Lumepirone tosylate orally disintegrating composition and preparation method thereof

Abstract

The invention belongs to the field of pharmaceutical preparations, and particularly relates to a lumepirone tosylate orally disintegrating composition and a preparation method thereof. The lumepirone tosylate orally disintegrating composition is obtained by taking lumepirone tosylate as an effective component and taking a methacrylic acid and methyl methacrylate (1: 2) copolymer or glyceryl monostearate and glyceryl distearate as a carrier to prepare drug-loaded nanoparticles and forming a highly porous loose structure which is rapidly disintegrated when meeting saliva through a freeze-drying process. The carrier effectively encapsulates the lumepirone tosylate through intermolecular force, bitter taste and numb taste of the medicine can be effectively shielded, meanwhile, the medicine is promoted to be rapidly dispersed in saliva and directly absorbed through oral mucosa by means of the small-size effect of nano-particles, the serious first-pass effect of the medicine absorbed through gastrointestinal tracts is avoided, and the bioavailability of the medicine is improved. The bioavailability of the medicine is obviously improved, the dosage of the medicine is reduced, and the medication compliance of a patient is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical preparations, in particular to a lumuperone tosylate orally disintegrating composition and a preparation method thereof. BACKGROUND

[0002] As a second-generation small molecule atypical antipsychotic drug, lumuperone exerts therapeutic effects by synergistically regulating the transmission of serotonin, dopamine and glutamine. It has been approved for the treatment of adult schizophrenia and bipolar I or II related depressive episodes. It also shows positive effects in improving sleep quality of patients, reducing negative symptoms of schizophrenia, relieving depression and anxiety, and improving impaired social function. Compared with traditional antipsychotic drugs, it has lower incidence of adverse reactions, better long-term safety and tolerability, and can be used alone or in combination with other drugs, providing more safe and reliable choices for clinical treatment. However, the current lumuperone capsule preparation has low bioavailability. Since the drug faces severe first-pass effect when absorbed through the gastrointestinal tract, a large amount of effective components are lost in liver metabolism, ultimately resulting in incomplete absorption and insufficient blood drug concentration, which directly restricts the full play of the therapeutic effect of lumuperone and makes it difficult to meet the clinical demand for high-efficiency treatment. SUMMARY

[0003] In view of the deficiencies of the prior art, the present application aims to provide a lumuperone tosylate orally disintegrating composition and a preparation method thereof. The lumuperone tosylate is used as the effective component, and the drug-loaded nanoparticles are prepared by using a copolymer of methacrylic acid and methyl methacrylate (1:2) or glycerol mono-diestearate as the carrier. A highly porous, loose structure that disintegrates rapidly in saliva is formed through a freeze-drying process, thereby obtaining a lumuperone tosylate orally disintegrating composition. The carrier effectively encapsulates the lumuperone tosylate through intermolecular forces, effectively masks the bitter and numbing taste of the drug, and promotes the rapid dispersion of the drug in saliva and direct absorption through the oral mucosa by virtue of the small size effect of the nanoparticles, thereby avoiding the severe first-pass effect of the drug absorbed through the gastrointestinal tract, significantly improving the bioavailability, reducing the drug dosage, and improving the patient's medication compliance.

[0004] To achieve the above-mentioned purposes, the present application adopts the following technical solutions:

[0005] In a first aspect, the present application provides lumuperone tosylate nanoparticles. The components of the lumuperone tosylate orally disintegrating composition, by mass percentage, are as follows: 54.5%-65.8% lumuperone tosylate nanoparticles and 34.2%-45.5% excipients.

[0006] The components of the lumerongan tosylate nanoparticles by mass percentage are: 5.6%-45.7% lumerongan tosylate, 31.3%-65.2% carrier and 8.6%-62.4% surfactant; the particle size of the lumerongan tosylate nanoparticles is 30-500nm;

[0007] The auxiliary materials are one or more of a structure stabilizer, a suspending agent, a freeze-drying support and a sweetening agent; the structure stabilizer is gelatin; the suspending agent is one or more of xanthan gum and sodium carboxymethyl cellulose; the freeze-drying support is one or more of mannitol and lactose; and the sweetening agent is sucralose.

[0008] The lumerongan tosylate nanoparticles are used as the core active carrier of the orally disintegrating composition, and the drug delivery efficiency is improved by relying on the nanoscale characteristics. After the active ingredient lumerongan tosylate is cooperatively wrapped by the carrier and the surfactant, the small size advantage of the nanoparticles can break the aggregation trend of the drug itself, the large specific surface area can significantly increase the contact area with saliva, the drug can be quickly dispersed into a uniform system in the oral cavity, and conditions are created for subsequent dissolution and absorption; meanwhile, as a protective shell of the active ingredient, the nanoparticle structure can isolate the influence of environmental factors such as humidity and temperature on the drug, ensure the chemical stability of the active ingredient during the storage, transportation and disintegration of the preparation, and avoid the loss of drug efficacy or the generation of degradation impurities.

[0009] The auxiliary materials are an important part of the orally disintegrating composition, and the overall performance of the preparation is ensured by the synergistic effect of different types of ingredients. The structure stabilizer can enhance the physical structure stability of the preparation, prevent the collapse of the loose structure after freeze-drying during storage, the suspending agent can improve the viscosity and uniformity of the preparation system, and prevent the stratification of the nanoparticles before filling and freeze-drying, the freeze-drying support can form a support skeleton during freeze-drying, and help to build a highly porous loose structure to provide a structural basis for rapid disintegration of the preparation, and the sweetening agent can further mask the bad taste of the drug residue, and the bitter taste masking effect of the carrier is complementary, and the palatability of the preparation is improved.

[0010] Further, the carrier is selected from one or more of a methacrylic acid and methyl methacrylate copolymer, a polyurethane resin, stearin, a monodiglyceride and a monostearate, a diglyceride and a distearate, a glycerol laurate and a glycerol behenate; the molar ratio of methacrylic acid and methyl methacrylate in the methacrylic acid and methyl methacrylate copolymer is 1: (1-2).

[0011] Further, the carrier is one or more of a methacrylic acid and methyl methacrylate copolymer, a monodiglyceride and a monostearate, and a glycerol behenate.

[0012] The carrier is preferably a methyl methacrylic acid and methyl methacrylate copolymer, a glycerin monostearate and distearate, and a glycerin behenate, wherein the methyl methacrylic acid and methyl methacrylate copolymer has both carboxyl and ester groups, the carboxyl group can be combined with the drug molecule through hydrogen bonding, the ester group can enhance the hydrophobicity of the encapsulation structure, realize the stable packaging of the drug, and the polymerization ratio can be adjusted to adapt to the release requirements of the drug; the glycerin monostearate and distearate, and the glycerin behenate lipid carrier has good biocompatibility, the lipophilic chain and hydrophilic group in the molecule can form a lipid bilayer structure, which can not only encapsulate the drug through hydrophobic interaction, but also promote the affinity of the drug and the oral mucosa, help the drug to penetrate and be absorbed, at the same time, the physical and chemical properties of such carrier are stable, it can maintain the integrity of the particle skeleton during the freeze-drying process, and has no obvious physiological toxicity, and is suitable for oral preparations. The carrier forms stable encapsulation through intermolecular forces, which not only masks the bitter taste of the drug, but also adjusts the release behavior, and provides structural support for the nanoparticles.

[0013] Further, the surfactant is selected from one or more of poloxamer 188, polyvinyl alcohol, polyethylene glycol-32-stearate, Tween-80, soybean phospholipid, sodium stearate, and sodium lauryl sulfate.

[0014] Further, the surfactant is one or more of poloxamer 188, polyethylene glycol-32-stearate, and Tween-80.

[0015] The surfactant is preferably poloxamer 188, polyethylene glycol-32-stearate, and Tween-80, which have suitable hydrophilic-lipophilic balance (HLB) values, can accurately reduce the interfacial tension between the water phase and the organic phase during preparation, break the dispersion barrier between the two phases, promote the rapid emulsification and uniform dispersion of the drug and carrier components in the organic phase in the water phase, and help form a nanoparticle system with uniform particle size and narrow distribution; at the same time, in their molecular structure, the hydrophilic segment can extend into the water phase, and the lipophilic segment can be adsorbed on the surface of the nanoparticles to form a stable double electric layer or a steric hindrance barrier, effectively preventing the aggregation and sedimentation of the particles, and maintaining the long-term stability of the system. More importantly, these surfactants are all pharmaceutical-grade excipients, have excellent biocompatibility, have no obvious irritation after oral administration, can be safely metabolized in the body, do not have adverse interactions with drugs, and ensure the safety of the drug.

[0016] In a second aspect, the present application provides a preparation method of a lumefanoxine mesylate orally disintegrating composition, comprising the following steps:

[0017] S1, dissolving the surfactant in pure water to obtain a water phase; mixing the carrier with an organic reagent and adding lumefanoxine mesylate to obtain an organic phase; dropping the organic phase into the water phase and performing first dispersion treatment or second dispersion treatment to obtain lumefanoxine mesylate nanoparticles;

[0018] S2, mixing the excipient and the first pure water, heating and dissolving to obtain a mixed solution; mixing the lumefantrine tosylate nanoparticles and the second pure water, adding to the mixed solution under stirring, vacuum degassing, filling into the bubble eyes of the bubble cap, freeze-drying to obtain the lumefantrine tosylate oral disintegration composition.

[0019] In a feasible implementation scenario, in the S1, the volume ratio of the pure water to the organic reagent is (70-99.2):(0.8-30); the organic reagent is one or more of methanol and dichloromethane; the first dispersion treatment step is: stirring at a rotation speed of 300-400 rpm for 12-18 h, then water bath heating at 45-55°C for 20-40 min, freeze-drying at -20°C for 20-28 h, and screening through a 65-mesh sieve to obtain the lumefantrine tosylate nanoparticles.

[0020] The dispersion treatment step in the preparation method is the core link for ensuring the quality of the nanoparticles, and full mixing and refinement of the drug and the carrier components are achieved through physical action. The first dispersion treatment mode takes stirring combined with subsequent heating, freeze-drying, and screening as the core, and the long-time stirring action can break the aggregation state of the drug and the carrier in the organic phase, promote uniform contact between the two in the aqueous phase, and create conditions for efficient encapsulation of the drug by the carrier; the subsequent water bath heating can assist in removing residual organic reagents in the system and improving the purity of the system; freeze-drying treatment can quickly solidify the structure of the nanoparticles, lock the encapsulation state of the drug and the carrier, and avoid structural damage of the particles in subsequent processing; and finally, grinding and screening are performed to precisely control the particle size, so that the lumefantrine tosylate nanoparticles with uniform particle size and meeting the subsequent preparation requirements are obtained.

[0021] In a feasible implementation scenario, in the S1, the second dispersion treatment step is: ultrasonic crushing at a power of 450-550 W for 8-12 min, cold storage at 4°C for 20-40 min, dialysis in a 50KD dialysis bag for 1-2 h, and centrifugation at 9500-10500 rpm for 8-12 min in a 10KD ultrafiltration tube to obtain the lumefantrine tosylate nanoparticles.

[0022] The second dispersion processing mode focuses on particle refinement and precise impurity removal by combining ultrasonic crushing with refrigeration, dialysis and centrifugation operations. The strong vibration and shear force generated by high-power ultrasound can directly break the aggregates of drugs and carriers into small units, helping to form a nanoscale dispersion system, and promoting the uniform wrapping of the carrier on the drug; refrigeration can reduce the molecular kinetic energy of the system, slow down the movement speed of the particles, and avoid the re-aggregation of the newly formed small particles; through dialysis, small molecular impurities and excess surfactants in the system that do not participate in encapsulation can be removed, improving the purity of the particles, and further separating small particles of unsatisfactory size through ultrafiltration centrifugation, ultimately obtaining nanoparticles with high purity, uniform particle size and excellent stability, providing protection for the subsequent preparation of drug efficacy.

[0023] In a feasible implementation scenario, in the S2, the volume ratio of the first pure water and the second pure water is 50:50; the temperature of the heating and dissolving is 55-65 DEG C; the stirring speed is 200-300 rpm; the vacuum degassing time is 15-20 min; the freeze-drying process is as follows: first, pre-freezing at-30 DEG C for 2.5-3.5 h within 40 min; then entering the main drying stage, under the condition of vacuum degree 10 Pa, the temperature is raised to-20 DEG C at the rate of 0.3-0.7 DEG C / min, maintaining 5-7 h, then the temperature is raised to-10 DEG C at the rate of 0.3-0.7 DEG C / min, maintaining 6.5-9.5 h, finally the temperature is raised to 0 DEG C at the rate of 0.3-0.7 DEG C / min, maintaining 8-12 h; finally, the terminal drying is carried out, controlling the vacuum degree to be less than 20 Pa, the temperature is raised to 10 DEG C at the rate of 0.3-0.7 DEG C / min, maintaining 0.8-1.2 h, then the temperature is raised to 25 DEG C at the rate of 0.6-0.9 DEG C / min, maintaining 1.5-2.5 h.

[0024] The freeze-drying process is a key technology for imparting excellent disintegration and absorption performance to the oral disintegration composition. The pre-freezing stage can quickly solidify the system to form small and uniform ice crystals, avoiding the destruction of the combination structure of the drug and the excipient by large ice crystals; the main drying stage gradually raises the temperature under vacuum conditions, allowing the ice crystals to sublimate and be removed, while retaining the porous skeleton structure formed by the drug and the excipient; the terminal drying stage further removes residual moisture to improve the stability of the preparation. The highly porous and loose structure formed ultimately can quickly absorb water and swell and disintegrate after contacting saliva, allowing the nanoparticles to be rapidly released and dispersed, promoting the direct absorption of the drug through the oral mucosa, thereby avoiding the first-pass effect of gastrointestinal absorption and significantly improving the bioavailability.

[0025] Compared with the prior art, the beneficial effects of the present application are as follows:

[0026] The present scheme is to prepare a lumefantrine tosylate oral disintegration composition by taking lumefantrine tosylate as an effective component, preparing drug-loaded nanoparticles by taking a copolymer of methacrylic acid and methyl methacrylate (1:2) or glycerol monostearate and glycerol distearate as a carrier, and combining a freeze-drying process, so as to realize stable drug dissolution, rapid effect and significantly improved bioavailability. The carrier forms effective encapsulation of lumefantrine tosylate through intermolecular forces, and can shield the bitter and numb taste of the drug itself. The highly porous loose structure formed through the freeze-drying process rapidly absorbs water and disintegrates upon contact with saliva, and does not need other water media for delivery, thereby improving patient compliance or helping special patients to passively take the medicine. Meanwhile, the small size effect of the nanoparticles promotes the rapid dispersion of the drug in saliva, promotes the direct absorption of the drug through the oral mucosa, thereby avoiding the serious first-pass effect of the drug absorption through the gastrointestinal tract, significantly improving the bioavailability of the drug from the absorption path, and solving the problem of low bioavailability of lumefantrine capsule preparation. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 A scanning electron microscope image of the lumefantrine tosylate oral disintegration composition prepared in Example 7 of the present application.

[0028] Figure 2 A radar chart of the lumefantrine tosylate oral disintegration composition prepared in Example 7, Example 14 and Example 15 of the present application and the lumefantrine tosylate electronic tongue.

[0029] Figure 3 A dissolution curve graph of the lumefantrine tosylate oral disintegration composition prepared in Example 7 of the present application in different media.

[0030] Figure 4 A dissolution curve graph of the marketed preparation (capsule) in different media.

[0031] Figure 5 A blood concentration change curve graph of the lumefantrine tosylate oral disintegration composition prepared in Example 7, Example 14 and Example 15 of the present application and the marketed preparation.

[0032] Figure 6 A nanoparticle size graph of the lumefantrine tosylate prepared in Example 4 of the present application.

[0033] Figure 7 A finished product graph of the lumefantrine tosylate oral disintegration composition prepared in Example 7 of the present application. DETAILED DESCRIPTION

[0034] In order to make the technical problems, technical solutions and beneficial effects of the present application clearer, the present application will be further described in detail below in conjunction with embodiments. However, it should not be understood as a limitation on the scope of the present application only to the following examples. Without departing from the above method idea of the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor shall fall within the scope of protection of the present application.

[0035] The singular forms "is", "are", "one", "any" and "the" used in the present application are intended to include the plural forms, unless the context clearly indicates otherwise. In addition, if the terms "first", "second" appear, they are only used for descriptive purposes and cannot be understood as indicating or implying relative importance.

[0036] Example 1

[0037] The prescription composition and preparation method of the toluenesulfonic acid lumefanum nanoparticles with methacrylic acid and methyl methacrylate (1:2) copolymer as the carrier material are as follows:

[0038] Prescription (by mass percentage): 45.7% toluenesulfonic acid lumefanum, 45.7% methacrylic acid and methyl methacrylate (1:2) copolymer, 8.6% poloxamer 188.

[0039] Preparation method:

[0040] S1, dissolve poloxamer 188 in 70% pure water based on the total volume of the liquid to obtain an aqueous phase; mix methacrylic acid and methyl methacrylate (1:2) copolymer with 30% methanol based on the total volume of the liquid, add toluenesulfonic acid lumefanum, and ultrasonic for 10 min at a power of 300 W to obtain an organic phase; drop the organic phase into the aqueous phase under the condition of 500 rpm, continue to stir under the condition of 350 rpm for 15 h after the drop is completed, heat to 50℃ for water bath heating for 30 min, freeze-drying at-20℃ for 24 h to obtain a powder, pass through a 65 mesh sieve to obtain toluenesulfonic acid lumefanum nanoparticles.

[0041] Evaluation results: the drug loading rate of the toluenesulfonic acid lumefanum nanoparticles prepared in Example 1 is 40.08%, the average particle size is 187.4 nm, and the polydispersity index (PDI) value is 0.218.

[0042] Example 2

[0043] The prescription composition and preparation method of the toluenesulfonic acid lumefanum nanoparticles with methacrylic acid and methyl methacrylate (1:2) copolymer as the carrier material are as follows:

[0044] Formulation (by mass percentage): 21.8% of lumirizine tosylate, 65.2% of methacrylic acid and methyl methacrylate (1:2) copolymer, 13.0% of poloxamer 188.

[0045] Preparation method:

[0046] S1, dissolve poloxamer 188 in 70% pure water based on the total volume of the liquid to obtain an aqueous phase; mix methacrylic acid and methyl methacrylate (1:2) copolymer with 30% methanol based on the total volume of the liquid, add lumirizine tosylate, and ultrasonic for 10 min at a power of 300 W to obtain an organic phase; drop the organic phase into the aqueous phase at a rotation speed of 500 rpm, continue to stir at a rotation speed of 300 rpm for 18 h after the drop is completed, heat to 45℃ for water bath heating for 40 min, freeze-drying at-20℃ for 20 h to obtain a powder, and sieve through a 65 mesh sieve to obtain lumirizine tosylate nanoparticles.

[0047] Evaluation results: the drug loading rate of the lumirizine tosylate nanoparticles prepared in Example 2 is 21.34%, the average particle size is 335.7 nm, and the PDI value is 0.178.

[0048] Example 3

[0049] The formulation composition and preparation method of lumirizine tosylate nanoparticles with methacrylic acid and methyl methacrylate (1:1) copolymer as a carrier material are as follows:

[0050] Formulation (by mass percentage): 45.7% of lumirizine tosylate, 47.7% of methacrylic acid and methyl methacrylate (1:1) copolymer, 8.6% of poloxamer 188.

[0051] Preparation method:

[0052] S1, dissolve poloxamer 188 in 70% pure water based on the total volume of the liquid to obtain an aqueous phase; mix methacrylic acid and methyl methacrylate (1:1) copolymer with 30% methanol based on the total volume of the liquid, add lumirizine tosylate, and ultrasonic for 10 min at a power of 300 W to obtain an organic phase; drop the organic phase into the aqueous phase at a rotation speed of 500 rpm, continue to stir at a rotation speed of 350 rpm for 12 h after the drop is completed, heat to 55℃ for water bath heating for 20 min, freeze-drying at-20℃ for 28 h to obtain a powder, and sieve through a 65 mesh sieve to obtain lumirizine tosylate nanoparticles.

[0053] Evaluation results: the drug loading rate of the lumirizine tosylate nanoparticles prepared in Example 3 is 35.67%, the average particle size is 298.2 nm, and the PDI value is 0.317.

[0054] Example 4

[0055] The prescription composition and preparation method of the toluenesulfonic acid lumefantrine nanoparticles with monostearin as the carrier material are as follows:

[0056] The prescription (in mass percentage): 8.6% toluenesulfonic acid lumefantrine, 34.3% monostearin, 57.1% polyethylene glycol-32-stearate.

[0057] The preparation method is as follows:

[0058] S1, dissolve polyethylene glycol-32-stearate in 99.2% pure water in total liquid volume to obtain an aqueous phase; heat monostearin in a water bath to liquefy, add toluenesulfonic acid lumefantrine and 0.8% dichloromethane in total liquid volume, and heat in a water bath for 30 min to obtain an organic phase; drop the organic phase into the aqueous phase at a rotation speed of 500 rpm, after the drop is completed, ultrasonic crushing is performed at a power of 500 W for 10 min, and then placed in a 4°C refrigerator for 30 min, put into a 50KD dialysis bag for dialysis for 1.5h, and then placed in a 10KD ultrafiltration tube, centrifuged at 10000 rpm for 10 min to obtain toluenesulfonic acid lumefantrine nanoparticles.

[0059] The evaluation results: the drug loading rate of the toluenesulfonic acid lumefantrine nanoparticles prepared in Example 4 is 13.74%, the average particle size is 52.4 nm, and the PDI value is 0.161.

[0060] Example 5

[0061] The prescription composition and preparation method of the toluenesulfonic acid lumefantrine nanoparticles with monostearin as the carrier material are as follows:

[0062] The prescription (in mass percentage): 8.6% toluenesulfonic acid lumefantrine, 34.3% monostearin, 57.1% polyethylene glycol-32-stearate.

[0063] The preparation method is as follows:

[0064] S1, dissolve polyethylene glycol-32-stearate in 99.2% pure water in total liquid volume to obtain an aqueous phase; heat monostearin in a water bath to liquefy, add toluenesulfonic acid lumefantrine and 0.8% dichloromethane in total liquid volume, and heat in a water bath for 30 min to obtain an organic phase; drop the organic phase into the aqueous phase at a rotation speed of 500 rpm, after the drop is completed, ultrasonic crushing is performed at a power of 500 W for 10 min, and then placed in a 4°C refrigerator for 30 min, put into a 50KD dialysis bag for dialysis for 1.5h, and then placed in a 10KD ultrafiltration tube, centrifuged at 10000 rpm for 10 min to obtain toluenesulfonic acid lumefantrine nanoparticles.

[0065] Evaluation results: the drug loading rate of the prepared lumefantrine tosylate nanoparticles in Example 5 is 8.13%, the average particle size is 68.7 nm, and the PDI value is 0.214.

[0066] Example 6

[0067] The prescription composition and preparation method of the lumefantrine tosylate nanoparticles with glyceryl behenate as the carrier material are as follows:

[0068] Prescription (by mass percentage): 5.6% lumefantrine tosylate, 38.9% glyceryl behenate, 55.5% polyethylene glycol-32-stearate.

[0069] Preparation method:

[0070] S1, dissolve polyethylene glycol-32-stearate in 99.2% pure water by volume of the total liquid to obtain an aqueous phase; heat the glyceryl behenate in a water bath to liquefy, add lumefantrine tosylate and 0.8% dichloromethane by volume of the total liquid, and heat in a water bath for 30 min to obtain an organic phase; drop the organic phase into the aqueous phase under the condition of 500 rpm, after the drop is completed, ultrasonic crushing is carried out under the condition of power 550W for 8 min, and then placed in a 4℃ refrigerator for 40 min, then placed in a 50KD dialysis bag for dialysis for 2h, then placed in a 10KD ultrafiltration tube, centrifuged at 10500 rpm for 8 min to obtain lumefantrine tosylate nanoparticles.

[0071] Evaluation results: the drug loading rate of the prepared lumefantrine tosylate nanoparticles in Example 6 is 6.32%, the average particle size is 112.4 nm, and the PDI value is 0.332.

[0072] Example 7

[0073] The prescription and preparation method of a lumefantrine tosylate oral disintegration composition are as follows:

[0074] Prescription (by mass percentage): 54.5% lumefantrine tosylate nanoparticles of Example 1, 31.1% mannitol, 2.1% xanthan gum, 10.4% gelatin, 1.7% sucralose, 0.2% sodium carboxymethyl cellulose.

[0075] Preparation method:

[0076] S2, the gelatin, xanthan gum, sodium carboxymethyl cellulose, mannitol, sucralose and 50% of the total volume of the liquid pure water mixed, heated to 60°C to dissolve, get the mixture of adjuvants; the toluene sulphonate lumefantrine nanoparticles of example 1 with 50% of the total volume of the liquid pure water mixed, under the condition of 250 rpm speed into the adjuvant mixture, vacuum degassing 17 min, filling to the bubble eye of bubble, placed in the freeze dryer, first in 40 min temperature to-30 DEG C pre-frozen 3h; then into the main drying stage, under the condition of 10 Pa vacuum, with 0.5 DEG C / min rate to-20 DEG C, maintain 6h, again with 0.5 DEG C / min rate to-10 DEG C, maintain 8h, finally with 0.5 DEG C / min rate to 0 DEG C, maintain 10h; finally, the final drying, control the vacuum is less than 20 Pa, with 0.5 DEG C / min rate to 10 DEG C, maintain 1h, again with 0.7 DEG C / min rate to 25 DEG C, maintain 2h, toluene sulphonate lumefantrine oral disintegration composition is obtained.

[0077] Evaluation results: the toluene sulphonate lumefantrine oral disintegration composition prepared in example 7, the tablet type is complete, the surface is smooth, the disintegration time is about 20s, and almost no bitter taste.

[0078] Example 8

[0079] A toluene sulphonate lumefantrine oral disintegration composition is prepared according to the following prescription and preparation method:

[0080] Prescription (in mass percent): 54.5% of the toluene sulphonate lumefantrine nanoparticles of example 2, 29.5% of mannitol, 2.4% of xanthan gum, 12.3% of gelatin, 1.2% of sucralose.

[0081] Preparation method:

[0082] S2, the gelatin, xanthan gum, mannitol, sucralose and 50% pure water by the total volume of liquid mixed, heated to 55°C to dissolve, get the mixture of adjuvants; the example 2 lumefantrine tosylate nanoparticles and 50% pure water by the total volume of liquid mixed, under the condition of 200 rpm speed into the adjuvant mixture, vacuum degassing 15 min, filling to the bubble eye of bubble, placed in the freeze dryer, first in 40 min to-30 ℃ pre-freezing 2.5 h; then into the main drying stage, under the condition of 10 Pa vacuum, with 0.3 ℃ / min rate to-20 ℃, maintain 5 h, again with 0.3 ℃ / min rate to-10 ℃, maintain 6.5 h, finally with 0.3 ℃ / min rate to 0 ℃, maintain 8 h; finally, the final drying, control the vacuum degree is lower than 20 Pa, with 0.3 ℃ / min rate to 10 ℃, maintain 0.8 h, again with 0.6 ℃ / min rate to 25 ℃, maintain 1.5, get lumefantrine tosylate oral disintegration composition.

[0083] Evaluation results: the preparation of lumefantrine tosylate oral disintegration composition of example 8, tablet type complete surface smooth, disintegration time is about 20 s, almost can not taste bitter.

[0084] Example 9

[0085] A kind of lumefantrine tosylate oral disintegration composition prescription and preparation method as follows:

[0086] Prescription (by mass percentage): 54.5% of example 1 lumefantrine tosylate nanoparticles, 31.1% mannitol, 2.1% xanthan gum, 10.4% gelatin, 1.7% sucralose, 0.2% sodium carboxymethyl cellulose.

[0087] S2, mix gelatin, xanthan gum, sodium carboxymethyl cellulose, mannitol, sucralose and 50% pure water based on the total volume of liquid, heated to 65℃ to dissolve, to obtain a mixture of adjuvants; the toluene sulphonate of lumebrolone nanoparticles of example 1 is mixed with 50% pure water based on the total volume of liquid, added to the mixture of adjuvants under the condition of 300 rpm, vacuum degassing for 20 min, filled into the bubble eye of the bubble cap, placed in the freeze dryer, first cooled to -30℃ in 40 min, pre-frozen for 3.5 h; then enter the main drying stage, under the condition of vacuum degree 10 Pa, heated to -20℃ at a rate of 0.7℃ / min, maintained for 7 h, then heated to -10℃ at a rate of 0.7℃ / min, maintained for 9.5 h, finally heated to 0℃ at a rate of 0.7℃ / min, maintained for 12 h; finally, the terminal drying is carried out, the vacuum degree is controlled to be lower than 20 Pa, heated to 10℃ at a rate of 0.7℃ / min, maintained for 1.2 h, then heated to 25℃ at a rate of 0.9℃ / min, maintained for 2.5 h, to obtain the toluene sulphonate of lumebrolone oral disintegration composition.

[0088] Evaluation result: the toluene sulphonate of lumebrolone oral disintegration composition prepared in example 9 is easy to break, the disintegration time is about 20 s, and almost no bitterness can be tasted.

[0089] Example 10

[0090] A prescription and preparation method of a toluene sulphonate of lumebrolone oral disintegration composition are as follows:

[0091] Prescription (in mass percentage): 62.1% toluene sulphonate of lumebrolone nanoparticles of example 4, 14.5% mannitol, 21.9% lactose, 1.5% sucralose.

[0092] S2, mix lactose, mannitol, sucralose and 50% pure water based on the total volume of liquid, heated to 60℃ to dissolve, to obtain a mixture of adjuvants; the toluene sulphonate of lumebrolone nanoparticles of example 4 is mixed with 50% pure water based on the total volume of liquid, added to the mixture of adjuvants under the condition of 250 rpm, vacuum degassing for 17 min, filled into the bubble eye of the bubble cap, placed in the freeze dryer, first cooled to -30℃ in 40 min, pre-frozen for 3 h; then enter the main drying stage, under the condition of vacuum degree 10 Pa, heated to -20℃ at a rate of 0.5℃ / min, maintained for 6 h, then heated to -10℃ at a rate of 0.5℃ / min, maintained for 8 h, finally heated to 0℃ at a rate of 0.5℃ / min, maintained for 10 h; finally, the terminal drying is carried out, the vacuum degree is controlled to be lower than 20 Pa, heated to 10℃ at a rate of 0.5℃ / min, maintained for 1 h, then heated to 25℃ at a rate of 0.7℃ / min, maintained for 2 h, to obtain the toluene sulphonate of lumebrolone oral disintegration composition.

[0093] Evaluation result: The prepared lumerongene tosylate orally disintegrating composition in Example 10 is in tablet form, the disintegration time is about 25 s, and the bitterness cannot be tasted completely.

[0094] Example 11

[0095] A prescription and preparation method of a lumerongene tosylate orally disintegrating composition are as follows:

[0096] Prescription (in mass percentage): 62.1% lumerongene tosylate nanoparticles in Example 4, 18.2% mannitol, 18.2% lactose, and 1.5% sucralose.

[0097] S2, the lactose, mannitol, sucralose, and 50% pure water based on the total volume of the liquid are mixed, heated to 55°C for dissolution, and a mixture of excipients is obtained; the lumerongene tosylate nanoparticles in Example 4 are mixed with 50% pure water based on the total volume of the liquid, added to the mixture of excipients under the condition of a rotation speed of 200 rpm, vacuum degassed for 15 min, filled into the bubble eyes of a bubble plate, placed in a freeze dryer, pre-frozen to -30°C for 2.5 h within 40 min; then enter the main drying stage, under the condition of a vacuum degree of 10 Pa, heated to -20°C at a rate of 0.3°C / min, maintained for 5 h, then heated to -10°C at a rate of 0.3°C / min, maintained for 6.5 h, and finally heated to 0°C at a rate of 0.3°C / min, maintained for 8 h; finally, perform final drying, control the vacuum degree to be lower than 20 Pa, heat to 10°C at a rate of 0.3°C / min, maintain for 0.8 h, then heat to 25°C at a rate of 0.6°C / min, maintain for 1.5 h, and obtain the lumerongene tosylate orally disintegrating composition.

[0098] Evaluation result: The prepared lumerongene tosylate orally disintegrating composition in Example 11 is in tablet form, the disintegration time is about 25 s, and the bitterness cannot be tasted completely.

[0099] Example 12

[0100] A prescription and preparation method of a lumerongene tosylate orally disintegrating composition are as follows:

[0101] Prescription (in mass percentage): 62.1% lumerongene tosylate nanoparticles in Example 4, 7.3% mannitol, 29.2% lactose, and 1.5% sucralose.

[0102] S2, lactose, mannitol, sucralose and 50% pure water based on the total volume of liquid were mixed, heated to 65°C to dissolve, and a mixture of excipients was obtained; the toluenesulfonic acid lumefantrine nanoparticles of Example 4 were mixed with 50% pure water based on the total volume of liquid, added to the mixture of excipients under the condition of a rotation speed of 300 rpm, vacuum degassed for 20 min, filled into the bubble eyes of a bubble cap, placed in a freeze dryer, first cooled to -30°C within 40 min to pre-freeze for 3.5 h; then entered the main drying stage, under the condition of a vacuum degree of 10 Pa, heated to -20°C at a rate of 0.7°C / min, maintained for 7 h, then heated to -10°C at a rate of 0.7°C / min, maintained for 9.5 h, and finally heated to 0°C at a rate of 0.7°C / min, maintained for 12 h; finally, the terminal drying was performed, the vacuum degree was controlled to be lower than 20 Pa, heated to 10°C at a rate of 0.7°C / min, maintained for 1.2 h, and then heated to 25°C at a rate of 0.9°C / min, maintained for 2.5 h, to obtain the toluenesulfonic acid lumefantrine orally disintegrating composition.

[0103] Evaluation result: the toluenesulfonic acid lumefantrine orally disintegrating composition prepared in Example 12 was easy to form a crust, the disintegration time was about 30 s, and the bitter taste could not be tasted completely.

[0104] Example 13

[0105] A prescription and preparation method of a toluenesulfonic acid lumefantrine orally disintegrating composition are as follows:

[0106] Prescription: 65.8% toluenesulfonic acid lumefantrine nanoparticles of Example 5, 13.2% mannitol, 19.7% lactose, and 1.3% sucralose.

[0107] S2, lactose, mannitol, sucralose and 50% pure water based on the total volume of liquid were mixed, heated to 65°C to dissolve, and a mixture of excipients was obtained; the toluenesulfonic acid lumefantrine nanoparticles of Example 4 were mixed with 50% pure water based on the total volume of liquid, added to the mixture of excipients under the condition of a rotation speed of 300 rpm, vacuum degassed for 20 min, filled into the bubble eyes of a bubble cap, placed in a freeze dryer, first cooled to -30°C within 40 min to pre-freeze for 3.5 h; then entered the main drying stage, under the condition of a vacuum degree of 10 Pa, heated to -20°C at a rate of 0.7°C / min, maintained for 7 h, then heated to -10°C at a rate of 0.7°C / min, maintained for 9.5 h, and finally heated to 0°C at a rate of 0.7°C / min, maintained for 12 h; finally, the terminal drying was performed, the vacuum degree was controlled to be lower than 20 Pa, heated to 10°C at a rate of 0.7°C / min, maintained for 1.2 h, and then heated to 25°C at a rate of 0.9°C / min, maintained for 2.5 h, to obtain the toluenesulfonic acid lumefantrine orally disintegrating composition.

[0108] Evaluation result: the raloxifene hydrochloride oral disintegration composition prepared in Example 13 has a tablet shape intact and a disintegration time of about 25 s, and no bitter taste can be tasted.

[0109] Example 14

[0110] The raloxifene hydrochloride oral disintegration composition was prepared by fluidized bed coating of the pellet core process, and the prescription is shown in Table 1

[0111] Table 1 raloxifene hydrochloride oral disintegration tablet

[0112]

[0113] Preparation of drug-containing pellets: raloxifene hydrochloride was added to water for micronization treatment and stirring to disperse, to obtain a raloxifene hydrochloride suspension; low-substituted hydroxypropyl cellulose and hydroxypropyl methyl cellulose were added to water and stirred until completely dissolved, talc was added and stirred evenly to obtain a talc-containing suspension; the talc-containing suspension was added to the raloxifene hydrochloride suspension to obtain a drug-containing suspension; the microcrystalline cellulose pellet core was placed in a fluidized bed, and the drug-containing suspension was sprayed by bottom spraying under the conditions of material temperature 35-45°C, air flow 20-25 m 3 -25m 3 , atomization pressure 0.6-1.0 bar, to obtain drug-containing pellets.

[0114] Preparation of taste-masking pellets: sodium dodecyl sulfate was added to water and stirred, and a copolymer of stearic acid and butyl methacrylate, dimethylaminoethyl methacrylate and methyl methacrylate (1:2:1) was added and stirred until a transparent colloidal solution was formed; low-substituted hydroxypropyl cellulose and talc were added to water and stirred evenly to prepare a talc-containing suspension; the talc-containing suspension was added to the colloidal solution to obtain a taste-masking suspension; the drug-containing pellets were placed in a fluidized bed, and the taste-masking suspension was sprayed by bottom spraying under the conditions of material temperature 25-35°C, air flow 20-25 m

[0115] Preparation of oral disintegration tablets: the taste-masking pellets were added to colloidal silicon dioxide, mixed evenly through a 40-mesh sieve to obtain mixture 1; mannitol, croscarmellose sodium, sucralose and strawberry flavor were added to mixture 1 and mixed evenly to obtain mixture 2; the total mixture was added to mixture 2, and the tablets were pressed to a hardness of 40 N to obtain a raloxifene hydrochloride oral disintegration tablet composition.

[0116] Example 15

[0117] The raloxifene hydrochloride oral disintegration composition was prepared by powder direct compression process, and the prescription is shown in Table 2.

[0118]

[0119] Preparation process: mix lumimolane tosylate and mannitol through 40 mesh sieve, crush sodium citrate through 80 mesh sieve, crush the rest of the excipients through 40 mesh sieve; add microcrystalline cellulose, calcium carboxymethyl cellulose, sucralose, sodium citrate and peppermint powder essence into the hopper mixer in turn, then add the lumimolane tosylate and mannitol blend into the hopper, set the mixer speed to 30 rpm, and the premixing time to 10 min to obtain the mixture; add magnesium stearate and colloidal silicon dioxide to the mixture, set the mixer speed to 30 rpm, and the mixing time to 5 min to obtain the total excipient mixture, which is added into the hopper of the rotary tablet press, the tablet press parameters are adjusted, the tablet weight is adjusted to 800 mg, the appropriate tablet pressing force is adjusted to make the hardness range 60-80 N, and the tablets are pressed using 18*9 mm oval punches to obtain the lumimolane tosylate orally disintegrating composition.

[0120] Performance test:

[0121] Basic property test: randomly take 10 tablets of the lumimolane tosylate orally disintegrating composition prepared in Example 7 for tablet weight and content test, and use the intelligent disintegration instrument to test the disintegration time, and the results are shown in Tables 3, 4 and 5.

[0122] Table 3 Lumimolane tosylate nanoparticle lyophilized orally disintegrating composition tablet weight measurement results

[0123]

[0124] Table 4 Lumimolane tosylate nanoparticle lyophilized orally disintegrating composition content test results

[0125]

[0126] Table 5 Lumimolane tosylate nanoparticle lyophilized orally disintegrating composition disintegration test results

[0127]

[0128] Taste test:

[0129] Taste evaluation of the lumimolane tosylate orally disintegrating composition of Example 7: 5 volunteers were asked to evaluate the taste of the lyophilized tablets, and all 5 volunteers indicated that the lumimolane tosylate orally disintegrating composition disintegrated rapidly in the mouth, had no bitter taste, and had a little bitter taste within the acceptable range, 3 volunteers indicated that the lumimolane tosylate orally disintegrating composition was slightly sweet, 1 volunteer indicated that it was almost not sweet, and 1 volunteer indicated that the sweet taste was obvious.

[0130] Electronic tongue evaluation: lumimolane tosylate and the lumimolane tosylate orally disintegrating compositions prepared in Examples 7, 14 and 15 were evaluated, and the results are shown in Table 6. Figure 2The taste-masking effect of the common oral disintegration composition prepared by the powder direct compression process in Example 15 is general, and the taste-masking effect of the compositions in Example 7 and Example 14, which are prepared by encapsulating the romifobine toluenesulfonate, is effective. The bitter taste and astringent taste values of the composition in Example 7 are both 0, and the taste-masking effect is better.

[0131] In-vitro dissolution test: the romifobine toluenesulfonate oral disintegration composition prepared in Example 7 and the marketed preparation are respectively taken, and the samples are taken at 5 mins, 10 mins, 15 mins, 20 mins, 30 mins, 45 mins, 60 mins, 90 mins and 120 mins by the basket method at a rotation speed of 75 rpm, and the dissolution rates are measured in 0.1M hydrochloric acid dissolution medium, pH 4.5 acetate dissolution medium, pH 6.8 phosphate dissolution medium and pH 7.4 phosphate dissolution medium, and the results are shown in Figure 3 and Figure 4

[0132] As shown in Figure 3 and Figure 4 , the romifobine toluenesulfonate oral disintegration composition is dissolved more slowly than the marketed preparation. The lower the pH value of the dissolution medium, the more the drug is dissolved, and the romifobine toluenesulfonate oral disintegration composition is completely dissolved in 0.1M hydrochloric acid medium.

[0133] Pharmacokinetic test: the romifobine toluenesulfonate oral disintegration compositions prepared in Example 7, Example 14 and Example 15 and the marketed preparation are subjected to oral instillation administration or gavage administration in rats, and the blood drug concentration changes are detected. 6-8 weeks old male SD rats with a body weight of 200-250g are selected in each group, and the rats are fasted for 12h (without water restriction) before administration. After administration, the blood samples are taken at 5 mins, 10 mins, 15 mins, 20 mins, 30 mins, 45 mins, 1h, 1.5h, 2h, 3h, 4h, 6h and 8h, the serum is taken by centrifugation, the romifobine concentration is detected by LC-MS, and the results are shown in Figure 5 .

[0134] Figure 5 ​​It can be seen that the bioavailability of the orally disintegrating combination of lumerigoxine tosylate is obviously higher than that of the marketed preparation. The average AUClast of the marketed preparation capsule administered by gavage is 56.04 ng*h / mL, the average AUClast of lumerigoxine tosylate administered by gavage is 52.44 ng*h / mL, and the average AUClast of the orally disintegrating combination of lumerigoxine tosylate administered by oral instillation is 310.87 ng*h / mL. It can be seen that the orally disintegrating combination of lumerigoxine tosylate of the application mainly avoids the first-pass effect by partially absorbing the drug in the oral cavity to improve the bioavailability. The average AUClast of the orally disintegrating combination of lumerigoxine tosylate of the application, which simulates oral disintegration and is taken orally (i.e., oral instillation in rats), is 5.55 times that of the marketed preparation. The Tmax of the orally disintegrating combination of lumerigoxine tosylate prepared by the application is significantly advanced, the absorption is faster, the onset is faster, and it is more suitable for patients to take during the acute stage. In addition, the bioavailability of the raw material drug lumerigoxine tosylate is significantly improved after nanocrystallization, the bioavailability of the non-nanocrystallized orally disintegrating combination is obviously improved, and the onset time is also faster. This is because the lumerigoxine tosylate nanoparticles have smaller particle sizes and good lipophilicity, which helps to penetrate the oral mucosa. The amount of effective ingredients penetrating the oral mucosa is more than that of ordinary orally disintegrating tablets, so the bioavailability is higher than that of ordinary orally disintegrating tablets.

[0135] In summary, the orally disintegrating combination of lumerigoxine tosylate of the application has significantly improved oral bioavailability, and a lower dose can achieve the blood concentration that can be achieved by the original dosage form. The absorption is fast, the onset is fast, and the pharmacokinetic properties are better. The dosage form of the application is more suitable for children, the elderly, and special patients (psychiatric patients, patients with Alzheimer's disease, patients with epilepsy, etc.). It can also reduce the deliberate behavior of patients to vomit medicine, ensure the therapeutic effect, and be more suitable for the acute stage of the disease. It truly improves patient medication compliance and has the advantages of simple preparation process and easy industrialization.

[0136] The above results show and describe the basic principles and main features of the application and the advantages of the application.

[0137] The embodiments of the application are described above with reference to the drawings, but the application is not limited to the specific embodiments described above, which are only illustrative rather than limiting. Those skilled in the art can make many forms under the inspiration of the application without departing from the purpose of the application and the scope protected by the claims.

Claims

1. A orally disintegrating composition of lumeriganer, characterized in that, The components of the lumerongene tosylate orally disintegrating composition by mass percentage are: 54.5%-65.8% lumerongene tosylate nanoparticles and 34.2%-45.5% excipients; The components of the lumerongene tosylate nanoparticles by mass percentage are: 5.6%-45.7% lumerongene tosylate, 31.3%-65.2% carrier and 8.6%-62.4% surfactant; the particle size of the lumerongene tosylate nanoparticles is 30-500nm; The excipients are one or more of a structure stabilizer, a suspending agent, a freeze-drying support and a sweetening agent; the structure stabilizer is gelatin; the suspending agent is one or more of xanthan gum and sodium carboxymethyl cellulose; the freeze-drying support is one or more of mannitol and lactose; the sweetening agent is sucralose.

2. A lurasidone tosylate orally disintegrating composition according to claim 1, wherein The carrier is selected from one or more of a methacrylic acid and methyl methacrylate copolymer, a polyurethane resin, stearin, glycerol monostearate, glycerol distearate, glycerol laurate and glycerol behenate; the molar ratio of methacrylic acid and methyl methacrylate in the methacrylic acid and methyl methacrylate copolymer is 1: (1-2).

3. A lurasidone tosylate orally disintegrating composition according to claim 2, wherein The carrier is one or more of a methacrylic acid and methyl methacrylate copolymer, glycerol monostearate and glycerol behenate.

4. A lurasidone tosylate orally disintegrating composition according to claim 1, wherein The surfactant is selected from one or more of poloxamer 188, polyvinyl alcohol, macrogol-32-stearate, Tween-80, soybean phospholipid, sodium stearate and sodium lauryl sulfate.

5. A lurasidone tosylate orally disintegrating composition according to claim 4, wherein The surfactant is one or more of poloxamer 188, macrogol-32-stearate and Tween-80.

6. A process for the preparation of a orally disintegrating composition of lumeraubanole mesylate according to any one of claims 1 to 5, characterized in that, The method comprises the following steps: S1, dissolving the surfactant in pure water to obtain an aqueous phase; mixing the carrier with an organic reagent and adding lumerongene tosylate to obtain an organic phase; dropping the organic phase into the aqueous phase and performing first dispersion treatment or second dispersion treatment to obtain lumerongene tosylate nanoparticles; S2, mixing the excipients and first pure water and heating to dissolve to obtain a mixed solution; mixing the lumerongene tosylate nanoparticles with second pure water and adding to the mixed solution under stirring, vacuum degassing, filling into the bubble eyes of a bubble plate, freeze-drying to obtain the lumerongene tosylate orally disintegrating composition.

7. A process for the preparation of a mouth disintegrating composition of lumerigotone tosylate as claimed in claim 6, wherein, In S1, the volume ratio of the pure water to the organic reagent is (70-99.2):(0.8-30); the organic reagent is one or more of methanol and dichloromethane; the steps of the first dispersion treatment are: stirring at a rotation speed of 300-400rpm for 12-18h, water bath heating at 45-55℃ for 20-40min, freeze-drying at-20℃ for 20-28h and passing through a 65-mesh sieve to obtain the lumerongene tosylate nanoparticles.

8. A process for the preparation of a mouth disintegrating composition of lumerigotone tosylate as claimed in claim 6, wherein, In the S1, the second dispersion treatment is performed by ultrasonic crushing for 8-12 min at a power of 450-550 W, refrigeration at 4℃ for 20-40 min, dialysis in a 50KD dialysis bag for 1-2 h, centrifugation at 9500-10500 rpm for 8-12 min in a 10KD ultrafiltration tube, to obtain the nanometer particles of lumefantrine toluenesulfonate.

9. A process for the preparation of a mouth disintegrating composition of lumerigotone tosylate as claimed in claim 6, wherein, In the S2, the volume ratio of the first pure water and the second pure water is 50:50; the temperature for heating and dissolving is 55-65℃; the stirring speed is 200-300 rpm; and the vacuum degassing time is 15-20 min.

10. A process for the preparation of a mouth disintegrating composition of lumerigotone tosylate as claimed in claim 6, wherein, In the S2, the freeze-drying is performed by pre-freezing at-30℃ for 2.5-3.5 h within 40 min, then entering the main drying stage, raising the temperature to-20℃ at a rate of 0.3-0.7℃ / min under a vacuum degree of 10 Pa, maintaining for 5-7 h, then raising the temperature to-10℃ at a rate of 0.3-0.7℃ / min, maintaining for 6.5-9.5 h, finally raising the temperature to 0℃ at a rate of 0.3-0.7℃ / min, maintaining for 8-12 h, and finally performing the final drying, raising the temperature to 10℃ at a rate of 0.3-0.7℃ / min under a vacuum degree of less than 20 Pa, maintaining for 0.8-1.2 h, then raising the temperature to 25℃ at a rate of 0.6-0.9℃ / min, maintaining for 1.5-2.5 h.

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