CN115894340A

CN115894340A - A crystal form of bupivacaine meloxicam salt, single crystal, preparation method and application thereof

Info

Publication number: CN115894340A
Application number: CN202211281061.0A
Authority: CN
Inventors: 唐田; 陈柏州; 梁东丽; 孙彦辉; 林锐彬; 蒋娟娟; 刘淑芬
Original assignee: Jiali Shenzhen Biotechnology Co ltd
Current assignee: Jiali Shenzhen Biotechnology Co ltd
Priority date: 2022-10-19
Filing date: 2022-10-19
Publication date: 2023-04-04

Abstract

The application relates to a crystal form A of bupivacaine meloxicam salt, a single crystal, a preparation method and an application thereof. The A crystal form of the bupivacaine meloxicam salt has characteristic diffraction peaks at the following 2 theta (DEG) angles in an X-ray powder diffraction pattern: 7.5 +/-0.2, 8.3 +/-0.2, 12.7 +/-0.2, 15.1 +/-0.2, 16.1 +/-0.2, 16.6 +/-0.2, 24.9 +/-0.2 and 25.0 +/-0.2. The crystal form A has stable form and determined melting point, good chemical stability, high temperature resistance and illumination resistance, has the performance required by preparing solid preparations, has higher administration concentration, better dissolution, better compressibility and disintegration, convenient storage, simpler and more convenient production operation of the preparation and easier quality control.

Description

A crystal form of bupivacaine meloxicam salt, single crystal, preparation method and application thereof

Technical Field

The application relates to the technical field of medicines, in particular to a A crystal form of bupivacaine meloxicam salt, a single crystal, a preparation method and an application thereof.

Background

Bupivacaine (Bupivacaine) is an amide local anesthetic, and the first product on the market is Bupivacaine hydrochloride injection which is developed and marketed by Hospira company in 1972 for surgical operation, oral operation, diagnostic and therapeutic operation and obstetrical operation, and has the trade name Marcaine, and is also called as cocaine. As a non-opioid, bupivacaine avoids opioid addiction, but is routinely administered at a concentration of 0.5%. Therefore, the clinical need is to improve the administration concentration, and the pharmaceutical need is to solve the solubility.

Meloxicam is a non-steroidal anti-inflammatory drug (NSAID) and antipyretic, currently used to relieve symptoms of arthritis, fever, and is useful as an analgesic for inflammatory conditions. Meloxicam was originally developed by boehringer ingelheim and sold in europe under the brand names Melox, movalis and Recoxa for the treatment of rheumatoid arthritis, for short-term use in osteoarthritis and ankylosing spondylitis. Meloxicam is commercialized as tablets, orally disintegrating tablets and capsules, 7.5 and 15 mg per dose, and 7.5 mg/5 ml per dose of oral suspension.

The bupivacaine/meloxicam sustained-release solution (trade name: zynrelef, research and development code: HTX-011) is developed by Heron therapeutics company, is approved by FDA to be marketed at 13.05.1.05.1.A patient with adult undergoes bunions after bunions excision, open inguinal hernia repair and total knee replacement, and is instilled on soft tissues or around joints in an operation area to generate postoperative analgesia. The product is approved by European Union as early as 09 months in 2020, and can be used for treating postoperative pain of body caused by small and medium-sized operation wound of adult.

Zynrelef is a fixed dose combination of bupivacaine and meloxicam, the ratio is 33.1, wherein, the dosage of meloxicam as a non-steroidal anti-inflammatory drug (NSAID) is less, which can reduce wound inflammation, relieve pain, increase the local pH value of wound, make the pH value tend to be normal and allow more bupivacaine to penetrate cell membrane, thus achieving good synergistic analgesic effect.

Zynrelef is the only double-acting local Anesthetic (DALA) currently on the market, but the preparation adopts

The technology takes fourth generation Polyorthoester (POE) prepared by condensation of diol and diketone acetal as material, and the polyorthoester material is actually composition material and comprises diethylene tetraoxaspiro undecane, triethylene glycol and triethylene glycol polyglycolide. The pharmaceutical stability of these novel excipients is to be investigated, and DMSO is required to be used as a preparation solvent, and the application in the process is to be further confirmed.

Disclosure of Invention

Based on the above, the application provides the A crystal form of the bupivacaine meloxicam salt, the single crystal, the preparation method and the application thereof, wherein the A crystal form of the bupivacaine meloxicam salt has higher administration concentration, better dissolution and stable property.

In a first aspect of the present invention, there is provided a crystalline form a of bupivacaine meloxicam salt having an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2 Θ (°) angles:

7.5±0.2、8.3±0.2、12.7±0.2、15.1±0.2、16.1±0.2、16.6±0.2、24.9±0.2、25.0±0.2。

2. the crystalline form a of a bupivacaine meloxicam salt according to claim 1 further comprising 1 or more than 2 characteristic diffraction peaks at angles 2 Θ (°) selected from the group consisting of: 18.9 plus or minus 0.2, 10.1 plus or minus 0.2, 17.3 plus or minus 0.2, 18.9 plus or minus 0.2, 21.0 plus or minus 0.2 and 25.2 plus or minus 0.2.

In a second aspect of the present invention, there is provided a method for preparing the crystalline form a of bupivacaine meloxicam salt according to the first aspect, comprising the steps of:

mixing bupivacaine, meloxicam and an organic solvent, heating and refluxing to dissolve, then concentrating the reaction solution until a solid is precipitated, adding an anti-solvent, continuing to precipitate the solid, collecting the solid, and drying to prepare the A crystal form of the bupivacaine meloxicam salt.

In a third aspect of the present invention, there is provided the monocrystals of form a of the bupivacaine meloxicam salt of the first aspect, wherein the monocrystals have a unit cell structure, and one unit cell comprises 4 monocrystals of the bupivacaine meloxicam salt.

In a fourth aspect of the present invention, a pharmaceutical composition is provided, which comprises one or more of the crystal form a of the bupivacaine meloxicam salt of the first aspect and the single crystal of the crystal form a of the bupivacaine meloxicam salt of the third aspect, and a pharmaceutically acceptable carrier and/or excipient.

In a fifth aspect of the present invention, there is provided a crystalline form a of a bupivacaine meloxicam salt according to the first aspect, a single crystal of crystalline form a of a bupivacaine meloxicam salt according to the third aspect, or a pharmaceutical composition according to the fourth aspect for use in the manufacture of a medicament having an effect in the treatment of local pain.

The inventor crystallizes the crude product of the bupivacaine meloxicam salt by a recrystallization method in the process of preparing the crystalline compound of the bupivacaine meloxicam salt to obtain a new crystal form, namely a crystal form A. The crystal obtained was confirmed to be a novel crystal by detecting and analyzing the crystal by X-ray powder diffraction, DSC, TG, IR, elemental analysis, etc., and was named as crystal form a of a crystalline compound of 1-N-butyl-2- (2, 6-dimethylaminoformyl) piperidine and 4-hydroxy-2-methyl-N- (5-methyl-2-thiazolyl) -2H-1, 2-benzothiazine-3-carboxamide 1, 1-dioxide. The crystal form A has stable form and determined melting point, good chemical stability, high temperature resistance and illumination resistance, has the performance required by preparing solid preparations, has higher administration concentration, better dissolution, better compressibility and disintegration, convenient storage, simpler and more convenient production operation of the preparation and easier quality control.

Meanwhile, the crystal form A is also found to have good bioavailability in the research process.

Drawings

FIG. 1 is an X-ray diffraction pattern of form A obtained in example 1 of the present invention;

FIG. 2 is a DSC pattern of the crystal form A obtained in example 1 of the present invention;

FIG. 3 is a TG spectrum of the form A obtained in example 1 of the present invention;

FIG. 4 is an IR spectrum of the form A obtained in example 1 of the present invention;

FIG. 5 is an HPLC chromatogram of form A obtained in example 1 of the present invention;

FIG. 6 is a graph comparing the stability tests of form A obtained in example 1 of the present invention;

FIG. 7 is a graph comparing the bioavailability of form A obtained in example 1 of the present invention and bupivacaine in animals;

FIG. 8 is a crystal morphology of a single crystal obtained in example 20 of the present invention;

FIG. 9 shows the molecular structure and atomic number (a ball-and-stick diagram) of a single crystal obtained in example 20 of the present invention;

FIG. 10 is a view showing the molecular structure and atomic number (line diagram) in a crystal of a single crystal obtained in example 20 of the present invention;

FIG. 11 is a unit cell structure (color by atomic species) of a single crystal obtained in example 20 of the present invention;

FIG. 12 shows a unit cell structure (colored in a symmetrical relationship) of a single crystal obtained in example 20 of the present invention;

FIG. 13 is the absolute configuration of a single crystal obtained in example 20 of the present invention (wherein black C is a chiral C atom).

Detailed Description

The following examples are provided to further illustrate the crystalline form a of the bupivacaine meloxicam salt of the present application and the single crystals, the preparation method and the use thereof. This application may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the present application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

As used herein, "one or more" refers to any one, any two, or any two or more of the listed items.

In this application, "first aspect", "second aspect", "third aspect", "fourth aspect", etc. are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or quantity, nor as implicitly indicating the importance or quantity of the technical feature indicated. Moreover, "first," "second," "third," "fourth," etc. are used merely as non-exhaustive lists and should not be construed as limiting the number of instances.

In the present application, the technical features described in the open manner include a closed technical solution including the listed features, and also include an open technical solution including the listed features.

In the present application, reference is made to numerical ranges which are considered to be continuous within the numerical ranges, unless otherwise specified, and which include the minimum and maximum values of the range, as well as each and every value between such minimum and maximum values. Further, when a range refers to an integer, each integer between the minimum and maximum values of the range is included. Further, when multiple range-describing features or characteristics are provided, the ranges may be combined. In other words, unless otherwise indicated, all ranges disclosed herein are to be understood to encompass any and all subranges subsumed therein.

The percentage contents referred to in the present application mean, unless otherwise specified, mass percentages for solid-liquid mixing and solid-solid phase mixing, and volume percentages for liquid-liquid phase mixing.

The percentage concentrations referred to in this application, unless otherwise specified, refer to the final concentrations. The final concentration refers to the ratio of the additive component in the system to which the component is added.

The temperature parameter in the present application is not particularly limited, and may be a constant temperature treatment or a treatment within a certain temperature range. The constant temperature process allows the temperature to fluctuate within the accuracy of the instrument control.

The room temperature in this application is generally 4 to 30 ℃ and preferably 22 to 27 ℃.

The "bupivacaine meloxicam salt" in the present application refers to a salt formed by reacting bupivacaine with meloxicam, which has the following structural characteristics:

the chemical names of the bupivacaine meloxicam salt are 4-hydroxy-2-methyl-N- (5-methyl-2-thiazolyl) -2H-1, 2-benzothiazine-3-carboxamide 1, 1-dioxide and 1-N-butyl-2- (2, 6-dimethylaminoformyl) piperidine.

The application provides a crystal form A of bupivacaine meloxicam salt, wherein an X-ray powder diffraction pattern of the crystal form A has characteristic diffraction peaks at the following 2 theta (DEG):

7.5 plus or minus 0.2, 8.3 plus or minus 0.2, 12.7 plus or minus 0.2, 15.1 plus or minus 0.2, 16.1 plus or minus 0.2, 16.6 plus or minus 0.2, 24.9 plus or minus 0.2 and 25.0 plus or minus 0.2.

Without limitation, the radiation source used in the X-ray powder diffraction pattern described above is a Cu radiation source. The X-ray powder diffraction pattern refers to a PXRD pattern.

Further, said form a has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2 Θ (°) angles:

7.5 plus or minus 0.1, 8.3 plus or minus 0.1, 12.7 plus or minus 0.1, 15.1 plus or minus 0.1, 16.1 plus or minus 0.1, 16.6 plus or minus 0.1, 24.9 plus or minus 0.1 and 25.0 plus or minus 0.1.

Further, the relative intensities (I/I) of the above-mentioned characteristic diffraction peaks ₀ ) Are all greater than or equal to 30 percent.

In one example, the form a further comprises 1 or 2 or more characteristic diffraction peaks at angles 2 θ (°) selected from: 18.9 plus or minus 0.2, 10.1 plus or minus 0.2, 17.3 plus or minus 0.2, 18.9 plus or minus 0.2, 21.0 plus or minus 0.2 and 25.2 plus or minus 0.2.

Further, said form a further comprises 1 or more than 2 characteristic diffraction peaks at angles selected from the following 2 Θ (°): 18.9 plus or minus 0.1, 10.1 plus or minus 0.1, 17.3 plus or minus 0.1, 18.9 plus or minus 0.1, 21.0 plus or minus 0.1 and 25.2 plus or minus 0.1.

Further, the relative intensity (I/I) of the above-mentioned characteristic diffraction peak ₀ ) Are all greater than or equal to 20%.

In one example, the form a further comprises 1 or 2 or more characteristic diffraction peaks at angles 2 θ (°) selected from: 18.1 + -0.2, 20.2 + -0.2, 22.7 + -0.2, 23.8 + -0.2, 24.0 + -0.2, 25.8 + -0.2, 26.4 + -0.2 and 26.5 + -0.2.

Without limitation, the radiation source used in the X-ray powder diffraction pattern is a Cu radiation source. The X-ray powder diffraction pattern refers to a PXRD pattern.

Further, said form a further comprises 1 or more than 2 characteristic diffraction peaks at angles selected from the following 2 Θ (°): 18.1 +/-0.1, 20.2 +/-0.1, 22.7 +/-0.1, 23.8 +/-0.1, 24.0 +/-0.1, 25.8 +/-0.1, 26.4 +/-0.1 and 26.5 +/-0.1.

Further, the relative intensities (I/I) of the above-mentioned characteristic diffraction peaks ₀ ) Are all greater than or equal to 10%.

In one example, the main parameters of the X-ray powder diffraction pattern of the crystal form A are as follows:

where the relative intensity values are close values, "close" refers to the uncertainty in the intensity measurements, it is understood by those skilled in the art that the uncertainty in the relative intensity is related to the measurement conditions, and may vary, for example, within ± 25% or preferably within ± 10%.

In one example, the form a has an X-ray powder diffraction pattern substantially as shown in figure 1. Without limitation, the radiation source used in the X-ray powder diffraction pattern described above is a Cu radiation source. The X-ray powder diffraction pattern refers to a PXRD pattern.

In one example, the differential scanning calorimetry curve of form a has an endothermic peak at 189.6 ℃ ± 3 ℃. Further, the differential scanning calorimetry curve of the crystal form A has an endothermic peak at 189.6 ℃ +/-1 ℃.

In one example, the differential scanning calorimetry curve of form a is substantially as shown in figure 2.

In one example, the thermogravimetric spectrum of form a has a weight loss of 57% ± 1% in the range of 210 ℃ to 300 ℃.

In one example, the thermogravimetric profile of the form a is substantially as shown in figure 3. Meanwhile, thermogravimetric analysis (TGA) shows a weight loss of 0.4% ± 1% in the range of 50 ℃ to 150 ℃, which is a loss due to solvent residue.

In one example, the infrared spectrum of form a includes the following absorption peaks:

3436cm ^-1 、3291cm ^-1 、2960cm ^-1 、1690cm ^-1 、1603cm ^-1 、1549cm ^-1 、1522cm ^-1 、1456cm ^-1 、1395cm ^-1 、1337cm ^-1 、1233cm ^-1 、1188cm ^-1 、1169cm ^-1 、1125cm ^-1 、1067cm ^-1 、941cm ^-1 、861cm ^-1 、768cm ^-1 、741cm ^-1 、619cm ^-1 、574cm ^-1 、524cm ^-1 and 464cm ^-1 。

In one example, the form a has an infrared spectrum substantially as shown in figure 4.

The present application also provides a method for preparing the form a of the bupivacaine meloxicam salt as described above, comprising the steps of:

In one example, the organic solvent is one or more of butanone, acetone, tetrahydrofuran, acetonitrile, dimethyl sulfoxide, ethyl acetate, and isopropyl acetate; further, the organic solvent is acetone.

In one example, the anti-solvent is one or more of water, isopropanol, and n-heptane. Further, the anti-solvent is water.

In one example, the volume ratio of the organic solvent to the anti-solvent is 1 to 4. Further, the volume ratio of the organic solvent to the anti-solvent is 1.5 to 2.5.

In one example, the molar ratio of meloxicam to bupivacaine is 1. Further, the molar ratio of the meloxicam to the bupivacaine is 1.

In one example, the ratio between the total mass of bupivacaine and meloxicam and the total volume of the organic solvent and the antisolvent is 5mg to 20mg. Further, the ratio is 7mg to 11mg.

In one example, the temperature of the heating reflux is 30 ℃ to 80 ℃. Further, the temperature of the heating reflux is 58 ℃ to 62 ℃.

In one example, the time for continuing to precipitate the solid is 2 to 8 hours. Further, the time for continuing to precipitate the solid was 3 to 5 hours.

In one example, the temperature at which the precipitation of solids continues is 0 to 40 ℃. Further, the temperature for continuously precipitating the solid is 20-30 ℃.

In one example, the temperature of drying is 30 ℃ to 60 ℃. Further, the drying temperature is 45 ℃ to 55 ℃.

In one example, the concentration is carried out at a temperature of 40 ℃ to 50 ℃ and a pressure of-0.2 MPa to-0.1 MPa.

The present application also provides a single crystal of form a of the bupivacaine meloxicam salt as described above, having a unit cell structure wherein one unit cell comprises 4 single crystal molecules of the bupivacaine meloxicam salt.

In one example, the single crystal has a unit cell structure in which the independent symmetric elements are a duplex helical axis and an n-slip plane.

In one example, the crystallographic structure parameters of the single crystal are as follows:

。

the application also provides a preparation method of the monocrystals of the A crystal form of the bupivacaine meloxicam salt, which comprises the following steps:

mixing bupivacaine, meloxicam and an organic solvent, heating and refluxing to dissolve, volatilizing the organic solvent in the reaction solution until a solid is precipitated, collecting the solid, and preparing the single crystal.

In one example, the organic solvent is one or more of butanone, acetone, tetrahydrofuran, acetonitrile, dimethyl sulfoxide, ethyl acetate, and isopropyl acetate. Further, the organic solvent is acetone.

In one example, the ratio between the total mass of the bupivacaine and the meloxicam and the volume of the organic solvent is 5mg to 20mg. Further, the ratio is 7mg to 11mg.

In one example, the temperature of the heating reflux is 30-80 ℃; further, the temperature of the heating reflux is 58 ℃ to 62 ℃.

In one example, the step of volatilizing the organic solvent in the reaction solution comprises the steps of: and (3) placing the reaction solution into a glass bottle, sealing the glass bottle by using a sealing layer, and arranging a vent hole on the sealing layer. Without limitation, the sealing layer is a sealing film.

Further, the vent hole is formed by pricking 1-2 holes with a needle of a 10mL syringe.

In one example, the organic solvent in the reaction solution is volatilized for 2 to 10 days at room temperature.

The present application also provides a pharmaceutical composition comprising one or more of the crystalline form a of the bupivacaine meloxicam salt described above and a single crystal of the crystalline form a of the bupivacaine meloxicam salt described above, and a pharmaceutically acceptable carrier and/or excipient.

It will be appreciated that the above pharmaceutical compositions may be further formulated for administration according to conventional formulation methods, including oral or parenteral administration. In the form for administration, a therapeutically effective amount of the crystalline form a of the bupivacaine meloxicam salt and/or a single crystal of the crystalline form a of the bupivacaine meloxicam salt should be included. By "therapeutically effective amount" is meant that at this dose, the single crystals of form a of bupivacaine meloxicam salt and of form a of bupivacaine meloxicam salt according to the present invention not only have improved therapeutic activity in pain relief, but also show greatly improved solubility when used for parenteral administration and also greatly improved transdermal penetration when used for topical treatment.

Without limitation, the pharmaceutical composition is suitable for use in topical treatments and its formulation form includes, but is not limited to, solutions, gels, milk gels, creams, ointments, lotions, skin patches or eye drops. The pharmaceutical composition is also suitable for parenteral injection.

The application also provides the application of the pharmaceutical composition in preparing the medicine with the effect of treating local pain. The local pain includes, but is not limited to, muscle pain, joint pain, pain associated with herpes infections, wound pain (e.g., surgical pain or burn pain), and the like.

The pharmaceutical compositions of the present application are particularly suitable for treating patients with localized pain. Without limitation, it is the topical and parenteral treatment of these patients by an effective amount of the pharmaceutical formulation.

Specific examples are as follows.

The meloxicam, bupivacaine and other agents referred to in the examples were all purchased commercially.

Test conditions of the examples samples:

PXRD:

a detection instrument: rigaku SmartLab SE

Detection conditions are as follows: the voltage of the X-ray light pipe is 40kV, the current of the X-ray light pipe is 40mA, the scanning range is 3-40 degrees (2 theta), the step length is 0.02 degrees, and the scanning speed is 5 degrees/min.

The detection basis is as follows: appendix IX F X ray powder diffraction method of the people's republic of China (2020 edition two parts)

And (3) detection results: as in fig. 1.

(II) DSC

A detection instrument: relaxation tolerant STA 449 F3

Detection conditions are as follows: nitrogen, 50mL/min

And (3) scanning procedure: 30-350 ℃, heating rate: 10 ℃/min

Detecting the quality of a sample: 2.906mg (alumina sample plate)

The detection basis is as follows: general rules for JY/T014-1996 thermal analysis method

And (3) detection results: as shown in fig. 2.

(III) TGA

A detection instrument: relaxation tolerant STA 449 F3

Detection conditions are as follows: nitrogen, 50mL/min

And (3) scanning procedure: 30-350 ℃, heating rate: 10 ℃/min

Detecting the quality of a sample: 1.68mg (alumina sample plate)

And (3) detection results: as shown in fig. 3.

(IV) Infrared Spectroscopy

A detection instrument: PE Fourier transform infrared microscope system (Spotlight 200 i)

Detection conditions are as follows: potassium bromide tableting method

The detection basis is as follows: general rule of GB/T6040-2002 infrared spectrum analysis method

And (3) detection results: as shown in fig. 4.

(V) HPLC

A detection instrument: waters ALLIANCE E2695

A chromatographic column: reversed C18 liquid chromatography column (octadecyl silane bonded silica gel as filler)

Column temperature: 30 deg.C

Mobile phase: methanol-0.1 mol/L ammonium acetate solution (1

Detection wavelength: 270nm

And (3) detection results: as shown in fig. 5.

(VI) SXRD

A detection instrument: bruker D8 VENTURE double micro focal spot single crystal X-ray diffractometer

Detection conditions are as follows: ambient temperature 193K, enhanced Cu light source, wavelength

The detection basis is as follows: JY/T0588-2020 general rules of Crystal and molecular Structure analysis method for measuring Small molecular Compounds by Single Crystal X-ray diffractometer

And (3) detection results: as shown in fig. 8 to 13.

Preparation of crystalline Compound A form of 1-N-butyl-2- (2, 6-dimethylaminobenzoyl) piperidine and 4-hydroxy-2-methyl-N- (5-methyl-2-thiazolyl) -2H-1, 2-benzothiazine-3-carboxamide 1, 1-dioxide

Example 1 a crystalline form of bupivacaine/meloxicam salt a was prepared in an enlarged scale by means of a method of elution crystallization.

10g of meloxicam and 8.2g of bupivacaine were added to 2L of acetone (organic solvent), heated under reflux at 60 ℃ and stirred to dissolve completely. And (3) concentrating the clear solution under reduced pressure (45 ℃ and-0.1 MPa) until solids are separated out, stopping concentrating, adding 1L of n-heptane (antisolvent), stirring for 4h (solidification time) at 25 ℃ (solidification temperature), filtering, and vacuum drying the solid sample at 50 ℃ (drying temperature) for 12h to obtain the bupivacaine meloxicam salt crystal form A. 15.8g of light yellow solid powder, the mass yield is 86.5%.

The compound traits were identified as shown in FIGS. 1 to 5.

[ examples 2 to 16]

A mixture of 10g of meloxicam and 8.2g of bupivacaine was added to each reaction flask, and the experiment was performed according to the same experimental procedure as in example 1 while changing the parameter conditions, which are designed as shown in the following Table 1:

TABLE 1

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[ example 17]

The stability of the crystalline form A of the bupivacaine meloxicam salt of example 1 under conditions of high temperature (50 ℃), light, acceleration (40 ℃,75% RH) was examined, and samples were taken for 5 days and 10 days for PXRD and HPLC analysis, respectively, and the results are shown in Table 2 and FIG. 6. The result shows that the moisture, the purity and the crystal form of the bupivacaine meloxicam salt A are compared with the data of 0 day under the conditions of high temperature, illumination and acceleration, and the result shows that the obtained crystal form A is stable.

TABLE 2 stability test results for bupivacaine/meloxicam salt A crystal form

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EXAMPLE 18 preparation of solid pharmaceutical preparation

Prescription 1:

composition (I)	Dosage of
		Crystalline form A of bupivacaine meloxicam salt (example 1)	10mg
Mannitol	108mg
		Lactose	50mg
Cross-linked polyvidone	6mg
		Silica gel micropowder	3mg
Glyceryl behenate	3mg
		Total amount of	180mg

The preparation method comprises the following steps: the above components are mixed according to conventional preparation method, and directly compressed into tablet.

Prescription 2:

composition (I)	Dosage of
		Crystalline form A of bupivacaine meloxicam salt (example 1)	10g
Mannitol	80g
		Lactose	65g
Cross-linked polyvidone	15g
		Silica gel micropowder	5g
Glyceryl behenate	5g
		3％HPMC	Proper amount of
Total amount of	180g

The preparation method comprises the following steps: the bupivacaine meloxicam salt A crystal form of the example 1 is evenly mixed with mannitol, lactose and crospovidone according to an equivalent multiplication method, HPMC solution prepared in advance is added to prepare soft materials, granulation is carried out through a 20-mesh sieve, drying is carried out for 30 minutes at 60 ℃, granulation is carried out through a 18-mesh sieve, superfine silica gel powder is added, and the mixture is filled into No. 2 capsules.

Comparison prescription:

EXAMPLE 19 factor control test

3 batches of samples are prepared according to the processes of the formulas 1 to 2 and the comparative formula in the embodiment 18, after basic project inspection is qualified, illumination, high temperature and high humidity tests are respectively carried out, the appearance, the content and the dissolution rate of the samples are inspected, and the results of influencing factors show that the samples have stable properties under the conditions of high temperature and illumination and can be used as a preparation reference formula and a preparation reference process.

The prescriptions 1-2 and the comparison prescription are tested for dissolution rate, compressibility and disintegration, the dissolution rate test method is a first method or a second method of < dissolution rate and release rate test method 0931> in the four parts of the 'Chinese pharmacopoeia' 2020 edition; the disintegration testing method is a disintegration time limit inspection method 0921 in the four parts of China pharmacopoeia 2020 edition; the compressibility is generally judged by the presence or absence of a crack in the tablet appearance, the presence or absence of a cap, hardness, and the like.

The test results are shown in table 3 below:

TABLE 3

Survey index	Prescription	1	Prescription 2	Comparison prescription
					Dissolution rate	Good taste	Good taste	Difference (D)
Compressibility	Good taste	/	Is better
				Degree of disintegration	Good taste	Is preferably used	Difference (D)

According to the test of dissolution rate in the pharmacopoeia 2020 edition, the dissolution rates of the prescriptions 1-2 in 15 minutes are all over 80 percent. The dissolution rate of the comparison prescription is only 70% in 15 minutes, and meanwhile, the compressibility and the disintegration are also superior to the comparison prescription.

Meanwhile, the melting point of the A crystal form of the bupivacaine meloxicam salt is 189.6 ℃, while the melting point of the bupivacaine is 106-110 ℃, and the solubility difference of the two is shown in the following table 4:

TABLE 4

Solvent(s)	Solubility of bupivacaine	Solubility of bupivacaine/meloxicam salt crystal form A
			H ₂ O	<1mg/ml	>1.5mg/ml
EtOH	<1mg/ml	>1.5mg/ml
			MeOH	<1mg/ml	>1.7mg/ml
IPA	<1mg/ml	<1mg/ml
			Acetone	>1.6mg/ml	>4.0mg/ml
ACN	<1mg/ml	>1.7mg/ml
			MTBE	<1mg/ml	<1mg/ml
1，4-Dioxane	>1.6mg/ml	>1.1mg/ml
			DMF	>35mg/ml	>100mg/ml
EA	<1mg/ml	<1mg/ml
			DMSO	>52mg/ml	>100mg/ml
DMA	>52mg/ml	>100mg/ml
			NMP	>51mg/ml	>100mg/ml
Toluene	<1mg/ml	<1mg/ml

Therefore, compared with a single bupivacaine product, the A crystal form of the bupivacaine meloxicam salt has the advantages that the preparation has better investigation indexes such as dissolution rate, compressibility, disintegration and the like, and has differences in the aspects of melting point, solubility, crystal solubility and the like.

EXAMPLE 20 bioavailability assay

Respectively carrying out intragastric administration on the bupivacaine, the crystalline form A of the bupivacaine meloxicam salt and the bupivacaine of the example 1 by adopting a rat with the dose of 1.5 mg/kg; each group of animals was administered plasma at different time points (0, 0.0833, 0.25, 0.5, 1,2, 3, 4, 6, 8, 12, 24, 30, 48 hours) and plasma concentrations at different time points were measured.

The test results are shown in table 5 below and fig. 7:

TABLE 5

The result shows that the bioavailability of the bupivacaine meloxicam salt A crystal form in an animal body is obviously improved, the AUC of the bupivacaine meloxicam salt A crystal form is about 10 times of that of bupivacaine, and the bupivacaine meloxicam salt A crystal form has a very obvious pharmacokinetic behavior improvement effect.

EXAMPLE 21 Single Crystal culture of bupivacaine/Meloxicam salt

Adding 20mg of meloxicam and 16.4mg of bupivacaine into 4mL of acetone, heating to 60 ℃ to completely dissolve the meloxicam and the bupivacaine, placing the clear solution into a glass bottle, pricking holes at 25 ℃ and volatilizing for 48h to obtain a single crystal of a bupivacaine meloxicam salt A crystal form, wherein the single crystal is a light yellow rod-shaped crystal, and the crystal morphology is shown in figure 8.

Subjecting the obtained single crystal to X-ray single crystal diffraction analysis to obtain crystallography data table and structure diagram

The basic structure is as follows:

the molecular structure of the unit cell of the bupivacaine meloxicam salt single crystal is presented to fig. 9 and 10, where the atoms in the molecule are numbered and for clarity, the hydrogen is not numbered. For ease of illustration, the present application refers to each atom by its atom number in the figures.

The unit cell structure:

the crystal unit cell structures are presented for fig. 11 and 12, with a single crystal molecule containing 4 bupivacaine/meloxicam salts in one unit cell. The independent symmetric elements in the crystal are a duplex spiral shaft and an n-slip plane.

Absolute configuration

The absolute configuration represents the true arrangement relationship of each group in the chiral molecule in space, namely the absolute spatial relationship. Fig. 13 presents the absolute configuration of the bupivacaine meloxicam salt single crystal molecules within the unit cell, each bupivacaine meloxicam salt single crystal molecule includes 1 chiral carbon atom (black label), the absolute configuration of the chiral carbon atom is C24 (R).

Specifically, the crystal data of the bupivacaine meloxicam salt single crystal are shown in tables 6 to 13:

TABLE 6 crystallographic structural parameters

TABLE 7 atomic coordinates (× 10) ⁴ ) And equivalent isotropic displacement parameter

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TABLE 8 bond Length and bond Angle of the bonding atoms

TABLE 9 bond angle and bond length of bonding atoms

TABLE 10 anisotropy Displacement factor index

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Note: form of anisotropy displacement factor index: -2 pi [ h ² a* ^2U11 +...+2h k a*b*U ¹² ]Wherein a, b, c are unit lengths of reciprocal lattice; u shape ¹¹ 、U ²² 、U ³³ 、U ²³ 、U ¹³ And U ¹² Is an anisotropic displacement parameter, also known as a temperature factor; h. k and l are diffraction indexes; parallel X-rays are emitted to a perfect monocrystal with proper size, and when the optical path difference meets the integral multiple of the conditional wavelength, diffraction is generated.

According to the three-dimensional laue equation:

a(cosα0-cosα)＝hλ

b(cosβ0-cosβ)＝kλ

c(cosγ0-cosγ)＝lλ

h. k and l are integers, and a group of h, k and l are called diffraction indexes and specify a specific diffraction direction.

TABLE 11 Hydrogen atom coordinates (× 10) ⁴ ) And equivalent isotropic displacement parameter

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TABLE 12 Single Crystal twist Angle (°)

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TABLE 13 Hydrogen bonding

All possible combinations of the technical features of the above embodiments may not be described for the sake of brevity, but should be considered as within the scope of the present disclosure as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present application, so as to understand the technical solutions of the present application in detail and in detail, but not to be construed as limiting the claims. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the concept of the present application, which falls within the scope of protection of the present application. It should be understood that the technical solutions obtained by logical analysis, reasoning or limited experiments based on the technical solutions provided by the present application are all within the scope of the claims appended to the present application. Therefore, the protection scope of the present patent application shall be subject to the content of the appended claims, and the description and the drawings shall be used for explaining the content of the claims.

Claims

1. A crystalline form a of bupivacaine meloxicam salt having an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2 Θ (°) angles:

7.5 +/-0.2, 8.3 +/-0.2, 12.7 +/-0.2, 15.1 +/-0.2, 16.1 +/-0.2, 16.6 +/-0.2, 24.9 +/-0.2 and 25.0 +/-0.2.

3. The crystalline form a of a bupivacaine meloxicam salt according to claim 2 further comprising 1 or more than 2 characteristic diffraction peaks at angles 2 Θ (°) selected from the group consisting of: 18.1 +/-0.2, 20.2 +/-0.2, 22.7 +/-0.2, 23.8 +/-0.2, 24.0 +/-0.2, 25.8 +/-0.2, 26.4 +/-0.2 and 26.5 +/-0.2.

4. The crystalline form A of bupivacaine meloxicam salt according to claim 3 wherein the X-ray powder diffraction pattern of said crystalline form A is substantially as shown in figure 1.

5. Crystal form A of bupivacaine meloxicam salt according to any of the claims 1 to 4, characterised in that the differential scanning calorimetry curve of crystal form A has an endothermic peak at 189.6 ℃ ± 3 ℃.

6. The crystalline form a of bupivacaine meloxicam salt according to any of the claims 1 to 4, characterized in that the thermogravimetric spectrum of the crystalline form a shows a weight loss of 57% ± 1% in the range of 210 ℃ to 300 ℃.

7. The crystalline form A of bupivacaine meloxicam salt according to any of the claims 1 to 4, characterized in that the IR spectrum of the crystalline form A comprises the following absorption peaks:

8. A process for preparing the crystalline form a of the bupivacaine meloxicam salt according to any of the claims 1 to 7, characterized in that it comprises the following steps:

9. The method of preparing crystalline form a of a bupivacaine meloxicam salt according to claim 8, characterized by one or more of the following features:

(1) The organic solvent is one or more of butanone, acetone, tetrahydrofuran, acetonitrile, dimethyl sulfoxide, ethyl acetate and isopropyl acetate; further, the organic solvent is acetone;

(2) The antisolvent is one or more of water, isopropanol and n-heptane; further, the anti-solvent is water;

(3) The volume ratio of the organic solvent to the anti-solvent is 1-4; further, the volume ratio of the organic solvent to the anti-solvent is 1.5 to 2.5;

(4) The molar ratio of the meloxicam to the bupivacaine is 1-5; further, the molar ratio of the meloxicam to the bupivacaine is 1;

(5) The proportion of the total mass of the bupivacaine and the meloxicam to the total volume of the organic solvent and the antisolvent is 5 mg-20mg; further, the ratio is 7mg to 11mg;

(6) The temperature of heating reflux is 30-80 ℃; further, the temperature of heating reflux is 58-62 ℃;

(7) The time for continuously precipitating the solid is 2 to 8 hours; further, the time for continuously precipitating the solid is 3 to 5 hours;

(8) The temperature for continuously separating out the solid is 0-40 ℃; further, the temperature for continuously precipitating the solid is 20-30 ℃;

(9) The drying temperature is 30-60 ℃; further, the drying temperature is 45-55 ℃;

(10) The concentration temperature is 40-50 ℃, and the pressure is-0.2 MPa to-0.1 MPa.

10. A single crystal of form A of the bupivacaine meloxicam salt according to any of the claims 1 to 7, wherein one unit cell of the unit cell structure of the single crystal comprises 4 single crystal molecules of the bupivacaine meloxicam salt.

11. The crystalline form a single crystal of a bupivacaine meloxicam salt of claim 10 wherein the single crystal has a unit cell structure wherein the independent symmetry elements are a duplex helical axis and an n-glide plane.

12. The single crystal of form a of bupivacaine meloxicam salt according to claim 10 or 11, wherein the crystallographic structure parameters of the single crystal are as follows:

。

13. a pharmaceutical composition comprising one or more of the crystalline form a of the bupivacaine meloxicam salt of any of claims 1 to 7 and the single crystal of the crystalline form a of the bupivacaine meloxicam salt of any of claims 10 to 12, and a pharmaceutically acceptable carrier and/or excipient.

14. Use of the crystalline form a of a bupivacaine meloxicam salt according to any of the claims 1 to 7, the single crystal of the crystalline form a of a bupivacaine meloxicam salt according to any of the claims 10 to 12, or the pharmaceutical composition according to claim 13 for the preparation of a medicament having an effect in the treatment of local pain.