in Table 8: a prescribed amount of ropivacaine was added to the pharmaceutical solvent, and the mixture was stirred to dissolve the drug to obtain a drug solution; the pharmaceutical phospholipid was added to the corresponding pharmaceutical oil, and the mixture was subjected to high-speed shearing until the pharmaceutical phospholipid was completely dissolved; the drug solution was added thereto, and the mixture was mixed well, and then the efficacy enhancer was added, and the mixture was stirred to obtain a drug-containing solution, and the solution was filtered and packed, filled with nitrogen, sealed, and then sterilized by moist heat to obtain a clear and transparent liquid.
TABLE-US-00008 TABLE 8 Formulation of Pharmaceutical Compositions
(w/w)
Drug Ropivacaine 2% 1% 1% 2% 3% 3.5% ropivacaine ropivacaine ropivacaine ropivacaine ropivacaine ropivacaine hydrochloride hydrochloride free base mesylate hydrochloride free base Pharmaceutical oil Castor oil 43% 57% 76% / 44% 56.5% Medium chain / / / 39% / / triglyceride Pharmaceutical Egg yolk / / / 40% 32% / phospholipid lecithin PC-98T Soya bean 35% 30% 10% / / 17% lecithin S100 Efficacy Fish oil 3322 5% 2% 5% / 4% 8% enhancer Purple perilla / / / 6% 2% / seed oil Pharmaceutical Benzyl alcohol / / 8% 13% 15% 15% solvent Propylene 15% 10% / / / / glycol Phenomenon clear and clear and clear and clear and clear and clear and transparent transparent transparent transparent transparent transparent
[0113] It can be seen from the results that all the compositions prepared by adding an appropriate amount of pharmaceutical phospholipid were clear and transparent, and the purpose of improving the solubility of the drug in the composition system was achieved.
[0114] The preparation process of the compositions in Table 9: a prescribed amount of ropivacaine was added to the pharmaceutical solvent to obtain a drug solution; the pharmaceutical phospholipid was added, and the mixture was stirred until the pharmaceutical phospholipid was completely dissolved; then the pharmaceutical oil and the efficacy enhancer were added, the mixture was stirred to mix well to obtain a drug-containing solution, and the solution was filtered and packed, filled with nitrogen, sealed, and then sterilized by moist heat to obtain a clear and transparent liquid.
TABLE-US-00009 TABLE 9 Formulation of Pharmaceutical Compositions (w/w)
Drug ropivacaine Ropivacaine 4% 3.5% 5% 4% hydrochloride Pharmaceutical oil Castor oil 40% 33.5%.sup. 22% 28% Pharmaceutical Soya bean 38% 42% 45% / phospholipid lecithin S100 Egg yolk / / / 45% lecithin PC-98T Efficacy enhancer Fish oil 3322 5% / 8% / Linseed oil / 6% / 5% Pharmaceutical Benzyl alcohol 13% 15% 20% 15% solvent Benzyl benzoate / / / 3% Phenomenon clear and clear and clear and clear and transparent transparent transparent transparent
[0115] The preparation process of the compositions in Table 10: The drug ropivacaine, the pharmaceutical oil, the pharmaceutical phospholipid, the pharmaceutical solvent, and the efficacy enhancer were weighted in the prescribed amounts, and placed into a pre-weighed round bottom flask, and weighed. An excess of anhydrous ethanol was added. Then the flask was placed in an ultrasonic wave water bath, and subjected to ultrasonication to completely disperse phospholipid and completely dissolve each component. At this time, the round bottom flask was connected to a suitable rotary evaporator, and subjected to evaporation under reduced pressure until the weight change of the round bottom flask indicated that the anhydrous ethanol had been completely removed. If necessary, ethanol was supplemented to the prescribed amount, and the mixture was mixed well. The resulting drug solution was filtered, packed, filled with nitrogen, sealed to obtain a clear and transparent liquid.
TABLE-US-00010 TABLE 10 Formulation of Pharmaceutical Compositions
(w/w)
Drug Ropivacaine 5% 2% 2% 3.5% 2.5% 3.5% ropivacaine ropivacaine ropivacaine ropivacaine ropivacaine ropivacaine hydrochloride hydrochloride hydrochloride free base mesylate hydrochloride Pharmaceutical oil Sesame oil / 54% / / / / Castor oil 34% / / 59.5% / 45.5% Soybean oil / / / / 42.5% / Medium chain / / 38% / / / triglyceride Pharmaceutical Soya bean 40% / / 15% 35% 35% phospholipid lecithin S100 Egg yolk / 32% 50% / / / lecithin PC-98T Efficacy Fish oil 3322 / 2% 4% / / / enhancer Laver oil 2% / / 2% / / Linseed oil / / / / / 6% Purple perilla / / / / 5% / seed oil Solvent Benzyl alcohol 19% 2% 6% 15% 10% 10% Ethanol / 8% / / 5% / Benzyl / / / 5% / / benzoate Phenomenon clear and clear and clear and clear and clear and clear and transparent transparent transparent transparent transparent transparent
[0116] The preparation process of the compositions in Table 11: The drug ropivacaine, the pharmaceutical oil, the pharmaceutical phospholipid, the pharmaceutical solvent, the antioxidant, the efficacy enhancer and the acid-base regulator were weighted in the prescribed amounts, and placed into a pre-weighed round bottom flask, and weighed. An excess of anhydrous ethanol was added. Then the flask was placed in an ultrasonic wave water bath, and subjected to ultrasonication to completely disperse the phospholipid and completely dissolve each component. At this time, the round bottom flask was connected to a suitable rotary evaporator, and subjected to evaporation under reduced pressure until the weight change of the round bottom flask indicated that the anhydrous ethanol had been completely removed. The resulting drug solution was filtered, packed, filled with nitrogen, sealed, and then sterilized by moist heat to obtain a clear and transparent liquid.
TABLE-US-00011 TABLE 11 Formulation of Pharmaceutical Compositions
(w/w)
Drug Ropivacaine 4.5% 3% 6% 3.5% 7% 4.5% ropivacaine ropivacaine ropivacaine ropivacaine ropivacaine ropivacaine hydrochloride hydrochloride hydrochloride hydrochloride hydrochloride hydrochloride Pharmaceutical oil Sesame oil / 47.8% / / / / Castor oil 29% / 17.6% 44% 15% 30% Pharmaceutical Soya bean / 35% 50% 30% 52% 35% phospholipid lecithin S100 Egg yolk 43% / / / / / lecithin PC-98T Efficacy Fish oil 3322 5% / 6% / 1% 8% enhancer Linseed oil / 2% / 2% / / Solvent Propylene 18% / / 15% / 21.7% glycol Benzyl alcohol / 12% 20% 5% 22.5% / Antioxidant α-tocopherol / / / / 0.4% / L-ascorbyl / / 0.2% / 0.1% / palmitate α-tocopheryl / / / 0.1% / / acetate Acid-base Meglumine 0.5% 0.2% / / / 0.8% regulator Diethanolamine / / / 0.4% 2% / Ethylenediamine / / 0.2% / / / Phenomenon clear and clear and clear and clear and clear and clear and transparent transparent transparent transparent transparent transparent
[0117] The preparation process of the compositions in Table 12: The drug ropivacaine, the pharmaceutical oil, the pharmaceutical phospholipid, the pharmaceutical solvent, the efficacy enhancer and the acid-base regulator were weighted in the prescribed amounts, and placed into a pre-weighed round bottom flask, and weighed. An excess of anhydrous ethanol was added. Then the flask was placed in an ultrasonic wave water bath, and subjected to ultrasonication to completely disperse the phospholipid and completely dissolve each component. At this time, the round bottom flask was connected to a suitable rotary evaporator, and subjected to evaporation under reduced pressure until the weight change of the round bottom flask indicated that the anhydrous ethanol had been completely removed. The resulting drug solution was filtered, packed, filled with nitrogen, sealed, and then sterilized by moist heat to obtain a clear and transparent liquid.
TABLE-US-00012 TABLE 12 Formulation of Pharmaceutical Compositions (w/w)
Drug Ropivacaine 4.5% 4.5% 4.5% ropivacaine ropivacaine ropivacaine hydrochloride hydrochloride hydrochloride Pharmaceutical oil 33.2% 33.2% 33.2% castor oil castor oil castor oil Pharmaceutical 42% soya bean 42% soya bean 42% soya bean phospholipid lecithin S100 lecithin S100 lecithin S100 Efficacy enhancer 5% fish 5% fish 5% algal oil 3322 oil 1812 oil Solvent 15% propylene 15% propylene 15% propylene glycol glycol glycol Acid-base regulator 0.3% 0.3% 0.3% meglumine meglumine meglumine Phenomenon clear and clear and clear and transparent transparent transparent
EXPERIMENTAL EXAMPLE
Experimental Example 1 Stability Investigation
[0118] The pharmaceutical compositions prepared in Example 1 were placed to observe the appearance to determine its stability. If there was no visible precipitate, it could be considered that the composition had good storage stability.
TABLE-US-00013 TABLE 13 Stability Investigation Result of Pharmaceutical Compositions
Comparative Example1 turbid and opaque / Comparative Example2 turbid and opaque / Composition 3 clear and transparent no significant change Composition 4 clear and transparent no significant change Composition 5 clear and transparent no significant change Composition 6 clear and transparent no significant change Composition 7 clear and transparent no significant change Composition 8 clear and transparent no significant change Composition 9 clear and transparent no significant change Composition 10 clear and transparent no significant change Composition 11 clear and transparent no significant change Composition 12 clear and transparent no significant change Composition 13 clear and transparent no significant change Composition 14 clear and transparent no significant change Composition 15 clear and transparent no significant change Composition 16 clear and transparent no significant change Composition 17 clear and transparent no significant change Composition 22 clear and transparent no significant change Composition 23 clear and transparent no significant change Composition 24 clear and transparent no significant change
[0119] It can be seen from the results that the prepared compositions were all clear and transparent, indicating that they had good storage stability. However, the preparation provided by CN109316602A (i.e., Comparative Example 6) had obvious precipitation after being placed for 5 hours, showing poor stability.
[0120] Some of the compositions were selected, placed under corresponding conditions, and detected for their changes in the content and the total amount of related substance according to the following methods.
[0121] Testing Equipment:
[0122] High performance liquid chromatograph 1260, Agilent Technologies Co., Ltd., USA.
[0123] Detection Conditions:
[0124] Chromatographic column: octadecylsilane-bonded silica gel column (4.6 mm×150 mm, 5 μm, Agilent Technologies Co., Ltd., USA); the mobile phase: a solution of acetonitrile-disodium hydrogen phosphate buffer (adjusted to pH=8.0 by phosphoric acid), the flow rate: 1 ml/min, the column temperature: 30° C., and the detection wavelength: 240 nm.
[0125] Preparation Method:
[0126] Test sample solution: an appropriate amount of the test sample was precisely weighed and placed into a volumetric flask. Methanol was added to dissolve the test sample, and the volume was set to the scale line. The mixture was vortexed and ultrasonicated to obtain the test sample solution.
[0127] Content reference substance solution: An appropriate amount of ropivacaine drug reference substance was weighed, placed into a volumetric flask, and precisely weighed. Methanol was used to dissolve the ropivacaine drug reference substance, and the volume was set to the scale line to obtain the content reference substance solution.
[0128] Related substance reference solution: 1 ml of the test sample solution was precisely pipetted into a 100 ml volumetric flask, and methanol was added to dilute to the scale line to obtain the related substance reference solution.
[0129] The above solution was precisely taken and injected into a liquid chromatograph. The external standard method was used to calculate the content. The self-dilution control method was used to calculate the related substance. The total amount of the related substance was calculated. The results were shown in Table 14.
TABLE-US-00014 TABLE 14 Determination result of the content and related substance of the compositions
Composition 98.12 99.32 98.34 98.76 99.09 0.11 0.12 0.12 0.13 0.14 3 Composition 101.45 100.98 101.34 100.32 99.96 0.13 0.14 0.15 0.14 0.14 4 Composition 99.66 99.87 100.32 99.95 98.08 0.11 0.12 0.13 0.12 0.12 5 Composition 98.54 98.35 97.67 97.89 98.33 0.11 0.12 0.13 0.13 0.14 6 Composition 100.87 100.39 99.87 99.65 99.03 0.11 0.12 0.13 0.13 0.13 7 Composition 99.42 98.59 98.12 100.09 98.80 0.13 0.14 0.15 0.14 0.14 8 Composition 101.83 101.29 100.69 99.97 100.05 0.12 0.13 0.13 0.12 0.14 9 Composition 101.08 100.65 100.75 101.01 100.69 0.12 0.13 0.12 0.13 0.14 14 Composition 99.13 98.92 99.02 98.55 98.13 0.13 0.12 0.13 0.14 0.13 16 Composition 99.56 100.02 99.72 99.49 100.59 0.12 0.13 0.12 0.13 0.13 17 Composition 100.07 99.42 100.34 99.31 99.05 0.13 0.14 0.13 0.13 0.14 22 Composition 100.12 100.03 99.91 99.87 100.11 0.14 0.13 0.14 0.15 0.13 23 Composition 99.08 99.82 100.04 99.29 99.56 0.13 0.14 0.13 0.15 0.12 24
[0130] It can be seen from the results that the content and related substance of the compositions of the present invention did not change significantly after being placed under accelerated and long-term conditions for 6 months.
[0131] It can be seen from the above research results that the compositions of the present invention had good stability.
Experimental Example 2 Syringeability Determination of Pharmaceutical Compositions
[0132] A 21G needle was used to measure the syringeability of the compositions, and the determination results were shown in Table 15.
TABLE-US-00015 TABLE 15 Viscometry results of pharmaceutical compositions Composition Injection syringeability Composition 3 to They had moderate viscosity, and could Composition 25 pass 21G needle smoothly. Comparative It was difficult to pass 21G needle, and had Example 4 high viscosity and poor injection syringeability. Comparative It had a low viscosity, and could Example 5 pass 21G needle smoothly.
[0133] It can be seen from the results that the compositions of the present invention had suitable injection syringeability; while the injection of Comparative Example 4 was difficult, and the injection syringeability was extremely poor.
Experimental Example 3
[0134] In the following experimental descriptions, the administration dose referred to the dose calculated according to the amount of hydrochloride of the active ingredient in the preparation (i.e., in the case of free base, converted to hydrochloride).
[0135] In Vivo Pharmacodynamic Investigation of Long-Acting Pharmaceutical Compositions (1)
[0136] Experimental animals: Male SD rats with 200-300 g were adaptively reared for 3 days, and the base threshold was measured every day.
[0137] Experimental grouping and administration dosage: Animals were randomly grouped according to the base threshold, with 6 animals in each group. The different compositions (or the compositions of the comparative examples) were respectively injected into the subcutaneous tissue of voix pedis in rats (the administration volume ranged from 0.10 to 0.35 ml). Pain threshold was measured by von Frey filament pain threshold detector. And motor block was investigated.
[0138] The doses administered in the pharmacodynamic test were seen in Table 16.
TABLE-US-00016 TABLE 16 Dosing information for in vivo pharmacodynamic studies of pharmaceutical compositions Composition Dose/mg/kg saline placebo / Comparative example 3 .sup. 13*.sup.1 Comparative example 4 45 Composition 6 45 Composition 7 45 Composition 9 45 Composition 16 45 Composition 17 45 Composition 22 45 *.sup.1In performing the pharmacodynamic test of the present invention, the dose of Comparative Example 3 was tried to be set higher, and it was found that the rats immediately experienced convulsion and tics after injected with 15 mg/kg of Comparative Example 3, and then died. The direct cause of this phenomenon was the toxic reaction that occurred when the plasma concentration was too high. However, for the compositions of the present invention, the rats did not appear the toxic reaction, indicating that the compositions of the present invention have improved safety compared with Comparative Example 3.
[0139] Sensory Nerve Block:
[0140] Detection method, von Frey acupuncture-foot-lifting method: The rat moved freely in the cage, and the activity space was fixed. After the rat was quiet, the rat's paw was stimulated with von Frey filaments to determine the rat's mechanical paw withdrawal threshold. The higher the mechanical paw withdrawal threshold was, the better the effect of sensory nerve block was.
[0141] Evaluation method: Paired t test was used to compare the mechanical paw withdrawal threshold of each group of animals with that before administration, and unpaired t test was used to compare the mechanical paw withdrawal threshold of each group of animals with Comparative Example 4, P<0.05 indicated a statistical difference, P<0.01 and P<0.001 indicated a significant statistical difference.
[0142] Measurement time: Since the sensory nerve block of Comparative Example 3 was found to be maintained for a short time during the experiment, up to about 3 hours, the measurement time of Comparative Example 3 was 0.5 h, 1 h, 2 h, 2.5 h, 3 h. However, if the measurement time of the long-acting composition was set according to this setting, the time points would be too dense, which could affect the behavior of rats. Therefore, the time point setting of the long-acting composition was sparser than that of Comparative Example 3. The results were shown in
[0143]
[0144]
[0145] According to the duration of sensory nerve block of the compositions, the effective therapeutic dose of each composition was calculated, and the results were shown in Table 17.
TABLE-US-00017 TABLE 17 Effective therapeutic dose calculation for drug Comparative Comparative Composition Composition Composition Composition Composition Composition Parameter Example3 Example4 6 7 9 16 17 22 R 6.5 1.5 0.9 0.9 0.6 0.8 0.6 0.8
[0146] Remarks: R=A/T.sub.t, wherein A is the administration dose of ropivacaine, T.sub.t is the effective treatment time of the drug in the body, and R is the effective therapeutic dose.
[0147] It can be seen that the effective therapeutic dose R of Compositions 6, 7, 9, 16, 17 and 22 can be reduced by at least 40% compared with Comparative Examples 3 and 4, indicating that when the same effective treatment time was achieved, there was a smaller drug dose consumed for the compositions of the present invention, which can avoid the waste of drug and the toxic reactions caused by excess drug in clinic. In addition, the pharmaceutical compositions of the present invention had a controllable pain relief time, which provided the beneficial value of on-demand selection for drug development.
[0148] Motor Nerve Block:
[0149] While the sensory nerve block was measured, the four-level scoring method was used to evaluate the motor nerve block of the rats in each group. The higher the score was, the more severe the motor block was.
[0150] Grade 1 (representing no motor block, expressed as a value of 1): the rat's paw can normally land on the ground, and the weight bearing was normal, and there was no weakness; Grade 2 (representing a slight motor block, expressed as a value of 2): the rat's paw can land normally, but there was slight weakness;
[0151] Grade 3 (representing complete motor block, expressed as a value of 3): the rat's paw occasionally touched the ground and there was weakness;
[0152] Grade 4 (representing complete motor block, expressed as a value of 4): The rat's paw did not touch the ground at all and there was severe weakness.
[0153] Evaluation method: Unpaired t test was used to compare the motor nerve block score of each group of animals with that of the normal saline placebo group. P<0.05 indicated a statistical difference, and P<0.01 and P<0.001 indicated a significant statistical difference. The results were shown in
[0154] The duration comparison of motor block and sensory block was shown in Table 18.
TABLE-US-00018 TABLE 18 Comparison result of sensory block and motor block for the composition Motor block duration/sensory Composition nerve block duration Comparative Example4 1.60 Composition 6 1.17 Composition 7 0.88 Composition 9 0.67 Composition 16 0.89 Composition 17 0.67 Composition 22 0.89
[0155] Based on the results of the ratio of the motor nerve block duration/sensory nerve block duration of the composition, it can be seen that the motor block duration of the compositions was at least 20% shorter than that of Comparative Example 4 if the same sensory nerve block duration was achieved. It can be seen that the compositions of the present invention had improved motor nerve block condition compared with Comparative Example 4, which meant that the compositions of the present invention did not increase the risk of nerve damage and had higher safety while meeting the clinical analgesic requirements.
[0156] In Vivo Pharmacodynamic Investigation of Long-Acting Pharmaceutical Compositions (2)
[0157] On the basis of Composition 9, the drug concentration and dosage were changed, and the method of pharmacodynamic investigation (1) was used to determine the sensory nerve block of the corresponding compositions. The compositions and dosing information were shown in Table 19, and the results of sensory nerve block were shown in
TABLE-US-00019 TABLE 19 Formulation and Dosing Information of Compositions Composition Composition (w/w) Dose mg/kg Composition 1.35% ropivacaine hydrochloride + 15 9-1 20% benzyl alcohol + 25.65% castor oil + 45% soybean phospholipid S100 + 8% fish oil 3322 Composition 2.25% ropivacaine hydrochloride + 25 9-2 20% benzyl alcohol + 24.75% castor oil + 45% soybean phospholipid S100 + 8% fish oil 3322 Composition 3.15% ropivacaine hydrochloride + 35 9-3 20% benzyl alcohol + 23.85% castor oil + 45% soybean phospholipid S100 + 8% fish oil 3322
[0158] The duration of sensory block of Compositions 9-1, 9-2 and 9-3 in vivo was 24 h, 36 h and 48 h, respectively. It can be seen that different pharmaceutical doses can achieve different pain relief time, which provided convenience for clinical on-demand selection.
[0159] After calculation, the effective therapeutic doses R of Compositions 9-1, 9-2 and 9-3 were 0.6, 0.7 and 0.7, respectively, which were significantly lower than that of the conventional sustained-release preparation of Comparative Example 4, indicating that the composition of the present invention improved the drug utilization.
[0160] In Vivo Pharmacodynamic Investigation of Long-Acting Pharmaceutical Compositions (3)
[0161] The ratio and type of the efficacy enhancer in the composition were changed, and the method of pharmacodynamic investigation (1) was used to determine the sensory nerve block of the corresponding composition in animals. The compositions and dosing information were shown in Table 20, and the results of sensory nerve block were shown in
TABLE-US-00020 TABLE 20 Formulation and Dosing Information of Composition Composition Dose mg/kg Comparative example 8 45 Comparative example 9 45 Composition 23 45 Composition 24 45 Composition 25 45
[0162]
TABLE-US-00021 TABLE 21 Mechanical Threshold Results for Compositions Time/h Composition 23 Composition 24 Composition 25 Before administration 12 ± 3 13 ± 3 13 ± 3 3 300 ± 0#& 300 ± 0#& .sup. 300 ± 0#& 6 240 ± 66# 260 ± 42# .sup. 240 ± 66# 24 153 ± 41#&& 200 ± 49###&&& 127 ± 41 30 113 ± 33##&& 180 ± 0###&&& 67 ± 16# 36 68 ± 28##&& 140 ± 44###&&& 49 ± 18##&& 48 54 ± 14###&&& 68 ± 28##&&.sup. 37 ± 18& 54 47 ± 21##&& 36 ± 19#& 14 ± 2 72 30 ± 15#& 18 ± 7 18 ± 7 Remarks: #p < 0.05, ##p < 0.01, ###p < 0.001 v.s. Comparative Example 8; &p < 0.05, &&p < 0.01, &&&p < 0.001 v.s. Comparative Example 9
[0163] It can be seen from the results in Table 21 that, compared with Comparative Examples 8 and 9, the mechanical paw retraction thresholds of Compositions 23 to 25 at 24 h or at different time points after 24 h had significant differences, indicating that the duration of action of the compositions had a significant advantage over those of Comparative Examples 8 and 9. It can be seen that the efficacy enhancer achieved an unexpected effect through a certain mechanism.
[0164] According to the sensory nerve block duration of the composition, the effective therapeutic dose of each composition was calculated, and the results were shown in Table 22.
TABLE-US-00022 TABLE 22 Effective therapeutic dose calculations for drugs Comparative Comparative Composition Composition Composition Parameter Example8 Example9 23 24 25 R 1.5 1.5 0.6 0.8 0.9 Remarks: R = A/T.sub.t, wherein A was the administration dose of ropivacaine, T.sub.t was the effective treatment time of the drug in the body, and R was the effective therapeutic dose.
[0165] It can be seen that the effective therapeutic dose of the compositions of the present invention was at least 40% lower than those of Comparative Examples 8 and 9, indicating that the compositions of the present invention can improve drug utilization.
Experimental Example 4
[0166] In the following experimental descriptions, the administered dose referred to the dose calculated according to the amount of hydrochloride of the active ingredient in the preparation (i.e., in the case of using free base, converted to hydrochloride).
[0167] In Vivo Pharmacokinetic Investigation of Long-Acting Pharmaceutical Compositions
[0168] The obtained pharmaceutical composition was subcutaneously injected into male SD rats. Blood was collected at predetermined time points. The content of the drug in plasma was determined by LC-MS method, and the pharmacokinetics in vivo was investigated.
[0169] Experimental animals: male SD rats with a weight of 200-300 g, source: SHANGHAI INSTITUTE OF MATERIA MEDICA, CHINESE ACADEMY OF SCIENCES;
[0170] Experimental grouping: Rats were randomly divided into groups according to the following table, with 6 rats in each group;
[0171] Administration formulations and dosing information are provided in Table 23.
TABLE-US-00023 TABLE 23 Dosing information for in vivo pharmacokinetic experiments of pharmaceutical compositions Composition Dose/mg/kg Comparative example 3 13 Comparative example 4 45 Composition 7 45 Composition 9 45 Composition 17 45
[0172] Experimental procedure: Rats were subcutaneously injected, and at a predetermined time point, 0.5 ml of blood was collected from the orbit of the rat and placed in a centrifuge tube with heparin sodium anticoagulation. The tube was centrifuged at 3000 rpm for 10 min to separate the plasma, and the plasma concentration was detected by LC-MS method.
[0173] The pharmacokinetic (PK) parameters of the compositions were calculated, and the results were statistically analyzed by t-test. The results were shown in Table 24 and
TABLE-US-00024 TABLE 24 PK Parameters of Pharmaceutical Compositions Comparative Comparative Pk parameters example 3 example 4 Composition 7 Composition 9 Composition 17 C.sub.max(ng/ml) 365.00 ± 91.73 283.38 ± 28.61 272.00 ± 29.10 209.82 ± 9.70 232.70 ± 15.49 AUC(h*ng/ml) 2332.60 ± 57.98 5164.67 ± 344.53 5755.06 ± 64.46** 6218.84 ± 395.54*** 6568.94 ± 235.99*** AUC.sub.t(h*ng/ml) / 4596.10 ± 110.34 5473.10 ± 153.89*** 6141.06 ± 163.89*** 6485.61 ± 232.89*** Remarks: AUC was bioavailability, and AUC.sub.t was effective bioavailability. Remarks: *p < 0.05 **p < 0.01 ***P < 0.001 v.s. Comparative Example 4.
[0174] It can be seen from the results in Table 24 that the C.sub.max of Compositions 7, 9 and 17 was at least 70% lower than that of Comparative Example 3 at the same dose, and compared with Comparative Example 4, the bioavailability and effective bioavailability of Compositions 7 and 9 were significantly improved.
[0175] Combined with the results of the drug pharmacodynamic investigation and the PK curve,
[0176] According to the results of the pharmacodynamic test of Experimental Example 3, because the composition of the present invention had a good correlation between plasma concentration and drug efficacy, and the duration of efficacy of compositions 7, 9, and 17 were 48 h, 72 h, and 72 h, respectively, it is inferred that the onset concentration of ropivacaine hydrochloride in rats was about 20 ng/ml (in Comparative Example 3, after injection into the body, the drug entered the blood rapidly, resulting in a poor correlation between its plasma concentration and local efficacy, so the onset concentration of 20 ng/ml was not applicable for Comparative Example 3). As shown in
[0177] To sum up, the composition of the present invention can reduce C.sub.max by at least 70% compared with Comparative Example 3 (ropivacaine hydrochloride injection) at the same dose. Compared with conventional sustained-release preparations, the composition of the present invention has more complete drug release, significantly improved bioavailability and effective bioavailability, can reduce the duration of drug retention in the body in an ineffective form, and improve drug utilization, and reduce the risk of toxicity due to prolonged exposure of nerve fibers to low drug concentrations. In addition, under the condition of having the same analgesic effect, the composition of the present invention can improve motor block and reduce neurotoxicity compared with conventional sustained-release preparations, which was of great significance in clinical use. In the case of a certain proportion, the efficacy enhancer in the present invention can exert unexpected effects.
Experimental Example 5
[0178] In the following experimental descriptions, the administered dose referred to the dose calculated according to the amount of the hydrochloride salt of the active ingredient in the preparation (i.e., in the case of using a free base, converted to hydrochloride salt).
[0179] Irritation Investigation at Injection Site
[0180] Composition 8 and Comparative Example 7 were respectively injected into the subcutaneous tissue of voix pedis in rats, and the irritation at the injection site was observed 24 hours after the administration. Dosing information was shown in Table 25, and results were shown in
TABLE-US-00025 TABLE 25 Dosing Information of Compositions Composition Dose Composition 8 45 mg/kg Comparative example 7 45 mg/kg
[0181] It can be seen from the results that at 24 h after administration, the skin at the injection site of Comparative Example 7 was obviously blackened or even slightly ulcerated, showing greater irritation, and the injection site of the composition of the present invention had no obvious abnormality except for slight redness and swelling. It can be seen that the composition of the present invention had better injection site tolerance and safety compared with Comparative Example 7.
[0182] Experimental Example 6
[0183] In the following experimental descriptions, the administered dose referred to the dose calculated according to the amount of the hydrochloride salt of the active ingredient in the preparation (i.e., in the case of using a free base, converted to hydrochloride salt).
[0184] Histopathological Observation of Injection Site
[0185] Rats were subjected to subcutaneous injection of 13 mg/kg of Comparative Example 3, 45 mg/kg of Comparative Example 5, Composition 7 and Composition 9, and at 48 h after administration, the rats were euthanized, and tissue samples at the injection site were dissociated using a scalpel. The sample size should cover the tissue size of the administration site. After flushing with normal saline, the sample was fixed in buffered formalin. After dehydration treatment with the dehydrating agent xylene, the tissue samples were embedded in paraffin, cut into thin sections (5 μm tissue sections) using a cryostat, placed on glass slides, and stained with hematoxylin-eosin to obtain tissue sections. Histomorphology and immunohistology were observed under microscope. The results were shown in
[0186] It can be seen from the results that there was no obvious abnormality in the tissue sections of Composition 7, Composition 9, non-injection group and Comparative Example 3, while obvious inflammatory cell infiltration occurred with serious irritation at the injection site in Comparative Example 5. It can be seen that the composition of the present invention had good safety and can reduce the irritation at the injection site compared with the conventional sustained-release preparation.
[0187] As reported in Acute Med Surg. 2017 April; 4(2): 152-160, for neurotoxicity, the inflammation and reactive fibrosis around nerve fibers near the administration site can be generally observed. In histopathological evaluation, it is mainly infiltration of infiltrating cells (including macrophages, foreign body giant cells, lymphocytes, and plasma cells). Compared with Comparative Example 5, the composition of the present invention significantly reduced the irritation reaction at the injection site, indicating that the composition of the present invention had a positive effect on reducing neurotoxicity.