CN102078593B - Application of toad peptide antibiotic for preparing medicines for curing diabetic foot - Google Patents
Application of toad peptide antibiotic for preparing medicines for curing diabetic foot Download PDFInfo
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- CN102078593B CN102078593B CN2011100072769A CN201110007276A CN102078593B CN 102078593 B CN102078593 B CN 102078593B CN 2011100072769 A CN2011100072769 A CN 2011100072769A CN 201110007276 A CN201110007276 A CN 201110007276A CN 102078593 B CN102078593 B CN 102078593B
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- peptide antibiotic
- toad peptide
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Abstract
The invention provides application of toad peptide antibiotic for preparing medicines for curing diabetic foot. With toad maggots as raw materials, the toad peptide antibiotic is extracted, separated and purified by adopting a solvent extraction method, an ultrasonic extraction method, an ion exchange resin absorption method, an ultrasonic standing wave liquid phase preparation spectrum method and the technical processes of vacuum freeze drying and the like. Various dosages of medicines are prepared by taking the toad peptide antibiotic as an active ingredient for curing diabetic foot and combining the toad peptide antibiotic with proper excipients. The ingredients for preparing the medicines are specific, the quality is stable and controllable, and the therapeutic effect on curing various diabetic foot is good. The invention has reasonable production technical process, consumes low energy and is suitable for industrial production.
Description
Technical field
The present invention relates to a kind of topical drug, the particularly application of toad peptide antibiotic in preparation treatment diabetic foot medicine.
Background technology
Diabetic foot is one of complication of diabetes; Clinically; Because diabetics receives the influence of hyperglycemia for a long time; Lower limb vascular be prone to form thrombosis and the inaccessible lower limb lesion tissue that also causes, and perhaps diabetics infects owing to reasons such as pedal skin abrade, chaps, beriberi cause, and causes foot or lower limbs edema, blackout, rots, necrosis and gangrenous.Now to the treatment of patient with diabetic feet injection of antibiotics commonly used, amputation, bridging and stem cell transplantation technology.
CN200510124025.3 discloses and has controlled vasculitis and diabetic foot Chinese medicine, is made up of Chinese herbal medicine such as Radix Et Rhizoma Rhei, Flos Lonicerae, Bufo siccus, Ramulus et folium taxi cuspidatae, Radix Scutellariae, Cortex Phellodendri, Radix Angelicae Pubescentis, the Rhizoma Pinelliae, Rhizoma Arisaematis, Radix Notoginseng, the Cortexs Eucommiae; CN200510088707.3 discloses the patented technology that lotion and method for preparing and application are put in a kind of health care that is used for diabetic foot on the skin, is made up of batchings such as Hirudo, Flos Carthami, Olibanum, Myrrha, Rhizoma Curcumae Longae, Rhizoma Curcumae, Rhizoma Polygoni Cuspidati, Ramulus Cinnamomi, ethyl hydroxybenzoates; CN200410092408.2 discloses a kind of patented technology of controlling diabetic foot Chinese medicine, is made up of Chinese herbal medicine such as the Radix Astragali, Radix Scrophulariae, Radix Salviae Miltiorrhizae, Radix Achyranthis Bidentatae and Caulis Loniceraes.
CN200410013742.4 discloses and has adopted Chinese medicine to cultivate the method for Bufo siccus maggot, can mass industrialized production Bufo siccus maggot.
In principle; Above-mentioned various medicines and treatment technology can be applied to treat the disease of diabetic foot; But each formulation of drug is various; Every kind of Chinese herbal medicine is because the difference of plantation region, weather, collecting time, the process of preparing Chinese medicine and processing method when suitability for industrialized production, is difficult to guarantee consistency of product and quality control.
Summary of the invention
The purpose of this invention is to provide the application of toad peptide antibiotic in preparation treatment diabetic foot medicine, its medicine prescription and technology design to the diabetic foot characteristic symptom specially, and composition is accurate, quality controllable, easy to use.
The objective of the invention is to realize through following technical scheme: (1) is with Bufo siccus maggot water or the alcohol dilute solution soaking and extracting is separated or ultrasonic extraction is separated; (2) remove protein and polysaccharide; (3) inject the acid cation exchange resin post and also use the alkaline aqueous solution eluting, collect toad peptide antibiotic part; (4) with ultrasonic standing wave liquid phantom preparing chromatogram separation and purification toad peptide antibiotic; (5) the toad peptide antibiotic is used in combination with suitable pharmaceutical necessities, is prepared into toad peptide antibiotic tincture, toad peptide antibiotic paste, toad peptide antibiotic ointment and toad peptide antibiotic respectively and sticks with paste membrane.
The specific embodiment
Below in conjunction with embodiment the present invention is done further description:
Embodiment:
1, (1) with the pure water of 10L-100L and add 0.02-0.10mol/L phosphoric acid-dipotassium hydrogen phosphate buffer and be heated to 35 ℃-90 ℃, keep 30min-300min and stir, extracts the bullion of toad peptide antibiotic with 1000g Bufo siccus maggot; (2) add the ethanol or the methanol of 5 times-10 times of extracting liquid volumes, quiescent setting 1h-24h under the room temperature, use then seperator under the 800r/min-16000r/min condition with polysaccharide and Separation of Proteins; (3) separating medium pumps in the HD-8 acid cation exchange resin post, room temperature held 1h-24h; Collect toad peptide antibiotic part with 1mol/L-10mol/L ammonia eluting then; (4) the toad peptide antibiotic of collecting is partially dissolved in the dicyandiamide solution of being made up of according to 5: 3: 4 proportioning chloroform-alcohol-water with the ultrasonic standing wave liquid phantom preparing chromatogram with the sound intensity 0.2kw-5.0kw separation and purification toad peptide antibiotic under flow velocity 0.1ml/min-1000ml/min, the UV-detector wavelength 220nm testing conditions; (5) with the toad peptide antibiotic of purification with vacuum freeze drier at-10 ℃--dry 1.0h-10h under 50 ℃, vacuum 0.01Mpa-0.1Mpa condition, content reaches 98.2%; (6) the toad peptide antibiotic of purification was pulverized 100 orders-200 mesh sieve and was collected powder.
2, (1) with the pure water of 5L-10L and add 10%-20% ethanol, to extract 10min-120min under the 1kw-10kw sound intensity ultrasound wave normal temperature condition, extracts toad peptide antibiotic crude product solution with 1000g Bufo siccus maggot in the ultrasonic extraction jar; (2) add 5 times-10 times ethanol in the extracting solution, under the room temperature, quiescent setting 1h-24h is with the seperator 800r/min-16000r/min above-mentioned solution of separation continuously; (3) with the 10%-1.0% of vacuum evaporator concentrated extracting solution to original volume under vacuum 0.01Mpa-0.5Mpa, 10 ℃ of-50 ℃ of conditions of temperature; (4) will extract concentrated solution and pump in the HZ001 acid cation exchange resin post,, collect toad peptide antibiotic part with 0.1mol/L-1.0mol/L NaOH eluant solution; (5) the toad peptide antibiotic of collecting is partly dissolved in the dicyandiamide solution of being made up of according to 5: 3: 4 proportioning chloroform-alcohol-water; With the ultrasonic standing wave liquid phantom preparing chromatogram with under sound intensity 0.1kw-5.0kw, flow velocity 0.1ml/min-1000ml/min, the UV-detector wavelength 220nm testing conditions, continuously separation and purification; (6) the toad peptide antibiotic of separation and purification is in that temperature-10 ℃--50 ℃, vacuum 0.01Mpa-0.1Mpa vacuum lyophilization 1h-10h, the toad peptide antibiotic content behind the purification reaches 98.9%.
3, get 20g lanoline and 30-35g vaseline heat fused after; Add and be pre-mixed uniform 10g-15g starch and 10g-15g zinc oxide; Stir become pasty state after, add 1g-5g toad peptide antibiotic powder mixing when being cooled to room temperature, toad peptide antibiotic paste.
4, with 1000g liquid paraffin and 1000g vaseline at xeothermic 100 ℃ of-200 ℃ of antibacterial 1h-3h; Be cooled to after the room temperature both mix homogeneously and grind 30min-180min; It is even to add 1.0g-50g toad peptide antibiotic powder mixes again, toad peptide antibiotic ointment.
5, get toad peptide antibiotic 2g-10g and be dissolved in the 1000ml 0.1%-95% alcoholic solution, and add 50ml-100ml glycerol, promptly be prepared into toad peptide antibiotic tincture with high pressure homogenizer homogenization processing under the 0.3Mpa-60Mpa normal temperature condition.
6, getting toad peptide antibiotic 10g is dissolved in the 1000ml 30%-95% alcoholic solution and adds the 1.0ml-100ml tween 80 and 10ml-50ml Oleum Ricini and 1.0ml-100ml propylene glycol; Add dissolving of 10g-200g hydroxypropyl emthylcellulose and mix homogeneously after stirring again, promptly be prepared into the toad peptide antibiotic and stick with paste membrane.
7, clinical treatment and result:
(1) patient who includes case in meets the patient of " diabetic foot (acromelic gangrene inspection method and diagnostic criteria (draft)) ", treatment time 10d-90d.
(2) after diagnosing the patient with diabetic feet state of an illness is pressed six grades of classifications of tcm clinical practice:
0 grade: skin does not have open focus.Show as the acra blood supply insufficiency, color cyanosis or pale, acra are sent out cold, numbness, insensitive or forfeiture, and acra twinge or causalgia are often with the deformity of toes or foot etc.;
One-level: acra skin has open focus.Vesicle, blood blister, clavus or callus, the caused shallow table ulcer of cold injury or scald and other skin injurys, but focus does not involve deep tissue as yet;
Secondary: infection focus has been invaded deep muscle tissue.Slight cellulitis is often arranged, and multiple pus kitchen range and sinus tract form, or infect along spatium intermusculare and enlarge, and cause vola, instep connectivity ulcer or gangrene, and purulent secretion is more, foot or refer to the skin of toe dry gangrene, but the tendon ligament does not still have destruction;
Three grades: the tendon ligament tissue destroys.Cellulitis merges and forms big vomica, and purulent secretion and slough increase, foot or minority toes dry gangrene, but bone destruction is still not obvious;
Level Four: severe infections has caused bone destruction.Osteomyelitis, osteoarthrosis destroys or has formed pseudarthrosis, and part toes or part brothers are taken place moist or the serious gangrene of dryness or downright bad;
Pyatyi: the big portion of foot or all infect or ischemia causes serious moist or dry gangrene, acra blackening, ordinary wave and ankle joint and shank.
(3) Therapeutic Method: all case all adopts strict blood sugar control; Dredge microcirculation simultaneously; With behind the necrotic tissue cleaning with 0.01% Hydrogen Peroxide Dilute Solution antibacterial, after drying, conventional treat with toad peptide antibiotic tincture coated; Reuse toad peptide antibiotic paste or toad peptide antibiotic ointment or toad peptide antibiotic are stuck with paste membrane, treatment cycle 10 days to 90 days.
(4) criterion of therapeutical effect is drafted with reference to " traditional Chinese medical science disease diagnosis criterion of therapeutical effect ", and recovery from illness: wound surface heals basically, produce effects: the wound surface area reduces the scope＞50%, effectively: the wound surface area reduces the scope＞and 30%; Invalid: wound surface area color is slightly improved or is not had a significant change.Therapeutic outcome is seen table
The invention has the beneficial effects as follows that its used toad peptide antibiotic composition can join in the various preparations by qualitative and quantitative through extracting purification, stable and controllable for quality.It is remarkable in treatment diabetic foot process effect that therapeutic outcome shows that the toad peptide antibiotic is used for preparing the medicine of controlling diabetic foot, and suitable suitability for industrialized production is also promoted the use of.
Claims (8)
1. the application of toad peptide antibiotic in preparation treatment diabetic foot medicine; It is characterized in that: 1. with 1000g Bufo siccus maggot with the pure water of 10L-100L and add 0.02mol/L-0.10mol/L phosphoric acid-dipotassium hydrogen phosphate buffer solvent and be heated to 35 ℃-90 ℃, keep 30min-300min and stir; Or, extract the bullion of toad peptide antibiotic with 1kw-10kw ultrasonic Treatment 10min-120min; 2. the ethanol or the methanol that add 5 times-10 times of extracting liquid volumes, quiescent setting 1h-24h under the room temperature, use then seperator under the 800r/min-16000r/min condition with polysaccharide and Separation of Proteins; 3. separating medium pumps in HZ001 or the HD-8 acid cation exchange resin post, and room temperature held 1h-24h uses 1mol/L-10mol/LNaOH solution or ammonia spirit eluting then, collects toad peptide antibiotic part; 4. the toad peptide antibiotic of collecting is partially dissolved in the dicyandiamide solution of being made up of according to 5: 3: 4 proportioning chloroform-alcohol-water with the ultrasonic standing wave liquid phantom preparing chromatogram with under sound intensity 0.2kw-5.0kw, injection flow velocity 0.1mL/min-1000ml/min, the UV-detector wavelength 220nm testing conditions separation and purification toad peptide antibiotic; 5. with the toad peptide antibiotic of purification with vacuum freeze drier at-10 ℃--dry 1.0h-10h under 50 ℃, vacuum 0.01Mpa-0.1Mpa condition, content reaches 98.2%-98.9%; 6. toad peptide antibiotic and pharmaceutic adjuvant are mixed and made into toad peptide antibiotic paste, toad peptide antibiotic ointment, toad peptide antibiotic tincture, toad peptide antibiotic paste membrane.
2. the application of toad peptide antibiotic as claimed in claim 1 in preparation treatment diabetic foot medicine; It is characterized in that: 1000g Bufo siccus maggot with the pure water of 20L-30L and add 0.02mol/L-0.05mol/L phosphoric acid-dipotassium hydrogen phosphate buffer solution and be heated to 50 ℃-60 ℃ and keep 30min-60min and stir, is extracted toad peptide antibiotic bullion.
3. the application of toad peptide antibiotic as claimed in claim 1 in preparation treatment diabetic foot medicine; It is characterized in that: separating medium injects in HZ001 or the HD-8 acid cation exchange resin post; Room temperature held 5h-10h; Use 2mol/L-5mol/LNaOH solution or ammonia spirit eluting then, collect toad peptide antibiotic part.
4. the application of toad peptide antibiotic as claimed in claim 1 in preparation treatment diabetic foot medicine is characterized in that: the toad peptide antibiotic of collection is partially dissolved in the dicyandiamide solution of being made up of according to 5: 3: 4 proportioning chloroform-alcohol-water with the ultrasonic standing wave liquid phantom preparing chromatogram with separation and purification toad peptide antibiotic under sound intensity 0.5kw-2.0kw, injection flow velocity 20ml/min-100ml/min, the UV-detector wavelength 220nm testing conditions.
5. the application of toad peptide antibiotic as claimed in claim 1 in preparation treatment diabetic foot medicine, it is characterized in that: toad peptide antibiotic paste is made up of 1g-2g toad peptide antibiotic, 20g lanoline, 30g vaseline, 10g starch and 10g zinc chloride.
6. the application of toad peptide antibiotic in preparation treatment diabetic foot medicine according to claim 1, it is characterized in that: toad peptide antibiotic ointment is made up of 5g-10g toad peptide antibiotic, 1000g liquid paraffin, 1000g vaseline.
7. the application of toad peptide antibiotic in preparation treatment diabetic foot medicine according to claim 1, it is characterized in that: toad peptide antibiotic tincture is made up of 2g-10g toad peptide antibiotic, 1000ml30%-60% ethanol and 50ml-100ml glycerol.
8. the application of toad peptide antibiotic in preparation treatment diabetic foot medicine according to claim 1, it is characterized in that: the toad peptide antibiotic is stuck with paste membrane and is made up of 10g toad peptide antibiotic, 1000ml30%-60% ethanol, 10ml-50ml tween 80,20ml-30ml Oleum Ricini, 10ml-50ml propylene glycol and 50g-150g hydroxypropyl emthylcellulose.
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| CN2011100072769A CN102078593B (en) | 2011-01-14 | 2011-01-14 | Application of toad peptide antibiotic for preparing medicines for curing diabetic foot |
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| CN2011100072769A CN102078593B (en) | 2011-01-14 | 2011-01-14 | Application of toad peptide antibiotic for preparing medicines for curing diabetic foot |
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| CN102676292A (en) * | 2012-06-02 | 2012-09-19 | 哈尔滨工业大学 | Separation process of toad maggot essential oil |
| CN113455470B (en) * | 2021-08-27 | 2022-03-08 | 上海瀚岳药源生物科技发展有限公司 | Method for artificially culturing toad feeding insects |
| CN114831927A (en) * | 2022-05-16 | 2022-08-02 | 西北工业大学 | Toad eating gel external use medicine composition and preparation method and application thereof |
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| CN1390851A (en) * | 2001-06-08 | 2003-01-15 | 中国科学院昆明动物研究所 | Bombesin of Bombina maxima and its preparing process and genes |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1390851A (en) * | 2001-06-08 | 2003-01-15 | 中国科学院昆明动物研究所 | Bombesin of Bombina maxima and its preparing process and genes |
| WO2003099842A2 (en) * | 2002-05-21 | 2003-12-04 | Wisconsin Alumni Research Foundation | Compositions and methods of use for a bombesin peptide |
| CN1568681A (en) * | 2004-05-10 | 2005-01-26 | 孔繁伟 | Method for cultivating toad evildoers using traditional Chinese medicine |
| CN1814091A (en) * | 2005-11-28 | 2006-08-09 | 吴银琼 | External-use Chinese medicine preparation for treating angeitis and diabetes foot |
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