OA19471A - Poorly soluble complex or solvate thereof, pharmaceutical composition, and application thereof. - Google Patents

Poorly soluble complex or solvate thereof, pharmaceutical composition, and application thereof. Download PDF

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Publication number
OA19471A
OA19471A OA1201900067 OA19471A OA 19471 A OA19471 A OA 19471A OA 1201900067 OA1201900067 OA 1201900067 OA 19471 A OA19471 A OA 19471A
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Prior art keywords
bupivacaine
solvaté
complex
préparation
pamoate
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OA1201900067
Inventor
Shanchun ZHANG
Yihua Wang
Jiashi PENG
Kaisheng CHENG
Xiao Wang
Shu Gao
Hongzhang Sun
Xiaorong LU
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Fruithy Holdings Limited
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Publication of OA19471A publication Critical patent/OA19471A/en

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Abstract

The present invention provides a complex of formula (I) or a solvate thereof (wherein n is 1 to 4), a pharmaceutical composition, and use of the pharmaceutical composition in the prevention or treatment of surgical pain, intraoperative pain and postsurgical pain. The technical solution according to the present invention provides a medicament which can be produced by a simple production process and can stably release a local anesthetic in body for a long period. The medicament can be released for at least three days or more, which can prolong the analgesic effect on the postsurgical pain, can be used conveniently by the physician and the patient, and has a good treatment compliance.

Description

The présent invention will be further described in detail below with reference to particular embodiments. It will be appreciated that other embodiments are contemplated and can be implemented without departing from the scope or spirit of the présent invention. Therefore, the following detailed description is non-limiting.
Unless otherwise indicated, ail numbers used in the description and claims to represent characteristic dimensions, amounts, and physical and chemical properties should be understood to be modified with the term “about” in ail cases. Therefore, unless contradictorily specified, ail numerical parameters listed in the above description and the appended claims are approximate values, and those skilled in the art can seek to attain the desired properties by appropriately changing these approximate values from the teaching of the présent invention. Use of a numerical range indicated with endpoints includes ail numbers and any range in the range. For example, a range of 1 to 5 includes 1, 1.1, 1.3, 1.5, 2, 2.75, 3, 3.80, 4, 5, and so forth.
The insoluble complex of the présent invention has a structure and composition according to formula (I).
In formula (I), n is 1 to 4.
The complex provided in the présent invention is consisted of bupivacaine and pamoic acid in a certain molecular proportion, including 1:1,2:1,3:1 and 4:1.
It is known in the art that bupivacaine is a chiral compound having two configurations of levorotatory bupivacaine and dextrorotatory bupivacaine. Particular chiral configurations of bupivacaine are not particularly limited according to the technical solutions of the présent invention. That is, a bupivacaine racemate (i.e., a mixture of levorotatory bupivacaine and dextrorotatory bupivacaine in a molar ratio of 1:1) may be used, and either of levorotatory bupivacaine and dextrorotatory bupivacaine or a mixture thereof in any ratio may also be used.
The présent invention provides not only a non-solvate of a bupivacaine-pamoic acid complex, but also a solvaté thereof, wherein the solvent includes, but not limited to, methanol, éthanol, acetone and water.
The “insoluble” of the présent invention means that the solubility thereof (calculated as bupivacaine) in pure water or a 0.01 M phosphate buffered saline at pH 7.4 is less than 0.01 g/mL.
The complex of the présent invention refers to a solid bound by a non-covalent action such as ionic bond, hydrogen bond, van de Waals force, π-π packing action and the like, which may be in a sait form or in a co-crystal form. The complex has a property significantly different from that of a single component thereof or a simple mixture with respect to physics, chemistry or mechanics. Reference can be made to Journal of China Pharmaceutical University 2012, 43(5): 475-480, for the définitions of co-crystal and a sait.
The présent invention provides bis(bupivacaine) pamoate in a solid State with a stable molecular composition proportion, wherein one molécule of pamoic acid is bound to two molécules of bupivacaine in a stable molecular proportion to form a sait.
Said bis(bupivacaine) pamoate may exist in a solvaté form, and the solvent of the solvaté is one or more selected from a group consisting of methanol, acetone, éthanol and water. A methanol solvaté, an éthanol solvaté, or a hydrate is préférable.
The bis(bupivacaine) pamoate or a solvaté thereof provided in the présent invention appears to be crystalline powders or amorphous powders, which hâve different X-ray powder diffraction characteristics.
The X-ray powder diffraction pattern, measured with Cu-Κα radiation, of typical crystalline powders of bis(bupivacaine) pamoate provided in the présent invention, has diffraction peaks at about 4.9±0.2, 9.8±0.2, 10.9±0.2, 12.0±0.2, 12.9±0.2, 13.7±0.2, 14.7±0.2, 15.6±0.2, 16.3±0.2, 17.6±0.2, 18.9±0.2, 19.7±0.2, 20.2±0.2, 24.7±0.2, and 26.1±0.2 represented by 2Θ. The crystalline powders of bis(bupivacaine) pamoate are defined as polymorph A, and the X-ray powder diffraction pattern thereof is substantially as shown in Fig. 1.
The X-ray powder diffraction pattern, measured with Cu-Κα radiation, of further crystalline powders of bis(bupivacaine) pamoate provided in the présent invention, has diffraction peaks at about 10.9±0.2, 12.6±0.2, 13.7±0.2, 14.2±0.2, 15.7±0.2, 16.7±0.2, 17.3±0.2, 18.3±0.2, 18.9±0.2, 19.4±0.2, 25.1±0.2, 26.4±0.2, 29.0±0.2, and 34.6±0.2 represented by 20. The crystalline powders of bis(bupivacaine) pamoate are defined as polymorph B, and the X-ray powder diffraction pattern thereof is substantially as shown in Fig. 2.
The présent invention also provides further typical crystalline powders of bis(bupivacaine) pamoate, wherein the X-ray powder diffraction, measured with Cu-Κα radiation, has diffraction peaks at about 10.8±0.2, 12.6±0.2, 13.7±0.2, 16.5±0.2, 18.2±0.2, 19.4±0.2, 20.0±0.2, and 27.0±0.2 represented by 2Θ. The crystalline powders of bis(bupivacaine) pamoate are defined as polymorph C, and the X-ray powder diffraction pattern thereof is substantially as shown in Fig. 3.
The présent invention also provides amorphous powders of bis(bupivacaine) pamoate, and the X-ray powder diffraction pattern thereof is substantially as shown in Fig. 16.
The X-ray powder diffraction patterns of the compounds provided in the présent invention are measured with a DX-27mini diffractometer (Dandong Haoyuan Instrument). The measurement parameters are as follows: wavelength = 1.5406 angstrom (Cu/καΐ); stepping measurement; incrément 0.02°; initial angle 4°; terminal angle 40°; scan rate 1.0 second/step;
tube voltage 35 KV; and tube current 15 mA.
It is well known by those skilled in the art that an error for the 20 value diffraction peak position within ±0.2° is acceptable. Further, since there are différences in peak identification in X-ray powder diffraction patterns due to sample préparation, instrument différence and software processing, for example, the amount of peaks may be more or less, polymorphs are considered to be the same if the amount of peaks differ by no more than 20%.
It can be determined by those skilled in the art that a single polymorph or a mixed polymorph comprising crystalline powders in different crystalline states or amorphous form also falls within the présent invention.
The embodiments of the présent invention provides methods for preparing bis(bupivacaine) pamoate in different solvent Systems, wherein 2.0 molar équivalents or more of bupivacaine and pamoic acid are heated in different solvent Systems to form a sait, and then the température is decreased to crystallize the sait to obtain bis(bupivacaine) pamoate. In order to stably obtain a sait in a molar ratio of 2:1, it is generally required to form a sait with 2.0 molar ratio or more of bupivacaine free base and 1 molar ratio of pamoic acid in the solvent System. The 2:1 sait formed is precipitated out from the solvent in a solid form, and the excess bupivacaine free base and a portion of the 2:1 sait remain in the solvent.
The présent invention provides methods for preparing different crystalline powders from the solvent Systems such as methanol/acetone, anhydrous methanol, methanol/water, éthanol, ethanol/dimethylsulfoxide, ethanol/dimethylsulfoxide/water, water and the like.
In the présent invention, it is préférable to préparé crystalline powders from a system of éthanol, ethanol/dimethylsulfoxide, methanol/acetone, methanol/water, methanol, or water. The range for the particle diameter of the solid powders, expressed as médian particle diameter D50, is 0.1 to 50 pm, preferably 1 pm to 50 pm, and more preferably 1 pm to 20 pm.
In the présent invention, it is préférable to préparé crystalline powders from a system of éthanol, ethanol/dimethylsulfoxide, methanol/acetone, methanol/water, methanol, or water.
In addition to the methods for preparing bis(bupivacaine) pamoate with a solvent crystallization process, the présent invention provides other methods for preparing a solid sait or a co-crystal well known by those skilled in the art, including, but not limited to, the methods for preparing a bupivacaine pamoate sait or a co-crystal thereof with a good solvent-poor solvent crystallization process, a spray drying process, a film évaporation process, and a solvent-free melting process, as well as a préparation method by hot melt extrusion.
The présent invention also provides a préparation method for converting crystalline powders containing an organic solvent solvaté of bis(bupivacaine) pamoate into a hydrate of bis(bupivacaine) pamoate without any organic solvent remained, wherein the organic solvent refers to one or more of methanol, éthanol, isopropanol, n-butanol, acetonitrile, diethyl ether, acetone, tetrahydrofuran, dichloromethane, dioxane, ethyl acetate, methyl t-butyl ether, toluene, n-hexane, petroleum ether, Ν,Ν-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, pyridine, and dimethylsulfoxide.
A simple physical mixture of bupivacaine and pamoic acid appears to hâve two endothermie peaks in Differential Thermal Analysis (DTA), which are consistent with the endothermie peak of a single component of bupivacaine or pamoic acid respectively. The complex provided in the présent invention has an endothermie peak which is not consistent with the endothermie peak of a single component of bupivacaine or pamoic acid, or has no obvious endothermie peak.
The polymorph A, polymorph B, polymorph C and amorphous form of bis(bupivacaine) pamoate provided in the présent invention hâve different endothermie characteristics and solvatés, appearing to hâve different endothermie peaks in Differential Thermal Analysis (DTA). In general, the polymorph A and the polymorph C each hâve a single endothermie peak, the polymorph B has two peaks, and the amorphous form has no obvious endothermie peak.
For comparison research, the embodiments of the présent invention also provide a method for preparing a bupivacaine-pamoic acid sait (1:1 sait) and a method for preparing an insoluble sait formed from bupivacaine with other acids, including dibenzoyl tartaric acid, di-p-toluoyl tartaric acid, (R)- l,l'-Binaphthyl-2,2'-diyl hydrogenphosphate, D- camphorsulfonic acid and the like. The présent invention also provides a method for preparing an insoluble sait formed from ropivacaine with different acids. These insoluble salts include a sait of basic group and acid radical in a molar ratio of 1:1 and a sait of basic group and acid radical in a molar ratio of 2:1.
The présent invention provides a method for determining the solubility of the insoluble sait in a simulated body fluid and data for the ratio of acid radical and basic group in the suspension to illustrate the solubility and stability of the insoluble sait in the suspension. Most of the insoluble salts provided in the présent invention hâve a low solubility, but the solubility of most insoluble salts is higher than that of bupivacaine free base or ropivacaine free base under the same condition. The suspensions of some insoluble salts are not stable, and the ratio of acid radical and basic group will vary over time.
It can be known by those skilled in the art that a lower solubility can enable a longer drug dissolution time in order to achieve the purpose of sustained release of a drug.
The bis(bupivacaine) pamoate provided in the présent invention has unexpected effects. It has a very low solubility (a saturated solubility of 0.3 mM in 0.01 M PBS at pH 7.4), and can be stably présent in a simulated body fluid medium. The ratio of acid radical and basic group in the solution remains stable (a ratio of 2:1). The bis(bupivacaine) pamoate is suitable to be formulated into a solid suspension injection for use.
The présent invention provides a pharmaceutical composition comprising bis(bupivacaine) pamoate. It contains bis(bupivacaine) pamoate and a pharmaceutically acceptable excipient, and releases the drug for at least 12 hours, preferably for at least 24 hours, and more preferably for at least 72 hours.
The pharmaceutical composition may be a solid, an aqueous suspension, or a solid obtained by drying the suspension with a suitable process. The suitable drying process includes a lyophilization process, a spray drying process or other drying process.
The pharmaceutical composition is preferably an injectable composition, and may be an injection. Such an injection can be used in a manner of subcutaneous injection, intracutaneous injection, or intramuscular injection, and can locally and slowly release bupivacaine to exert a long-term analgésie effect.
The présent invention provides that the active ingrédient of the bis(bupivacaine) pamoate injection exists in a physical form of a solid microparticle suspension, which may be prepared from any crystalline powders of bis(bupivacaine) pamoate.
The solid microparticles in the bis(bupivacaine) pamoate injection provided in the présent invention hâve a particle size (expressed as médian particle size (D5q)) in a range of 0.2 pm to 50 pm, and preferably 1 pm to 20 pm.
The bis(bupivacaine) pamoate provided in the présent invention can cooperate with a suitable solvent and an additive commonly used in the injection to be formulated into corresponding compositions for subcutaneous or intramuscular injection. The pharmaceutically acceptable excipient is one or more of the following: (1) a suspending agent, (2) a surfactant, (3) a filler, (4) a preservative, (5) an isoosmotic adjusting agent, (7) a buffer, and (8) water. The suspending agent is one or more selected from a group consisting of carboxymethyl cellulose or a sodium sait thereof, hydroxypropyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, sodium hyaluronate, and polyvinylpyrrolidone, preferably one or more of sodium carboxymethyl cellulose and polyvinylpyrrolidone; the surfactant is one or more selected from a group consisting of polysorbate-20 (Tween-20), polysorbate-40 (Tween-40), polysorbate-60 (Tween-60), polysorbate-65 (Tween-65), polysorbate-80 (Tween-80), polysorbate-85 (Tween-85), polyoxyethylated castor oil, polyoxyethylated hydrogenated castor oil, lecithin, polyvinylpyrrolidone, polyethylene glycols, polyethylene oxide and polypropylene oxide ethers (Poloxamer 188, Poloxamer 407, and the like), and polyethylene glycol 15-hydroxystearate, preferably one or more of Tween-20, Tween-80, polyethylene glycol 15-hydroxystearate and Poloxamer 188; the filler is one or more selected from a group consisting of mannitol, sucrose, maltose, xylitol, lactose, glucose, starch, sorbitol and analogs thereof, preferably, mannitol, lactose and sucrose; the preservative is one or more selected from a group consisting of benzoic acid, benzyl alcohol, butylated hydroxytoluene ether, butylated hydroxytoluene, chlorobutanol, gallate, hydroxybenzoate, ethylenediamine tetraacetic acid (EDTA) and a sait thereof, phénol, chlorocresol, m-cresol, methylbenzethonium chloride, myristyl-Y-methylpyridine chloride, phenylmercuric acetate, and thimerosal, preferably one or more of benzyl alcohol and hydroxybenzoate; the isoosmotic adjusting agent is one or more selected from a group consisting of mannitol, sorbitol, sodium chloride, glucose, sucrose, fructose, and lactose, preferably one or more of mannitol, sodium chloride and glucose; and the buffer is one or more selected from a group consisting of a phosphate, an acetate, a citrate, and a tris(hydroxymethyl)aminomethane (TRIS) buffer solution, preferably a phosphate.
The injection composition provided in the présent invention contains 1 to 300 mg, and preferably 5 to 100 mg of bis(bupivacaine) pamoate per 1 mL suspension, based on the total volume of the aqueous composition; or contains not less than 10 wt%, and preferably not less than 20 wt% of bis(bupivacaine) pamoate, wherein the weight percentage of each component is calculated based on the total weight of the composition containing no water.
The présent invention also provides a test on the dissolution of the bis(bupivacaine) pamoate injection and the results thereof, indicating the features of stable dissolution and drug sustained release thereof.
The injection composition provided in the présent invention has an analgésie effect lasting for not less than 12 hours, preferably not less than 24 hours, and more preferably not less than 72 hours.
One preferred embodiment of the présent invention provides the in vivo pharmacokinetic test and the results thereof, indicating that the complex of the présent invention has a long-acting release feature exceeding 72 hours.
Therefore, the présent invention also provides use of bis(bupivacaine) pamoate and an injection thereof in the prévention or treatment of surgical pain, intraoperative pain, and postsurgical pain, preferably, postsurgical pain. The typical postsurgical pain includes, but not limited to, postsurgical pains after surgical operations such as hemorrhoidectomy, colectomy, cyst resection and the like.
Examples
The examples provided below facilitate the understanding of the présent invention, but not intended to limit the présent invention.
Ail drugs or reagents used in the présent invention are conventional commercial products, unless specifically indicated.
In the présent invention, ail conditions for High Performance Liquid Chromatography related to bupivacaine are as follows, unless specifically indicated.
The conditions for High Performance Liquid Chromatography:
HPLC-a: Stationary phase: octadecylsilyl silica gel, 250x4.6 mm, 5 pm; mobile phase A: methanol, mobile phase B: 0.1% trifluoroacetic acid, eluting gradient: as follows, flow rate: 1.0 mL/min, column température: 35°C, and UV détection wavelength: 216 nm.
Time/min Mobile phase A (%) Mobile phase B (%)
0.01 55 45
10 55 45
14 90 10
23 90 10
30 55 45
35 55 45
36 Stop
HPLC-b: Stationary phase: octadecylsilyl silica gel, 250x4.6 mm, 5 pm; mobile phase: a 10 mmol/L phosphate buffered solution at pH 2.5-acetonitrile (50:50), isocratic elution, flow rate: 1.0 mL/min, column température: 40°C, and détection wavelength: 216 nm.
Préparation Example 1. Préparation of mono(bupivacaine) dibenzoyl tartarate
Bupivacaine (1 g, 3.47 mmol) and dibenzolyl tartaric acid (DBTA, 1.3 g, 3.64 mmol) were weighed and added into ethyl acetate (30 mL). The reaction mixture was stirred and heated to become clear gradually. The reaction mixture was heated and stirred for another 2 hours after a solid was precipitated, then cooled, and filtered. The filter cake was washed with ethyl acetate twice, and dried in vacuum at 60°C for about 8 hours, to obtain 2.2 g of a white solid, i.e., mono(bupivacaine) dibenzoyl tartarate, with a yield of 95%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to dibenzolyl tartaric acid was 1:1.
Endothermie Peak: 161.2°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
‘H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 9.81(br,lH, NH), 7.95 (d, 4H, PhCO), 7.67 (m, 2H, PhCO), 7.53 (t, 4H, PhCO), 7.10 (m, 3H, MePh), 5.73 (s, 2H, CHOBz), 3.60 (m, 1H), 3.35 (m, 1H), 2.6-3.0 (m, 3H), 2.14 (s, 6H, CH3), 2.05 (m, 1H), 1.3-1.8 (m, 7H), 1.27 (m, 2H, Et), 0.88 (t, 3H, Et).
Préparation Example 2. Préparation of bis(bupivacaine) dibenzoyl tartarate
Bupivacaine (2 g, 6.98 mmol) and dibenzoyl tartane acid (1 g, 2.79 mmol) were dissolved in 20 mL acetone. The reaction mixture was heated to clear, slowly cooled to room température, then subjected to crystallization for 1 h, and filtered. The filter cake was dried in vacuum at 50°C to obtain 1.1 g of a solid, i.e., bis(bupivacaine) dibenzoyl tartarate, with a yield of 42%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to dibenzolyl tartaric acid was 2:1.
Endothermie Peak: 110.1°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
‘H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 7.94 (d, 4H, PhCO), 7.64 (m, 2H, PhCO), 7.53 (t, 4H, PhCO), 7.09 (m, 6H, MePh), 5.68 (s, 2H, CHOBz), 3.25-3.52 (m, 8H), 2.78 (m, 2H), 2.14 (s, 12H, CH3), 1.8-2.05 (m, 2H), 1.3-1.8 (m, 14H), 1.29 (m, 4H, Et), 0.89 (t, 6H, Et).
Préparation Example 3. Préparation of mono(bupivacaine) di-p-toluoyl tartarate
Bupivacaine (1 g, 3.47 mmol) and di-p-toluoyl tartaric acid (DTTA, 1.34 g, 3.47 mmol) were dissolved in 14 mL ethyl acetate. The reaction mixture was heated to reflux to become turbid gradually, slowly cooled to room température, then subjected to crystallization for 1 h, and filtered. The filter cake was dried in vacuum at 50°C to obtain 1.3 g of a white solid, i.e., mono(bupivacaine) di-p-toluoyl tartarate, with a yield of 55.6%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to di-p-toluoyl tartaric acid was 1:1.
Endothermie Peak: 161.1°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
^-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 9.87 (br, IH, NH), 7.85 (d, 4H, PhCO), 7.34 (t, 4H, PhCO), 7.10 (m, 3H, MePh), 5.69 (s, 2H, CHOBz), 3.66 (m, IH), 3.37 (m, IH), 2.6-3.0 (m, 3H), 2.38 (s, 6H, Toi), 2.14 (s, 6H, CH3), 2.05 (m, IH), 1.3-1.8 (m, 7H), 1.27 (m, 2H, Et), 0.88 (t, 3H, Et).
Préparation Example 4. Préparation of bis(bupivacaine) di-p-toluoyl tartarate
Bupivacaine (2.5 g, 8.67 mmol) and di-p-toluoyl tartaric acid (1.34 g, 3.47 mmol) were dissolved in 20 mL ethyl acetate. The reaction mixture was heated to reflux to become turbid gradually, heated and refluxed for another 20 min to precipitate a large amount of solid, then slowly cooled to room température, and filtered. The filter cake was dried in vacuum at 50°C to obtain 2.2 g of a white solid, i.e., bis(bupivacaine) di-p-toluoyl tartarate, with a yield of 65.8%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to di-p-toluoyl tartaric acid was 2:1.
Endothermie Peak: 160.9°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
'H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 9.83 (br, 2H, NH), 7.86 (d, 4H, PhCO), 7.35 (t, 4H, PhCO), 7.08 (m, 6H, MePh), 5.69 (s, 2H, CHOBz), 3.64 (m, 2H), 3.39 (m, 2H), 2.65-2.95 (m, 6H), 2.37 (s, 6H, Toi), 2.14 (s, 12H, CH3), 1.95-2.05 (m, 2H), 1.3-1.8 (m, 14H), 1.28 (m, 4H, Et), 0.86 (t, 6H, Et).
Préparation Example 5. Préparation of mono(ropivacaine) di-p-toluoyl tartarate
Ropivacaine (823 mg, 3 mmol) and di-p-toluoyl tartaric acid (1.22 g, 3 mmol) were dissolved in 20 mL acetone. The reaction mixture was heated to reflux to become turbid gradually, slowly cooled to room température, then placed in an ice water bath for crystallization for 1 h, and filtered. The filter cake was dried in vacuum at 50°C to obtain 1.7 g of a white solid, i.e., mono(ropivacaine) di-p-toluoyl tartarate, with a yield of 83.3%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of ropivacaine to di-p-toluoyl tartaric acid was 1:1.
Endothermie Peak: 174.7°C (decomposed at the same time, Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
’H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 9.88 (br, 1H, NH), 7.85 (d, 4H, PhCO), 7.33 (t, 4H, PhCO), 7.10 (m, 3H, MePh), 5.69 (s, 2H, CHOBz), 3.67 (m, 1H), 3.36 (m, 1H), 2.6-2.9 (m, 3H), 2.37 (s, 6H, Toi), 2.13 (s, 6H, CH3), 2.08 (m, 1H), 1.4—1.9 (m, 7H), 0.88 (t, 3H, Et).
Préparation Example 6. Préparation of bis(ropivacaine) di-p-toluoyl tartarate
Ropivacaine (1.375 g, 5 mmol) and di-p-toluoyl tartaric acid (773 mg, 2 mmol) were dissolved in 10 mL acetone, and heated to be dissolved to clear. The reaction mixture was slowly cooled to room température, then stirred ovemight, and filtered. The filter cake was dried in vacuum at 50°C to obtain 500 mg of a white solid, i.e., bis(ropivacaine) di-p-toluoyl tartarate, with a yield of 26.7%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of ropivacaine to di-p-toluoyl tartaric acid was 2:1.
Endothermie Peak: 147.4°C and 162.1°C (decomposed at the same time, Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
’H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 9.78 (br, 2H, NH), 7.85 (d, 4H, PhCO), 7.32 (t, 4H, PhCO), 7.07 (m, 6H, MePh), 5.66 (s, 2H, CHOBz), 3.51 (m, 2H), 3.30 (m, 2H), 2.60-2.79 (m, 6H), 2.37 (s, 6H, Toi), 2.12 (s, 12H, CH3), 1.95-2.05 (m, 2H), 1.3-1.8 (m, 14H), 0.83 (t, 6H, Et).
Préparation Example 7. Préparation of bupivacaine binaphthol phosphate
Bupivacaine (290 mg, 1 mmol) and binaphthol phosphate (l,r-binaphthyl-2,2'-diyl hydrogenphosphate, 350 mg, 1 mmol) were dissolved in 15 mL methanol, and heated to be dissolved to clear. The reaction mixture was slowly cooled to room température, placed in an ice water bath for crystallization for 1 h, and filtered. The filter cake was dried in vacuum at 50°C to obtain 320 mg of a solid, i.e., bupivacaine binaphthol phosphate, with a yield of 50%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to binaphthol phosphate was 1:1.
Endothermie Peak: 280.0°C (decomposed at the same time, Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
*H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 10.42 (s, 1H), 9.70 (br, 1H, NH), 8.04 (m, 4H), 7.41 (m, 4H), 7.31 (m, 2H), 7.20 (m, 2H), 7.13 (m, 3H, MePh), 4.05 (m, 1H), 3.38 (m, 1H), 2.9-3.1 (m, 3H), 2.20 (m, 1H), 2.14 (s, 6H, CH3), 1.3-1.8 (m, 7H), 1.28 (m, 2H, Et), 0.86 (t, 3H, Et).
Préparation Exampie 8. Préparation of bupivacaine camphorsulfonate
Bupivacaine (1 g, 3.46 mmol) and D-camphorsulfonic acid (850 mg, 3.66 mmol) were dissolved in 30 mL acetone, and heated to be dissolved to clear. The reaction mixture was slowly cooled to room température, placed in an ice water bath for crystallization for 1 h, and filtered. The filter cake was dried in vacuum at 60°C to obtain 760 mg of a solid, i.e., bupivacaine camphorsulfonate, with a yield of 41%. It was analyzed and identified through Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to camphorsulfonic acid was 1:1.
Endothermie Peak: 224.8°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
'H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 10.2 (s, 1H), 9.70 (s, 1H), 7.14 (m, 3H, MePh), 4.16 (m, 1H,), 3.55 (m, 1H), 2.9-3.25 (m, 3H), 2.9 (d, 1H), 2.68 (m, 1H), 2.40 (d, 1H), 2.20-2.30 (m, 2H), 2.16 (s, 3H), 1.5-2.0 (m, 8H), 1.1-1.4 (m, 4H), 1.04 (s, 3H), 0.88 (t, 3H), 0.74 (s, 3H).
Préparation Example 9. Préparation of mono(ropivacaine) pamoate
Ropivacaine (3.02 g, 11 mmol) and pamoic acid (1.94 g, 5 mmol) were added into a mixed solvent of 30 mL methanol and 6 mL acetone, heated to clear, then distilled at normal pressure, and supplemented with 100 mL ethyl acetate gradually. About 50 mL solvent was remained, and a large amount of solid was precipitated. The reaction mixture was filtered. The filter cake was rinsed with ethyl acetate and dried in vacuum at 50°C to obtain 2.7 g of a solid, i.e., mono(ropivacaine) pamoate, with a yield of 40.9%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of ropivacaine to pamoic acid was 1:1.
Endothermie Peak: 247.7°C (decomposed at the same time, Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
‘H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 10.23 (s, 1H, NH), 8.34 (s, 2H), 8.17 (d, 2H), 7.78 (d, 2H), 7.26 (m, 2H), 7.12 (m, 5H), 4.75 (s, 2H), 4.10 (m, 1H), 3.53 (m, 1H), 2.9-3.1 (m, 3H), 2.26 (m, 1H), 2.17 (s, 6H, Me), 1.4-1.9 (m, 7H), 0.91 (t, 3H, Et).
X-ray powder diffraction characteristic peak (wavelength = 1.5406 angstrom, Cu/καΐ):
20 (°) d (angstrom) 20 (°) d (angstrom)
7.08 12.475 16.20 5.467
8.22 10.747 16.92 5.236
10.24 8.632 19.36 4.581
10.76 8.215 20.66 4.296
12.42 7.1200 21.56 4.118
13.20 6.702 23.58 3.770
14.42 6.137 24.66 3.607
15.14 5.847 26.52 3.358
15.66 5.654
Préparation Example 10. Préparation of mono(bupivacaine) pamoate
Bupivacaine (262 g, 0.91 mol) and pamoic acid (160 g, 0.41 mol) were added into a mixed solvent of 2 L methanol and 2 L acetone, heated to clear and refluxed for 2 h, then distilled at normal pressure, and supplemented with 4 L ethyl acetate gradually. About 2 L solvent was remained, and a large amount of solid was precipitated. The reaction mixture was filtered. The filter cake was rinsed with ethyl acetate and dried in vacuum at 60°C to obtain 250 g of a light yellow solid, i.e., mono(bupivacaine) pamoate, with a yield of 90%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic
Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 1:1.
Endothermie Peak: 256.7°C (decomposed at the same time, Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
'H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 10.42 (s, 1H, NH), 8.37 (s, 2H), 8.19 (d, 2H), 7.79 (d, 2H), 7.27 (m, 2H), 7.13 (m, 5H), 4.77 (s, 2H), 4.21 (m, 1H), 3.51 (m, 1H),
3.07 (m, 3H), 2.30 (m, 1H), 2.17 (s, 6H, Me), 1.4-1.9 (m, 7H), 1.29 (m, 2H, Et), 0.91 (t, 3H, Et).
X-ray powder diffraction characteristic peak (wavelength = 1.5406 angstrom, Cu/καΐ):
20 (°) d (angstrom) 20 (°) d (angstrom)
7.04 12.549 16.00 5.535
8.14 10.854 16.40 5.401
10.22 8.649 20.60 4.308
10.68 8.277 21.44 4.141
14.18 6.241 23.68 3.754
15.08 5.871 24.40 3.645
15.38 5.757
Préparation Example 11. Préparation of bis(bupivacaine) pamoate
Bupivacaine (7.21 g, 0.025 mol) and pamoic acid (3.88 g, 0.01 mol) were added into a mixed solvent of 50 mL methanol and 50 mL acetone, and heated to obtain a clear solution (about 100 mL, a small portion of which was used for single crystal cultivation). About 98 mL thereof was slowly cooled and left standing for 2 days for crystallization, and filtered. The filter cake was rinsed with a little amount of a mixed solvent of methanol/acetone (1:1, V/V), and then dried in vacuum at 60°C, to obtain 3.82 g of a light yellow crystalline solid, i.e., bis(bupivacaine) pamoate, with a yield of 39.6%. It was analyzed and identified through High Performance Liquid 5 Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1.
Endothermie Peak: 117.2°C and 145.4°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
‘H-NMR (DMSO-d6, 300M, BRUKER AV-300): δ (ppm) 10.36 (s, 2H, NH), 8.22 (m, 4H), 10 7.68 (d, 2H), 7.15 (m, 8H), 7.06 (m, 2H), 4.71 (s, 2H), 4.11 (m, 2H), 3.50 (m, 2H), 3.01 (m, 6H),
2.25 (m, 2H), 2.18 (s, 12H, Me), 1.4-1.9 (m, 14H), 1.30 (m, 4H, Et), 0.89 (t, 6H, Et).
The above clear solution (about 2 mL) obtained by heating was diluted with acetone/methanol (1:1, V/V) by a factor of two, and then left standing at room température for crystallization for about 10 days, to obtain a bis(bupivacaine) pamoate single crystal. The single 15 crystal test data were determined through Single Crystal X-ray diffraction (Bruker Kappa Apex Duo), and shown in the table below. The Single Crystal X-ray diffraction pattern is as shown in Fig. 4. The crystal cell packing diagrams are as shown in Figs. 5, 6 and 7. The results indicated that the product was a methanol solvaté of bis(bupivacaine) pamoate.
Chemical formula C6iH8oN4Oio, i.e., 2(Ci8H28N2O)-C23Hi6O6-2(CH3OH)
Formula weight 1029.29
Température 296(2)K
Wavelength 0.71073 Â
Crystal System Monoclinic
Space group P2/c
a = 18.23(2) Â a= 90°.
Unit cell parameter b = 9.517(12) Â β= 103.44(2)°.
c= 18.40(2) Â
γ = 90°.
Volume 3104(7) Â3
Z 2
Calculated density 1.101 Mg/m3
Absorption coefficient 0.074 mm'1
F(000) 1108
Crystal size 0.280 x 0.260 x 0.220 mm3
0 range for data collection 2.276 to 25.008°.
Index range -19<=h<=21, -1 l<=k<=10, -21< :=1<=21
Collected diffraction points 16083
Independent diffraction points 5475 [R(int) = 0.0336]
The integrity of 0=25.008° 99.8 %
Fine tuning method Full-matrix least-squares on F2
Data/Limitation/Parameter 5475 / 1 /350
F2 fitting degree 1.037
R index [I>2sigma(I)] RI = 0.0805, wR2 = 0.2436
R index (full data) RI =0.1109, wR2 = 0.2791
Extinction coefficient n/a
Maximal différence peak and hole 1.317 and -0.316 e.Â-3
Préparation Example 12. Préparation of bis(bupivacaine) pamoate, polymorph B
Bupivacaine (216 g, 0.75 mol) and pamoic acid (116 g, 0.3 mol) were added into a mixed 5 solvent of 1000 mL methanol and 1000 mL acetone, and heated to clear. The solution was filtered while it was hot, then slowly cooled to room température, stirred and subjected to crystallization for 4 h, and filtered. The filter cake was washed in slurry with 500 mL of a mixed solvent of methanol/acetone (1:1, V/V), filtered, and then dried in vacuum at 60°C, to obtain 231 g of a light yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 79.9%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear
Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, with methanol residue. The content of methanol was analyzed to be 5.26% through Gas
Chromatography (GC).
Endothermie Peak: 119.0°C and 138.5°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
The X-ray powder diffraction patterns are as shown in Fig. 2 (polymorph B) and BS 149 in Fig. 13.
Préparation Example 13. Préparation of bis(bupivacaine) pamoate, polymorph B
Bupivacaine (5.04 g, 17.5 mol) and pamoic acid (1.94 g, 5 mol) were added into 70 mL methanol, heated to clear, slowly cooled to room température, stirred and subjected to crystallization ovemight, and filtered. The filter cake was rinsed with a little methanol, dried in vacuum at 50°C, to obtain 3.7 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 76.67%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, with methanol residue.
Endothermie Peak: 121.4°C and 138.5°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
It is shown from the X-ray powder diffraction pattern that the product has a polymorph B, as shown in BS 166 of Fig. 13.
Préparation Example 14. Préparation of bis(bupivacaine) pamoate, polymorph B
Bupivacaine (5.04 g, 17.5 mol) and pamoic acid (1.94 g, 5 mol) were added into 47.5 mL methanol, heated to clear, supplemented with 2.5 mL water (corresponding to 95% methanol), slowly cooled to room température, stirred and subjected to crystallization ovemight, and filtered. The filter cake was rinsed with a little methanol, dried in vacuum at 50°C, to obtain 3.9 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 80.8%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, with methanol residue.
Endothermie Peak: 120.3°C and 140.3°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
It was shown from the X-ray powder diffraction pattern that the product had a polymorph B, as shown in BS 157 of Fig. 13.
Préparation Example 15. Préparation of bis(bupivacaine) pamoate, polymorph A
7.21 g (25 mmol) of bupivacaine was dissolved in 200 mL anhydrous éthanol, and heated to reflux. A solution of pamoic acid (3.88 g, 10 mmol) of pamoic acid dissolved in 10 mL dimethyl sulfoxide) was slowly added thereto dropwise. After that, the reaction mixture was kept refluxing for 2 h, then slowly cooled to 30°C, and filtered. The filter cake was washed with a little éthanol, and dried in vacuum at 50°C, to obtain 7.1 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 74%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1. It was analyzed through GC that the content of éthanol was 8.85%.
Endothermie Peak: 149.3°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
Weight loss when melted (from 105 to 188°C): 7.712% (Thermalgravimetric Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min). The TGA-DTA diagram is as shown in Fig. 8. The results indicated that the product was an éthanol solvaté of bis(bupivacaine) pamoate.
The X-ray powder diffraction patterns are as shown in Fig. 1 (polymorph A) and BS 178 of Fig. 12.
Préparation Example 16. Préparation of bis(bupivacaine) pamoate, polymorph A
50.5 g (175 mmol) of bupivacaine was dissolved in 1400 mL anhydrous éthanol, and heated to reflux. A solution of pamoic acid in dimethyl sulfoxide (27.2 g, 70 mmol) of pamoic acid dissolved in 76 mL dimethyl sulfoxide) was slowly added thereto dropwise. After that, the reaction mixture was kept refluxing for 2 h, then slowly cooled to 30°C, and filtered. The filter cake was washed with a little éthanol, and dried in vacuum at 50°C, to obtain 51.3 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 75.9%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1. It was analyzed through GC that the content of éthanol was 7.48%.
Endothermie Peak: 149.7°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
Weight loss when melted (from 105 to 180°C): 7.137% (Thermalgravimetric Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
It was shown from the X-ray powder diffraction pattern that the product had a polymorph A, as shown in BS181 of Fig. 12.
Préparation Example 17. Préparation of bis(bupivacaine) pamoate, polymorph A
Bupivacaine (10.08 g, 35 mol) and pamoic acid (3.88 g, 10 mol) were added into 150 mL anhydrous éthanol, heated to reflux for 2 h, slowly cooled to room température in 18h with stirring, and filtered. The filter cake was rinsed with a little éthanol, dried in vacuum at 50°C, to obtain 7.18 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 74.4%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1. The content of éthanol residue was 6.5%.
Endothermie Peak: 142.4°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
It was shown from the X-ray powder diffraction pattern that the product had a polymorph A (as shown in BS 174 of Fig. 12).
Préparation Example 18. Préparation of bis(bupivacaine) pamoate, polymorph A
7.21 g (25 mmol) of bupivacaine was dissolved in 200 mL of 95% éthanol, and heated to reflux. A solution of pamoic acid (3.88 g, 10 mmol) in dimethyl sulfoxide (20 mL dimethyl sulfoxide) was slowly added thereto dropwise. After that, the reaction mixture was kept refluxing for 2 h, then slowly cooled to 30°C, and filtered. The filter cake was washed with a little 95% éthanol, and dried in vacuum at 60°C, to obtain 5.4 g of a light yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 56%. It was analyzed and identified through High
Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1. The content of éthanol residue was 5.2%.
Endothermie Peak: 144.2°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
From the X-ray powder diffraction pattern, the product was shown to be polymorph A (as shown in BS183 ofFig. 12).
Préparation Example 19. Préparation of bis(bupivacaine) pamoate, polymorph C
1.5 g of bis(bupivacaine) pamoate (polymorph A) powders obtained in Préparation Example 16 were added into 25 mL purified water, stirred at room température for 20 hours, and filtered. The filter cake was rinsed with a little purified water, and the wet product was dried in vacuum at 60°C, to obtain 1.4 g of light yellow solid powders with a yield of 93.3%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, without éthanol residue. It was analyzed through Gas Chromatography that the content of éthanol was 0.19%.
Endothermie Peak: 136.2°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
Weight loss when melted (from 105 to 185°C): 3.465% (Thermalgravimetric Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
The product was a hydrate of bis(bupivacaine) pamoate. The TGA-DTA diagram is as shown in Fig. 9.
The X-ray powder diffraction patterns are as shown in Fig. 3 and Fig. 14 (designated with BS 189-1), and the product is defined as polymorph C.
Préparation Example 20. Préparation of bis(bupivacaine) pamoate, polymorph C
10.3 g of bis(bupivacaine) pamoate (polymorph A) powders obtained in Préparation Example 16 were added into 110 mL purified water, stirred at room température for 12 hours, and filtered. The filter cake was rinsed with a little purified water, and the wet product was dried in vacuum at 60°C, to obtain 9.3 g of light yellow solid powders with a yield of 90.3%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and
Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, without éthanol residue. It was analyzed through Gas Chromatography that the content of éthanol was 0.10%.
Endothermie Peak: 136.7°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
Weight loss when melted (from 105 to 185°C): 3.674% (Thermalgravimetric Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min). The product was a hydrate of bis(bupivacaine) pamoate.
The X-ray powder diffraction pattern is as shown in Fig. 14 (designated with BS 189-2), and the product is defined as polymorph C.
Préparation Example 21. Préparation of bis(bupivacaine) pamoate, polymorph C
10.0 g of bis(bupivacaine) pamoate (polymorph B) powders obtained in Préparation Example 12 were added into 100 mL purified water, stirred at room température for 12 hours, and filtered. The filter cake was rinsed with a little purified water, and the wet product was dried in vacuum at 60°C, to obtain 9.0 g of light yellow solid powders with a yield of 90.0%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, without methanol residue. It was analyzed through Gas Chromatography that the content of methanol was 0.15%.
Endothermie Peak: 137.8°C (Differential Thermal Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
Weight loss when melted (from 105 to 185°C): 3.575% (Thermalgravimetric Analysis, SHIMADZU DTG-60A, température increasing rate: 10°C/min).
The product was a hydrate of bis(bupivacaine) pamoate.
The X-ray powder diffraction pattern is as shown in Fig. 14 (designated with BS189-3), and the product is defined as polymorph C.
The diffraction angle data in the X-ray powder diffraction patterns of Examples 19 to 21 are compared as follows. The results about polymorph are consistent with each other, but there is a différence between the peak numbers. There are peaks at ail of the diffraction angles (20) of 10.8, 12.6, 13.7, 16.5, 18.2, 19.4, 20.0 and 27.0, the maximum peak is generally at the diffraction angle of 10.8, and three peaks at the diffraction angles of 10.8, 12.6 and 13.7 are typical characteristic peaks.
BS189-1 BS 189-2 BS189-3 Minimum Average Maximum
10.801 10.800 10.820 10.800 10.8 10.820
12.619 12.560 12.620 12.560 12.6 12.620
13.701 13.640 13.700 13.640 13.7 13.701
14.199 14.199 14.2 14.199
16.502 16.502 16.542 16.502 16.5 16.542
17.600 17.359 17.601 17.359 17.5 17.601
18.200 18.221 18.240 18.200 18.2 18.240
19.360 19.359 19.360 19.359 19.4 19.360
20.000 19.980 20.000 19.980 20.0 20.000
21.020 21.040 21.021 21.020 21.0 21.040
21.739 21.740 21.739 21.739 21.7 21.740
22.899 22.879 22.961 22.879 22.9 22.961
23.920 23.801 23.899 23.801 23.9 23.920
24.639 24.699 24.738 24.639 24.7 24.738
25.618 25.482 25.539 25.482 25.5 25.618
27.040 27.060 27.060 27.040 27.1 27.060
28.221 28.143 28.143 28.2 28.221
29.338 29.282 29.340 29.282 29.3 29.340
30.220 30.177 30.177 30.2 30.220
31.023 31.059 31.023 31.0 31.059
33.522 33.581 33.520 33.520 33.5 33.581
34.819 34.159 36.678 34.159 35.2 36.678
37.258 37.258 37.3 37.258
38.420 38.499 38.358 38.358 38.4 38.499
Préparation Example 22. Other préparations and comparison between the X-ray powder diffraction patterns
Polymorphs A were prepared in different manners such as different solvent ratio, different material amounts, different crystallization rate and the like. A comparison between the diffraction angles (2Θ) in the X-ray powder diffraction patterns indicated that the diffraction patterns were substantially consistent with each other. The averages (bold) of characteristic peak angles common in 6 groups of above patterns were 4.9, 9.8, 10.9, 12.0, 12.9, 13.7, 14.7, 15.6, 10 16.3, 17.6, 18.9, 19.7, 20.2, 20.8, 21.5, 21.9, 22.7, 24.2, 24.7, 26.1, 27.3, 27.9, 31.0, 33.7, and
36.5 respectively. The underlined angle values existed in ail 6 patterns, and had a certain abondance and a certain resolution. The value for the maximum peak was generally 9.8. The batch information and X-ray powder diffraction pattern for each batch are as shown in Fig. 12 and the table below. The peaks at the diffraction angles of 21.9, 22.7, 24.2 and 27.9 for BS163 were not identified, but the comparison between the patterns indicated that the polymorphs were consistent with each other. Four unidentified peaks represented 16% of total 25 peaks. Here, the peak at the diffraction angle of 4.9 is the characteristic peak mostly distinct from other polymorphs; and three peaks at the diffraction angles of 4.9, 9.8 and 12.0 are typical characteristic peaks.
Batch Préparation method Solvent system
BS163 Same as Préparation Example 17 Anhydrous éthanol
BS 174 Préparation Example 17 Anhydrous éthanol
BS175 Same as Préparation Example 15 Ethanol/dimethylsulfoxide
BS178 Préparation Example 15 Ethanol/dimethylsulfoxide
BS181 Préparation Example 16 Ethanol/dimethylsulfoxide
BS183 Préparation Example 18 Ethanol/dimethylsulfoxide/water
Peak No. BS163 BS17 4 BS175 BS17 8 BS18 1 BS18 3 Minimu m Average Maximu m
1 5.041 4.901 4.918 4.841 4.900 4.919 4.841 4.9 5.041
2 9.940 9.820 9.820 9.741 9.800 9.820 9.741 9.8 9.940
3 10.980 10.86 0 10.859 10.84 0 10.89 9 10.86 10.840 10.9 10.980
4 12.160 12.00 0 12.019 11.94 1 12.00 0 12.00 1 11.941 12.0 12.160
5 13.039 12.93 9 12.958 12.88 0 12.94 0 12.93 9 12.880 12.9 13.039
6 13.760 13.68 0 13.662 13.68 1 13.75 9 13.65 9 13.659 13.7 13.760
7 14.839 14.76 0 14.741 14.64 1 14.71 9 14.77 7 14.641 14.7 14.839
8 15.721 15.59 9 15.481 15.54 15.61 9 15.47 8 15.478 15.6 15.721
9 16.360 16.20 2 16.259 16.23 8 16.29 9 16.29 8 16.202 16.3 16.360
10 17.660 17.56 0 17.579 17.50 0 17.56 0 17.46 2 17.462 17.6 17.660
11 19.079 18.91 9 18.841 18.85 9 18.90 0 18.90 0 18.841 18.9 19.079
12 19.800 19.70 0 19.739 19.65 9 19.68 0 19.74 0 19.659 19.7 19.800
13 20.200 20.22 0 20.260 20.20 0 20.26 0 20.26 0 20.200 20.2 20.260
14 20.960 20.80 1 20.800 20.80 0 20.82 1 20.74 1 20.741 20.8 20.960
15 21.700 21.50 0 21.501 21.40 0 21.46 1 21.52 0 21.400 21.5 21.700
16 21.84 1 21.959 21.92 0 21.94 0 21.92 1 21.841 21.9 21.959
17 22.80 0 22.620 22.67 9 22.66 0 22.73 9 22.620 22.7 22.800
18 24.25 9 24.200 24.12 24.18 0 24.20 0 24.120 24.2 24.259
19 24.640 24.69 8 24.699 24.65 9 24.66 0 24.68 0 24.640 24.7 24.699
20 26.141 26.17 9 26.100 26.06 1 26.18 0 26.14 0 26.061 26.1 26.180
21 27.462 27.26 4 27.281 27.16 0 27.34 1 27.42 2 27.160 27.3 27.462
22 27.86 0 27.903 27.90 0 27.88 1 27.96 0 27.860 27.9 27.960
23 31.322 31.26 1 31.340 31.24 1 31.33 9 29.70 1 29.701 31.0 31.340
24 33.819 33.72 1 33.760 33.73 9 33.72 0 33.72 0 33.720 33.7 33.819
25 36.482 36.55 6 36.560 36.42 3 36.54 0 36.51 7 36.423 36.5 36.560
Polymorphs B were prepared in different manners such as different solvent ratio, different material amounts, different crystallization rate and the like. A comparison between the diffraction angles (20) in the X-ray powder diffraction patterns indicated that the diffraction 5 patterns were substantially consistent with each other. The averages (bold) of characteristic peak angles common in 4 groups of above patterns were 10.9, 12.6, 13.7, 14.2, 15.7, 16.7, 17.3, 18.3, 18.9, 19.4, 20.4, 22.1, 25,1, 26.4, 27.1, 29.0, 33.6, 34,6 and 39.0 respectively. The underlined angle values existed in ail 4 patterns, and had a certain abundance and a certain resolution. The value for the maximum peak was generally 10.9. The batch information and X-ray powder 10 diffraction pattern for each batch are as shown in Fig. 13 and the table below.
It was found from a comparison with polymorph C that ail patterns had common characteristic peaks at the diffraction angles of 10.9, 12.6 and 13.7, but the intensity of the characteristic peak at the diffraction angle of 10.9 is low. The relative intensity ratio between three peaks was different from that having a polymorph C. In combination with the 15 Thermogravimetric Analysis and Gas Chromatograph (Préparation Example 12), the product was a methanol solvaté, which had a Chemical composition different from that having a polymorph C.
Batch Préparation method Solvent system
BS156 Same as Préparation Example 12 Methanol/acetone
BS157 Préparation Example 14 Methanol/water
BS149 Préparation Example 12 Methanol/acetone
BS166 Préparation Example 13 Methanol
Peak No. BS156 BS157 BS149 BS166 Minimum Average Maximum
1 10.96 10.76 10.94 10.84 10.76 10.9 10.96
2 12.58 12.52 12.66 12.62 12.52 12.6 12.66
3 13.66 13.62 13.679 13.641 13.62 13.7 13.68
4 14.179 14.14 14.22 14.14 14.14 14.2 14.22
5 15.679 15.642 15.679 15.621 15.62 15.7 15.68
6 16.201 16.16 16.181 16.16 16.2 16.20
7 16.661 16.621 16.68 16.66 16.62 16.7 16.68
8 17.28 17.201 17.278 17.319 17.20 17.3 17.32
9 18.341 18.3 18.321 18.36 18.30 18.3 18.36
10 18.901 18.881 18.901 18.90 18.88 18.9 18.90
11 19.38 19.32 19.379 19.419 19.32 19.4 19.42
12 20.44 20.361 20.439 20.46 20.36 20.4 20.46
13 21.601 21.6 21.634 21.60 21.6 21.63
14 22.041 22.082 22.081 22.201 22.04 22.1 22.20
15 22.581 22.58 22.62 22.58 22.6 22.62
16 25.14 25.04 25.18 25.22 25.04 25.1 25.22
17 26.379 26.321 26.341 26.361 26.32 26.4 26.38
18 27.08 27.069 27.161 27.261 27.07 27.1 27.26
19 28.98 28.979 28.96 29.290 28.96 29.0 28.98
20 33.456 33.516 33.779 33.48 33.46 33.6 33.78
21 34.581 34.56 34.619 34.498 34.50 34.6 34.62
22 35.703 35.719 35.7 35.70 35.7 35.72
23 38.982 39.002 39.059 39.021 39.00 39.0 39.06
Préparation Example 23. Préparation of bis(bupivacaine) pamoate
6.34 g (22 mmol) of bupivacaine and 3.88 g (10 mmol) of pamoic acid were dissolved in 5 20 mL dimethyl sulfoxide at room température. Anhydrous éthanol (200 mL) was slowly added thereto. A large amount of solid was precipitated, and filtered. The filter cake was washed with a little éthanol, and dried in vacuum at 50°C, to obtain 8.80 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 91.3%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that 10 the molar ratio of bupivacaine to pamoic acid was 2:1.
Préparation Example 24. Préparation of bis(bupivacaine) pamoate
6.34 g (22 mmol) of bupivacaine and 3.88 g (10 mmol) of pamoic acid were dissolved in 30 mL Ν,Ν-dimethyl formamide at room température. Anhydrous éthanol (200 mL) was slowly added thereto. A large amount of solid was precipitated, filtered and filtered. The filter cake was washed with a little éthanol, and dried in vacuum at 50°C, to obtain 8.97 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 93.05%. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1.
Préparation Example 25. Préparation of bis(bupivacaine) pamoate
5.76 g (20 mmol) of bupivacaine and 3.88 g (10 mmol) of pamoic acid were dissolved in 20 mL dimethyl sulfoxide at room température. Water (100 mL) was slowly added thereto. A large amount of solid was precipitated, filtered and filtered. The filter cake was washed with a little water, and dried in vacuum at 50°C, to obtain 9.6 g of a yellow solid, i.e., bis(bupivacaine) pamoate, with a yield of 99.6%. It was analyzed and identified through High Perfonnance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1.
Préparation Example 26. Préparation of bis(bupivacaine) pamoate
Bupivacaine (17.3 g, 0.06 mol) and pamoic acid (11.6 g, 0.03 mol) were added into a mixed solvent of 100 mL methanol and 100 mL acetone, and heated to clear. The solution was filtered while it was hot, and then spray dried, to obtain 29 g of a light yellow solid, i.e., bis(bupivacaine) pamoate. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, with methanol residue.
Préparation Example 27. Préparation of bis(bupivacaine) pamoate
Bupivacaine (5.76 g, 20 mmol) and pamoic acid (3.88 g, 10 mmol) were added into a mixed solvent of 33 mL methanol and 33 mL acetone, and heated to clear. The solution was film evaporated to remove solvent, to obtain 9.64 g of a light yellow solid, i.e., bis(bupivacaine) pamoate. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1, with methanol residue.
Préparation Example 28. Préparation of bis(bupivacaine) pamoate
Bupivacaine (5.76 g, 20 mmol) and pamoic acid (3.88 g, 10 mmol) were added into a reaction flask under the protection of argon, heated (150°C) to be melted, cooled to be solidified, and then ground, to obtain 9.64 g of a light yellow solid, i.e., bis(bupivacaine) pamoate. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1. No obvions characteristic peak existed in the X-ray powder diffraction pattern, indicating that the product was substantially amorphous. From the TG/DSC, the product was shown to be a single material, rather than a simply physical mixture of two mixed materials. It was also shown from the weight loss calculation that there was no solvaté. The above material was substantially consistent with the product obtained in Formulation Example 30.
Préparation Example 29. Préparation of bis(bupivacaine) pamoate, polymorph C
7.73 kg (26.8 mol) of bupivacaine was dissolved in 160 kg anhydrous éthanol, and heated to reflux and to be dissolved to clear. A solution of pamoic acid in dimethyl sulfoxide (4.16 kg (10.7 mol) of pamoic acid dissolved in 22.9 kg dimethyl sulfoxide) was slowly added thereto dropwise. After that, the reaction mixture was kept refluxing for 0.5 h, then slowly cooled to room température and stirred ovemight, and filtered. The filter cake was rinsed with éthanol, and then rinsed with water for injection. The wet filter cake was then transferred to a reaction kettle, supplemented with 220 kg water for injection, stirred at room température ovemight, filtered, rinsed with water for injection, and sucked to dryness. The wet product was blow dried at 60°C to weight loss on drying less than 5%. 9.3 kg of a light yellow solid was obtained with a yield of 86.6%, which was a hydrate of bis(bupivacaine) pamoate. It was analyzed and identified through High Performance Liquid Chromatography (HPLC-a) and Nuclear Magnetic Résonance (NMR) that the molar ratio of bupivacaine to pamoic acid was 2:1. It was analyzed through GC that the content of éthanol was <0.1%. From the TG/DSC analysis in combination with the X-ray powder diffraction pattern, the product was shown to be polymorph C.
Préparation Example 30. Préparation of non-solvate of bis(bupivacaine) pamoate
5g of the solid having a polymorph C obtained in Préparation Example 29 was placed into an oven at 150°C for 1 h such that the solid was melted, cooled to room température, and ground into fine powders.
From the X-ray powder diffraction pattern (Fig. 16), the product was shown to be amorphous, and there was no obvious diffraction peak.
The TG/DSC analysis (Fig. 17) showed that the melting point was about 112°C; the melting endothermie peak was at 123°C; and there were no obvious weight loss (<1%) in two température ranges of 25 to 105°C and 105 to 180°C, indicating that the product was a non-solvate.
The fine powders obtained was supplemented with water, and magnetically stirred for 24 hours, filtered and dried. From the TG/DSC analysis in combination with the X-ray powder diffraction pattern, the product was shown to be polymorph C.
The above results indicated that the polymorph and the amorphous form were interconvertible.
Préparation Example 31. Préparation of non-solvate of bis(bupivacaine) pamoate
5g of the solid having a polymorph B obtained in Préparation Example 12 was placed into an oven at 150°C for 1 h such that the solid was melted, cooled to room température, and ground into fine powders. From the TG/DSC analysis in combination with the X-ray powder diffraction pattern, the product was shown to be amorphous, which was consistent with the non-solvate obtained in Example 30.
Préparation Example 32. Préparation of non-solvate of bis(bupivacaine) pamoate
5g of the solid having a polymorph A obtained in Préparation Example 16 was placed into an oven at 150°C for 1 h such that the solid was melted, cooled to room température, and ground into fine powders. From the TG/DSC analysis in combination with the X-ray powder diffraction pattern, the product was shown to be amorphous, which was consistent with the non-solvate obtained in Example 30.
Préparation Example 33. Co-crystal of bupivacaine and pamoic acid
Bupivacaine and pamoic acid were mixed in a molar ratio of 1:1, 2:1 and 4:1, respectively. A portion of each mixture was taken as the physical mixture. The remaining portion was placed into an oven, heated (150°C) to be melted, cooled to be solidified, and then ground, to obtain a light yellow solid as co-crystal by melting. Additionally, the single component of bupivacaine and that of pamoic acid were heat treated in the same manner respectively. Their X-ray powder diffraction and TG/DSC were tested respectively (Fig. 19), and the results are as shown in the table below.
Bupivacaine: pamoic acid (1:1) Bupivacaine: pamoic acid (2:1) Bupivacaine: pamoic acid (4:1) Bupivacaine Pamoic acid
Physical mixture X-ray powder diffraction The diffraction peaks of two components were overlapped with each other The diffraction peaks of two components were overlapped with each other The diffraction peaks of two components were overlapped with each other
Co-crystal by melting X-ray powder diffraction The characteristic peaks of bupivacaine disappeared, and some of the characteristic peaks of pamoic acid remained Amorphous form, without obvious characteristic peak Amorphous form, without obvious characteristic peak Waxy, undetected Crystalline, without change in peaks
Physical mixture TG/DSC Bupivacaine peak was at 93.53C; pamoic acid peak was interfered, and has a gentle négative peak at 200 to 300C; and the intégration was not accurate (about 260.97) Bupivacaine peak was at 104.52C; pamoic acid peak was interfered, and has a gentle négative peak at 200 to 300C; and no intégration could be carried out Bupivacaine peak was at 106.34C; pamoic acid peak was interfered, and has a gentle négative peak at 200 to 300C; and no intégration could be carried out 105.98C 326.83C (batch 20171208)
Co-crystal by melting TG/DSC Bupivacaine has no endothermie peak; pamoic acid has a gentle négative peak at 200 to 300C; and no intégration could be carried out Bupivacaine has no endothermie peak; pamoic acid peak was interfered, and has a gentle négative peak at 200 to 300C; and no intégration could be carried out Bupivacaine has no endothermie peak; pamoic acid peak was interfered, and ahnost has no négative peak of endothermie peak; and no intégration could be carried out 99.61C, (there was a small endothermie peak at 50.34C) 326.04 (batch 20171208, dried)
It can be seen that there are several différences between the co-crystal and a single material or a physical mixture thereof. The X-ray powder diffraction pattern for the physical mixture equals to a simple addition of those of two single materials. In the TG/DSC diagram, bupivacaine has an endothermie peak, and the pamoic acid peak shifts to an earlier time. The reason may be that bupivacaine has been melted with time increasing in the test, which has a certain co-melting effect on pamoic acid before pamoic acid is melted, thereby affecting the melting peak of pamoic acid. The X-ray powder diffraction pattern for the melting co-crystal almost has no obvious diffraction peak (some of the characteristic peaks of pamoic acid remain for the 1:1 co-crystal), and the endothermie peak of bupivacaine substantially disappears in the TG/DSC diagram. Fig. 19 is an X-ray powder diffraction pattern of 8 samples, wherein the désignations 1 to 8 from bottom to top represent bupivacaine, pamoic acid, a physical mixture of bupivacaine and pamoic acid (in a ratio of 2:1), a co-crystal of bupivacaine and pamoic acid (in a ratio of 2:1), a physical mixture of bupivacaine and pamoic acid (in a ratio of 4:1), a co-crystal of bupivacaine and pamoic acid (in a ratio of 4:1), a physical mixture of bupivacaine and pamoic acid (in a ratio of 1:1), and a co-crystal of bupivacaine and pamoic acid (in a ratio of 1:1), respectively.
Formulation Example 1 g of the compound of Préparation Example 10 and 20 g of mannitol were added in a 10 mmol/L sodium phosphate buffered solution (pH7.4), stirred appropriately for suspending, and homogenized with a Panda Plus 2000 homogenizer. The effects of the particle size of the raw materials by homogenization pressure and number of cycles were investigated. The volume was set to 100 mL, and the solution after treatment was a first suspension. The particle size was determined with a laser particle size analyzer (BT9300, Liaoning Dandong Bettersize Instrument Ltd.), and the results were as follows:
Pressure and number of cycles Dio (pm) D50 (pm) D90 (pm)
untreated 11.870 34.800 78.610
6 cycles at 800 bar 0.892 4.244 8.767
2 cycles at 1200 bar 0.932 3.028 6.922
4 cycles at 1200 bar 0.776 2.770 6.108
6 cycles at 1200 bar 0.610 1.343 6.579
Further, a long-acting suspension injection was prepared according to the table below:
la 1b
First suspension (1200 bar, 6 cycles, homogenization) 100 mL 100 mL
Sodium carboxymethyl cellulose (CMC 7L2P) 1-5 g 2g
Tween-80 0.5 g 1 g
Mannitol 15 g 20 g
10 mmol/L sodium phosphate buffer (pH 7.4) to 200 mL to 200 mL
Formulation Example 2
10 g of the compound of Préparation Example 12 and 20 g of mannitol were added in a 10 mmol/L sodium phosphate buffered solution (pH7.4), stirred appropriately for suspending, and homogenized with a Panda Plus 2000 homogenizer. The effects of the particle size of the raw materials by homogenization pressure and number of cycles were investigated. The volume was set to 100 mL, and the solution after treatment was a first suspension. The particle size was 10 determined with a laser particle size analyzer (BT9300, Liaoning Dandong Bettersize Instrument Ltd.), and the results were as follows:
Particle size Dio D50 D90
untreated 3.767 18.767 42.287
2 cycles at 400 bar 1.990 8.306 17.280
4 cycles at 400 bar 1.586 7.107 14.890
6 cycles at 400 bar 1.524 4.885 11.199
2 cycles at 800 bar 1.374 4.221 8.196
4 cycles at 800 bar 1.218 4.088 8.107
6 cycles at 800 bar 1.268 3.502 6.994
2 cycles at 1200 bar 1.418 4.450 9.324
4 cycles at 1200 bar 1.338 4.238 8.798
6 cycles at 1200 bar 1.245 3.807 8.744
Further, a long-acting suspension injection was prepared according to the table below:
2a 2b
First suspension (6 cycles, homogenization) (1200 bar), 100 mL (800 bar) 100 mL
Sodium carboxymethyl cellulose (CMC 7M31F PH) 4g 0.5 g
Tween-80 1 g 0.5 g
Mannitol 20 g 15 g
mmol/L sodium phosphate buffer (pH 7.4) to 200 mL to 200 mL
Formulation Example 3 g of the compound of Préparation Example 12 and 0.1 g of Tween-80 were added in a 10 mmol/L sodium phosphate buffered solution (pH 7.4) and diluted to 100 mL with the buffer solution, stirred for suspending, and homogenized with a Panda Plus 2000 homogenizer, this suspention was named “first suspension”. The particle size of the compound after homogenization was as follows: D[0 was 1.18 pm, D50 was 4.06 pm, and D90 was 15.29 pm.
Further, a suspension was prepared according to the table below. The suspension was dispensed into vials in 10 mL per bottle, and lyophilized (LGJ-18S lyophilizer). 9 mL of water for injection was added for reconstruction and suspending before use.
First suspension (1200 bar, 6 cycles, homogenization) 100 mL
Sodium carboxymethyl cellulose (CMC 7M31F PH) 1 g
Tween-80 0.4 g
Mannitol 35 g
10 mmol/L sodium phosphate buffer (pH 7.4) to 200 mL
Formulation Example 4 g ofthe compound of Préparation Example 12 and 0.5 g of Tween-80 were added in a 10 mmol/L sodium phosphate buffered solution (pH 7.4) and diluted to 100 mL with the buffer solution, stirred for suspending, and homogenated with a Tl8 digital homogenizer, this suspention was named “first suspension”. The particle size of the compound after homogenization (measured for three times) was as follows: Dio was between 3.70 and 4.08 pm, D50 was between 13.28 and 16.80 pm, and D90 was between 28.44 and 49.01 pm.
Further, a long-acting suspension injection was prepared according to the table below. The suspension was dispensed into vials in 10 mL per bottle, and lyophilized by using a LGJ-18S lyophilizer according to the lyophilization température increasing procedure as shown in the table below. 9 mL of water for injection was added for reconstruction and suspending before use.
First suspension 100 mL
Sodium carboxymethyl cellulose (CMC 7M31F PH) 1 g
Tween-80 0-5 g
Mannitol 35 g
10 mmol/L sodium phosphate buffer (pH 7.4) to 200 mL
Lyophilization température increasing procedure:
Température Maintaining time
pre-freezing -40°C 2h
First drying -20°C 2h
-13°C 15 h
Second drying -5°C 2h
5°C 2 h
30°C 15 h
Formulation Examples 5-7 g of the compound of Préparation Example 29 was pulverized with a jet mill (J-20 type jet mill, Tecnologia Meccanica Sri, Italy).
0.1 g of Tween-80, 0.6 g of sodium carboxymethyl cellulose, 5.0 g of mannitol, and 0.28 g of sodium dihydrogen phosphate dihydrate were added in 90 mL water, and stirred to 10 dissolution, to obtain a matrix solution.
4.82 g of pulverized or unpulverized compound (Préparation Example 29) was added in 90 mL of the matrix solution, stirred to make a uniform suspension, adjusted to pH 6.5 to 7.5 with 1 mol/L sodium hydroxide, then diluted to 100 mL with water, and stirred for suspending, to obtain a long-acting suspension injection.
Formulation Example 5 Formulation Example 6 Formulation Example 7
Raw material Prescribed amount (g) Prescribed amount (g) Prescribed amount (g)
compound of Préparation Example 29 (HYR-PB21) 4.82 4.82 4.82
Treatment method Jet milling Jet milling Unpulverized
Pulverization parameter: Feeding pressure: 4 kg, Pulverization pressure: 4 kg, Rotation rate of feeder motor: 500 rpm Feeding pressure: 3kg, Pulverization pressure: 3 kg, Rotation rate of feeder motor: 500 rpm —
Particle size Dio (pm) 0.997 1.689 5.314
D5o (gm) 2.845 5.873 25.088
D90 (pm) 5.963 11.466 70.305
Excipients
Tween-80 0.10 0.10 0.10
Sodium carboxymethyl cellulose (CMC 7M31FPH) 0.60 0.60 0.60
Mannitol 5.00 5.00 5.00
NaH2PO4-2H2O 0.28 0.28 0.28
Sodium hydroxide q.s. q.s. q.s.
Water to 100 mL to 100 mL to 100 mL
pH 7.06 7.04 7.05
Needle passing ability 0.45 mm 0.45 mm 0.7 mm
Content 88.31% 91.83% 99.02%
Sample State Suspension ______Suspension______ Suspension
Formulation Examples 8-10
The compound of Préparation Example 29 was weighed in the amount as shown in the table 5 below to préparé a first suspension and a second solution respectively. The first suspension was homogenized with a Panda Plus 2000 homogenizer, added the second solution in the first suspension , and stirred to make a uniform suspension. The suspension was adjusted to pH 6.5 to 7.5 with 1 mol/L sodium hydroxide, and diluted to 1000 mL with water, mixed to make a uniform suspention. The blank excipient was formulated once in the second suspending manner
The suspension or blank excipient solution was filled into vials in 10 mL per bottle, and lyophilized according to the lyophilization procedure in Example 4. The lyophilization was tested and the results were as follows.
Formulation Example 8 Formulation Example 9 Formulation Example 10
Strength (stated amount of bupivacaine) 100 mg 300 mg Excipient blank
Raw material Prescribed amount (g) Prescribed amount (g) Formulated once and unhomogenized
First suspension compound of Préparation Example 29 (HYR-PB21) 17.36 52.09 —
Tween-80 LO LO 1.0
Mannitol 20 20 —
Water 150 150 —
Second solution Sodium carboxymethyl cellulose (CMC 7M31F PH) 6.0 6.0 6.0
Mannitol 25.0 25.0 45.0
Sodium dihydrogen phosphate (dihydrate) 1.56 1.56 1.56
Water 800 800 900
Sodium hydroxide S.q S.q S.q
Water to 1000 mL to 1000 mL to 1000 mL
Tested after yophilization
Quality properties pH 7.15 7.25 6.80
Needle passing ability Passing through a (p0.5mm needle Passing through a (p0.5mm needle
Content 93.02% 98.30% —
Sample State Pale yellow mass Pale yellow mass White mass
Water content 2.47% 1.52% 2.98%
Reconstruction time 30 seconds 40 seconds 40 seconds
Particle size distribution Dio (pm) D5o (pm) 0.850 2.232 0.825 2.050 —
D90 (pm) 4.447 3.917 —
Formulation Example 11 g of the compound of Préparation Example 16 was added in 30 mL water, stirred for 5 suspending, and homogenized with a Panda Plus 2000 homogenizer (1000 bar, 3 cycles). The partiele size was determined with a laser particle size analyzer (BT9300, Liaoning Dandong Bettersize Instrument Ltd.), and Di0, D50 and D90 were 0.923, 3.887 and 8.025 pm respectively. This suspension was named “first suspension”. Further, a long-acting suspension injection was prepared according to the table below:
First suspension (1000 bar, 3 cycles, homogenization) As described above
Sodium carboxymethyl cellulose (CMC 7M31FPH) 1-2 g
Tween-80 0.2 g
Mannitol 9 g
NaH2PO4.2H2O 0.312 g
10 mmol/L sodium phosphate buffer (pH 7.4) to 200 mL
The above suspension was lyophilized according to the lyophilization procedure in Example 4. The powder X-ray diffractions of the product and Préparation Example 11 (excipients blank) were determined. It was found from the comparison between them and an excipients blank sample, polymorph A and polymorph C (see Fig. 18 (four X-ray diffraction patterns are attributed to polymorph A, polymorph C, the excipients blank, and Formulation Example 11 from top to bottom respectively)) that the characteristic peak having a polymorph A at the diffraction angle of 4.9°/9.8° substantially disappeared, while the characteristic peak having a polymorph C (10.8°/12.6°) was obvious, indicating that polymorph A was converted into polymorph C during the suspension préparation and the lyophilization.
Formulation Example 12
2.17 g of the compound of Préparation Example 30, 0.045 g of Tween-80, and 2.25 g of mannitol were added in 15 mL water and mixed uniformly. Zirconia pellets were added thereto, and the mixture was milled with a bail mill (puiverisette 7 bail mill, PRITSCH). The parameters for bail milling were as follows: rotation rate: 1200 rpm, time: 3 min, interval time: 15 min, and number of cycles: 10. A first suspension was obtained. The particle size of the compound after bail milling was as follows: Di0 was 2.050 pm, D50 was 6.795 9 pm, and D90 was 12.480 pm.
1.0 g of sodium carboxymethyl cellulose (CMC 7MF PH) and 0.128 g of sodium dihydrogen phosphate were added in 27 mL water, stirred to dissolution, mixed with the bail milled first suspension, stirred for uniformly suspending, adjusted to pH 6.5 to 7.5 with 1 mol/L sodium hydroxide, then diluted to 45 mL with water, and stirred for suspending, to obtain a long-acting suspension injection.
Formulation Example 13
100 g of the compound of Préparation Example 29 was pulverized with a jet mill (J-20 type jet mill, Tecnologia Meccanica Sri, Italy). The parameters for pulverization were as follows: feeding pressure: 4 kg, pulverization pressure: 4 kg, and feeder motor rotation rate: 500 rpm. The particle size after pulverization was as follows: Dio = 1.125 pm, D50 = 3.017 pm, D90 = 6.224 pm.
0.1 g of Tween-80, 1.0 g of sodium carboxymethyl cellulose (7L2P), 2.5 g of mannitol, 2.0 g of polyethylene glycol 400, and 0.28 g of sodium dihydrogen phosphate dihydrate were placed into 100 mL water, stirred to dissolution, and adjusted to pH 6.5 to 7.5 with 1 mol/L sodium hydroxide, to obtain a dedicated solvent.
0.174 g of pulverized or unpulverized compound and 10 mL of the dedicated solvent were filled into a package container respectively, and formulated immediately before use, to préparé a long-acting suspension injection.
Formulation Example 14
100 g of the compound of Préparation Example 29 was pulverized with a jet mill (J-20 type jet mill, Tecnologia Meccanica Sri, Italy). The parameters for pulverization were as follows: feeding pressure: 4 kg, pulverization pressure: 4 kg, and feeder motor rotation rate: 500 rpm. The particle size after pulverization was as follows: Dio = 1.125 pm, D50 = 3.017 pm, D90 = 6.224 pm.
0.1 g of propylene glycol, 1.0 g of sodium carboxymethyl cellulose (7L2P), 2.0 g of polyethylene glycol 400, and 0.16 g of sodium dihydrogen phosphate dihydrate were placed into 100 mL water, stirred to dissolution, and adjusted to pH 6.5 to 7.5 with 1 mol/L sodium hydroxide, to obtain a dedicated solvent.
0.174 g of pulverized or unpulverized compound was mixed with 2.5 g of mannitol to obtain solid powders. 0.314 g of the mixed solid powders and 10 mL of the dedicated solvent were filled into a package container respectively, and formulated immediately before use, to préparé a long-acting suspension injection.
Tests on the properties of the compound
In the présent application, the insoluble complex represented by formula (I) or a solvaté thereof and a formulation thereof according to the présent invention were tested for the in vitro solubility, dissolution, systemic pharmacokinetics in animal body, and the like.
Test Example 1
Test on the solubility in a simulated body fluid
About 200 mg of solid powders of the example was suspended in a 50 mL phosphate buffered saline at pH 7.4 (0.01 M PBS, containing 8 mM Na2HPO4, 2 mM KH2PO4, 136 mM NaCl, and 2.6 mM KO), and stirred at 37°C for 24 hours. Appropriate amounts of the suspension were taken out at 5 min, 15 min, 30 min, 1 h, 2h, 6 h, and 24 h respectively, quickly 10 filtered, and diluted with methanol by a factor of two. The concentration of the drug dissolved in the PBS buffered solution was determined with an HPLC-a method. The results for the compounds ofthe Préparation Examples are as shown in the table below (table 1) and Fig. lOa-c. Table 1. Data for the solubility of the compounds of the examples in a simulated body fluid
Compound concentration (mM) 5 min 15 min 30 min 1 h 2 h 6h 24 h
Bupivacaine free base Saturated solubility: 1.45 mM
Ropivacaine free base Saturated solubility: 1.36 mM
Préparation Example 1 4.8846 6.9872 7.7552 8.1960 7.9117 7.0061 5.7679 Bupivacaine
4.9920 7.1077 7.8666 8.2846 8.0155 7.5905 7.3037 DBTA
Préparation Example 2 0.8190 1.5937 2.1332 2.8487 3.5654 6.5770 6.3896 Bupivacaine
0.7696 1.5504 2.1838 3.0359 3.9533 6.3363 6.1227 DBTA
Préparation Example 3 7.9476 7.8626 7.6622 7.5190 7.7193 7.8832 7.9178 Bupivacaine
7.3163 7.2446 7.0122 6.9006 7.0481 7.2117 7.2490 DTTA
Préparation Example 4 11.3894 12.0091 11.4269 11.2567 11.278 1 11.851 8 11.6933 Bupivacaine
5.1610 5.3995 5.1350 5.0848 5.0683 5.3120 5.2197 DTTA
Préparation Example 5 7.2687 7.2155 7.2872 7.3437 7.6304 7.6041 7.6489 Ropivacaine
6.6768 6.6128 6.7040 6.7521 6.9880 6.9718 7.0738 DTTA
Préparation Example 6 26.0585 29.8888 38.6576 41.9836 41.011 1 40.777 8 47.4453 Ropivacaine
13.3808 15.0173 18.5864 19.9901 19.679 0 19.617 2 22.8439 DTTA
Préparation Example 7 0.2630 0.2714 0.3105 0.2807 0.3148 0.3177 0.2684 Bupivacaine
0.2935 0.2756 0.2643 0.2633 0.2789 0.2884 0.2646 naphthol phosphate
Préparation Example 8 47.6783 57.1254 68.3149 68.1958 68.902 6 71.253 9 66.0200 Bupivacaine
— — — — — — — Camphorsulf onic acid
Préparation Example 9 0.3479 0.6026 0.8870 1.2856 1.6608 1.9440 1.9720 Ropivacaine
0.3459 0.6015 0.8803 1.2808 1.6684 1.9539 2.0523 Pamoic acid
Préparation Example 10 0.7220 0.8330 0.8859 0.9387 0.9097 0.0833 0.0670 Bupivacaine
0.7427 0.8579 0.9168 0.9816 0.9957 1.8673 2.5605 Pamoic acid
Préparation Example 12 0.1475 0.2101 0.2474 0.2672 0.2802 0.2908 0.2848 Bupivacaine
0.0736 0.1070 0.1274 0.1375 0.1400 0.1497 0.1429 Pamoic acid
Préparation Example 15 0.1128 0.1538 0.2394 0.2574 0.2875 0.2901 0.2903 Bupivacaine
0.0572 0.0779 0.1202 0.1278 0.1415 0.1467 0.1430 Pamoic acid
Préparation Example 29 0.1598 0.1969 0.2415 0.2699 0.2713 0.2751 0.3050 Bupivacaine
0.0782 0.0973 0.1221 0.1195 0.1400 0.1450 0.1516 Pamoic acid
Préparation Example 30 0.1423 0.2188 0.2337 0.2805 0.2773 0.2872 0.2869 Bupivacaine
0.0757 0.1116 0.1285 0.1356 0.1308 0.1379 0.1402 Pamoic acid
Conclusion:
It can be seen from the results that since different insoluble salts hâve different solubility, and the solubility of most salts is larger than that of the free base, so the préparation of the insoluble salts cannot be determined by reasoning from conventional technology, and the ratio between the acid radical and the basic group in the suspension is not stable for some of the insoluble salts in a simulated body fluid, which cannot be predicted from any technology and principle. In comparison, the compound of Préparation Example 7 (naphthol phosphate, about 0.3 mM), the compound of Préparation Example 9 (ropivacaine pamoate, about 2.0 mM) and the compounds of Examples 12, 15, 29 and 30 (bis(bupivacaine) pamoate, about 0.3 mM) hâve a very low solubility, and the suspensions thereof are stable.
Test Example 2
A good insoluble sait also should hâve a stable dissolution property at different pH values.
Test on the solubility in media at different pH
About 200 mg of solid powders of Préparation Example 10 and Préparation Example 15 were added in a 500 mL phosphate buffered solution at different pH values (50 mmol/L, pH 5.5, pH 6.5, pH 7.4, and pH 8.0) respectively, placed into a dissolution tester, kept at a constant température of 37°C, and paddle stirred at 50 rpm for 72 hours. Appropriate amounts of the suspension were taken out at 15 min, 30 min, 45 min, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h, 32 h, 48 h, and h respectively, and centrifuged immediately (15000 rpm, 5 min). The supematant was diluted with methanol by a factor of two. The concentration of the drug dissolved in the PB buffer solution was determined with an HPLC-b method. The results of the solubility of the examples are as shown in the table below:
The solubility table of mono(bupivacaine) pamoate (in a ratio of 1:1)
Molar concentratio n (mmol/L) of Compound of Préparation Example 10 pH5.5 pH 6.5 pH7.4 pH8.0
Bupivacai ne Pamoic acid Bupiva caine Pamoic acid Bupiva caine Pamoic acid Bupiva caine Pamoi c acid
Time /h 0.25 0.047 0.046 0.05 0.053 0.0555 0.0605 0.0575 0.063
0.5 0.078 0.076 0.084 0.09 0.095 0.105 0.099 0.110
0.75 0.106 0.103 0.115 0.124 0.1315 0.1465 0.1375 0.154
1 0.136 0.132 0.148 0.160 0.170 0.190 0.178 0.200
2 0.188 0.182 0.274 0.296 0.288 0.320 0.308 0.344
4 0.222 0.218 0.384 0.414 0.386 0.430 0.416 0.468
6 0.234 0.232 0.396 0.424 0.428 0.476 0.494 0.554
8 0.234 0.23 0.404 0.448 0.440 0.486 0.558 0.624
24 0.246 0.244 0.186 0.440 0.426 0.550 0.610 0.688
32 0.248 0.248 0.15 0.426 0.402 0.592 0.622 0.702
48 0.198 0.236 0.14 0.428 0.204 0.498 0.592 0.694
72 0.142 0.224 0.128 0.416 0.166 0.458 0.44 0.7
The solubility table of bis(bupivacaine) pamoate (in a ratio of 2:1)
Molar concentratio n (mmol/L) of Compound of Préparation Example 15 pH5.5 pH6.5 pH7.4 pH8.0
Bupivacain e Pamo ic acid Bupivac aine Pamo ic acid Bupivac aine Pamo ic acid Bupivac aine Pamo ic acid
Time/ h 0.25 0.050 0.020 0.046 0.024 0.038 0.020 0.036 0.020
0.5 0.070 0.034 0.076 0.038 0.080 0.042 0.092 0.050
0.75 0.140 0.070 0.138 0.076 0.192 0.100 0.252 0.138
1 0.196 0.092 0.202 0.106 0.240 0.134 0.328 0.182
2 0.198 0.094 0.204 0.108 0.246 0.134 0.336 0.186
4 0.198 0.092 0.202 0.106 0.242 0.134 0.338 0.192
6 0.198 0.094 0.198 0.104 0.234 0.13 0.338 0.190
8 0.198 0.092 0.198 0.106 0.232 0.128 0.334 0.186
24 0.192 0.088 0.198 0.106 0.226 0.124 0.326 0.182
32 0.196 0.092 0.202 0.106 0.226 0.124 0.328 0.186
48 0.198 0.092 0.200 0.106 0.230 0.126 0.336 0.186
72 0.190 0.084 0.196 0.100 0.224 0.120 0.336 0.184
Conclusion:
The molar ratios of the solubility of the sait comprised of bupivacaine and pamoic acid in a molar ratio of 1:1 (the compound of Préparation Example 10) in the media at pH 5.5, pH 7.4, and pH 8.0 remained around 1.0 for 48 hours or less, while the molar ratio in the medium at pH 6.5 only remained for 8 hours, and decreased after a longer time, during which the concentration of bupivacaine decreased and the concentration of pamoic acid increased, that is, the acid and the base were separated from each other. However, the molar ratios of the solubility of the sait comprised of bupivacaine and pamoic acid in a molar ratio of 2:1 (the compound of Préparation Example 15) in the media at pH 5.5, pH 6.5, pH 7.4, and pH 8.0 remained around 2.0 for at least 72 hours, and the concentrations of bupivacaine and pamoic acid did not change substantially, that is, the acid and the base would not be separated from each other.
Test Example 3
Test on the dissolution of the injection
A chromatographie column (150*4.6mm) was washed to remove the Piller, serving as a flow through cell. 1. 5 mL of samples obtained by redissolving Préparation Examples 3, 4, 6 and 8 were injected into the column. Two ends of the column were screwed tightly. The column was eluted with a high performance liquid phase. The elution medium was an aqueous solution of 1% Tween-80 and 10 mmol/L PBS. The elution flow rate was controlled to be 0.5 mL/min. The eluents were collected at 1 h, 2 h, 3 h, 4 h, 21 h, 27 h, 44 h, 51 h, and 69 h respectively, and diluted with methanol by a factor of two, and shaken up, to serve as a test sample solution. An appropriate amount of bupivacaine control sample was weighed precisely, supplemented with methanol for dissolution, and diluted to a metered volume, to préparé a solution containing 500 pg of bupivacaine per 1 mL as a control stock solution. The control stock solution was diluted to a control solution of 50 pg/mL. 20 pL of each of the control solution and the test sample solution was metered precisely and injected into a liquid chromatograph respectively. The chromatogram was recorded. The results were calculated from the peak area with an extemal standard method.
The results of the release rate of Préparation Examples 3, 4, 6 and 8 are as shown in the table below:
Release rate (%) 0 0.5 h 1 h 2h 4 h 20 h 24 h 30 h 51 h 69 h
Formulation Example 3 0 23.9 42.6 55.1 67.7 86.7 88.7 92.0 93.2 97.2
Formulation Example 4 0 15.0 21.0 27.0 32.6 59.3 64.2 70.1 - -
Formulation Example 6 0 20.2 35.4 50.0 59.7 77.7 85.4 89.3
Formulation Example 8 0 31.3. 50.4 63.3 75.5 94.0 95.2
Test Example 4
Systemic pharmacokinetic study of a bis(bupivacaine) pamoate suspension after a single subcutaneously injection at three points in a hind limb in SD rats
The systemic absorption and exposure of bis(bupivacaine) pamoate in a rat body was further evaluated by performing a systemic pharmacokinetic research on SD rats which were subcutaneously injected with a bis(bupivacaine) pamoate suspension (Formulation Example 3) once at three points in a left hind limb and comparing with a commercial bupivacaine hydrochloride injection. The long-acting sustained release feature of bis(bupivacaine) pamoate was verified by comparing the pharmacokinetic parameters of bis(bupivacaine) pamoate with a commercial formulation.
In the example, 20 healthy SD male rats (Beijing Vital River Laboratory Animal Technology Co., Ltd., 190 to 210 g) were chosen and randomly divided into 2 groups, i.e., a group for Formulation Example 3 and a group for the commercial bupivacaine hydrochloride injection (Wuhu Kangqi Pharmaceutical Co., Ltd.) group, with 10 animais in each group. Detailed administration regimen is as shown in the table below:
Table 2. The administration regimen for pharmacokinetic comparison research on SD rats which are subcutaneously injected with a bis(bupivacaine) pamoate suspension
Group Method and frequency of administration Dosage (mg/kg) Number of animal
Group for formulation Example 3 (suspension) Subcutaneous injection (single) 15 10
Bupivacaine hydrochloride injection Subcutaneous injection (single) 5 10
About 0.5 mL of venous blood was collected at 30 min, 1 h, 2 h, 6 h, 8 h, 24 h, 48 h, 72 h and 96 h respectively after administration for both groups of laboratory animais for determining the blood drug concentration of bupivacaine.
The blood drug concentration-time curve and pharmacokinetic parameters for the group for Formulation Example 3 (suspension) and the group for commercial bupivacaine hydrochloride injection are as shown in Fig. 11 and table 3, respectively. In comparison with the group for 5 commercial formulation (5 mg/kg), the Cmax value of triple dosages of bis(bupivacaine) pamoate suspension injection (15 mg/kg) was only about 12% of that of the group for commercial formulation, while the half life was as long as 32 hours, more than 30 times longer than that of the group for commercial formulation, and the AUC calculated with respect to the dosage was only 70% of that of the commercial formulation. The average blood drug concentration ofthe 10 group for Formulation Example 3 (suspension) was more than that of the group for bupivacaine hydrochloride at 6 hours after administration, and the blood drug concentration even at 72 hours after administration was still more than that of the group for bupivacaine hydrochloride at the time point of 6 hours.
The results of the présent study indicated that the bis(bupivacaine) pamoate solid 15 suspension formulation has the pharmacokinetic advantages as a sustained-release formulation for the long-acting local postsurgical analgésie development.
Table 3. Main pharmacokinetic parameters of the comparison research on SD rats which are subcutaneously injected with a bis(bupivacaine) pamoate suspension and bupivacaine hydrochloride respectively once at three points in left hind limb
Group AUCo.t (ng/mL*h) c vmax (ng/mL) T max (h) Ti/2 (h)
Group for formulation Example 3 (Suspension) 1496.3±132.1 36.2+2.3 0.63+0.2 32.2+2.4
Group for bupivacaine hydrochloride 711.1+75.3 329.6+39.1 0.63+02 1.0+0.3
Test Example 5
Test on the needle passing ability of the injection
The needle passing ability of the example formulation was investigated with different types of syringe needles. The needle passing ability was investigated by drawing the suspension injection with a larger needle, fitting 18—22G needle, and injecting the suspension injection by pushing it through the 18-22G needles. The results showed that ail the suspension injections were suitable for injection.
Scores were evaluated according to the following rating system.
Score Resuit
0 Blocked
1 Passing through a 18G needle (with an inner diameter of 0.9 mm)
2 Passing through a 19G needle (with an inner diameter of 0.7 mm)
3 Passing through a 20G needle (with an inner diameter of 0.6 mm)
4 Passing through a 21G needle (with an inner diameter of 0.5 mm)
5 Passing through a 22G needle (with an inner diameter of 0.4 mm)
The investigation results are as follows:
Formulation Example la Formulation Example 1b Formulation Example 2a Formulation Example 2b Formulation Example 3 Formulation Example 4
Score 5 2 4 4 5 3
Test Example 6
The effect of the injection dosage and the particle size on the rat pharmacokinetics healthy SD male rats (Beijing Vital River Laboratory Animal Technology Co., Ltd., 190 to 210 g) were chosen and divided into 5 groups, with 3 animais in each group. The drug of each Formulation Example was subcutaneously injected in a single dosage once at multiple points (3 points). About 0.5 mL of venons blood was collected at 5 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h, 48 h, 72 h, and 96 h respectively after administration for determining the blood drug concentration of bupivacaine. The detail information of group, administration regimen, and pharmacokinetic parameters are summarized as follows.
Group Group 17A(20 mg/kg) Group 17B (40 mg/kg) Group 17C (60 mg/kg) Group 18 (20 mg/kg) Group 19 (20 mg/kg)
Administration Formulation Example 5 Formulation Example 6 Formulation Example 7
Particle size D50 (pm) 2.845 5.873 25.088
Time (h) Blood drug level (ng/mL)
0.083 20.258 36.224 38.012 16.271 12.749
0.5 51.296 86.249 84.761 36.075 23.761
1 53.705 71.847 93.665 37.707 26.668
2 42.89 47.841 89.415 37.868 27.97
4 53.908 60.683 102.787 52.39 39.204
6 97.798 106.777 168.019 64.722 47.734
8 79.585 120.815 170.296 72.501 42.648
24 43.069 69.681 126.884 43.545 24.552
48 21.628 35.42 65.019 21.055 14.999
72 9.333 17.16 32.632 11.156 11.679
96 5.378 13.147 18.626 7.251 7.554
Pharmacokinetic parameter
AUC(O-t) 2821.85 4409.805 7431.822 2723.385 1855.703
AUC(O-œ) 2966.329 4707.339 7951.607 2990.26 2322.823
MRT(O-t) 26.744 29.466 30.781 30.448 33.38
tl/2z 21.21 23.969 23.062 27.642 43.217
Tmax 6 7.333 6.667 8 6.667
Cmax 97.798 134.091 175.224 72.501 48.847
The results indicated that ail of the particle sizes and each of the dosages had obviously 5 sustained-release pharmacokinetic characteristics, AUCs were substantially in a dose-response linear relationship. However, the linear relationship for Cmax was weaker, that is, the blood drug concentration was more stable, avoiding the adverse effect due to excessive high blood drug concentration at a large dosage. Additionally, ti/2 prolonged along with the particle size increased, and the time of sustained release and the time of maintaining the analgésie efficacy could be controlled by adjusting the particle size.
Test Example 7
The pharmacokinetic study of bupivacaine pamoate after a single subcutaneously injection in rabbit hernia models
Normal rabbits and hernia surgery model rabbits were administrated with bupivacaine pamoate (Formulation Examples 8 and 9) respectively. In comparison with the hernia model surgery model rabbits injected with the commercial bupivacaine hydrochloride injection, the différences regarding the systemic absorption and exposure of bupivacaine in the hernia surgery model animais injected with bupivacaine pamoate were evaluated, and the différences regarding the absorption and exposure of bupivacaine between normal and the hernia surgery model rabbits after bupivacaine pamoate administration were also investigated The long-acting sustained release feature of bupivacaine pamoate was verified by comparing the pharmacokinetic parameters of bupivacaine pamoate formulation with the commercial formulation.
healthy New Zealand white rabbits (Yizheng Anlimao Biological Technology Co., Ltd., 2.5 to 3.5kg) were chosen and divided into 4 groups, i.e., three groups for Formulation Examples and one group for the commercial bupivacaine hydrochloride injection (Wuhu Kangqi Pharmaceutical Co., Ltd.), with 4 animais in each group, half male and half female. Detailed administration regimen is as shown in the table below:
The administration regimen for pharmacokinetic comparison research on rabbits which are subcutaneously injected with bupivacaine pamoate
Group Drug Method and frequency of administration Dosage (mg/kg) Number of Animal
Group 1 Postsurgi cal Formulation Example 8 group (suspension) Subeutaneous injection (single) 10 4
Group 2 Postsurgical Formulation Example 9 group (suspension) Subeutaneous injection (single) 30 4
Group 3 Non-operative Formulation Example 9 group (suspension) Subeutaneous injection (single) 30 4
Group 4 Postoperative bupivacaine Subeutaneous injection (single) 10 4
hydrochloride injection
About 0.3 mL of venons blood was collected at 1 h, 2 h, 4 h, 8 h, 24 h, 48 h, 72 h and 96 h, respectively, after drug administration from ail experimental animais for determining the blood drug concentration of bupivacaine.
The blood drug concentration-time curve and pharmacokinetic parameters are shown in the table below. In comparison with the commercial formulation group(10 mg/kg), the Cmax value of triple dosages of the groups for Formulation Examples (30 mg/kg) was only up to about 15.85% of that of the group for commercial formulation, while the half life was as long as 98 hours, more than 16 times longer than that of the group for commercial formulation. The AUC of Group 1 for Example was about 106% of that of the commercial formulation, and the AUCs of Group 2 and Group 3 for high concentration formulation Examples were about 55% and 50% of that of the commercial formulation, respectively. The average blood drug concentration of the groups for Formulation Examples (suspension) was more than that of the group for bupivacaine hydrochloride at 8 hours after administration. The blood drug concentration of the low dosage group even at 72 hours after administration was still close to that of the group for bupivacaine hydrochloride at the time point of 8 hours, and the blood drug concentration of the high dosage group even at 96 hours after administration was still about 2 times greater than that of the group for bupivacaine hydrochloride at the time point of 8 hours.
The above research results indicated that the solid suspension formulation of bupivacaine pamoate has a good sustained-release pharmacokinetic feature, maintaining an active drug concentration for 96 hours or longer, and thus has a good prospect of being developed to a long-acting sustained release and local postsurgical analgésie; and the plasma bupivacaine exposure in postsurgical animais were not significantly increased than those in non-surgery animais, which ensures the safety of postsurgical administration effectively.
Table. Time-dependent drug concentration data of pharmacokinetic comparison research on rabbits which are subcutaneously injected with a bupivacaine pamoate suspension
Group 1 Group 2 Group 3 Group 4
Time Blood drug
h level —
0 0.0 0.0 0.0 0.0
1 97.8 91.4 55.5 757.4
2 99.5 89.8 83.0 516.0
4 97.0 92.4 96.4 303.8
8 92.7 93.5 99.9 169.6
24 76.6 108.5 88.0 26.2
48 42.0 69.5 67.4 2.8
72 23.1 64.4 54.6
96 14.1 47.0 45.4
Pharmacokinetic parameter
AUCo-t (ng/mL*h) 4730 7386 6692 4442
Cmax (ng/mL) 109 120 104 757
Tmax (h) 9 18 6 1
T i/2 (h) 34 72 ________98 6
Test Example 8
Local anesthetic efficacy study of intradermal administrations of bupivacaine pamoate fomulation in Guinea pigs
The local anesthetic and analgésie effects of bupivacaine pamoate on the injection site and the intensity were investigated by intradermally injecting bupivacaine pamoate to Hartley-based Guinea pigs. The long-acting local analgésie effect of bupivacaine pamoate was verified by comparing it with the commercial bupivacaine hydrochloride injection.
In this example, 6 healthy Guinea pigs (Qinglong Mountain Breeding Ground, Jiangning 10 District, Nanjing) were chosen and divided into 3 groups, i.e., low concentration group of bupivacaine pamoate (Formulation Examples 8), high concentration group of bupivacaine pamoate (Formulation Examples 9), and commercial formulation group (bupivacaine hydrochloride injection, Shanghai Zhaohui Pharmaceutical Co., Ltd.), with 2 in each group. Detailed administration regimen is shown in the table below:
Table 1. Dosage information for each group and animal group information
Concentration Dose Number of Animal
Group Sample (mg/mL) volume
1 (low concentration Formulation Example 8 (mL/indivi dual) (n)
group) 10 0.4 2
2 (high concentration Formulation Example 9 30 0.4 2
group) (commercial formulation control group)
Bupivacaine hydrochloride injection
0.4 2
Before administration, the skin in the middle 1/3 région of the back on the left side ofthe animal vertébral column was depilated, and the corresponding drug was intracutaneously injected with a 5 gauge needle in the depilated région (close injection sites were chosen at different positions as far as possible). The papule after injection was made round as far as possible. At 0.5, 3, 6, 12, 24, and 48 h after administration, the administration papule région of the Guinea pig was acupunctured with a 3 gauge needle (the acupuncture sites of different animais were made close as far as possible). The acupuncture was performed 9 times for each test. A pain response was recorded when the skin of the Guinea pig contracted or the Guinea pig brayed, or otherwise a painless response was recorded. The total painless response number was recorded to calculate the index of the painless response occurring rate for subséquent comparison of the analgésie effects.
The painless response number-time curves for the low and high concentration groups for bupivacaine pamoate and the group for bupivacaine hydrochloride for injection are shown in Fig. 15. At 0.5 h after administration, the painless response occurring rates of the low concentration group (10 mg/mL) and high concentration group (30 mg/ mL) for bupivacaine pamoate and the group for bupivacaine hydrochloride injection (5 mg/ mL) were comparable, between 8 and 9 times (a painless response occurring rate of 89%~100%); At 12 h after administration, the painless response number of the low concentration group and high concentration group for bupivacaine pamoate were between 7.5 and 9 times (a painless response occurring rate of 83%~100%); and at 24 h after administration, ail the painless response numbers were maintained 4 times (a painless response occurring rate of 44%); and at 48 h after administration, the painless response numbers were maintained once (a painless response occurring rate of 11%); The painless response numbers of the group for bupivacaine hydrochloride injection reduced to once (a painless response occurring rate of 11%) at 6 h after administration.
The above research results indicated that bupivacaine pamoate had a potentially long-acting local analgésie effect, and the local analgésie efficacy could be maintained for up to 48 hours.
The preferred embodiments of the présent invention are described in detail above with reference to the drawings. However, the présent invention is not limited to the particular details of the above embodiments. Various simple variations can be made to the technical solutions of the présent invention within the technical concept of the présent invention, and ail these simple variations fall within the protection scope of the présent invention.
Further, it should be noted that various particular technical features described in the above particular embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary répétition, various possible combinations are not further described in the présent invention.
In addition, various different embodiments of the présent invention can also be combined in any manner, as long as they do not départ from the spirit of the présent invention, and the combinations should also be regarded as being within the présent invention.

Claims (15)

1. A complex of formula (I) or a solvaté thereof:
wherein n is 1 to 4.
2. The complex or the solvaté thereof according to claim 1, wherein n is 2.
3. The complex or the solvaté thereof according to claim 2, wherein the solvaté is a 5 methanol solvaté, an éthanol solvaté, or a hydrate.
4. The complex or the solvaté thereof according to claim 2, wherein the complex or the solvaté is an éthanol solvaté having a polymorph A, wherein an X-ray powder diffraction pattern thereof, measured with Cu-Κα radiation, has diffraction peaks at about 4.9+0.2, 9.8+0.2, and 12.0+0.2 represented by 2Θ, or the X-ray powder diffraction pattern of the θ polymorph A is substantially as shown in Fig. 1.
5. The complex or the solvaté thereof according to claim 2, wherein the complex or the solvaté thereof is a methanol solvaté having a polymorph B, wherein an X-ray powder diffraction pattern thereof, measured with Cu-Κα radiation, has diffraction peaks at about 10.9+0.2, 12.6+0.2, and 13.7+0.2 represented by 20, or the X-ray powder diffraction pattern 11 of the polymorph B is substantially as shown in Fig. 2.
6. The complex or the solvaté thereof according to claim 2, wherein the complex or the solvaté thereof is a hydrate having a polymorph C, wherein an X-ray powder diffraction pattern thereof, measured with Cu-Κα radiation, has diffraction peaks at about 10.8+0.2, 12.6+0.2, and 13.7+0.2 represented by 2Θ, or the X-ray powder diffraction pattern of the 2 polymorph C is substantially as shown in Fig. 3.
7. The complex or the solvaté thereof according to claim 1 or 2, wherein the complex or the solvaté thereof is in an amorphous form.
8. The complex or the solvaté thereof according to any one of daims 1 to 7, wherein the complex or the solvaté thereof has a médian particle size D50 in a range of 0.1 to 50 pm.
9. A method for preparing the complex or the solvaté thereof according to any one of 5 daims 1 to 8, comprising mixing bupivacaine and pamoic acid in a molar ratio of greater than 1:1 and Iess than or equal to 4:1, or greater than or equal to 2:1 and Iess than or equal to 4:1, in a solvent and heating the résultant mixture, wherein the solvent is selected from a group consisting of methanol, acetone, éthanol, dimethylsulfoxide, Ν,Ν-dimethylformamide, water and a mixed solvent thereof.
Ί θ
10. A method for preparing the complex or the solvaté thereof according to claim 4, comprising mixing bupivacaine and pamoic acid in a molar ratio of greater than or equal to 2:1 in a solvent and heating the résultant mixture, wherein the solvent comprises éthanol and optionally comprises one or more selected from a group consisting of methanol, acetone, dimethylsulfoxide, Ν,Ν-dimethylformamide and water.
15
11. A method for preparing the complex or the solvaté thereof according to claim 5, comprising mixing bupivacaine and pamoic acid in a molar ratio of greater than or equal to 2:1 in a solvent and heating the résultant mixture, wherein the solvent comprises methanol and optionally comprises one or more selected from a group consisting of acetone, dimethylsulfoxide, Ν,Ν-dimethylformamide and water.
20
12. A method for preparing the complex or the solvaté thereof according to claim 6, comprising converting the complex or the solvaté thereof according to claim 4, 5 or 7 into a bis(bupivacaine) pamoate hydrate in water.
13. A method for preparing the complex according to claim 7, comprising converting the complex or the solvaté thereof according to claim 4, 5 or 6 into amorphous powders by heating it to remove the solvent; or preparing amorphous powders from bupivacaine and pamoic acid by a melting method.
14. A pharmaceutical composition comprising a pharmaceutically effective amount of the complex or the solvaté thereof according to any one of daims 1 to 8 and a fi pharmaceutically acceptable excipient.
15. The complex or the solvaté thereof according to any one of claims 1 to 8 for use in the prévention or treatment of surgical pain, intraoperative pain, and postsurgical pain, wherein the complex or the solvaté thereof is administrated via subcutaneous injection, intracutaneous injection, or intramuscular injection.
OA1201900067 2017-03-27 2018-03-26 Poorly soluble complex or solvate thereof, pharmaceutical composition, and application thereof. OA19471A (en)

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