CN107137381B - Gefitinib @ PLLA microsphere dry powder inhalant and preparation method thereof - Google Patents
Gefitinib @ PLLA microsphere dry powder inhalant and preparation method thereof Download PDFInfo
- Publication number
- CN107137381B CN107137381B CN201710293236.2A CN201710293236A CN107137381B CN 107137381 B CN107137381 B CN 107137381B CN 201710293236 A CN201710293236 A CN 201710293236A CN 107137381 B CN107137381 B CN 107137381B
- Authority
- CN
- China
- Prior art keywords
- gefitinib
- plla
- dry powder
- solvent
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/007—Pulmonary tract; Aromatherapy
- A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/145—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/146—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Otolaryngology (AREA)
- Pulmonology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a gefitinib @ PLLA microsphere dry powder inhalant based on a supercritical anti-solvent technology and a preparation method thereof. Dissolving gefitinib and a carrier material PLLA in an organic solvent to prepare a mixed solution of the PLLA and the gefitinib; introducing supercritical carbon dioxide into a settling kettle as an anti-solvent, spraying the mixed solution into the settling kettle when the temperature in the settling kettle is 33-48 ℃ and the pressure is 90-120 bar, so that the medicine is embedded in the PLLA carrier and is separated out in the settling kettle as spherical particles, collecting the spherical particles in the settling kettle, and uniformly mixing the spherical particles with lactose to prepare the dry powder inhalant. The preparation method based on the supercritical anti-solvent technology has the advantages of simple process, mild conditions and good reproducibility, and the obtained gefitinib @ PLLA microspheres have high drug loading rate and encapsulation efficiency, small and uniform particle size, controlled slow release effect and stronger cytotoxicity.
Description
Technical Field
The invention relates to gefitinib, in particular to a slow release microsphere lung targeting dry powder inhalant of gefitinib and a preparation method thereof.
Background
Gefitinib (Gefitinib), also known as iressa, is a semisynthetic derivative of camptothecin, a novel selective Epidermal Growth Factor Receptor (EGFR) tyrosine kinase inhibitor; the chemical name is N- (3-chloro-4-fluorophenyl) -7-methoxy-6- (3-morpholine-4-propoxy) quinazoline-4-amine, and the molecular formula is C22H24ClFN4O3Molecular weight is 446.9, modifiedThe chemical Abstract accession number (CAS number) is 184475-35-2, which has the chemical structure shown below:
the gefitinib tablet produced by Aslicon in 2011 is officially approved by the food and drug administration of China. Gefitinib is a drug indicated for the treatment of locally advanced or metastatic non-small cell lung cancer (NSCLC) that has previously been treated with chemotherapy or is otherwise not suitable for chemotherapy. The action mechanism is mainly as follows: competition for Mg-ATP binding sites on the catalytic region of EGFR-TK, and blocking signal transmission; inhibiting activation of mitogen-activated protein kinase, and promoting apoptosis; inhibiting tumor angiogenesis.
But gefitinib also has some disadvantages, such as poor water solubility and high binding degree with human plasma protein, thereby limiting the free drug from entering tumor tissues and causing low bioavailability of the drug; furthermore, oral gefitinib tablets cause adverse physiological reactions such as diarrhea, nausea, vomiting, etc. Experiments and clinical researches show that the preparation of the novel dosage form of the drug delivery system based on microspheres, liposomes and the like can improve the water solubility of the drug, improve the utilization rate of the drug, improve the stability and realize controlled and sustained release.
The lung is an open organ and constitutes, with the respiratory tract, the main site where the human body continuously exchanges gas with the outside world. Gas enters the alveolar region from the nasal and oral cavities through the pharynx, larynx, trachea, bronchi, bronchioles and terminal branches. The total surface area of the alveoli is large and the distance between the surface of the alveoli to the capillaries is only about 1 μm, a good site for drug absorption; the huge absorption area, the abundant capillary vessels and the extremely small transport distance determine the rapid absorption of the pulmonary drug delivery, and the absorbed drug directly enters the blood circulation without the liver first pass effect.
The dry powder inhalant is an important pulmonary administration form, and after passing through a specific administration device, most patients actively inhale the dry powder to reach the lung for exerting the drug effect. The medicine microspheres and lactose (screened by a 200-mesh sieve) are uniformly mixed in different proportions to serve as a dry powder inhalant, the addition of the lactose plays a role in diluting the medicine microspheres, meanwhile, the medicine microspheres with smaller particle sizes are adsorbed on the surface of the lactose with larger particle sizes, the flowability of the medicine microspheres is improved, the sedimentation of the medicine microspheres in oral cavities, throats and respiratory tracts is reduced, and more medicines directly reach the lung to generate medicine effect; when the inhalant is inhaled into the oral cavity, due to the large particle size and large momentum of the lactose particles, the inhalant is deposited in the oral cavity and the throat by inertial collision and finally swallowed into a human body and cannot enter the lung, but small particles adsorbed on the surface of the lactose can be separated from the lactose under the action of air flow and continue to move along with the air flow; particles with smaller particle size (generally, the diameter is less than 0.5 μm) exist in the air in the form of Brownian motion, can enter the lung, but are not easy to deposit, and most of the particles are exhaled along with the air; only when the particle size is 1-5 μm can the particles be deposited in the lung. The inhalation therapy belongs to local administration, and the medicine directly enters the pathological change part of the lung without systemic blood circulation, so the required medicine dose is small, the effect is fast, the safety is good, the side effect is small, and the generated adverse medicine reaction is less. The solubility of the gefitinib in water is increased due to the smaller particle size, so that the water solubility of the gefitinib is improved; in addition, the gefitinib microsphere medicine entering the lung can reach the lesion part without systemic blood circulation, and the gefitinib is wrapped by the PLLA, so that the gefitinib microsphere medicine can be less combined with plasma protein, side effects are reduced, and the bioavailability is improved. The method has a profound significance for improving the medicinal value of the gefitinib, so that the gefitinib is embedded in the carrier PLLA by the supercritical anti-solvent technology, the contact of the gefitinib and the external environment is reduced, and the stability of the medicament can be enhanced; the obtained medicinal microspheres with small and uniform particle size and controllable particle size and lactose are uniformly mixed in different proportions to be used as a dry powder inhalant, so that the water solubility of the gefitinib can be improved, the bioavailability is improved, and the medicinal dose is reduced, so that the side effect of the gefitinib on human bodies is reduced.
Chinese patent CN103536537 discloses a preparation method of a lung targeting therapeutic drug gefitinib PLGA microsphere, the particle size of the obtained microsphere is 8-12 μm, the encapsulation rate is 85-90%, and the drug-loading rate is 9-15%. The preparation method comprises the steps of mixing gefitinib and PLGA, dissolving in dichloromethane or chloroform to serve as a dispersed phase (oil phase), and taking 0.5-2% of sodium dodecyl sulfate aqueous solution as a continuous phase (water); injecting the dispersed phase into the continuous phase under the stirring action to form an oil-in-water O/W type emulsion, continuously stirring until the organic solvent is fully volatilized, centrifugally separating gefitinib to obtain PLGA microspheres, washing with distilled water, and drying to obtain the final PLGA microspheres.
The gefitinib PLGA microspheres with high drug loading capacity and encapsulation efficiency can be obtained by the existing method, but organic solvents used by the method are difficult to completely remove, a large amount of residues are remained, a large amount of deionized water is needed for washing, the preparation process involves long-time stirring, the drugs can be degraded, the product contains more water, the particle size of the product is large, the product is not suitable for being used as a dry powder inhalant, the product can only be used for oral administration, and the preparation process is long and complicated.
Disclosure of Invention
The invention aims to provide a gefitinib @ PLLA dry powder inhalant which has uniform and controllable microsphere particle size, high stability and slow release and can be prepared into a dry powder inhalant and a preparation method thereof, so that the water solubility, the bioavailability and the cytotoxicity of the gefitinib are further improved, and the side effects of the medicine are reduced.
The preparation method has the advantages of simple operation, good reproducibility, small dissolving residual quantity, small environmental pollution and easy industrialization.
The purpose of the invention is realized by the following technical scheme:
the preparation method of the gefitinib @ PLLA microsphere dry powder inhalant based on the supercritical anti-solvent technology is characterized in that gefitinib and a carrier material PLLA are dissolved in an organic solvent to prepare a mixed solution of the PLLA and the gefitinib; introducing supercritical carbon dioxide into a settling kettle as an anti-solvent, spraying the mixed solution into the settling kettle when the temperature in the settling kettle is 33-48 ℃ and the pressure is 90-120 bar, so that the medicine is embedded in the PLLA carrier and is separated out in the settling kettle as spherical particles, collecting the spherical particles in the settling kettle, and uniformly mixing the spherical particles with lactose to prepare a dry powder inhalant; the organic solvent is DCM or a mixed solvent of DCM and EtOH.
To further achieve the object of the present invention, preferably, the introducing of the supercritical carbon dioxide into the settling tank as the anti-solvent is introducing the supercritical carbon dioxide into the settling tank at a constant rate by using a high-pressure pump, and the flow rate of the supercritical carbon dioxide is 15g/min to 25 g/min.
Preferably, the step of spraying the mixed solution into the settling tank is to spray the mixed solution of the PLLA and the gefitinib into the settling tank at a constant speed by using a high-pressure pump, wherein the spraying flow rate of the mixed solution is 0.5 ml/min-1.6 ml/min.
Preferably, the concentration of gefitinib in the mixed solution of PLLA and gefitinib is 0.8mg/ml-1.2 mg/ml.
Preferably, the concentration of the carrier material PLLA in the mixed solution of PLLA and gefitinib is 4mg/ml-14.4 mg/ml.
Preferably, the volume of EtOH in the mixed solvent of DCM and EtOH is 10-50% of the volume of the mixed solvent.
Preferably, the spherical particles in the settling tank are finally dried with supercritical carbon dioxide for 50-60 min.
Preferably, the lactose is lactose sieved with a 200 mesh sieve.
Preferably, the mass ratio of the drug microspheres to the auxiliary material carrier lactose in the dry powder inhalant is 1:1-1: 3.
The gefitinib @ PLLA microsphere dry powder inhalant is characterized in that gefitinib medicaments of the gefitinib @ PLLA microsphere dry powder inhalant are embedded in a carrier material to form microspheres, and the particle size of the microspheres formed by the gefitinib encapsulated in the carrier is 1-5 mu m.
The supercritical anti-solvent method is mainly used for preparing micro-nano particles or microspheres. When the PLLA entrapped gefitinib microsphere is prepared by adopting a supercritical anti-solvent granulation method, the solute, the solvent and the anti-solvent play a synergistic effect, and the process involves complex processes such as a phase equilibrium process, mass transfer, fluid dynamics and the like. Gefitinib is a drug for treating lung cancer, and the Gefitinib is prepared into a dry powder inhalant which can be used for lung local treatment, but no related Gefitinib dry powder inhalant is reported at present. The prior art does not find reports of preparing gefitinib microsphere dry powder inhalant by using supercritical carbon dioxide. The invention discovers that the supercritical carbon dioxide anti-solvent granulation technology utilizes the characteristic that the solubility of the gefitinib medicament in the mixed solvent is lower than that of the PLLA carrier material, so that the medicament is firstly separated out and nucleated and then is quickly wrapped by the carrier material, the medicament loss is effectively avoided, and the medicament loading capacity and the encapsulation efficiency are improved.
The supercritical anti-solvent technology is a new method for preparing the drug sustained-release microspheres, and can ensure that the drug is micronized, thereby increasing the solubility of the insoluble drug and improving the stability of the drug. The principle is that the raw material medicine is dissolved in an organic solvent to form a solution, the solution is sprayed in a supercritical fluid through a micropore nozzle, the solvent and the solute in the solution can be dissolved in supercritical carbon dioxide, and the solute is not dissolved in the supercritical carbon dioxide, so that the solute in the solution forms a great supersaturation degree in a short time, and nanoparticles are separated out to form microspheres. The gefitinib @ PLLA microsphere powder is prepared by adopting a supercritical anti-solvent technology, so that the cytotoxicity of gefitinib can be obviously improved, the particle size is reduced, the operation method is simple, and the residual amount of an organic solvent is small; then the microsphere powder is mixed with lactose which passes through 200 meshes to prepare the dry powder inhalant which has higher evacuation rate and in vitro deposition rate.
Compared with the prior art, the invention has the following advantages:
1. the particle size of the PLLA-encapsulated gefitinib microspheres obtained by the invention is obviously smaller than that of the original drug, the morphology of the PLLA-encapsulated gefitinib microspheres is spherical, the particle size of the original drug is 8-10 mu m, the particle size of the PLLA-encapsulated gefitinib microspheres is 1-5 mu m, and the average particle size is 2.6 mu m. The gefitinib dry powder inhalant prepared by the invention for the first time can be directly used for medicine in the lung, so that the medicine use effect and efficiency are improved, the medicine use amount is reduced, and the side effect of gefitinib is lightened.
2. The supercritical anti-solvent granulation technology utilizes the characteristic that the solubility of the medicine gefitinib in a solvent is lower than that of a carrier material PLLA, so that the medicine is firstly separated out and nucleated in a supercritical process, and is quickly wrapped by the carrier material separated out later to form microspheres with higher encapsulation efficiency; the Gefitinib PLLA dry powder inhalant sustained-release microspheres obtained by the technical scheme of the invention have the encapsulation rate of about 90 percent, the emptying rate of more than 90 percent after being uniformly mixed with lactose which passes through a 200-mesh sieve, the first-level distribution in vitro is 30-50 percent, and the second-level distribution is 15-22 percent.
3. The supercritical anti-solvent granulation technology used in the invention is a process that gefitinib and carrier PLLA are in a dissolved state, and nucleation is rapidly precipitated along with the solvent being taken away by supercritical carbon dioxide, and the process is a microscopic process that nucleation is instantly precipitated after dissolution, so that the particle size of microspheres is uniform.
4. The supercritical anti-solvent granulation technology of the invention does not use any additive, and the used solvent is also removed by supercritical carbon dioxide, so that the additive and the solvent have very little residue and contain no water.
5. The supercritical anti-solvent granulation technology is one-step operation, and the whole operation condition is accurate and controllable and has good reproducibility.
6. The preparation method of the technical scheme of the invention is simple and convenient to operate, the raw materials and the used reagents are commercially available, the environmental pollution is small, and the industrial production is easy.
Drawings
Figure 1 is an SEM image of the parent gefitinib and PLLA of examples 1 and 3 of the present invention;
figure 2 is a graph of the particle size distribution of the technical gefitinib of example 1 of the present invention;
fig. 3 is a PXRD pattern of gefitinib prodrug and PLLA of examples 1 and 2 of the present invention;
FIG. 4 is an infrared spectrum of the technical gefitinib and PLLA of example 1 of the present invention;
fig. 5 is a graph showing the cytotoxicity results of the gefitinib prodrug and example 1 of the present invention on a549 cells after 48 hours of culture.
Detailed Description
For a better understanding of the present invention, the present invention will be further described with reference to the following drawings and examples, but the embodiments of the present invention are not limited thereto.
Example 1
A method for preparing gefitinib @ PLLA microspheres of dry powder inhalant comprises the following steps: accurately measuring 40mL of EtOH and 160mL of LCDMM as solvents, accurately weighing 50mg of gefitinib powder and 250mg of PLLA, dissolving the gefitinib powder and the PLLA in 50mL of solvents to prepare a mixed solution containing 1mg/mL of gefitinib and 5mg/mL of PLLA, and using the rest of solvents for later use; the temperature of the high-pressure settling kettle is set to be 48 ℃ and the pressure is set to be 90bar before starting. Opening a steel cylinder, introducing carbon dioxide from the top end of the settling kettle at a flow rate of 20g/min by using a high-pressure pump, introducing the rest mixed solvent of DCM and EtOH into the settling kettle at a speed of 1.2mL/min by using another high-pressure pump when the temperature and the pressure in the kettle reach the set values, stopping introducing the mixed solvent after a system in the reaction kettle reaches an equilibrium state after 15min, and injecting the mixed solution containing 1mg/mL of gefitinib and 5mg/mL of PLLA at the same speed. And after the sample introduction is finished, continuously introducing carbon dioxide for 60min to dry the microsphere powder, then stopping a carbon dioxide pump, reducing the pressure of the settling kettle to the atmospheric pressure, and taking out the dry microsphere powder, wherein a large amount of the gefitinib microsphere powder encapsulated by the PLLA can be collected on the kettle wall and the kettle bottom of the settling kettle.
Scanning Electron Microscope (SEM) characterization is performed on the PLLA-entrapped gefitinib microsphere powder obtained in example 1, and a morphology chart shown in fig. 1(c) is obtained, wherein (a) and (b) in fig. 1 are morphology charts of technical gefitinib and PLLA, respectively. Figure 1 shows that the gefitinib powder entrapped in PLLA is substantially spherical, while the morphology of the bulk drug is substantially rectangular. The microspheres of the powder were relatively uniform in size, and the gefitinib microsphere powder encapsulated by PLLA obtained in example 1 was characterized by a malvern particle size analyzer, and the particle size distribution thereof was as shown in fig. 2, and the average particle size of this case was 2.6 μm and the average particle size of the original drug gefitinib was 6.2 μm.
The PLLA-encapsulated gefitinib microsphere powder obtained in example 1 was subjected to drug loading and encapsulation efficiency measurements, where the drug loading is the percentage of the encapsulated substance (e.g., gefitinib) in the total microsphere, and the drug loading is (mass of drug in microsphere/total mass of microsphere); the encapsulation efficiency is the percentage of the encapsulated substance in the microspheres based on the total amount of the drug, and is (the total amount of the drug actually encapsulated in the microspheres/the theoretical encapsulated drug in the microspheres) × 100%, and the determination method is as follows: preparing a series of DCM mixed solutions with gefitinib concentration of 10-30 mu g/ml and the like, measuring the maximum absorbance at 328nm by adopting an ultraviolet spectrophotometer, and then obtaining a regression equation of Y-0.04893X after linear fitting-0.07545(R20.9986, Y is the absorbance value, X is the concentration of gefitinib, μ g/mL), then accurately weighed gefitinib microspheres of example 1 are dissolved in DCM, the absorbance is measured after 1min of ultrasound, and the content of gefitinib is calculated, repeated 3 times, and the drug loading of example 1 is calculated to be 15.82%, and the encapsulation efficiency is 94.91% by taking the average value.
The PLLA-encapsulated gefitinib microsphere powder obtained in example 1 was characterized by X-ray diffraction (PXRD), the radiation source was copper target, the obtained PXRD pattern is shown in fig. 3(c), and (a) and (b) in fig. 3 are PXRD patterns of original gefitinib and PLLA, respectively. The microsphere has the characteristic peaks of gefitinib at diffraction angles 2 theta of 6.5, 13.4 and 16.7 degrees. Gefitinib is seen to be entrapped in the PLLA.
Infrared spectroscopic analysis of the PLLA-entrapped gefitinib microsphere powder obtained in example 1, mixing the microspheres with KBr, grinding, tabletting and measurement from 4000cm-1To 500cm-1Characteristic infrared absorption of (1). The infrared absorption spectrum shown in figure 4 is obtained, and the infrared absorption spectrum has characteristic absorption peaks of gefitinib and PLLA at positions 1629.0, 1579.0, 1570.0 and 1439.0, which shows that the chemical structures of gefitinib and PLLA are not changed after being treated by the supercritical elution technology, namely the curative effect of gefitinib is not changed.
The cytotoxicity test of the PLLA-encapsulated gefitinib microsphere powder obtained in example 1 with a549 cells showed that the IC50 concentration was less than that of the original gefitinib after 48 hours, and no IC50 concentration appeared after 48 hours, which indicates that the encapsulated gefitinib had improved drug efficacy, probably due to the fact that the gefitinib particle size became smaller, the water solubility increased and the PLLA-encapsulated had a controlled sustained release effect.
The gefitinib microsphere powder encapsulated in PLLA obtained in example 1 and lactose were uniformly mixed at a ratio of 1:1, and then the mixture was measured by a method for measuring evacuation of inhaled aerosol powder (appendix IL in second part of chinese pharmacopoeia 2010), and a double-layer liquid impactor was used to measure the distribution of inhaled aerosol particles (appendix XH in second part of chinese pharmacopoeia 2010), and the experimental apparatus was the same as the apparatus for measuring the evacuation rate. The average emptying rate, average first-order distribution (in receiver bottle D), and average second-order distribution (in receiver bottle H) were found to be 97.0%, 32.08%, and 20.06%, respectively. When the microsphere powder was mixed with lactose at a ratio of 1:2, an average emptying rate of 96.4%, a primary average distribution of 35.12% and a secondary average distribution of 18.57% was obtained. When the microsphere powder was mixed with lactose at a ratio of 1:3, an average emptying rate of 97.4%, a primary average distribution of 43.06% and a secondary average distribution of 17.96% was obtained. Since the second-order average distribution is larger than 10% of scalar quantity and the second-order distribution is maximum when the ratio of microsphere powder to lactose is 1:1, the smaller the amount of carrier introduced when the dry powder inhalant is made, the better the technical scheme of the ratio of microsphere powder to lactose is, the better the technical scheme is.
Example 2
A method for preparing gefitinib @ PLLA microspheres of dry powder inhalant comprises the following steps: accurately weighing 50mg of gefitinib powder and 250mg of PLLA, and dissolving the gefitinib powder and the 250mg of PLLA in a 50mL solvent in case 1 to prepare a mixed solution containing 1mg/mL of gefitinib and 5mg/mL of PLLA; the temperature of the high-pressure settling kettle is set to be 48 ℃ and the pressure is set to be 110bar before starting. Opening a steel cylinder, introducing carbon dioxide from the top end of the settling kettle at a flow rate of 20g/min by using a high-pressure pump, introducing the rest mixed solvent of DCM and EtOH into the settling kettle at a speed of 0.8mL/min when the temperature and the pressure in the kettle reach the set values, stopping introducing the mixed solvent after a system in the reaction kettle reaches an equilibrium state after 15min, and injecting the mixed solution containing 1mg/mL of gefitinib and 5mg/mL of PLLA at the same speed. And after the sample introduction is finished, continuously introducing carbon dioxide for 60min to dry the microsphere powder, then stopping a carbon dioxide pump, reducing the pressure of the settling kettle to the atmospheric pressure, and taking out the dry microsphere powder, wherein a large amount of the gefitinib microsphere powder encapsulated by the PLLA can be collected on the kettle wall and the kettle bottom of the settling kettle.
Scanning Electron Microscope (SEM) characterization, malvern particle size analyzer, ultraviolet spectrophotometer test, and fourier transform infrared spectroscopy (FT-IR) analysis were performed on the PLLA-encapsulated gefitinib microsphere powder obtained in example 2, the specific steps are the same as those in example 1, and the results show that: the average particle diameter of the PLLA-coated gefitinib microsphere powder is 1.8 mu m, the drug loading is 12.32 percent, and the coating is carried outThe sealing rate is 87.76%, and the infrared absorption is 1629.0cm-1、1579.0cm-1、1570.0cm-1And 1439.0cm-1Both have characteristic peaks, which indicates that the chemical structures of gefitinib and PLLA are not changed after the supercritical elution technology, i.e. the curative effect of gefitinib is not changed.
After the gefitinib microsphere powder encapsulated by the PLLA obtained in the example 2 and lactose are uniformly mixed in a ratio of 1:1, a capsule shell No. 3 is arranged, the determination is carried out according to a method for determining the evacuation of inhaled fog powder (appendix IL in the second part of the 2010 edition of Chinese pharmacopoeia), and a double-layer liquid collider is adopted; the experimental apparatus was the same as the apparatus for measuring the evacuation rate, as measured by the aerosol particle distribution measuring method (2010 version, appendix XH of China pharmacopoeia, second division). The average emptying rate, average primary distribution (in receiver bottle D), and average secondary distribution (in receiver bottle H) were found to be 95.0%, 31.32%, and 19.45%, respectively. When the microsphere powder was mixed with lactose at a ratio of 1:2, an average emptying rate of 96.8%, a first order average distribution of 34.37% and a second order average distribution of 15.02% was obtained. When the microsphere powder was mixed with lactose at a ratio of 1:3, an average emptying rate of 98.7%, a first order average distribution of 44.79% and a second order average distribution of 14.59% were obtained. The ratio of microsphere powder to lactose is 1:1 optimal since the secondary average distribution is greater than 10% of the scalar and is greatest when the ratio of microsphere powder to lactose is 1:1, while the smaller the amount of carrier introduced in the dry powder inhaler the better.
Example 3
A method for preparing gefitinib @ PLLA microspheres of dry powder inhalant comprises the following steps: accurately measuring 25mL of EtOH and 25mL of DCM as solvents, weighing 50mg of gefitinib powder and 350mg of PLLA, and dissolving the gefitinib powder and the PLLA in 50mL of solvents to prepare a mixed solution with 1mg/mL of gefitinib concentration and 7mg/mL of PLLA concentration; the temperature of the high-pressure settling kettle is set to be 43 ℃ and the pressure is set to be 90bar before starting. Opening a steel cylinder, introducing carbon dioxide from the top end of the settling kettle at a flow rate of 20g/min by using a high-pressure pump, introducing the rest mixed solvent of DCM and EtOH into the settling kettle at a speed of 1.6mL/min by using another high-pressure pump when the temperature and the pressure in the kettle reach the set values, stopping introducing the mixed solvent after a system in the reaction kettle reaches an equilibrium state after 15min, and injecting the mixed solution containing 1mg/mL of gefitinib and 7mg/mL of PLLA at the same speed. And after the sample introduction is finished, continuously introducing carbon dioxide for 60min to dry the microsphere powder, then stopping a carbon dioxide pump, reducing the pressure of the settling kettle to the atmospheric pressure, and taking out the dry microsphere powder, wherein a large amount of the gefitinib microsphere powder encapsulated by the PLLA can be collected on the kettle wall and the kettle bottom of the settling kettle.
Scanning Electron Microscope (SEM) characterization, malvern particle size analyzer, ultraviolet spectrophotometer test, and fourier transform infrared spectroscopy (FT-IR) analysis were performed on the PLLA-encapsulated gefitinib microsphere powder obtained in example 3, the specific steps are the same as those in example 1, and the results show that: an SEM image of the PLLA-encapsulated gefitinib microsphere powder is shown in figure 1(d), and the particle size is 3.6 μm, the drug loading is 10.87%, the encapsulation efficiency is 91.66%, and the infrared absorption is 1629.0cm-1、1579.0 cm-1、1570.0 cm-1And 1439.0cm-1Both have characteristic peaks, which indicates that the chemical structures of gefitinib and PLLA are not changed after the supercritical elution technology, i.e. the curative effect of gefitinib is not changed.
After the gefitinib microsphere powder encapsulated by the PLLA obtained in the example 3 and lactose are uniformly mixed in a ratio of 1:1, a capsule shell No. 3 is arranged, and the mixture is measured according to a method for measuring the evacuation of inhaled mist powder (2010 version, appendix IL of the second part of Chinese pharmacopoeia), and a double-layer liquid collider is adopted; the experimental apparatus was the same as the apparatus for measuring the evacuation rate, as measured by the aerosol particle distribution measuring method (2010 version, appendix XH of China pharmacopoeia, second division). The average emptying rate, average primary distribution (in receiver bottle D), and average secondary distribution (in receiver bottle H) were found to be 96.0%, 33.43%, and 19.89%, respectively. When the microsphere powder was mixed with lactose at a ratio of 1:2, an average emptying rate of 96.8%, a primary average distribution of 33.56% and a secondary average distribution of 18.81% were obtained. When the microsphere powder was mixed with lactose at a ratio of 1:3, an average emptying rate of 96.7%, a primary average distribution of 42.16% and a secondary average distribution of 16.75% was obtained. The ratio of microsphere powder to lactose is 1:1 optimal since the secondary average distribution is greater than 10% of the scalar and is greatest when the ratio of microsphere powder to lactose is 1:1, while the smaller the amount of carrier introduced in the dry powder inhaler the better.
A summary of the drug loading, encapsulation and particle size for gefitinib prodrugs and examples 1-3 is shown in table 1.
TABLE 1
From the above table 1 and SEM images, it can be seen that PLLA-encapsulated gefitinib microsphere powder with uniform particle size, high drug loading and high encapsulation efficiency can be prepared by using the supercritical anti-solvent technology. Analysis in infrared spectroscopy shows that the positions of characteristic peaks of gefitinib and PLLA are unchanged after the supercritical anti-solvent technology, which indicates that the chemical structure is unchanged, i.e. the curative effect of gefitinib is unchanged. The particle size of the obtained PLLA-entrapped gefitinib microsphere is smaller than that of the gefitinib raw material, the solubility is improved, and the PLLA-entrapped gefitinib microsphere has a controlled slow release effect; the cytotoxicity result of A459 cells shows that no IC50 value appears in the original drug after 48 hours, and the microsphere obtained after entrapment has IC50 in 48 hours, so that the entrapped drug microsphere has stronger cytotoxicity.
Embodiment 1-3 gefitinib microsphere powder encapsulated by PLLA and lactose are uniformly mixed according to a ratio of 1:1 and then are loaded in a No. 3 capsule shell, and the mixture is measured according to a suction mist powder emptying measurement method (appendix IL of second part of Chinese pharmacopoeia 2010 edition) by adopting a double-layer liquid collider; the experimental apparatus was the same as the apparatus for measuring the evacuation rate, as measured by the aerosol particle distribution measuring method (2010 version, appendix XH of China pharmacopoeia, second division). The measured average emptying rate, average primary distribution (in receiver bottle D), and average secondary distribution (in receiver bottle H) are summarized in table 2:
TABLE 2
Table 2 shows that the emptying rate of the microspheres prepared by the invention mixed with lactose as a dry powder inhalant is over 90 percent, the requirement of the dry powder inhalant on the emptying rate is met, the secondary distribution is over 15 percent, and the requirement of the dry powder inhalant is also met.
The process for preparing the gefitinib dry powder inhalant has the advantages of simple process, mild condition, no solvent residue and good reproducibility, and greatly reduces the particle size of gefitinib medicaments; the emptying rate, the primary distribution and the secondary distribution of the gefitinib @ PLLA microsphere powder obtained from the settling kettle after SAS and the auxiliary material carrier lactose are mixed uniformly in a proper proportion can meet the regulations of Chinese pharmacopoeia. Experiments and clinical researches show that the preparation of the novel preparation based on the drug delivery systems such as microspheres, liposomes and the like can improve the water solubility of the drug, improve the utilization rate of the drug, improve the stability and realize controlled slow release; and the particle size of the gefitinib is reduced, so that the solubility of the gefitinib is increased, the defect of poor water solubility of the gefitinib is overcome, and the bioavailability of the gefitinib is improved.
The dry powder inhalant is actively inhaled by a patient, the gefitinib @ PLLA microsphere medicine entering the lung through the oral cavity, the throat, the bronchus and the like can reach the pulmonary disease part without systemic blood circulation, and the liver first-pass effect is avoided, so that the medicinal dose of gefitinib can be reduced, and the gefitinib coated in the PLLA can reduce the combination of the gefitinib medicine and plasma protein and improve the bioavailability of gefitinib by controlling the slow release effect. The gefitinib @ PLLA microsphere dry powder inhalant prepared by the invention can reduce the medicinal dose of a patient by pulmonary administration, simultaneously improve the water solubility of the gefitinib and improve the bioavailability of the gefitinib, and the dose reduction, the increase of the water solubility of the medicament and the improvement of the bioavailability can greatly reduce the dose of the gefitinib under the condition of reaching the same medicinal effect, thereby reducing the side effect of the gefitinib on a human body.
The particle size of the PLLA-encapsulated gefitinib microspheres obtained by the invention is obviously smaller than that of the original drug, the morphology of the PLLA-encapsulated gefitinib microspheres is spherical, the particle size of the original drug is 8-10 mu m, the particle size of the PLLA-encapsulated gefitinib microspheres is 1-5 mu m, and the average particle size is 2.6 mu m. The gefitinib dry powder inhalant prepared by the invention for the first time can be directly used for medicine in the lung, so that the medicine effect and efficiency are improved, the medicine consumption is reduced, and the side effect of gefitinib is reduced; the invention has good application prospect in the aspects of preparing drug sustained-release microspheres, dry powder inhalant and the like.
Claims (9)
1. The preparation method of the gefitinib @ PLLA microsphere dry powder inhalant based on the supercritical anti-solvent technology is characterized in that gefitinib and a carrier material PLLA are dissolved in an organic solvent to prepare a mixed solution of the PLLA and the gefitinib; introducing supercritical carbon dioxide into a settling kettle as an anti-solvent, spraying the mixed solution into the settling kettle when the temperature in the settling kettle is 33-48 ℃ and the pressure is 90-120 bar, so that the medicine is embedded in the PLLA carrier and is separated out in the settling kettle as spherical particles, collecting the spherical particles in the settling kettle, and uniformly mixing the spherical particles with lactose to prepare a dry powder inhalant; the organic solvent is a mixed solvent of DCM and EtOH; the volume of EtOH in the mixed solvent of DCM and EtOH is 20-50% of the volume of the mixed solvent.
2. The preparation method of gefitinib @ PLLA microsphere dry powder inhaler based on the supercritical anti-solvent technology as claimed in claim 1, wherein the supercritical carbon dioxide is introduced into the settling tank as the anti-solvent by a high pressure pump at a constant rate, and the flow rate of the supercritical carbon dioxide is 15g/min-25 g/min.
3. The preparation method of gefitinib @ PLLA microsphere dry powder inhaler based on the supercritical anti-solvent technology as claimed in claim 1, wherein the step of spraying the mixed solution into the settling tank is to spray the mixed solution of PLLA and gefitinib into the settling tank at a constant rate by using a high pressure pump, and the spraying flow rate of the mixed solution is 0.5ml/min to 1.6 ml/min.
4. The preparation method of gefitinib @ PLLA microsphere dry powder inhaler based on the supercritical anti-solvent technology as claimed in claim 1, wherein the concentration of gefitinib in the mixed solution of PLLA and gefitinib is 0.8mg/ml-1.2 mg/ml.
5. The preparation method of gefitinib @ PLLA microsphere dry powder inhaler based on the supercritical anti-solvent technology as claimed in claim 1, wherein the concentration of carrier material PLLA in the mixed solution of PLLA and gefitinib is 4mg/ml-14.4 mg/ml.
6. The method for preparing gefitinib @ PLLA microsphere dry powder inhaler based on the supercritical anti-solvent technology as claimed in claim 1, wherein the spherical particles in the settling tank are finally dried with supercritical carbon dioxide for 50-60 min.
7. The preparation method of gefitinib @ PLLA microsphere dry powder inhaler based on the supercritical anti-solvent technology as claimed in claim 1, wherein the lactose is lactose sieved with 200 mesh sieve.
8. The preparation method of the gefitinib @ PLLA microsphere dry powder inhalant based on the supercritical anti-solvent technology as claimed in claim 1, wherein the mass ratio of the drug microspheres to the lactose as the adjuvant carrier in the dry powder inhalant is 1:1-1: 3.
9. The gefitinib @ PLLA microsphere dry powder inhaler prepared by the preparation method of any one of claims 1-8, wherein gefitinib drug of the gefitinib @ PLLA microsphere dry powder inhaler is embedded in a carrier material to form microspheres, and the particle size of the microspheres formed by the gefitinib encapsulated in the carrier is 1-5 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710293236.2A CN107137381B (en) | 2017-04-28 | 2017-04-28 | Gefitinib @ PLLA microsphere dry powder inhalant and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710293236.2A CN107137381B (en) | 2017-04-28 | 2017-04-28 | Gefitinib @ PLLA microsphere dry powder inhalant and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107137381A CN107137381A (en) | 2017-09-08 |
| CN107137381B true CN107137381B (en) | 2020-01-14 |
Family
ID=59774020
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710293236.2A Active CN107137381B (en) | 2017-04-28 | 2017-04-28 | Gefitinib @ PLLA microsphere dry powder inhalant and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107137381B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106083739A (en) * | 2016-05-31 | 2016-11-09 | 华南理工大学 | New gefitinib crystal form and preparation method based on super-critical anti-solvent technology thereof |
-
2017
- 2017-04-28 CN CN201710293236.2A patent/CN107137381B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106083739A (en) * | 2016-05-31 | 2016-11-09 | 华南理工大学 | New gefitinib crystal form and preparation method based on super-critical anti-solvent technology thereof |
Non-Patent Citations (1)
| Title |
|---|
| Insulin-loaded poly-l-lactide porous microspheres prepared in supercritical CO2 for pulmonary drug delivery;Ai-Zheng Chen等;《J. of Supercritical Fluids》;20150328;第101卷;第117-123页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107137381A (en) | 2017-09-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2932978A1 (en) | Application of silicon dioxide aerogel in pharmacy | |
| CN103086346B (en) | A kind of preparation method of mesoporous carbon and application | |
| Chen et al. | Inhaled curcumin mesoporous polydopamine nanoparticles against radiation pneumonitis | |
| Nolan et al. | Particle engineering of materials for oral inhalation by dry powder inhalers. II—Sodium cromoglicate | |
| Gallo et al. | Development of porous spray-dried inhalable particles using an organic solvent-free technique | |
| CN102283851B (en) | Imperialine-BETA-N-oxide and the new application of isoverticine-BETA-N-oxide | |
| Xu et al. | Formulation and characterization of spray-dried powders containing vincristine-liposomes for pulmonary delivery and its pharmacokinetic evaluation from in vitro and in vivo | |
| Carr et al. | Particle formation of budesonide from alcohol-modified subcritical water solutions | |
| CN102961341B (en) | Nanoscale doxorubicin hydrochloride and preparation method thereof | |
| Wang et al. | Preparation of inhalable quercetin-β-cyclodextrin inclusion complexes using the supercritical antisolvent process for the prevention of smoke inhalation-induced acute lung injury | |
| EP3362040B1 (en) | Method of production of inhalable composite particles using a three-fluid nozzle | |
| CN101747305A (en) | Five crystal forms of nicousamide compound and preparation method, pharmaceutical composition and usage thereof | |
| CN106361724B (en) | A sustained release nanometer microsphere composition of 20(R) -ginsenoside Rg3 and its preparation method | |
| CN107137381B (en) | Gefitinib @ PLLA microsphere dry powder inhalant and preparation method thereof | |
| Wang et al. | Enhancing bioavailability of natural extracts for nutritional applications through dry powder inhalers (DPI) spray drying: technological advancements and future directions | |
| CN106265624B (en) | Pharmaceutical composition for treating breast cancer, drug delivery system and preparation method thereof | |
| US20240050450A1 (en) | Inhalable Cannabinoid Formulations | |
| Wang et al. | Stiripentol enteric solid dispersion-loaded effervescent tablets: Enhanced dissolution, stability, and absorption | |
| WO2014090169A1 (en) | Nanoscale paclitaxel and preparation method thereof | |
| CN106913528A (en) | Eliquis micropill and preparation method thereof | |
| CN104622850A (en) | Mini-pill type nicergoline capsule and preparation method thereof | |
| WO2014090174A1 (en) | Nanoscale capecitabine and preparation method therefor | |
| CN108553445B (en) | A kind of preparation method of taxol powder spray | |
| CN115124532A (en) | Eutectic crystal of rhein and matrine, preparation method, composition and application thereof | |
| US20230372345A1 (en) | Inhaled PDE-V Inhibitor Drugs |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |