A kind of ambroxol hydrochloride oral solution and preparation method thereof and new application
Technical field:
The present invention relates to chemicals technical field, more particularly to a kind of ambroxol hydrochloride medicine and its new pharmaceutical use.
Technical background:
Ambroxol hydrochloride, chemical name are:Trans -4- [(2- amino -3,5- dibromo-benzyl) amino] cyclohexanol HCI,
Molecular formula is C13H18Br2N2OHCL, structural formula are as follows:
Ambroxol hydrochloride is white to yellowish crystalline powder;It is almost odorless.It can promote sticky secretion inside respiratory tract
The exclusion of thing and the delay for reducing mucus, thus remarkably promote expectoration.Suitable for secreting abnormal and expectoration function with sputum
Undesirable acute, chronic respiratory disease.
Ambroxol hydrochloride is expectorant, has good glutinous phlegm dissolution and lubrication respiratory tract effect, can promote lung surface
The secretion of active material, the secretion for breathing liquid and ciliary movement etc..It is abnormal etc. for acute and chronic breathing problem, bronchial secretion
Treatment.Ambroxol hydrochloride can also be used to treat in Bronchopneumonia, chronic obstructive pulmonary disease, bronchiolitis, secretory
The diseases such as otitis, chronic bronchitis.
Inventor chances in clinic diagnosis practice, to Diabetic Nephropathy patients, especially diabetic nephropathy kidney
Loose patient gives ambroxol hydrochloride therapy, achieves highly significant curative effect.
After specification, science, rigorous clinical test and pharmacodynamic experiment is carried out, technical scheme has been obtained.
The content of the invention:
The object of the present invention is to provide a kind of ambroxol hydrochloride medicine and preparation method thereof.
It is a further object of the present invention to provide ambroxol hydrochloride medicine to prepare treatment diabetes, diabetes mellitus encephalopathy, glycosuria
Application in sick nephrosis, myocarditis, vital myocarditis medicine.
The purpose of the present invention is what is be accomplished by the following way:
A kind of ambroxol hydrochloride medicine, the medicine are ambroxol hydrochloride oral solution, and its preparation method is:Take hydrochloric acid ammonia bromine
Rope 3g, adds Sucralose 0.4g, Steviosin 0.7g, citric acid 0.22g, sodium citrate 0.16g, menthol 0.1g, ethanol
7.5mL, flavoring pineapple essence 0.1mL, add purified water to add to 1000mL, mix, filter, is filling, sterilizing, lamp inspection obtains ambroxol hydrochloride mouth
Take solution.
A kind of ambroxol hydrochloride medicine is preparing the application in treating diabetes medicament, which is hydrochloric acid
Ambroxol oral liquid, its preparation method are:Ambroxol hydrochloride 3g is taken, adds Sucralose 0.4g, Steviosin 0.7g, citric acid
0.22g, sodium citrate 0.16g, menthol 0.1g, ethanol 7.5mL, flavoring pineapple essence 0.1mL, adds purified water to add to 1000mL, mixes
Even, filtering, filling, sterilizing, lamp inspection obtain ambroxol hydrochloride oral solution.
A kind of application of ambroxol hydrochloride medicine in treatment diabetes mellitus encephalopathy medicine is prepared, the ambroxol hydrochloride medicine are
Ambroxol hydrochloride oral solution, its preparation method are:Ambroxol hydrochloride 3g is taken, adds Sucralose 0.4g, Steviosin 0.7g, Chinese holly
Rafter acid 0.22g, sodium citrate 0.16g, menthol 0.1g, ethanol 7.5mL, flavoring pineapple essence 0.1mL, adds purified water to add to
1000mL, mixes, filters, is filling, sterilizing, lamp inspection obtains ambroxol hydrochloride oral solution.
A kind of application of ambroxol hydrochloride medicine in treatment medicine for treating diabetic nephropathy is prepared, the ambroxol hydrochloride medicine are
Ambroxol hydrochloride oral solution, its preparation method are:Ambroxol hydrochloride 3g is taken, adds Sucralose 0.4g, Steviosin 0.7g, Chinese holly
Rafter acid 0.22g, sodium citrate 0.16g, menthol 0.1g, ethanol 7.5mL, flavoring pineapple essence 0.1mL, adds purified water to add to
1000mL, mixes, filters, is filling, sterilizing, lamp inspection obtains ambroxol hydrochloride oral solution.
The ambroxol hydrochloride is preparing the application in treating diabetic nephropathy renal hypertrophy medicine.
A kind of ambroxol hydrochloride medicine is preparing the application in treating myocarditis medicine, which is hydrochloric acid
Ambroxol oral liquid, its preparation method are:Ambroxol hydrochloride 3g is taken, adds Sucralose 0.4g, Steviosin 0.7g, citric acid
0.22g, sodium citrate 0.16g, menthol 0.1g, ethanol 7.5mL, flavoring pineapple essence 0.1mL, adds purified water to add to 1000mL, mixes
Even, filtering, filling, sterilizing, lamp inspection obtain ambroxol hydrochloride oral solution.
The ambroxol hydrochloride medicine is preparing the application in treating vital myocarditis medicine.
Brief description of the drawings:
Fig. 1 cuts into slices for normal rat Pathological
Fig. 2 is 12 weeks pathological sections of Diabetic nephropathy animal model group
Fig. 3 is 12 weeks pathological sections of diabetic nephropathy rats treatment group
Embodiment:
Embodiment 1:The pharmacodynamic experiment research of ambroxol hydrochloride therapy diabetic nephropathy rats
1 materials and methods
1.1 reagents and instrument
Streptozotocin (STZ) is produced by Beijing Suo Laibao Science and Technology Ltd;Urine detection tetrad reagent strip, by Anhui
Dark blue medical science and technology limited company production;Coomassie brilliant blue reagent (CBB), is given birth to by Beijing Suo Laibao Science and Technology Ltd
Production;Metal metabolism cage, is produced by Suzhou Feng Shi experimental animal equipment Co., Ltd;755 type ultraviolet-uisible spectrophotometers, by upper
Hai Aoxi scientific instrument Co., Ltd produces;Blood glucose meter, is produced by Beijing is happy into biological electronic technology limited company.Hydrochloric acid
Ambroxol oral liquid:Ambroxol hydrochloride 3g is taken, adds Sucralose 0.4g, Steviosin 0.7g, citric acid 0.22g, citric acid
Sodium 0.16g, menthol 0.1g, ethanol 7.5mL, flavoring pineapple essence 0.1mL, add purified water to add to 1000mL, mix, filter, is filling,
Sterilizing, lamp inspection obtain ambroxol hydrochloride oral solution, are produced by Heilungkiang Zhong Gui pharmaceutical Co. Ltds, specification 5mL:15mg.
1.2 experimental animal
Bull Wislar rats 30, SPF grades, 200 ± 20g of weight, purchased from Heilongjiang University of Chinese Medicine animal
The heart provides.Animal quality certification number:The dynamic word the 1024-03rd of doctor.Experimental animal be raised precedently in advance 24 it is small when, adapt to laboratory raise
The condition of supporting.All animals watch material with high pressure and feed, and arbitrarily drinking-water, ingests, and room temperature control is at 25 DEG C or so, natural lighting.
1.3 animal models make and packet
30 rats are randomly divided into 3 groups, normal group, model treatment group and model non-treatment group, every group 10.Model group
Fasting 12h (can't help water) before rat experiment.Streptozotocin is dissolved in 4.2 citrate buffers of pH, by 50mg/Kg once
Property intraperitoneal injection, monitor glucose in urine daily, the blood sugar concentration of animal is more than 16.7mmoL/L after 3d, and glucose in urine is positive, is determined as sugar
Urine disease model is formed.Ambroxol hydrochloride oral solution 0.5mg/Kg gives in treatment group, and gavage, 2 times a day, is treated 4 weeks altogether.Glycosuria
Disease substantially increases into Urine proteins after mould 3 weeks, and diabetic nephropathy model is set up.During the experiment rat show as more drinks, more foods,
Diuresis, the symptom such as become thin.Normal group gives isometric citrate buffer.
1.4 collections of specimens and detection
Rat is put into clean metal metabolism cage, each group rat 24h Urine specimens is collected in pretherapy and post-treatment, during staying urine
Fasting, can't help water, take 4mL after recording urine volume, 2000r/min centrifugation 10min, remove sediment and preserved in -20 DEG C of refrigerators and treated
Survey.Urine proteins are detected with Coomassie Brilliant Blue.After experiment, rat is anesthetized with ether, eyeball is plucked and takes blood, for blood glucose, blood
The detection of creatinine, urea nitrogen.Blood glucose blood glucose meter and blood sugar test paper detection, renal function are detected using Beckman automatic biochemical analyzers.
1.5 Renal Paphologies detect
After anesthetized rat takes blood, kidney is taken out immediately, is removed coating, is weighed, 10% formalin is fixed, and alcohol takes off step by step
Water, dimethylbenzene is transparent, paraffin embedding.Slice thickDo conventional H E dyeing to dye with PAS, om observation.
1.6 statistical method
Using SPSS10.0 software data processings, all data are with average value standard deviationRepresent, comparison among groups is examined with t
Test.
2 results
2.1 changes of each group rat urine protein content afterwards before the treatment
After diabetes model is set up 4 weeks, the increase of rat Urine proteins, after ambroxol hydrochloride oral solution is treated, Urine proteins
Reduce, compared with pre-treatment compared with difference significance (P<0.05).And non-treatment group's Urine proteins have the trend increased, but it is poor
Different nonsignificance (P>0.05), it is shown in Table 1.
The change of the 1 pretherapy and post-treatment Urine proteins of each group rat of table
Note:Compared with before treatment, * P<0.05;Compared with before treatment, #P>0.05.
The change of each group rat blood serum, blood biochemistry index and kidney weight/weight after 2.2 treatments
Model group blood glucose is apparently higher than normal group of (P<0.01), but serum creatinine, urea nitrogen difference are without clear meaning (P>
0.05) after, ambroxol hydrochloride oral solution treatment is given by treatment group, kidney weight/body mass index is significantly lower than non-treatment group (P<
0.05), with normal group difference without clear meaning (P>0.05), illustrate that ambroxol hydrochloride oral solution can mitigate diabetic nephropathy
The renal hypertrophy of rat, is shown in Table 2.
Blood glucose, blood urea nitrogen, creatinine and kidney weight/changes of weight after table 2 is treated
Compared with normal group, * P<0.05, * * P<0.01.
Influence of 2.3 ambroxol hydrochloride oral solutions to diabetic nephropathy tectology
HE dyeing can see normal rats extracellular matrix under light microscopic and mesangial cell distribution is normal, capillary lumen
It is open, see that attached drawing 1, rather than the extracellular matrix for the treatment of group increase, system cells hyperplasia, while capillary vessel member collapses, and sees
Attached drawing 2.Treatment group's extracellular matrix slightly increases, and proliferation of mesangial cells also mitigates.Glomerular basement membrane thickening mitigates, and sees attached drawing
3, illustrate that ambroxol hydrochloride oral solution can mitigate glomerulus hypertrophy to a certain extent, so as to have to diabetic nephropathy necessarily
Therapeutic effect.
3 conclusions
This experiment shows that ambroxol hydrochloride oral solution can reduce the urinary protein excretion of diabetic nephropathy rats, reduces
Kidney weight/body mass index, pathological data confirm that model treatment group extracellular matrix build-up is reduced, and mesentery expansion and mesangial cell increase
It is raw to mitigate, illustrate that ambroxol hydrochloride oral solution has certain effect to treatment diabetic nephropathy renal hypertrophy.
Embodiment 2:Ambroxol hydrochloride oral solution studies the pharmacodynamic experiment of diabetes mellitus encephalopathy rat
Diabetes are one group of lifelong participation metabolic diseases as caused by multi-pathogenesis characterized by chronic hyperglycemia.Long-term blood
Sugar increases, and big blood vessel, capilary are damaged and jeopardize the heart, brain, kidney, peripheral nerve, eyes, foot etc., according to the statistics of the World Health Organization,
Diabetic complication is up to kind more than 100, is to be currently known a kind of most disease of complication, common are diabetic nephropathy, sugar
Urinate characteristic of disease retinopathy and the relevant uveitis of diabetes, diabetic cataract, diabetes, diabetes mellitus encephalopathy, sugar
The sick cardiovascular complication of urine, diabetic cerebrovascular disease, diabetic neuropathy etc..Diabetes Death person has more than half to be
Caused by cardiovascular and cerebrovascular, caused by 10% is nephropathy.Because the patient of diabetes amputation is 10~20 times of non-diabetic.Clinical data
It has been shown that, 10 years or so after onset diabetes, a kind of complication, and complication one can at least be occurred into for the patient that have 30%~40%
Denier produces, and drug therapy is difficult to reverse, therefore emphasizes to prevent diabetic complication as early as possible.Ambroxol hydrochloride oral solution can be controlled
Treat diabetic nephropathy, then predict that it also has diabetes mellitus encephalopathy certain therapeutic effect.Inventor is to ambroxol hydrochloride mouth
Take Solution In The Treatment diabetes mellitus encephalopathy and carry out pharmacodynamic experiment research, probe into its therapeutic effect.
1 material and method
1.1 experimental animals and reagent
8 week old SD rats of healthy male, 200~250g of weight, provides purchased from Heilongjiang University of Chinese Medicine's animal center.
Animal quality certification number:The dynamic word the 1024-03rd of doctor.Watermaze protocol auto-controller, by the limited public affairs of Hefei four seas instrument science and technology
Department's production;Streptozotocin (STZ), is produced by Beijing Suo Laibao Science and Technology Ltd;Ambroxol hydrochloride oral solution:Take hydrochloric acid
Ambroxol 3g, adds Sucralose 0.4g, Steviosin 0.7g, citric acid 0.22g, sodium citrate 0.16g, menthol 0.1g, second
Alcohol 7.5mL, flavoring pineapple essence 0.1mL, add purified water to add to 1000mL, mix, filter, is filling, sterilizing, lamp inspection obtains ambroxol hydrochloride
Oral administration solution, is produced, specification 5mL by Heilungkiang Zhong Gui pharmaceutical Co. Ltds:15mg.
The foundation and packet of 1.2 diabetes rat models
Rat adaptability is raised 1 week, fasting 12h, after diluting STZ as 4.5 0.1moL/L sodium citrate buffers using pH,
Lucifuge simultaneously quickly establishes diabetes rat model with the injection of 60mg/Kg amounts disposable celiac.After 3d, modeling Rat Septal curfew eats not
Prohibit drink more than 8h, take rat tail vein blood examination to survey fasting blood sugar, by fasting blood-glucose>It is big that the rat of 18mmoL/L is set to diabetes
Mouse, rejects the below standard person of blood glucose.Model successfully after a week, diabetes rat is randomly divided into diabetes mellitus encephalopathy group and hydrochloric acid ammonia bromine
Rope oral administration solution treatment group, every group 10;Normal rat is randomly divided into Normal group and ambroxol hydrochloride oral solution control
Group, every group 10.Ambroxol hydrochloride oral solution treatment group, ambroxol hydrochloride oral solution control group give ambroxol hydrochloride mouth
Solution 10mg/ (Kgd) gavage is taken, Normal group gives Isodose physiological saline gavage, continues 14 weeks.In experimentation
In, each group rat weight and blood glucose value are detected weekly.
1.3 Morris water maze laboratories
At the 14th week, using the observation Cognition Function in Rats change of Morris water mazes.Morris water mazes are a diameter
The round pool of 200cm, is built-in with the circular platform (diameter 15cm) of a moveable position, and platform is located at underwater 2cm, and water temperature is permanent
It is scheduled on (25 ± 2) DEG C.Water maze laboratory has 6d altogether, and the 1st day is the water maze environment laundering period.2nd~5 day is the Experiment Training phase,
Record is a little 120s to the time used in platform i.e. escape latency, each testing time from being put into, will be big if not finding platform
Mouse leads to platform and it is stopped 10~15s to remember gate position.Tested for space search within 6th day, withdraw platform, surveyed every time
The examination time is 60s, records every rat in the platform traversing times of platform position and hovering the time in purpose quadrant.
1.4 heart perfusions and collection of specimens
After water maze laboratory, 1 rat is randomly selected from every group, with 10% hydration chlorine wake up 3mL/Kg be injected intraperitoneally into
Row anesthesia, fixes and cuts off thoracic cavity exposure heart, after separating pericardium, catheter needle insertion left ventricle and fixation, while cut off the right side
Atrium.First got express developed with physiological saline, then slowly irrigated with 4% paraformaldehyde.After fully being fixed in rat body, cut
Skull is opened, whole brain is completely moved in 4% paraformaldehyde and soaks 3d waits embedding for H-E dyeing.Remaining rat is broken
Head takes brain, is placed in quick separating on ice and goes out hippocampus, is put into cryopreservation tube and is stored in -120C and is used for western blot analysis.
1.5 H-E are dyed
Row serial section after the tissue of paraffin embedding is fixed with Coronal, by the section containing hippocampal tissue paste through
It is placed in baking 30~60min of piece in 60~65 DEG C of ovens on 3- aminopropyl -3- methoxy silanes (APES) processed glass slide.
Section dewaxing is to water, and hematoxylin solution dip dyeing, rinses unnecessary dye liquor, be then dehydrated with graded ethanol, last mounting.Optical microphotograph
Microscopic observation Rat hippocampus CA1 area's neuron morphologies and its pathological change.
1.6 statistical procedures
All numerical value using SPSS10.0 softwares carry out statistical analysis, data with (Form represents, with multifactor and
One-way analysis of variance compares group difference.Inspection level (a) is 0.05.
2 results
The ordinary circumstance of 2.1 each group rats
There is obvious more drink, diuresis, polyphagia shapes in diabetes rat, and rat hair color sallow is apathetic, and rat-tail occurs
It is different degrees of to fester and the phenomenon that docks.When testing 2 months, diabetes rat is because diarrhea and subcutaneous abscess occurs in infection, warp
After antibiotic intraperitoneal injection treatment, infection symptoms are controlled.Diabetes rat is slow in action, and autonomous actions are reduced, and occurs easy
Excitation or to the behavior expression such as extraneous stimulate the reaction is indifferent.During the experiment, blood glucose in diabetic rats value are maintained at (27 ± 6)
MmoL/L, while normal rat blood glucose value is maintained at (6 ± 2) mmoL/L.
2.2 water maze laboratory results
After rat blood sugar value persistently raises 3 months, during water maze laboratory is trained, diabetes mellitus encephalopathy group and hydrochloric acid ammonia
Xiu Suo oral administration solutions treatment group escape latency substantially increases (P compared with Normal group<0.01), experimental result is shown in Table 3.
The formal during the experiment of water maze, diabetes mellitus encephalopathy group and ambroxol hydrochloride oral solution treatment group platform traversing times
With the time of hovering in purpose quadrant (P is significantly reduced compared with Normal group<0.01), experimental result is shown in Table 4.
3 each group rat water maze of table tests the comparison of escape latency
Note:Compared with normal control, * * P<0.01;
4 each group rat water maze space search experiment porch traversing times of table and purpose quadrant are hovered time n=6,
Note:Compared with normal control, * * P<0.01;
3 conclusions
Originally experimental studies have found that, ambroxol hydrochloride oral solution successive administration observes that ambroxol hydrochloride oral is molten after 14 weeks
Liquid treatment group Cognition Function in Rats obstacle without be improved significantly, show ambroxol hydrochloride oral solution to diabetes rat all
Brain does not have significant protective effect, it is impossible to effectively treats diabetes mellitus encephalopathy.