CN105640924B - Alendronate sodium powder spray and its preparation method and application for respiratory tract administration - Google Patents
Alendronate sodium powder spray and its preparation method and application for respiratory tract administration Download PDFInfo
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/66—Phosphorus compounds
- A61K31/662—Phosphorus acids or esters thereof having P—C bonds, e.g. foscarnet, trichlorfon
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Abstract
The present invention provides a kind of Alendronate sodium powder spray and its preparation method and application for respiratory tract administration.It is single Alendronate sodium micro mist that the Alendronate sodium powder spray, which includes composition, does not contain auxiliary material.It is not related to other auxiliary materials or solvent in the production preparation process of Alendronate sodium micro mist, is directly crushed and be prepared by Alendronate sodium bulk pharmaceutical chemicals.The Alendronate sodium Alevaire of the present invention can improve the respiratory function about 40% of COPD rats, better than the effect of the salbutamol sulfate of conventional amount used, and can overcome the drug resistance problems of the β 2 receptor agonists such as salbutamol sulfate.It is mainly deposited in the lung airways such as trachea-bronchial epithelial cell and tiny bronchus after powder spray administration sucking and directly plays effect, the respiratory tract diastolic function speed that takes effect is fast, lasting medicine；In into alveolar and to be absorbed into blood volume relatively low, can avoid generating system side effect, be suitble to develop into COPD and treating asthma medication.
Description
Technical field
The invention belongs to materia medica and galenic pharmacy field, are related to a kind of Alendronate sodium powder spray for respiratory tract administration
And its preparation method and application, mainly it is deposited on the Alendronate sodium dust cloud that drug effect is directly played in respiratory tract after especially sucking
Agent.
Background technology
Alendronate sodium (Alendronate) is a kind of small molecule combination drug, molecular weight 249.97, similar in construction to
Amido pyrophosphoric acid sodium salt.Existing dosage form is oral for tablet, and primary efficacy is to hinder bone reabsorption, for treating and preventing menopause
The osteoporosis of women afterwards.Its pharmacological mechanism acted on is the affinity based on Alendronate sodium Yu bone hydroxyapatite crystal,
Selectively inhibit osteoclastic reabsorption in bone reforming cycle.Nineteen ninety-five U.S. FDA ratifies Alendronate sodium piece oral medication and wears
Osteoporosis after lucky Te Shi diseases and women's menopause.Clinical dosage is oral about 0.2mg/kg (10mg/ days).Oral alendronic acid
For sodium piece under hydrochloric acid in gastric juice effect, being formed has irritating alendronic acid, can cause gastrointestinal discomfort, it is also possible to cause stomach and food
Pipe damages and ulcer；The cancer of the esophagus is even resulted in, therefore, the patient of gastrointestinal problems or esophagus disorder such as cardiospasm is presented,
Alendronate sodium piece cannot be taken.After Alendronate sodium enters blood, most (about 99.3%) passes through kidney excretion quickly
Fall, only fraction (about 0.7%) is deposited into bone, and therefore, bioavilability is very low.The local absorption of Alendronate sodium compared with
It is good：Subcutaneous and intramuscular injection can be implemented.Sutton et al. has tested the nasal absorption of Alendronate sodium with dog and rat -- knot
Fruit shows that nasal absorption degree is bigger than oral, but they do not provide new form formula.In rat Germicidal efficacy to A Lun phosphines
The height of sour sodium enters blood and is such as injected intravenously Alendronate sodium, has the risk for causing Toxicity of Kidney, and cause Alendronate sodium in non-calcium
It is accumulated in the tissue of change.
Chronic Obstructive Pulmonary Disease (referred to as " chronic obstructive pulmonary disease " or " COPD "), is characterized by not fully reversible flow limitation
Chronic lung disease, clinically often show as the symptoms such as the cough, expectoration, expiratory dyspnea of recurrent exerbation, typically exhibit out into
The characteristics of row is in progress, includes most chronic bronchitis and pulmonary emphysema.As disease develops, lead to airway remodeling,
Irreversibility airflow obstruction finally is developed into, or Chong Die with asthma generation coexists.Asthma (full name：Bronchial asthma) it is a kind of slow
It is narrow that property respiratory tract disease, the usually flow limitation caused by airway inflammation are divided into acute bronchus, cause to stridulate, is uncomfortable in chest,
Expiratory dyspnea, the recurrent exerbation coughed.COPD and asthma seriously threaten the health of the public in the world.The current whole world is
There are 2.1 hundred million COPD patients and about 300,000,000 asthmatic patients.COPD and asthmatic patient respectively reach 40,000,000 people and 30,000,000 in China
People.The epidemiological survey presided over by Zhong Nanshan academician is the results show that the overall illness rate of China COPD is at present
8.2%, wherein male's illness rate is 12.4%, and women illness rate is 5.1%.Estimate according to the World Health Organization, in social environment
With treatment be delayed under the influence of, COPD by be mankind's disablement one of Etiological.COPD is in global disease death
In reason, it is only second to heart disease, cerebrovascular disease and acute pulmonary infection, 4th arranged side by side together with AIDS.And the death rate is
Increase year by year, because the number of COPD death just reached 3,000,000 in 2004.
Broxaterol (including salbutamol Salbutamol, salmeterol Salmeterol etc.) is that most have at present
The bronchodilators of effect can release rapidly bronchial spasm and clinical symptoms, be that the first choice of institute has age COPD and asthma is controlled
Treat drug.(1~2 week or for more time) can lead to drug resistance using β 2 receptor agonist and weaken or even lose however, long-term
Go curative effect.Such as：It was reported respectively in 2010 1996 and Bonini et al. by Giannini et al., for respectively by taking exercise
Or the asthma that anaphylactogen induces, β 2 receptor agonist Salmeterol's, the application of 1-2 hour, curative effect highly significant,
FEV1% (one second forced expiratory volume/forced vital capacity, normal value 83%；Asthma causes the value to reduce or be substantially reduced)
Maximum reduction amount is reduced to about 4%-8%, but after continuous use 7-14 days, maximum reduction amount is returned to about by about 30%
24%, curative effect seriously reduces.Since there are Commpensation And Adaptation, the use of β 2 receptor agonist clinically is restricted, at present
Mainly as drug is alleviated, i.e. interim application as needed, or be combined with glucocorticoid.Glucocorticoid be it is current most
Common control drug (drug all used daily), but glucocorticoid cannot cure asthma, need largely to use for a long time,
Many adverse reactions can be caused to patient, including：Burden of liver aggravates, thinning of skin and to be easy scratch, adrenal suppression, bone close
Leaf-hypothalamic pituitary adrenal axis inhibits, is fertile after degree reduces (osteoporosis), cataract, hypertension, diabetes, Hypothalamus-pituitary
Fat, myasthenia, the resistance decline etc. to herpesvirus infection.Wherein, osteoporosis and caused by the incidence fractured it is outstanding
It is high, therefore clinically recommends, to no matter the patient of systemic glucocorticoid treatment asthma being used for a long time in what manner, to answer
Implement preventative anti-osteoporosis disease treatment, and regularly X-ray sclerotin inspection.This not only causes prodigious warp to patient
Ji and psychological burden, and long-term X-ray examination itself also carries radiation sex chromosome mosaicism.
Studies have shown that the tolerance principle of β 2 receptor agonist is to lead to beta 2 receptor after β 2 receptor agonist is combined with beta 2 receptor
It follows endolysosome degradation pathway and degrades.Japanese Sasaki etc. and Ueno etc. verify Alendronate sodium by mouse experiment respectively
There is anti-inflammatory effect to asthma and COPD mouse.But these effects are to be directly appended to culture solution by cell culture or take orally
Mode is not tested without suggesting being directly administered by respiratory tract yet.Katsumi etc., Cruz etc., Sultana etc.,
EzzatiNazhadDolatabadiab etc. reports that Alendronate sodium inhalant includes atomized drop, powder spray, nanometer respectively
Grain etc., experiment show that about 200 times of bioavilability can be improved, are likely to be suitable for treating osteoporosis or antidote, so
And it does not test without suggesting whether being possible to be suitble to COPD or treating asthma yet.The Alendronate sodium of this several document reports
Powder spray, micro mist preparation are to use spray drying process, and Alendronate sodium is needed to be dissolved in the organic solvents such as ethyl alcohol in preparation process,
And using auxiliary materials such as ammonium hydrogen carbonate, leucines, not only preparation process is relatively complicated, but also due to the powder spray of Chinese Pharmacopoeia approval
Auxiliary material only has lactose one kind, so the more difficult acquirement Chinese food Drug Administration of powder spray prepared using this kind of method
(CFDA) approval.Meanwhile (d100≤12 μm, air are dynamic for the granularity very little of the Alendronate sodium micro mist of this several document reports
Mechanics granularity≤2.5 μm), blood is largely absorbed by alveolar after sucking.Enter the characteristic after blood due to Alendronate sodium
It is that by kidney excretion, small part is deposited in bone major part quickly, is played in this way by being connected to respiratory tract between absorbed into serum
The efficiency of effect is very low, and increases the risk of the side effects such as Toxicity of Kidney, the small Alendronate sodium micro mist of such granularity and
Its powder spray is not ideal COPD or asthma medication.
Invention content
The present invention is provided and a kind of preparing a kind of Alendronate sodium dust cloud to overcome at least one deficiency of the prior art
Agent reduces sucking in alveolar, to reduce into blood, avoids causing to body system while in improving respiratory tract locally by rate
The side effect of system.
To achieve the goals above, the present inventor has done numerous studies and practical work, and the specific present invention adopts
With following technical scheme：
The present invention provides a kind of Alendronate sodium powder spray for respiratory tract administration, the Alendronate sodium powder sprays
Composition be single Alendronate sodium micro mist, do not contain auxiliary material.
Further, the Alendronate sodium powder spray is directly crushed by Alendronate sodium bulk pharmaceutical chemicals and is prepared.
Preferably, the present invention proposes to prepare and using the larger Alendronate sodium powder spray of granularity, this belongs to international pioneering.
In the dosage form of Pulmonary inhalation, the drugloading rate of powder spray is big, and production technology is simpler.According to the designed use of powder spray, medicine
The size distribution of powder, dissolubility, heap density, angle of repose, atomization, moisture absorption variation reach curative effect and must take into consideration.Often
The administration target for advising the powder spray used is into alveolar absorbed into serum, and medicinal powder will pass through tracheae, branch from oral cavity to lung
Considerably long one section of moist respiratory tract such as tracheae, and granularity 5 μm or more of particle be easy to be deposited in respiratory tract without into
Enter alveolar, thus granularity (d90) need to generally control at 5 μm hereinafter, and be frequently necessary to selection good water solubility, irritation it is small,
The auxiliary material of good biocompatibility reduces and avoids to be deposited in respiratory tract in favor of diffusing into alveolar.These auxiliary materials such as surface
Activating agent etc. is used for a long time, and may generate damage to lung.It is contemplated that being all tracheae to the cardinal symptom of COPD and asthma
And bronchial obstruction, the medicinal powder of granularity moderate (5~20 μm) can be deposited on trachea-bronchial epithelial cell and tiny bronchus directly plays
Effect is equivalent to respiratory tract local application, than enter alveolar absorbed into serum effect more rapidly, and can to avoid or mitigate may
System side effect.According to particle in respiratory tract sedimentary characteristic：5~20 μm are mainly deposited on trachea-bronchial epithelial cell and tiny branch gas
Pipe ,≤2.5 μm can enter tiny bronchus and be deposited in alveolar, the powder of trachea-bronchial epithelial cell and tiny bronchus local application
The granularity (d90) of mist agent is relatively suitble to control in 5~20 μm of this ranges.
Preferably, the powder size of the Alendronate sodium micro mist is distributed as d90≤20 μm, and d50 >=5 μm, and d10≤
2.5μm。
Further, Alendronate sodium micro mist is packed into capsule with the explosive payload of 0.6mg~5mg, daily medication is primary.
It is the present invention also provides a kind of preparation method of above-mentioned Alendronate sodium powder spray, Alendronate sodium bulk pharmaceutical chemicals are straight
It connects crushing to be prepared, does not add auxiliary material or solvent.
Further, it is 50g~1000g to crush applied sample amount, and crushing pressure is 0.7Mpa, and grinding time is 2.5h~3.5h,
It is 1 time or 1 time or more to crush number.
The present invention also provides a kind of above-mentioned Alendronate sodium powder sprays to prepare treatment Chronic Obstructive Pulmonary Disease and asthma
Purposes in drug.
Further, the present invention also provides a kind of above-mentioned Alendronate sodium powder sprays to prepare treatment to β 2 receptor agonist production
Purposes in the Chronic Obstructive Pulmonary Disease drug of raw drug resistance.
Compared with prior art, the present invention has the advantage that：
The Alendronate sodium powder spray of the present invention is only directly crushed by Alendronate sodium main ingredient and is prepared, do not need profit
With or any auxiliary material of addition, compared with the spray drying process used by the document report, not only process for producing is simple for this law, holds
Easily implement, and avoid spray drying process it is possible because using auxiliary material and solvent caused by side effect.
The Alendronate sodium powder spray of the present invention, which is mainly deposited in respiratory tract directly to play, promotees bronchiectasis effect.By
It is larger in granularity, it is smaller into alveolar part, therefore it is low to enter blood volume, it avoids and deposits in its hetero-organization and kidney pressure
Power.
The Alendronate sodium powder spray of the present invention, overcomes its tolerance at the effect of can not only enhancing β 2 receptor agonist
The problem of, also, because the usage amount of β 2 receptor agonist can be reduced, reduce and avoid the side effect of β 2 receptor agonist
(mainly in terms of heart).In addition, there is Alendronate sodium anti-inflammatory effect, the dosage of glucocorticoid can accordingly reduce,
The side effect of glucocorticoid can be mitigated.Meanwhile Alendronate sodium powder spray of the invention includes the small energy of a small amount of granularity
Enough enter the ingredient that alveolar absorbs, this partially absorbs the Alendronate sodium into blood, it is possible to play anti-inflammatory and pre- preventing bone rarefaction
The effect of, the side effect for preventing and overcoming inhaled glucocorticoid is helped, therefore, Alendronate sodium powder spray of the invention is only
It needs to control d90≤20 μm, and d50 >=5 μm, d10≤2.5 μm, granularity need not be excluded 2.5 μm of parts below.From
And the Alendronate sodium powder spray of the present invention is expected to develop into COPD and asthma medications, is suitble to COPD and asthmatic patient long
Phase uses.
Description of the drawings
Fig. 1 is the SEM electron microscopes of ALE-4 samples.
Fig. 2 is the SEM electron microscopes of ALE-6 samples.
Fig. 3 is the high-efficient liquid phase chromatogram of Alendronate sodium micro mist.
Fig. 4 is lung tissue of rats H&E colored graphs.
Fig. 5 is the relative parameter change figure of special airway resistance after each rat sucking gradient concentration salbutamol.
Fig. 6 is the relative parameter change figure of special airway resistance after each rat sucking gradient concentration Alendronate sodium.
Fig. 7 is that four normal rats schemed the reactivity of salbutamol at the 0th day.
Fig. 8 is that four COPD rats schemed the reactivity of salbutamol at the 0th day.
Fig. 9 is four normal groups and four COPD groups rats in the 0th day reactive mean value figure to salbutamol.
Figure 10 is salbutamol after tolerance-induced 21 days, and 4 normal rats scheme the reactivity of salbutamol.
Figure 11 is salbutamol after tolerance-induced 21 days, and 4 COPD groups rats scheme the reactivity of salbutamol.
Figure 12 is salbutamol after tolerance-induced 21 days, 4 normal groups and reactions of 4 COPD groups rats to salbutamol
The mean value of property compares figure.
Figure 13 is that salbutamol is tolerance-induced forward and backward, and 4 COPD groups rats compare the reactive mean value of salbutamol
Figure.
Figure 14 is salbutamol after tolerance-induced 21 days, and 4 normal rats scheme the reactivity of Alendronate sodium.
Figure 15 is salbutamol after tolerance-induced 21 days, response diagram of 4 COPD groups rats to Alendronate sodium.
Figure 16 is salbutamol after tolerance-induced 21 days, reactive mean value figure of 4 COPD groups rats to Alendronate sodium.
Figure 17 is that salbutamol is tolerance-induced forward and backward, and 4 normal rats and 4 COPD group rats are to Alendronate sodium
Reactive mean value compares figure.
Specific implementation mode
For above and other objects of the present invention, feature and advantage can be clearer and more comprehensible, preferred embodiment cited below particularly,
And coordinate attached drawing, it is described in detail below.
1. formula
The present invention uses the Alendronate sodium bulk pharmaceutical chemicals for meeting national standard for raw material, and production system is carried out with mechanical crushing method
It is standby, other auxiliary materials are not used or are related in the production preparation process of micro mist.
1 Alendronate sodium domestic raw material medicine situation of table
| Name of product | Authentication code | Production unit | Drug one's own department or unit code |
| Alendronate sodium | Chinese medicines quasi-word H20054858 | Shaanxi Hanjiang Pharmaceutical Group Co., Ltd. | 86902372000018 |
| Alendronate sodium | Chinese medicines quasi-word H20123067 | Chongqing Carelife Pharmaceutical Co., Ltd. | 86901012000210 |
| Alendronate sodium | Chinese medicines quasi-word H20057738 | Yangzijiang Pharmaceutical Group Co., Ltd | 86901749000026 |
| Alendronate sodium | Chinese medicines quasi-word H10980108 | Shijiazhuang Hua Sheng pharmaceutical Co. Ltds of pharmacy group | 86902769000010 |
| Alendronate sodium | Chinese medicines quasi-word H20067673 | Hainan Zhonghe Pharmaceutical Co., Ltd | 86905842000013 |
| Alendronate sodium | Chinese medicines quasi-word H20058936 | BeiJing WanSheng Pharmacy Co., Ltd | 86900177000011 |
| Alendronate sodium | Chinese medicines quasi-word H20093202 | Zhejiang Subo'er Pharmaceutical Co., Ltd. | 6904710000070 |
| Alendronate sodium | Chinese medicines quasi-word H20093276 | Hangzhou Aoyi Baoling Pharmaceutical Co., Ltd | 86904471000166 |
| Alendronate sodium | Chinese medicines quasi-word H19980100 | The graceful jinx pharmaceutical factory in Hainan | 86905796000039 |
| Alendronate sodium | Chinese medicines quasi-word H20059934 | Hainan Haishen Tongzhou Pharmaceutical Co., Ltd. | 86905777000027 |
From the point of view of each producer's bulk pharmaceutical chemicals situation explanation, specification standards are identical, are all the symbols of Alendronate sodium purity >=98%
Close the bulk pharmaceutical chemicals of national standard.We have selected the bulk pharmaceutical chemicals that Shaanxi Hanjiang Pharmaceutical Group Co., Ltd. produces, quality inspection knot
Fruit is qualified.
2. micro mist preparation process.The present invention uses mechanical crushing method.
Mechanical crushing method is to prepare one of common method of powder spray, but be spray dried method gradually and substituted.It is main
Want the reason is that compared with spray drying process, the powder size that mechanical crushing method is prepared is larger, 2~3 μm be the method crushing
The limit, and micro mist shape is irregular；Pulverization can also generate electrostatic, and drug has higher surface certainly after micronizing
By energy, powder is easily assembled agglomerating, causes drug powder viscosity to enhance, poor fluidity influences drug and discharged from micro mist inhalator
Redisperse afterwards.The violent pulverization of mechanical crushing method is also possible to make drug that changing for unfavorable physicochemical properties occur
Become.Especially in the plane of crystal of fracture, there may be indefinite forms.However, inventor's research is thought, for respiratory tract part
Medication, mechanical crushing method are advantageous in that：Powder size is larger and shape is irregular, and it is locally heavy in respiratory tract to be conducive to
Product；Preparation process is simple, and the production cycle is shorter, and the chance of microbial contamination is very low；Because the auxiliary materials such as solvent need not be used, only
It crushes pressure and time control is proper, then main ingredient is less likely to occur rotten etc..It is contemplated that being to this powder spray purpose of design
Respiratory tract local application is carried out, does not need very strong mobility, powder size is larger more advantageous, and Alendronate sodium is structure
Simple small-molecule drug, physicochemical properties are more stable, can be with the process of withstanding mechanical comminuting method, therefore, we
Select mechanical crushing method.The operating condition of pulverization conditions Primary Reference inhaled tiotropium bromide powder preparation, be respectively adopted QS100 and
2 kinds of pulverizers such as AO types are tested.Pressure control is crushed in 0.7MPa, the continuous grinding time of single is no more than 3.5 hours.?
Under the pulverization conditions, Alendronate sodium does not go bad.Micro mist grain is carried out using Mastersizer2000 laser granulometries
Degree measures, as a result such as the following table 2.
Sample parameters under 2 different condition of table after mechanical crushing
By upper table 2 as it can be seen that crushing pressure control in 0.7MPa, the continuous grinding time of single is no more than 3.5 hours powder
Under the conditions of broken, size distribution is related to applied sample amount and crushing number.Applied sample amount 1000g crushes 3 times or more (3.0~3.5h),
It can be granularity D90 controls less than 20nm (19.58~19.59 μm).Wherein 4 times crushing reduce seldom with 3 grinding particle sizes,
Show under conditions of 1000g applied sample amounts, 3 times and 3 times or more crushing effect is substantially suitable, therefore the applied sample amount of 1000g, crushes
Number can select 3 times.Applied sample amount 50g crushes 1 time or more (2.5h), can be granularity D90 controls at 12 μm or so
(11.43~12.32 μm).Wherein 2 times crushing reduce seldom with 1 grinding particle size, show under conditions of 1000g applied sample amounts, 1
Secondary and 1 time or more crushing effect is substantially suitable, therefore the applied sample amount of 50g, and crushing number can select 1 time.According to the literature,
The granularity of 7~20nm belongs to the range for being deposited on respiratory tract (trachea and bronchus), therefore, this 2 kinds of condition (1000g applied sample amounts
Crush 3 times or more, 50g applied sample amounts crush 1 time or more) all meet our requirement.We take ALE-4 and ALE-6 respectively
This 2 kinds of samples carry out SEM electron microscope analysis, as depicted in figs. 1 and 2 respectively as the representative sample of 2 kinds of preparation conditions.Wherein
Fig. 1 is the SEM electron microscopes of ALE-4 samples, and Fig. 2 is the SEM electron microscopes of ALE-6 samples.
We to ALE-4 and ALE-6 this 2 kinds of representative samples carried out solubility, ninhydrin method identification, HPLC identifications,
Molybdenum blue colorimetric method measurement and microbial contamination and various physicochemical properties measure.
2.1 dissolubility：ALE-4 the and ALE-6 each 10mg of Alendronate sodium micro mist for weighing trial-production respectively, are put into equipped with 2ml
It in the test tube with ground stopper of water, vibrates 1 minute, observation dissolving situation.2 kinds of powder can be completely dissolved and solution is clarified, and display is molten
Solution property is all qualification.
Property Identification：Each 10mg of ALE-4 and ALE-6 after adding water 2ml to dissolve, then add 0.2% ninhydrin solution 1ml, mix
It closes, heating is boiled 10 minutes, equal displaing amaranth.Absorbance is measured in 568nm, absorbance is very close with bulk pharmaceutical chemicals.
2.2HPLC identification：
The strategy of Alendronate sodium is measured using pre-column derivatization.Alendronate sodium molecular structure is relatively simple, lacks phase
The chromophore answered.Using the amino in alendronic acid sodium molecule fluorophor is formed with FMOC reaction bondeds.
High pressure liquid chromatograph and its model：Japanese Shimadzu Shimadzu LCsolution
Chromatographic condition：
Chromatographic column:Shiseido Capcell Pak C18 (150mm × 4.6mm, 5 μm, Japanese Shiseido company).
Mobile phase:
(1) A liquid：Acetonitrile；B liquid：0.01M disodium hydrogen phosphates, gradient elution program:0~24min, 86%A；24~
29min, 30%A；29~30min, 86%A；Column temperature:35℃；Flow velocity:1.0mL·min-1.
(2) A liquid：Acetonitrile；B liquid：25mM citric acids, 25mM pyrophosphoric acids.Gradient elution program:0~24min, 86%A；24
~29min, 30%A；29~30min, 86%A；Column temperature:35℃；Flow velocity:1.0mL·min-1.
Loading volume：1uL or 2uL.
Fluorescence detector：
Fluorescence exciting wavelength:260nm, fluorescence emission wavelengths:310nm.
With acetonitrile -25mM citric acids, it is as shown in Figure 3 to obtain chromatogram for 25mM pyrophosphoric acid gradient elutions.Wherein 3.5min-
It is Alendronate sodium peak at 4min, is the peaks Fmoc at 13min-14.5min.ALE-4 and ALE-6 is in Alendronate sodium peak, peak
High suitable with bulk pharmaceutical chemicals, there is no main ingredient degradations with ALE-6 by display ALE-4.
2.3 molybdenum blue colorimetric methods measure：
Each 8mg of ALE-4 and ALE-6 are taken, appropriate amount of water is added to dissolve, adds moisture time to move into 100ml measuring bottles, sets in 40 DEG C of water-baths
Heating 20 minutes, and constantly shake, it lets cool to room temperature, is diluted with water to scale, as test solution；Another precision weighs A Lun
Alendronate bulk pharmaceutical chemicals 10mg, sets in 100ml measuring bottles, appropriate amount of water is added to dissolve, and be diluted with water to scale, shake up, as a contrast product
Solution.Precision measures test solution and each 5ml of reference substance solution, sets respectively in 25ml measuring bottles, respectively adds ammonium persulfate solution
(1%) 8ml sets in water-bath and heats 20 minutes, let cool to room temperature, ammonium molybdate solution is added (to take ammonium molybdate 7.5g, add water 100ml molten
Solution, adds 5mol/L sulfuric acid solution 100ml, mixing) 2.0ml, it shakes up, after placing 15 minutes, adds paramethylaminophenol sulfate solution
(take paramethylaminophenol sulfate 0.5g, add 15% solution of sodium bisulfite 195ml, add 20% sodium sulfite solution 5ml, shake up)
2ml shakes up, and after placing 15 minutes, adds 34% sodium acetate solution 5ml, adds water to scale, shake up.Distinguish at the wavelength of 710nm
Absorbance is measured, as a result 2 test solutions are suitable with reference substance solution absorbance, and main ingredient is not present in display ALE-4 and ALE-6
Degradation.
2.4 Micro biological Tests：
Each 3mg of ALE-4 and ALE-6 are taken respectively, are dissolved in respectively in 10ml purified waters.It presses《Pharmacopoeia of People's Republic of China》
Second annex XI J Microbe restriction test method is tested within 2010.Inspection result bacteria colony count≤2/ml；Mould,
Saccharomycete clump count=0/ml.It is qualification.
2.5 heap density, angle of repose, porosity, critical relative moisture：
Heap density, porosity：Heap density refers to the density that volume of a container V is acquired shared by powder quality divided by the powder.
When filling powder, the density measured after certain rule is vibrated or is touched claims tap density (tap density) ρ tap.Porosity
It refer to the voidage ratio of the total volume formed in matrix.After drug carries out micronization processes, heap density, porosity are equal
It has greatly changed, the density contrast of drug and auxiliary material may be caused, cause the difficulty on mixing uniformity.So micronizing
Drug should carry out heap density and porosity measurement.It includes powder true volume, grain that powder, which is fitted into volume measured in container,
Sub- internal pore, inter-particle voids etc., therefore the influences such as shape, size, the packing speed of material and type of feed for measuring container
Powder volume.It is most bulk density that powder filling is not applied density measured by any external force when measuring container, applies external force
And powder is made to be under most tight filling state measured density most tight heap density.Tap density is with oscillation (tapping) number
And change, the final tap density measured when constancy of volume that vibrates is most tight heap density.
A certain amount of powder sample is filled in the 5ml graduated cylinders of restructuring, at the beginning of reading initial volume V, touches and observes graduated cylinder
The volume change of middle powder (will generally tap 2500 times until volume is without significant change！), it is whole to record final volume V.Add
Quality difference value before and after sample is the quality w of powder be added.
At the beginning of heap density pb=w/V, tap density ptap=w/V is whole, porosity=1- heaps density/tap density
Angle of repose：The measurement method at angle of repose is to take bore about 6cm, one, the small funnel of caliber (internal diameter) about 0.4cm,
It is fixed on iron stand, a blank sheet of paper is spread below funnel, the lower end of funnel and the height of paper are 4-5cm, by medicinal powder from funnel
Side is poured slowly into funnel, when the medicinal powder to leak down is close to hopper outlet, measures the height and lower end diameter of the medicinal powder cone, according to
This calculates the tangent of an angle that stops, and further calculates angle of repose.
The measurement of critical relative moisture.Drug is after carrying out micronization processes, and due to the increase of specific surface area, hygroscopicity can
It can obviously change, and the inspection item that moisture, which is powder spray, strictly to be controlled, so facing for micronized medicine should be measured
Boundary's relative humidity (Critical Relative Humidity, CRH).
Critical relative moisture (CRH) assay method is in two steps：
The first, sucting wet curve is made：
Drug 2mg is weighed, is laid in dry to (thickness about 2mm) in the flat measuring cup of constant weight, drying to constant weight, precision title
It is fixed, bottle cap is opened, is set in the glass desicator for filling concentrated sulfuric acid solution or other salting liquids (relative humidity 75%), in 25 DEG C
It is preserved in insulating box, measuring cup is taken out respectively at 2,4,8,12,24 ... or other times section, it is accurately weighed, calculate moisture absorption hundred
Divide rate, is plotted against time with hydroscopicity and obtains sucting wet curve, find T days corresponding time (or hour) when moisture equilibrium at dry side.
The second, the measurement of CRH：
Weigh 6 parts of drug (powder or particle) 1-2mg, ibid operate, set the concentrated sulfuric acid solution for filling various concentration or its
In the glass desicator of his salting liquid, in being preserved T days in 25 degrees Celsius of insulating box, measuring cup is taken out, it is accurately weighed, it calculates
Moisture percentage makees two tangent lines, tangent line focus is corresponding with the moisture equilibrium at dry side curve that hygroscopic capacity maps to relative humidity in inflection point
Relative humidity, as CRH.
By measuring, the various properties difference for the Alendronate sodium micro mist that 2 kinds of process conditions are prepared is little.Cause
This, it is contemplated that it is attained by the demand of design object.
3. Alendronate sodium dust cloud agent capsules
Alendronate sodium powder spray of the present invention can be applied in a capsule.The size of capsule used can be with
It is any model, preferred model is less than No. 0, usually selects No. 1-4.Alendronate sodium capsule can not only be carried out easily
Industrialized production, and it is easy to carry.The amount of Alendronate sodium powder is certain in capsule, it is ensured that inhalation dose when use
Accurately, patient can be effectively prevented to inhale more or inhale less.Micro mist is fitted by the single dose of 0.6mg~5.0mg in capsule, sealing is protected
It deposits.It is taken by single capsule when use.Anti- moisture sorption effect is good.Still it can reach in the environment that relative humidity reaches 90% satisfied
Emptying Rate and atomization.Residual quantity after atomizing effect and use generates good synergistic effect, can especially reduce Ah
The hygroscopicity of logical sequence Alendronate powder spray.
The content uniformity of 3.1 capsules.Method：30 capsules of sampling, balance weigh
The Emptying Rate of 3.2 capsules measures.Method：It is carried out by 2010 editions Chinese Pharmacopoeias (two) annex IL methods.Emptying Rate
It is required that >=90%
Capsule stability is primarily upon the influence that capsule softgel shell absorbs water to preparation performance
Method：-- accelerated test：Capsule 3 batches (30/batches) is taken, being placed in climatic chamber, (40 ± 2 DEG C, 75 ± 5% is opposite
Humidity) in preserve, respectively at 0,1,2,3 the end of month sample, investigate appearance, Emptying Rate, deposition, content
3.3 powder retentions and atomization：By the Alendronate sodium inhalation powder spray precise weighing (W1) of trial-production, it is put into
In special inhalator (production of Shanghai balance pharmaceutical factory), capsule is punched, then inhalator is connected with 5000ml vials, is connected
Place is equipped with switching knob, this knob begins in the closed position.It is vacuumized with 60l/min throughputs, opens above-mentioned knob, in capsule
Medicinal powder i.e. sprayed from inhalator, continuously three times.If powder forms uniform smog, exist without big particle after deposition, explanation
Atomization is excellent；If most of powder is atomized, bottom of bottle only has a small amount of particle, and atomization is medium.If most of powder
It is not atomized, it is deposited into blocky in bottom of bottle, illustrates that atomization is poor.Used capsule shells are taken out from suction apparatus,
Precise weighing (W2), is wiped the residual powder of inside capsule wall only with small brushes, then claims capsulae vacuus weight (W3).Capsule 's content powder
The computational methods of last residual quantity are：[(W2-W3)/(W1-W3)] × 100%
Humidity influences and capsule wall attaches experiment：Alendronate sodium inhalation powder spray is taken, accurately weighed, 25 DEG C of room temperatures are underlying
In 75% environment of relative humidity, is taken out after 24 hours, weigh capsules weight again.Then powder is poured out, observes characters powder
Variation, and measure the residual quantity of powder in capsule by upper method and attach situation to understand capsule wall.
Following table is to screen the experimental conditions of Alendronate sodium dust cloud agent prescription：
The prescription screening of 3 Alendronate sodium inhalation powder spray of table
The nature difference for mixing the capsule formed with ALE-6 by ALE-4, ALE-6 or ALE-4 above is little.
Alendronate sodium micro mist of the content of capsule provided by the present invention by granularity less than 30 μm forms.More preferably
The granularity of micro mist be less than 20 μm, the control of the granularity of most preferred micro mist is in d90≤20 μm, and d50 >=5 μm, the μ of d10≤2.5
m.The dissolubility of micro mist, heap density, angle of repose, Emptying Rate are all preferable, and atomization is medium, during processing, storage, use
It is not easy to moisture absorption.General treatment COPD and the medicinal powder of asthma are drawn into air flue compared with can generate predetermined curative effect at center.Because by this
The atomizing effect for inventing the Alendronate sodium powder spray prepared is good, it is possible to utilize the sucking of commercially available treatment COPD and asthma
Powder unit.Such as the inhalator using Shanghai balance pharmaceutical factory.What the device actually play a part of is to rack capsule.Institute
It states and racks the device of capsule it can be used repeatedly.Ghost can be abandoned after one capsule 's content extinction, next time in use,
A capsule is further filled with into the device.The Alendronate sodium capsule of the present invention can also be struck with the hand open, and micro mist therein can
Airflow function when by breathing enters respiratory tract and plays a role.This be conducive to it is outdoor or in emergency circumstances patient voluntarily to
Medicine.
4.HPLC measure Alendronate sodium powder spray respiratory tract medication after Alendronate sodium lung tissue of rats distribution and
Metabolism
4.1 operating procedure
4.1.1 tissue sample handles experimental procedure：
Precision weighs 0.5g lung tissues, mark-on solution 300uL, grinding.
Add the TCA (pH=1) of 1ml 6%, 4000rpm to centrifuge 10 minutes, pours out supernatant.
Sequentially add 0.1mol/L potassium dihydrogen phosphates 200uL, 0.1mol/L calcium chloride solution 200uL, 1mol/L hydrogen
Sodium hydroxide solution 400uL.
3000rpm 5 minutes removes supernatant.
After 200uL acetums (0.2mol/L) dissolving of gained precipitation, diluted with 0.6mL water, then with 1mol/L hydrogen-oxygens
Change sodium 50uL solution precipitation.
Above-mentioned sodium hydroxide pellets are repeated to react 1 time.
Gained precipitation 0.2mol/L sodium acetate buffers (pH 4.5) 1mL dissolves.
4.1.2Bond Elut-DEA Solid Phase Extraction column purification and derivatization label
Bond Elut-DEA (company Agilent, article No. 12102016), bed volume：300uL
Solid-phase extraction column is activated with 1ml methanol, then washes solid-phase extraction column 2 times with 1ml pure water.
Sample liquid 0.5mL is taken to cross column.Efflux samples 10uL.
It is washed once with 1ml pure water.Efflux samples 10uL.
It is washed once with 1ml pure water.
It is eluted with eluent 1mL sodium pyrophosphates.300ul is taken directly pH to be adjusted to do derivative reaction
Respectively take 300uL, eluent directly to take 300ul, add 1mol/L sodium carbonate buffers (pH 11.9) 100uL and
FMOC working solution 50uL, shake up rapidly, 5min.
Again plus 1mol/L citric acids 100uL adjusts pH value 6.5 or so, (optional：Solution centrifuges, 12 000r/min*
3min), taking supernatant 450uL, sample introduction is analyzed.
HPLC loadings, applied sample amount 4uL.
4.1.3HPLC testing conditions
Chromatographic condition
Chromatographic column:Shiseido Capcell Pak C18 (150mm × 4.6mm, 5 μm, Japanese Shiseido company)；
Mobile phase:A:Acetonitrile B：25mM sodium pyrophosphates, 25mM citric acids (pH 5.05)
Elution program:0~22min, 70%B
Column temperature:35℃；Flow velocity:1.0mL·min-1；
Fluorescence exciting wavelength:260nm, fluorescence emission wavelengths:310nm.
Applied sample amount：4uL
Distribution after 4.2 Alendronate sodium powder spray respiratory tract administrations in induced lung
Take SD rats two, monthly age August.Isoflurane Anesthesia machine anesthesia after, using rat lung dust cloud delivery device give Ah
Logical sequence Alendronate 0.32mg after placing 5min, puts to death rat, takes out lung tissue, isolate left lung:Upper and lower two lobes of the lung；Right lung:
The lobe of the lung of upper, middle and lower three, tissue grinder is carried out after weighing respectively.Lapping liquid alendronic acid sodium content such as table 4 is measured using HPLC
It is shown.
4 lapping liquid alendronic acid sodium content of table
First spray is calculated by equal 0.32mg
The result shows that：After the administration of Alendronate sodium powder spray transtracheal, it can be distributed in each lobe of the lung, especially with close to main branch
The lobe of the lung distribution of tracheae is higher.5min or so, total dose of the lobe of the lung account for the 87.5% of input dose.Show alendronic acid sodium powder
Mist agent has preferable air dispersing characteristic, is easy to the uniform pickup of pulmonary administration in granularity, dispersion degree etc..
HPLC collection of illustrative plates：3.4-3.9min peak be Alendronate sodium appearance.
5. the non-clinical pharmacology pharmacodynamic research (pharmacodynamics zoopery) of Alendronate sodium powder spray (ALN)
Test material
(1) animal：Cleaning grade SD rats 72, wherein female 36,176 ± 22g of average weight；Each 36 with male,
195 ± 25g of average weight.Shanghai Pulmonary Hospital's experimental animal center provides.Animal quality certification number：Shanghai dynamic circuit connector demonstrate,proves word XXX.
(2) drug：1) Alendronate sodium powder spray.Hangzhou Dan Jie medical science and technologies Co., Ltd entrusts Zhejiang celestial being jade pendant pharmacy stock
The trial-production of part Co., Ltd.Capsule 's content is poured out when use, the agent of experiment is pressed with the assay balance that precision is ten a ten thousandths
Amount requires accurately weighed, is carefully fitted into another blank capsules.2) salbutamol sulfate capsule.XXX pharmaceutical factories produce, specification：
XXX。
(3) test method
Sootiness combination elastoser perfusion structure rat COPD model
Using toxicant exposure cabinet, closed air-flow.Rat carries out sootiness modeling by 18/batch.Cigarette brand：Ha Demen.Quilt
Dynamic smoking, holds light 8 cigarettes on cigarette device every time, and the cigarette combustion time is about 5-6 minutes, and closing is kept for 30 minutes.On daily
Noon (four batches) and afternoon (four batches) are each smoked primary.Totally 12 weeks modelling period.In order to further protrude COPD symptoms, using bullet
Property protease Intratracheal instillation, rat anesthesia, tracheal strips elastoser spray (4.8Units/100g weight), rat press 300g
It calculates and sprays into 15units/0.5mL.It is checked after about 7 days or drug treatment.
It is as follows to inspect situation by random samples：
Picking weight close to (348g ± 5%) normal rat 2, male, female it is each one；Rat model 2, it is male
Property, each one of female.The breath state of rat is measured using respirometer.
Rats breathing parametric measurement, as a result see the table below 5.
5 rats breathing parameter of table
The result shows that：
In terms of respiration parameter, there is notable difference with normal mouse in the part basis respiration parameter of model mice, and model is big
The maximum expiratory flow volume PEFb and peak inspiratory flow PIFb of mouse increase, and prompt lung excessively to inflate symptom apparent.Rat model
Respiratory rate is accelerated, and breathing time (Te) and inspiratory duration (Ti) are shortened.The breathing flow velocity (EP50) of 50% capacity into
One step reduces.Show that COPD symptoms are more obvious.Further by pathologic finding, the pathological change degree of lung tissue is observed.
Lung tissue of rats pathological section
After above-mentioned four rats are put to death, lung tissue is taken, carries out paraffin embedding, H&E dyeing.After dyeing as shown in Figure 4.As a result
Show：Rat model lung tissue is shown in a large amount of inflammatory cells, endothelial cell proliferation, cladding, the notable hyperplasia of goblet cell.Alveolar space expands
Greatly, most of rupture of alveoli merges to form pulmonary belb, and alveolar number substantially reduces.With typical COPD features pathological characteristic.
Alendronate sodium powder spray weigh and the HPLC of dust cloud dosage calibration
According to the dosage of rat powder spray pulmonary administration, about in 100-1000ug Alendronate sodiums/rat, in order to
Enough accurate weighing Alendronate sodium powder sprays, use reversed standardization.
Small medicine is made by oneself using hard alloy foil to copy, is weighed for pulmonary administration, and it is 1.3mm that medicine, which copies fixed capacity,3,
Continuous copy takes Alendronate sodium powder spray medicinal powder 10 times, is dissolved in 1mL 0.2M citrate buffer solutions respectively, uses
HPLC methods measure alendronic acid sodium content in solution.The results are shown in Table 6：
Alendronic acid sodium content in 6 solution of table
Conclusion：Alendronate sodium powder spray is often copied greatly containing about Alendronate sodium 1.53mg ± 0.13mg.
In order to carry out the dust cloud dosed administration of trace drug to rat, it is necessary to rat lung dust cloud delivery device into rower
Dosage fixed, that primary calibration first sprays.
It is copied with the small medicine of self-control and takes 1 part of Alendronate sodium powder spray medicinal powder, be loaded into rat lung dust cloud delivery device, used
Usual dusting speed carries out the first spray, sprays into the 50mL centrifuge tubes of the citrate buffer solutions of 0.2M containing 1mL, is adopted after whole mixings
Alendronic acid sodium content is demarcated with HPLC.It does altogether 10 times.The results are shown in Table 7：
Alendronic acid sodium content in 7 solution of table
Conclusion：The dosage of rat lung dust cloud administration the first spray of suit is 0.32 ± 0.12mg.It will be as radix
Carry out rat lung administration.
Therapeutic response (by property of salbutamol sulfate compare) of the rat COPD model to Alendronate sodium
Respiration parameter explanation：
sRaw:Special airway resistance.
Rpef:Exhalation amount before PEF peak values
EF50：It exhales when breathing out 50% tidal volume speed
TV：Tidal volume
Frc:Function residual capacity
PIF：Inspiration peak
PEF：Expiration peak value
Effect of the salbutamol Neulized inhalation to COPD rats (S+E) respiratory function
With COPD rats (S+E：Smoking+elastoser modeling) it is material, distinguish Neulized inhalation 0,0.125mg/ml,
Salbutamol-normal saline solution of 0.25mg/ml, 0.5mg/ml, 1mg/ml, 2mg/ml are monitored using Buxco animal breaths
The respiration parameters such as system detectio airway resistance.Minute is to measure 150-250 respiratory cycle 5 minutes after atomization medication.With
The respiration parameter of physiological saline Neulized inhalation is baseline (own control), and the opposite respiration parameter for calculating each rat changes, and
It is averaged (n=4).The results are shown in Figure 5.
Brief summary：After COPD rats suck salbutamol, special airway resistance sRaw is under the raising gradually of drug concentration
Drop, final fall are~20%.Active drug concentration range：0.25mg/mL-2mg/mL.
Effect of the Alendronate sodium Neulized inhalation to COPD rats (S+E) respiratory function
With COPD rats (S+E：Smoking+elastoser modeling) it is material, distinguish Neulized inhalation 0,0.32mg,
The Alendronate sodium powder spray of 0.625mg, 1.25mg, 2.5mg, 5mg monitor system detectio air flue using Buxco animal breaths
The respiration parameters such as resistance.Minute is to measure 150-250 respiratory cycle 5 minutes after atomization medication.It is atomized with physiological saline
The respiration parameter of sucking is baseline (own control), and the opposite respiration parameter for calculating each rat changes, and is averaged (n=4),
The results are shown in Figure 6.
Brief summary：COPD rats suck Alendronate sodium after, special airway resistance sRaw with drug concentration raising gradually
Decline, final fall is~40%.Active drug concentration range：0.625mg/mL-5mg/mL.Its single medication effect is better than sulphur
Sour salbutamol.
6. therapeutic effect of the Alendronate sodium powder spray to salbutamol drug resistance COPD rats
Alendronate sodium (ALN) is resistant to salbutamol the therapeutic effect of COPD rats
Experimental design：Self-control method is mainly used, in the special airway resistance of forward and backward measurement of salbutamol drug resistance induction
SRAW investigates the COPD rats after salbutamol tolerance to avoid influence of the modeling individual difference to experimental result to greatest extent
Whether there is therapeutic response to Alendronate sodium.It is also equipped with normal rat control group simultaneously, so as to monitoring model quality.
Rat is grouped：4 (the ear tags number of COPD rats for taking SE (smoking+Elastase) method to model：S-1(#53),S-
2 (#52), S-3 (#51), S-4 (#74)) it is used as experimental group, it is female.Another 4 (ear tags number of normal rat for taking the same monthly age
Control-1 (#70), control-2 (#71), control-3 (#72), control-4 (#73), female, as a contrast.
Drug：Salbutamol sulfate aerosol (Wan Tuolin, GlaxoSmithKline PLC, 200 sprays/bottle, 50ug/ sprays), salbutamol
Active compound (nine ancient cooking vessels chemistry), Alendronate sodium powder spray (the outstanding medicine of denier).
Specific experiment flow is：
0th day：Above-mentioned 4 control rats and 8 COPD rats are measured using BUXCO BFL0500 toy respirometers
To the air flue sRaw values of the salbutamol of gradient concentration.Salbutamol concentration gradient：0,0.125mg/ml, 0.25mg/ml,
0.5mg/ml, 1mg/ml, 2mg/ml.
- the 21 day 1st day：Drug resistance induces
4 control rats, 4 COPD rats respectively put a cage, give that albuterol aerosol is primary, and Saturday, day removes daily
Outside.Administering mode：Cage is covered with large size polybag, 1 spray (50ug) is given on alignment cage top, and rat is allowed freely to suck,
Sack is rolled over, 1 spray is given in holding again after five minutes, then is kept for 5 minutes.
22nd day：Each rat airway sRaw values under salbutamol Neulized inhalation are measured, compared with the 0th day result, investigate warp
Whether rat generates tolerance to salbutamol after crossing 21 days application salbutamols.
23rd day：Each rat airway sRaw values under Alendronate sodium Neulized inhalation are measured, salbutamol tolerance is investigated
Whether COPD rats have drug response to Alendronate sodium.Alendronate sodium concentration gradient：0,0.31mg/ml, 0.625mg/ml,
1.25mg/ml, 2.5mg/ml, 5mg/ml.
Experimental result
(1) before drug resistance normal rat and COPD rats (the 0th day) to the reactivity of salbutamol
Four normal rats are in the 0th day reactivity to salbutamol as shown in fig. 7, four COPD rats are right at the 0th day
The reactivity of salbutamol is as shown in Figure 8.Four normal groups and four COPD groups rats were in the 0th day reactivity to salbutamol
(mean value) is as shown in Figure 9.
Conclusion：Normal rats also have salbutamol slight response, and average sRaw values decline 5-8%.COPD rats
There is apparent response to salbutamol, average sRaw values decline 16-19%.Show that the COPD rats that this experiment uses are suitable for
The assessment of this curative effect of medication.
(2) salbutamol is after tolerance-induced 21 days, and normal group and COPD groups rat are to the reactivity of salbutamol.
After tolerance-induced 21 days of salbutamol, 4 normal rats are as shown in Figure 10 to the reactivity of salbutamol, Sha Ding
After tolerance-induced 21 days of amine alcohol, 4 COPD groups rats to salbutamol reactivity as indicated at 11, salbutamol tolerance-induced 21
After it, 4 normal groups and 4 COPD groups rats compare as shown in 12 the reactive mean value of salbutamol.Salbutamol is resistance to
Forward and backward by inducing, 4 COPD groups rats are more as shown in figure 13 to the reactive mean value of salbutamol.
Brief summary:Salbutamol by 21 days is tolerance-induced, and COPD rats are resistant to salbutamol, does not induce preceding sand
The sucking of butylamine alcohol causes average sRaw values to decline 16-19%, and salbutamol, which sucks, after induction only results in average sRaw values decline 3-
5%, COPD rats generate drug resistance to salbutamol after illustrating induction in 21 days.
(2) salbutamol is after tolerance-induced 21 days, and normal group and COPD groups rat are to the reactivity of Alendronate sodium.
After tolerance-induced 21 days of salbutamol, 4 normal rats are as shown in figure 14 to the reactivity of Alendronate sodium, husky
After tolerance-induced 21 days of butylamine alcohol, 4 COPD groups rats are as shown in figure 15 to the reactivity of Alendronate sodium, salbutamol tolerance
After 21 days of induction, 4 COPD groups rats are as shown in figure 16 to the reactive mean value of Alendronate sodium, and salbutamol is tolerance-induced
Forward and backward, 4 normal rats and 4 COPD groups rats are more as shown in figure 17 to the reactive mean value of Alendronate sodium.
Brief summary:COPD rats are resistant to salbutamol after 21 days salbutamols are tolerance-induced, but to Ah
Logical sequence Alendronate still has apparent reactivity.Salbutamol is resistant to COPD rats, can still result in using effect of oral administration alendronate sodium for treatment flat
Equal sRaw values decline 28-30%, illustrate that Alendronate sodium has different pharmacological mechanisms from salbutamol, can be used for salbutamol
It is resistant to the treatment of COPD rats.
Embodiment
Embodiment 1
Alendronate sodium bulk pharmaceutical chemicals 1000g is taken, is loaded in QS100 type pulverizers.Pulverization conditions are that 0.7MPa crushes pressure
Power 3 hours, crushes 4 times, obtains Alendronate sodium micro mist.
Measure alendronic acid sodium content, with 6mg/ capsule subpackages in hard capsule to obtain the final product.Grain size analysis with laser grain size analyzer result：Britain
MalvernInstrumentsLtd. laser granulometry has carried out granulometry to the capsule 's content powder of different lot numbers,
As a result above-mentioned sample granularity is less than 20 μm of part percentages more than 90%, and the percentage shared by part of the granularity less than 5 μm
Than being less than 50%.Capsule 's content powder is observed with Electronic Speculum, this visible powder under 10000 times of SEM electron microscopes
End is single tiny particle, and no adhesion, no agglomerate is well dispersed, is a kind of irregular stereochemical structure.
In order to investigate whether preparation process causes the destruction of Alendronate sodium to be degraded, with molybdenum blue colorimetric method to A Lun before crushing
Alendronate bulk pharmaceutical chemicals are determined with the powder after crushing.Method is to take bulk pharmaceutical chemicals or each 8mg of powder, and Alendronate sodium is made
Concentration is equivalent to the solution of 0.08mg/ml, and the absorbance measurement of respective wavelength is carried out by analysis method.As a result, the powder after crushing
Last absorbance (0.071 ± 0.007) is suitable with the bulk pharmaceutical chemicals (0.067 ± 0.006) before crushing, it is seen that the content of Alendronate sodium
Change without apparent.
Embodiment 2
Alendronate sodium bulk pharmaceutical chemicals 50g is taken, is loaded in AO type pulverizers.Pulverization conditions are that 0.7MPa crushes pressure,
It 2.5 hours, crushes 2 times.Obtain Alendronate sodium micro mist.Alendronic acid sodium content is measured, with 2mg/ capsule subpackages in hard capsule
In to obtain the final product.Grain size analysis with laser grain size analyzer result：Britain's MalvernInstrumentsLtd. laser granulometries are to different lot numbers
Capsule 's content powder has carried out granulometry, and as a result above-mentioned sample granularity is less than 20 μm of part percentages and is more than 90%,
And the percentage shared by part of the granularity less than 5 μm is less than 50%.Capsule 's content powder is observed with Electronic Speculum,
This visible powder is single tiny particle under 10000 times of SEM electron microscopes, and no adhesion, no agglomerate is well dispersed,
For a kind of irregular stereochemical structure.
In order to investigate whether preparation process causes the destruction of Alendronate sodium to be degraded, with molybdenum blue colorimetric method to A Lun before crushing
Alendronate bulk pharmaceutical chemicals are determined with the powder after crushing.Method is to take above-mentioned raw materials medicine or each 8mg of powder, and A Lun phosphines are made
Sour na concn is equivalent to the solution of 0.8mg/ml, and the absorbance measurement of respective wavelength is carried out by analysis method.As a result, after crushing
Powder absorbance (0.069 ± 0.008) is suitable with the bulk pharmaceutical chemicals (0.067 ± 0.006) before crushing, it is seen that Alendronate sodium contains
Amount changes without apparent.
Embodiment 3
By Alendronate sodium Alevaire obtained in embodiment 1, Pcnten-1 yne-4 is measured using atomization absorption device, 12.7%
It is deposited in level-one distribution bottle；78.2% is deposited in two level distribution bottle.The Alendronate sodium powder spray of the present embodiment is applied
Afterwards, it takes rat lung to carry out HPLC analyses respectively, as a result shows into lung amount 83.40%~92.2%,
Embodiment 4
Rat COPD model treatment results show that Alendronate sodium powder spray inhalant in 0.6~5.0mg of dosage, is applied daily
With 1 time, COPD rats breathing function~40% can be improved, be better than the effect (~20%) of salbutamol sulfate.The present invention's
Alendronate sodium powder spray continuous administration 3 weeks remains able to improve COPD rats breathing function~30%, shows that there is no resistance to
Pharmacological property problem.
Embodiment 5
It is significant in efficacy to decline the big of (respiratory function only improves 3%~5%) for salbutamol sulfate continuous administration 3 weeks
Mouse, Alendronate sodium powder spray of the invention are applied 1 time in 0.6~5.0mg of dosage, remain able to further improve it daily
Respiratory function 28%~30% shows the drug resistance problems that can overcome salbutamol sulfate.
Although the present invention is disclosed above by preferred embodiment, however, it is not intended to limit the invention, this any known skill
Skill person can make some changes and embellishment without departing from the spirit and scope of the present invention, therefore protection scope of the present invention is worked as
Subject to claims range claimed.
Claims (7)
1. the Alendronate sodium powder spray for respiratory tract administration, which is characterized in that the composition of the Alendronate sodium powder spray
For single Alendronate sodium micro mist, auxiliary material is not contained；
The powder size of the Alendronate sodium micro mist is distributed as d90≤20 μm, and d50 >=5 μm, d10≤2.5 μm.
2. Alendronate sodium powder spray according to claim 1, which is characterized in that the Alendronate sodium powder spray by Ah
Logical sequence Alendronate bulk pharmaceutical chemicals are directly crushed and are prepared.
3. Alendronate sodium powder spray according to claim 1, which is characterized in that by Alendronate sodium micro mist with 0.6mg
The explosive payload of~5mg is packed into capsule, and daily medication is primary.
4. the preparation method of the Alendronate sodium powder spray as described in claim 1-3 any one, which is characterized in that by A Lun
Alendronate bulk pharmaceutical chemicals are directly crushed and are prepared, and do not add auxiliary material or solvent.
5. the preparation method of Alendronate sodium powder spray according to claim 4, which is characterized in that crushing applied sample amount is
50g~1000g, crushing pressure are 0.7Mpa, and grinding time is 2.5h~3.5h, and it is 1 time or more to crush number.
6. Alendronate sodium powder spray as described in claim 1-3 any one prepare treatment Chronic Obstructive Pulmonary Disease and
Purposes in asthmatic medicament.
7. the Alendronate sodium powder spray as described in claim 1-3 any one is preparing treatment to β 2 receptor agonist generation
Purposes in the Chronic Obstructive Pulmonary Disease drug of drug resistance.
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| CN100345549C (en) * | 2001-05-02 | 2007-10-31 | 诺瓦提斯公司 | Method of administration of bisphosphonates by inhalation in the treatment or prevention of bone resorption and osteoporosis |
| CN101522032A (en) * | 2006-11-21 | 2009-09-02 | 帝国制药美国公司 | Biphosphonate inhalant formulations and methods for using the same |
| CN103655524A (en) * | 2013-12-23 | 2014-03-26 | 南京中医药大学 | Nose powder inhalation for treating epilepsy and preparation method thereof |
-
2016
- 2016-01-18 CN CN201610032682.3A patent/CN105640924B/en active Active
- 2016-04-01 WO PCT/CN2016/078257 patent/WO2017124640A1/en active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100345549C (en) * | 2001-05-02 | 2007-10-31 | 诺瓦提斯公司 | Method of administration of bisphosphonates by inhalation in the treatment or prevention of bone resorption and osteoporosis |
| CN101522032A (en) * | 2006-11-21 | 2009-09-02 | 帝国制药美国公司 | Biphosphonate inhalant formulations and methods for using the same |
| CN103655524A (en) * | 2013-12-23 | 2014-03-26 | 南京中医药大学 | Nose powder inhalation for treating epilepsy and preparation method thereof |
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| Title |
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| Formulation and in vivo evaluation of sodium alendronate spray-dried microparticles intended for lung delivery;Leticia Cruz，et al.;《Journal of controlled release》;20110308;第152卷;370-375 * |
| Inhalation of alendronate nanoparticles as dry powder inhaler for the treatment of osteoporosis;Shaheen Sultana,et al.;《Journal of microencapsulation》;20121231;第29卷(第5期);445-454 * |
| Pulmonary delivery of nanosized alendronate for decorporation of inhaled heavy metals:formulation development,characterization and gamma scintigraphic evaluation;Shaheen Sultana，et al.;《Pharmaceutical development and technology》;20130724;第19卷(第5期);623-633 * |
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| Publication number | Publication date |
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| CN105640924A (en) | 2016-06-08 |
| WO2017124640A1 (en) | 2017-07-27 |
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