CN107569532A - A kind of dimension medicine compound of leucoderma and preparation method thereof - Google Patents
A kind of dimension medicine compound of leucoderma and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to dimension medicine compound of a kind of leucoderma and preparation method thereof, the compound is made up of Carum carvi, psoralea corylifolia, puncture vine, ajowan-caraway seed, turpeth medicinal material, using solvent extraction, removal of impurities, concentration, drying means, the peroral dosage forms such as oral liquid, syrup, particle, hard shell capsules, tablet are made plus proper auxiliary materials.The present invention is theoretical according to uighur medicine, with reference to national medical drugs feature and long-term clinical practice among the people, obtains the dimension medicine compound of leucoderma using modern extracting method, strengthens drug effect, improve bioavilability.External activity experiment display：The dimension medicine compound different dosage forms of the leucoderma obtained using the method for the invention have different degrees of influence to Murine B 16 Melanoma Cells form, have concentration dependent activation to cytoactive and B16 cell amount.Test result indicates that the dimension medicine compound different dosage forms of the leucoderma show very strong activation B16 melanoma cells, melanin is promoted to be formed, available for treating leucoderma.
Description
Technical field
The present invention relates to a kind of dimension medicine compound and preparation method for being used to treat leucoderma, belong to uighur medicine and nationality
Pharmaceutical technology field.
Background technology
Leucoderma is the depigmentation that a kind of acquired, gradual, multifactor related chronic epidermal melanocytes are lost in
Property Complex Diseases, it is characterised in that the progressive of Functional Surface melanocyte is lost.The pathogenesis of leucoderma is not yet bright
Really.But, it has been proposed that various mechanisms of causing a disease, including genetic association, cell and body fluid autoimmunity, oxidative stress, nerve
Hormone imbalances, the exception of melanocyte surrounding microenvironment and the latent defect of melanocyte.Some report leucoderma is complete at present
The ball incidence of disease is 0.5%~1%, also it was reported that the local incidence of disease is even up to 8% individually in the world.More recently by the screening whole world
The illness rate of individual research report leucoderma is between 0.5 and 2% more than 50.Asexuality and racial difference, typically from infancy or green grass or young crops
Phase in spring falls ill.Leucoderma is easily to examine refractory disease, is not infected, and does not also have big influence to body, but influence is attractive in appearance, gives people to bring
Spiritual pressure.China's prevalence of vitiligo is in the trend for gradually having rising in recent years, and it is more especially to invade person between twenty and fifty.Mesh
Before, treating the method for leucoderma mainly has to add light treatment etc. using corticosteroid parahormone or photochemistry class medicine.Although
Western medical treatment short term effect is preferable, but toxic side effect is big, it is impossible to which long-term use of, easily recurrence aggravates after drug withdrawal, and late result is poor.
Leucoderma Uygur medical science is referred to as " white heat this ", and the Complex Diseases Uygur medicine such as treatment leucoderma has its original
Part and significant curative effect, the especially discussion to leucoderma and treatment have long history, there is the understanding of comparison system, at the same have compared with
Complete treatment method and means, Uygur medicine classical works《Medicinal total storehouse》In to this with regard on the books.Successive dynasties Uygur medicine name
Doctor constantly brings forth new ideas on the basis of leucoderma traditional remedies are summarized, and have accumulated rich experience, at home and abroad generates extensive
Influence.
Medically treatment leucoderma is still difficult point at present, but classification of the Uygur medicine to leucoderma, morbidity machine
Exclusive characteristic in terms of system, principle of reatment, characteristic treatment and treatment, makes some progress.Leucoderma Uygur medicine morbidity machine
Reason：(1) raw environment is chronically at, the physiological dysfunction of liver generation normal body fluid, makes mucus caused by natural force declines
Anomalous variation generation abnormal phlegm occurs for matter.Abnormal phlegm contains excessive moist and excessively cold, and infringement digests first
With second and third digestion (normal physiological function of the organ such as stomach, small intestine, liver, blood circulation), vicious circle is formed, is weakened
Energy and natural force needed for material cyclic process, the colouring function of exhaustion blood matter, further result in the production of abnormal humour
It is raw；(2) long-term use of raw property food is that raw property makings and raw property body fluid are vigorous in human body, causes the exhaustion of liver heat energy
Cause the physiological dysfunction of liver generation normal body fluid, generate abnormal phlegm；(3) abnormal phlegm of surplus deposits in vivo
In skin histology, the normal blood circulation of skin is destroyed, nutritional ingredient can not be transported to the 4th digestion (skin part),
Cause " natural force " (breathing power, pinching power, eupepsy, voiding force) growing power (transformation power, forming power) dysfunction of skin,
Melanocyte is unable to normal growth and development, so as to depigmentation of skin, leucoderma stove occurs；(4) skin is pressurized for a long time, microcirculation barrier
Hinder, skin cannot get required nutriment, and melanocyte is unable to normal development, so as to depigmentation of skin, leucoderma stove occurs；
(5) there is relation with inherent cause.Therefore, advocate to apply abnormal phlegm maturing agent first during clinical treatment leucoderma." ripe,
Remove abnormal humour, correct abnormal makings, promote the generation of normal body fluid ", then " functional status for recovering each internal organs " so as to
Recover body natural force.After body body fluid and makings restore balance, leucoderma is treated using hot medicine dedicated.Uygur medicine is treated
Leucoderma mainly to improve the natural heating power of body, dissolves the abnormal humour of precipitation, promoting blood circulation and removing blood stasis is purpose medication, so as to improve
The microcirculation of skin and nutrition condition, promote the pigment generation at leukoderma.
The content of the invention
Present invention aims at a kind of, there is provided dimension medicine compound of leucoderma and preparation method thereof, the compound be by Carum carvi,
Psoralea corylifolia, puncture vine, ajowan-caraway seed, the taste medicinal material of turpeth 6 composition, using solvent extraction, removal of impurities, concentration, dry the methods of, add
Oral liquid, syrup, particle, hard shell capsules or tablet is made in upper proper auxiliary materials.The compound medicine is theoretical according to uighur medicine, with reference to
The clinical practice among the people of national medical drugs feature and the Uygur nationality for a long time, is obtained in the compound medicine using reasonable drawing method
Active ingredient, drug effect is enhanced, improve bioavilability.External activity experiment display：Obtained using the method for the invention
Extract has different degrees of influence to Murine B 16 Melanoma Cells form, has to cytoactive and B16 cell amount dense
Spend dependence activation.Test result indicates that the extract of the compound various concentrations shows very strong activation B16 black
Plain oncocyte, melanin is promoted to be formed, available for treating leucoderma.
A kind of dimension medicine compound of leucoderma of the present invention, it by bulk drug is Carum carvi 100-5 parts that the compound, which is, Psoralen
Fat 5-50 parts, puncture vine 5-50 parts, ajowan-caraway seed 5-50 parts, turpeth 5-50 parts, auxiliary material are Steviosin 10-100 parts, sorbic acid
10-100 parts, sucrose 10-100 parts, lactose 10-100 parts, microcrystalline cellulose 10-100 parts, mannitol 10-100 parts, starch 10-
100 parts, dextrin 10-100 parts, magnesium stearate 1-10 parts, sodium carboxymethyl starch 5-40 parts or low-substituted hydroxypropyl methylcellulose 5-40
Oral liquid, syrup, granule, hard shell capsules or tablet is made in part according to a conventional method.
The dimension medicine compound preparation method of the leucoderma, concrete operations follow these steps to carry out：
A, the Carum carvi 100-5 parts screened are crushed, crosses 10-80 mesh sieves, the water immersion 4- for adding 4-20 times of mass ratio to measure
12 hours, volatile oil 2-8 hours are extracted with volatile oil extractor, collect volatile oil and aqueous extract respectively；
B, the volatile oil in step a is included according to a conventional method with beta-schardinger dextrin, obtained inclusion compound is dried, standby
With；
C, the psoralea corylifolia 5-50 parts screened, puncture vine 5-50 parts, ajowan-caraway seed 5-50 parts, turpeth 5-50 parts are mixed
Broken or crushing, crosses 10-40 mesh sieves, the alcohol reflux extraction for being 10-95% with the 4-20 times of water measured or concentration or Soakage extraction
1-4 times, each 1-4 hours, filtration, merging filtrate, respectively obtain water or ethanol extract；
D, Aqueous extracts in step c are merged with Aqueous extracts in step a, is concentrated under reduced pressure, dries, crushes, obtain extract powder；
Or will merge when alcohol extract is concentrated under reduced pressure into no ethanol flavor in step c with Aqueous extracts in step a, it is concentrated under reduced pressure, does
Dry, crushing, obtains extract powder；
E, inclusion compound in step b is merged with extract powder in step d, add Steviosin 10-100 parts, sorbic acid 10-100 parts,
Pure water, after being uniformly dissolved, plate-frame filtering, dispense, sterilizing, oral liquid is made according to a conventional method；
Or merge inclusion compound in step b with extract powder in step d, add sucrose 10-100 parts, pure water, be uniformly dissolved
Afterwards, filter, dispense, sterilizing, syrup is made according to a conventional method；
Or merge inclusion compound in step b with extract powder in step d, add lactose 10-100 parts, mannitol 10-100 parts, sugarcane
Sugared 10-100 parts or dextrin 10-100 parts, fully mix, granule is made according to a conventional method；
Or merge inclusion compound in step b with extract powder in step d, add lactose 10-100 parts, microcrystalline cellulose 10-100
Part, sodium carboxymethyl starch 5-40 parts, low-substituted hydroxypropyl methylcellulose 5-40 parts, magnesium stearate 1-10 parts, fully mix, routinely
Hard shell capsules or tablet is made in method.
Dimension medicine compound of a kind of leucoderma of the present invention and preparation method thereof, the pharmacological property of the compound medicine
For：Uygur medicine thinks that this prescription commonly uses hot, mucilaginous substance by Uygur medicine and adjusts the Uygur medicine groups such as class, mucilaginous substance removing class
Into clinical experience side, Carum carvi is main key medicine in side, and psoralea corylifolia, ajowan-caraway seed adjuvant, puncture vine, turpeth are regulation
Medicine.Preparation made of the preparation method has dysregulation mucilaginous substance, removes abnormal phlegm, corrects abnormal makings, promotes
The generation of normal body fluid ", then " functional status for recovering each internal organs " is so as to recovering body natural force.Body body fluid and makings are extensive
The natural heating power of body is improved on the basis of multiple equilibria, dissolves the abnormal humour of precipitation, it is promoting blood circulation and removing blood stasis.Improve the microcirculation of skin
And nutrition condition, promote the pigment generation at leukoderma, so as to reach treatment leucoderma effect.
The pharmacological property of each flavour of a drug is in a kind of dimension medicine compound of leucoderma of the present invention：
Carum carvi：Uygur's name can fall tooth, alias storehouse wood Buddhist nun such as rice or cumin such as rice, be samphire Caraway
Carum carvi Linn. dry fruit, its plant are perennial herb, and two level or three-level are xeothermic, have raw dry heat, temperature
Stomach eliminating dampness, increase pigment, de-swelling analgesic, diuresis is felt at ease, and drives away helminth.Cure mainly leucoderma, stomach cold receives difference, abdominal distension stomachache, urine
Obstructed, palpitaition is had palpitation, and enteral is infested.It is for oral administration：3~5g；Suitable amount used externally.
Psoralea corylifolia：Uygur's name enters gram Wei Zi Europe as closed, and other Bark itself or Ba Biqi, are legumes psoraleae
Psoralea corylifolia L. ripening fruits.Two level is xeothermic, has raw dry heat, understands abnormal phlegm, red skin hair
Bubble, increase pigment, net blood is understood, and dispelling wind and arresting itching, eliminating dampness removes rash, kills anthelmintic etc..Raw property or mucilaginous substance disease are cured mainly, such as
Leucoderma, leprosy, pruitus, all kinds of eczemas, Enterozoa etc..1~3g for oral administration；Suitable amount used externally.
Tribulus terrestris：Uygur name Ou Hutikan, alias Hai Saike or Ha Erhaisaike, it is puncture vine (Tribulus
Terrestris L.) it is zygophyllaceae (Zygophy-laceae) Tribulus (Tribulus) plant.One-level is xeothermic or fever and chills, tool
There is raw dry heat, diuresis dysmenorrhoea, stone decomposing and discharging is relieving cough and asthma, strengthens pigment.Raw property or mucilaginous substance disease are cured mainly, it is such as small
Just it is obstructed, irregular menstruation, kidney stone, vesical calculus, gonorrhoea, cough, asthma, leucoderma.It is for oral administration：5~7g；Suitable amount used externally.
Ajowan-caraway seed：Uygur name Ju Weina, alias difficulty flower, capsule are colored or open wooden Nimu Luke.For the rough fruit of samphire
Trachyspermum ammi (L.) Sprague dry mature fruits of celery.Harvest, dry during autumn fruit maturation.Three-level is done
Heat.The dry heat of function life, warm stomach are disappeared, and dissipate gas analgesic, wind dispelling cold dispelling, and dysmenorrhoea pain is urinated, diuresis and stone expeling, replenishing essence lactagogue, life of warming up
Brightness.Cure mainly raw property or mucilaginous substance disease, as stomach void receive it is poor, stomachache intestines twist, paralyse, tremble, muscle muscular relaxation, menstruation is obstructed,
Oedema, difficult urination, lithangiuria, impotence defective ejaculation, pruitus, leucoderma, eczema, cold grain sore.3-5g for oral administration；External application is fitted
Amount.
Turpeth：Uygur's name Bristol cloth moral, alias dash forward Shandong must, white prominent Shandong must or answer youngster not, be Convolvulaceae
Plant box fruit rattan Opercalina turperthum L. dry rhizome and stem.Two level is xeothermic, raw dry heat, removes abnormal viscous
Liquid matter, eliminating dampness detumescence, sweat of dispelling analgesic, wind-dispelling nourishing the muscle and tendon is relieving cough and asthma, and perspiration removing solution is strongly fragrant, and softening hard masses disappear hemorrhoid, and it is skilful to open ear.Cure mainly raw
Property or mucilaginous substance disease, such as oedema, arthralgia, Minor articulus pain, sciatica, facial paralysis, cough, asthma, depression,
Hemorrhoid, tinnitus, deafness etc..It is for oral administration：3~5g.
The dimension medicine compound of leucoderma of the present invention is the prescription under the guidance of Uygur medicine traditional theory, and in clinic
It is upper to be used for for many years, it is curative for effect.
Embodiment
Embodiment 1 (using 1000ml as radix, prepares oral liquid)
A, by 50 parts of crushing of Carum carvi through having screened, 10 mesh sieves is crossed, add mass ratio to be soaked 4 hours for the water of 20 times of amounts,
Volatile oil is extracted with volatile oil extractor 8 hours, collects volatile oil and aqueous extract respectively；
B, take beta-schardinger dextrin 10g to put in round-bottomed flask, 25 times of amount distilled water are added, in the magnetic agitation of temperature 50 C constant temperature
Stirring and dissolving, it is 1 by volume by the volatile oil absolute ethyl alcohol in step a on device:1 amount dilution, be slow added into β-
In cyclodextrin solution, in continuing stirring 2h on the magnetic stirring apparatus of temperature 50 C constant temperature, room temperature cooling, put and 24h is refrigerated in refrigerator
Afterwards, take out, filter, (30-60 DEG C) is rinsed 3 times, each 10mL with petroleum ether, and 4h, get Gan Bao are dried in temperature 50 C baking oven
Compound, it is standby；
C, 20 parts of 50 parts of the psoralea corylifolia through having screened, 50 parts of puncture vine, 20 parts of ajowan-caraway seed, turpeth mixing were crushed
10 mesh sieves, add the water retting of 20 times of amounts to extract 4 times, every time extraction 1 hour, filtration, merging filtrate, obtain aqueous extract；
D, aqueous extract in step c is merged with Aqueous extracts liquid in step a, is concentrated under reduced pressure, dry, pulverize, obtain extract powder；
E, inclusion compound in step b is merged with extract powder in step d, adds 30 parts of Steviosin, 30 parts of sorbic acid, more than pure water
Amount, after being uniformly dissolved, plate-frame filtering, packing, every or every bottle packaging 10ml, 20ml, 100ml or 200ml sterilize, routinely square
Oral liquid is made in method；
Embodiment 2 (using 1000ml as radix, prepares syrup)
A, by 20 parts of crushing of Carum carvi through having screened, 50 mesh sieves is crossed, add mass ratio to be soaked 5 hours for the water of 10 times of amounts,
Volatile oil is extracted with volatile oil extractor 5 hours, collects volatile oil and aqueous extract respectively；
B, take beta-schardinger dextrin 15g to put in round-bottomed flask, 25 times of amount distilled water are added, in the magnetic agitation of temperature 50 C constant temperature
Stirring and dissolving, it is 1 by volume by the volatile oil absolute ethyl alcohol in step a on device:1 amount dilution, be slow added into β-
In cyclodextrin solution, in continuing stirring 2h on the magnetic stirring apparatus of temperature 50 C constant temperature, room temperature cooling, put and 24h is refrigerated in refrigerator
Afterwards, taking out, filter, inclusion compound is rinsed 3 times, each 10mL for (30-60 DEG C) with petroleum ether, and 4h is dried in temperature 50 C baking oven,
Inclusion compound must be done, it is standby；
C, by 50 parts of 20 parts of the psoralea corylifolia through having screened, 20 parts of puncture vine, 50 parts of ajowan-caraway seed, turpeth co-grindings,
40 mesh sieves are crossed, add the alcohol reflux that 15 times of amount concentration are 50% to extract 1 time, extraction time is 4.0 hours, obtains alcohol extract；
D, by when alcohol extract is concentrated under reduced pressure into no alcohol taste in step c, merge with Aqueous extracts liquid in step a, be concentrated under reduced pressure,
It dry, pulverize, obtain extract powder；
E, step d medicinal extract and step b dry inclusion compound are taken, adds 40 parts of distilled water, 15 parts of sucrose, after being uniformly dissolved, mistake
Filter, surplus distilled water is added, packing, every or every bottle packaging 10ml, 20ml, 100ml, 200ml or 300ml, is sterilized, routinely
Syrup is made in method.
Embodiment 3 (using 1000ml as radix, prepares syrup)
A, by 10 parts of the Carum carvi crushing through having screened, 40 mesh sieves is crossed, add the water of 6 times of amounts of mass ratio to soak 6 hours, with waving
Hair oil extractor extraction volatile oil 3 hours, collects volatile oil and aqueous extract respectively；
B, take beta-schardinger dextrin 15g to put in round-bottomed flask, 25 times of amount distilled water are added, in the magnetic agitation of temperature 50 C constant temperature
Stirring and dissolving, it is 1 by volume by the volatile oil absolute ethyl alcohol in step a on device:1 amount dilution, be slow added into β-
In cyclodextrin solution, in continuing stirring 2h on the magnetic stirring apparatus of temperature 50 C constant temperature, room temperature cooling, put and 24h is refrigerated in refrigerator
Afterwards, taking out, filter, inclusion compound is rinsed 3 times, each 10mL for (30-60 DEG C) with petroleum ether, and 4h is dried in temperature 50 C baking oven,
Inclusion compound must be done, it is standby；
It is c, 20 parts of 10 parts of the psoralea corylifolia through having screened, 10 parts of puncture vine, 20 parts of ajowan-caraway seed, turpeth mixing are broken,
24 mesh sieves are crossed, add the ethanol that 8 times of amount concentration are 75%, Soakage extraction 1 time, extraction time is 4.0 hours, obtains alcohol extract；
D, by when alcohol extract is concentrated under reduced pressure into no alcohol taste in step b, merge with Aqueous extracts liquid in step a, be concentrated under reduced pressure,
It dry, pulverize, obtain extract powder；
E, step d medicinal extract and step b inclusion compound are taken, adds 30 parts of distilled water, 100 parts of sucrose, after being uniformly dissolved, mistake
Filter, surplus distilled water is added, packing, every or every bottle packaging 10ml, 20ml, 100ml, 200ml or 300ml, is sterilized, routinely
Syrup is made in method.
Embodiment 4 (using 1000g as radix, prepares granule)
A, by 100 parts of crushing of Carum carvi through having screened, 80 mesh sieves is crossed, adds the water of 15 times of amounts of mass ratio to soak 8 hours, uses
Volatile oil extractor extraction volatile oil 3 hours, collects volatile oil and aqueous extract respectively；
B, take beta-schardinger dextrin 15g to put in round-bottomed flask, 25 times of amount distilled water are added, in the magnetic agitation of temperature 50 C constant temperature
Stirring and dissolving, it is 1 by volume by the volatile oil absolute ethyl alcohol in step a on device:1 amount dilution, be slow added into β-
In cyclodextrin solution, in continuing stirring 2h on the magnetic stirring apparatus of temperature 50 C constant temperature, room temperature cooling, put and 24h is refrigerated in refrigerator
Afterwards, taking out, filter, inclusion compound is rinsed 3 times, each 10mL for (30-60 DEG C) with petroleum ether, and 4h is dried in temperature 50 C baking oven,
Inclusion compound must be done, it is standby；
C, by 25 parts of 25 parts of the psoralea corylifolia through having screened, 25 parts of puncture vine, 25 parts of ajowan-caraway seed, turpeth co-grinding mistakes
24 mesh, the alcohol reflux that the concentration for adding 15 times of amounts is 30% extract 2 times, and extraction 2.0 hours, obtain alcohol extract every time；
D, by when alcohol extract is concentrated under reduced pressure into no alcohol taste in step c, merge with Aqueous extracts liquid in step a, be concentrated under reduced pressure,
It dry, pulverize, obtain extract powder；
E, step d extract powder and step b inclusion compound are taken, 50 parts of sucrose is added, 50 parts of dextrin, fully mixes, by normal
Granule is made in regulation prescription method, per packed 0.5g, 1g or 2g.
Embodiment 5 (using 1000g as radix, prepares granule)
A, by 5 parts of the Carum carvi crushing through having screened, 50 mesh sieves is crossed, add the water of 4 times of amounts of mass ratio to soak 4 hours, with waving
Hair oil extractor extraction volatile oil 2 hours, collects volatile oil and aqueous extract respectively；
B, take beta-schardinger dextrin 15g to put in round-bottomed flask, 25 times of amount distilled water are added, in the magnetic agitation of temperature 50 C constant temperature
Stirring and dissolving, it is 1 by volume by the volatile oil absolute ethyl alcohol in step a on device:1 amount dilution, be slow added into β-
In cyclodextrin solution, in continuing stirring 2h on the magnetic stirring apparatus of temperature 50 C constant temperature, room temperature cooling, put and 24h is refrigerated in refrigerator
Afterwards, taking out, filter, inclusion compound is rinsed 3 times, each 10mL for (30-60 DEG C) with petroleum ether, and 4h is dried in temperature 50 C baking oven,
Inclusion compound must be done, it is standby；
C, by 5 parts of 5 parts of the psoralea corylifolia through having screened, 5 parts of puncture vine, 5 parts of ajowan-caraway seed, turpeth co-grindings, 10 are crossed
Mesh sieve, the alcohol dipping that 4 times of amount concentration are 95% is added to extract 4 times, extraction 4.0 hours, obtain alcohol extract every time；
D, by when alcohol extract is concentrated under reduced pressure into no alcohol taste in step c, merge with Aqueous extracts liquid in step a, be concentrated under reduced pressure,
It dry, pulverize, obtain extract powder；
E, step d extract powder and step b inclusion compound are taken, 20 parts of lactose, 30 parts of mannitol is added, fully mixes, press
Granule is made in conventional pharmaceutical method, per packed 0.5g, 1g or 2g.
Embodiment 6 (with 1000 for radix, prepares tablet)
A, by 50 parts of the Carum carvi crushing through having screened, 80 mesh sieves is crossed, add the water of 8 times of amounts of mass ratio to soak 4 hours, with waving
Hair oil extractor extraction volatile oil 4 hours, collects volatile oil and aqueous extract respectively；
B, take beta-schardinger dextrin 15g to put in round-bottomed flask, 25 times of amount distilled water are added, in the magnetic agitation of temperature 50 C constant temperature
Stirring and dissolving, it is 1 by volume by the volatile oil absolute ethyl alcohol in step a on device:1 amount dilution, be slow added into β-
In cyclodextrin solution, in continuing stirring 2h on the magnetic stirring apparatus of temperature 50 C constant temperature, room temperature cooling, put and 24h is refrigerated in refrigerator
Afterwards, taking out, filter, inclusion compound is rinsed 3 times, each 10mL for (30-60 DEG C) with petroleum ether, and 4h is dried in temperature 50 C baking oven,
Inclusion compound must be done, it is standby；
It is c, 25 parts of 25 parts of the psoralea corylifolia through having screened, 25 parts of puncture vine, 25 parts of ajowan-caraway seed, turpeth mixing are broken,
20 mesh sieves are crossed, the alcohol reflux that the concentration for adding 15 times of amounts is 75% extracts 3 times, 2.5 hours every time, obtains alcohol extract；
D, by when alcohol extract is concentrated under reduced pressure into no alcohol taste in step c, merge with Aqueous extracts liquid in step a, be concentrated under reduced pressure,
It dry, pulverize, obtain extract powder；
E, step d extract powder and step b inclusion compound are taken, adds 30 parts of lactose, 20 parts of microcrystalline cellulose, CMS
5 parts of 10 parts of sodium, 20 parts of low-substituted hydroxypropyl methylcellulose and magnesium stearate, fully mix, tablet, every weight are made according to a conventional method
For 0.3g, 0.4g or 0.5g.
Embodiment 7 (using 1000g as radix, prepares hard capsule)
A, by 70 parts of crushing of Carum carvi through having screened, 10 mesh sieves is crossed, adds the water of 15 times of amounts of mass ratio to soak 5 hours, uses
Volatile oil extractor extraction volatile oil 5 hours, collects volatile oil and aqueous extract respectively；
B, take beta-schardinger dextrin 15g to put in round-bottomed flask, 25 times of amount distilled water are added, in the magnetic agitation of temperature 50 C constant temperature
Stirring and dissolving, it is 1 by volume by the volatile oil absolute ethyl alcohol in step a on device:1 amount dilution, be slow added into β-
In cyclodextrin solution, in continuing stirring 2h on the magnetic stirring apparatus of temperature 50 C constant temperature, room temperature cooling, put and 24h is refrigerated in refrigerator
Afterwards, taking out, filter, inclusion compound is rinsed 3 times, each 10mL for (30-60 DEG C) with petroleum ether, and 4h is dried in temperature 50 C baking oven,
Inclusion compound must be done, it is standby；
C, by 5 parts of 25 parts of the psoralea corylifolia through having screened, 35 parts of puncture vine, 5 parts of ajowan-caraway seed, turpeth co-grindings, mistake
10 mesh sieves, the alcohol reflux that the concentration for adding 15 times of amounts is 95% extract 2 times, and each extraction time is 2.0 hours, obtains alcohol extracting
Liquid；
D, by when alcohol extract is concentrated under reduced pressure into no alcohol taste in step c, merge with Aqueous extracts liquid in step a, be concentrated under reduced pressure,
It dry, pulverize, obtain extract powder；
E, step d extract powder and step b inclusion compound are taken, adds 10 parts of lactose, 10 parts of microcrystalline cellulose, CMS
2 parts of 10 parts of sodium and magnesium stearate, fully mix, routinely pharmaceutical methods are made hard shell capsules, every capsule weight 0.25g, 0.3g,
0.35g or 0.4g.
Embodiment 8
The evaluating drug effect of the dimension medicine compound different dosage forms of leucoderma of the present invention：
1. experiment purpose：This research is intended utilizing modern medicine study theory, and the dimension medicine compound different dosage forms of leucoderma are entered
Row mouse melanin tumor cell (B16 cells) screening active ingredients are tested, so as to confirm the dimension medicine Compound Resisting leucoderma of leucoderma biology
Activity；
2. experiment material：
Instrument and consumptive material：CO2gas incubator, electric pipettor, the pipette of different ranges, ice machine, inversion are micro-
Mirror, 6 porocyte culture plates, micropipettor (suction nozzle), Biohazard Safety Equipment, carbon dioxide canister, cell counter, centrifuge,
Vortex oscillation instrument, cell are scraped, Tissue Culture Dish (different size), 96 hole elisa Plates, M5 ELIASAs, 1.5mL EP pipes, 0.5mL
EP is managed；Disposable Thin film glove, emgloves；
Reagent：Hyclone (REF：04-001-1ACS, Lot：1544459), high glucose medium (REF：
C11995500BT, Lot：8116188), trypsase (REF：25200-056, Lot：1742078), mycillin is dual anti-
(REF：15140-122, Lot：1523693), dimethyl sulfoxide (DMSO) (Sigma, numbering D4540), phosphate buffer (PBS),
75% alcohol, sodium hydroxide, ultra-pure water；
2.2 solution are prepared：
2.2.1 the preparation of phosphate buffer (PBS) storing liquid：
Sodium chloride 80g, disodium hydrogen phosphate dodecahydrate 32.5g, two hypophosphite monohydrate sodium dihydrogen 4.5g to 900mL are weighed respectively
In ultra-pure water, it is 7.4 that pH is adjusted after shaking dissolving, adds ultra-pure water to 1000ml, in 4 DEG C of preservations of temperature；
Face the used time, take storing liquid 100mL to add 900mL ultra-pure waters, shake up, used after sterilizing；
2.2.2 the preparation of sodium hydroxide lysate：
4g sodium hydroxides are weighed, after adding 80mL ultra-pure waters fully to dissolve, add 10mL dimethyl sulfoxide (DMSO)s, ultra-pure water is settled to
100mL；
2.2.3 the preparation of 8-methoxypsoralen (8-MOP)：
0.0108g 8-methoxypsoralen is weighed, 1mL dimethyl sulfoxide (DMSO)s is added and fully dissolves, 100 μ L/ pipe lucifuges
Freeze；
The 2.2.4 cell pyrolysis liquids RIPA of table 1 preparation (liquid storage)：
2.2.5 the preparation of protease inhibitors
4-NPP disodium salt hexahydrate (pNPP), 1M liquid storages are prepared with ultra-pure water：Weigh 0.371g 4-
Nitrophenyl phosphate disodium salt hexahydrate, 1mL ultra-pure waters, fully dissolving are added, -20 DEG C of temperature freezes standby；
Sodium fluoride (NaF), 1M liquid storages are prepared with ultra-pure water：419.9mg NaF is weighed, adds 10mL ultra-pure waters, it is fully molten
Solution, -20 DEG C of temperature freezes standby；
Phenylmethylsulfonyl fluoride (PMSF), 10mM liquid storages are prepared with isopropanol：174.19mg phenylmethylsulfonyl fluoride is weighed, is added
10mL ultra-pure waters, fully dissolving, -20 DEG C of temperature freezes standby；
DL- dithiothreitol (DTT)s (DTT), 100mM liquid storages are prepared with ultra-pure water：154.25mg DL- dithiothreitol (DTT)s are weighed,
1mL ultra-pure waters, fully dissolving are added, -20 DEG C of temperature freezes standby；
Benzenecarboximidamide prepares 100mM liquid storages with ultra-pure water：120.15mg benzenecarboximidamide is weighed, adds 10mL ultra-pure waters, fully
Dissolving, -20 DEG C of temperature freezes standby；
Sodium orthovanadate (OV) prepares 200mM storing liquids with ultra-pure water：800.26mg sodium orthovanadate is weighed, 10mL is added and surpasses
Pure water fully dissolves, and -20 DEG C of temperature freezes standby；
Table 2
According to the how much matching while using for collecting cell concentration；
2.2.6 the inactivation and packing of serum：
Every bottle of 500mL serum places 4 DEG C of defrostings of temperature, and 56 DEG C of water-baths placement 30min of temperature, are gone out after thawing completely
Living, -20 DEG C of temperature is frozen after the packing of 50mL/ pipes, and packaging operation is carried out in iuntercellular superclean bench；
2.2.7 dual anti-packing：
4 DEG C of the dual anti-placement temperature of 100mL/ bottles is carried out after thawing completely, and -20 DEG C of temperature freezes after the packing of 5.6mL/ pipes,
Packaging operation is carried out in iuntercellular superclean bench；
2.2.8 complete medium prepares (matching while using)：
Dual anti-containing 10% hyclone and 1% mycillin in culture medium, specific compound method is high glucose medium
(DMEM) it is dual anti-that hyclone and the 5.6mL mycillins of 50mL packing are added in 500mL, is used after well mixed, it is current existing
Match somebody with somebody；
2.3 experimental method：
2.3.1 cell culture：
Recovery mouse black-in tumor cell B16 (passage of this room freezes).With cell culture complete medium culture to cell attachment face
When product accounts for more than 80%, pancreatin digestion, cell culture complete medium terminates digestion.Cell density is adjusted to 2.0 × 105cells/mL
It is inoculated in 15cm Tissue Culture Dish, puts 37 DEG C of temperature, concentration 5%CO2Cultivated in incubator, contain 10% tire in culture medium
Cow's serum and 1% mycillin are dual anti-；
2.3.2 passage and inoculation：
Take the logarithm the B16 melanoma cells of phase, discard nutrient solution, cleaned 2 times with phosphate buffer, after pancreatin digestion,
2.5 × 105/hole of cell concentration is adjusted, is accessed in 6 orifice plates, 2mL, overnight incubation (about 12h-16h) are added per hole；
2.3.3 drug-treated：
The culture medium 2mL/ holes that next day more renews, cleaned 1 time with phosphate buffer before changing.Monomeric compound according to
12.5 μM, 25 μM and 50 μM concentration administrations, crude extract are administered according to 12.5 μ g/mL, 25 μ g/mL and 50 μ g/mL concentration, volatile oil
According to 50ul//mL, 100ul//mL, 200ul//mL and (50 μM of solvent control group (dimethyl sulfoxide (DMSO)) and positive controls are set
8-MOP), each concentration group sets 3 multiple holes, in 37 DEG C of temperature, concentration 5%CO248h is cultivated in constant incubator；(note：
If cell inoculum concentration is higher, culture medium its colour changed into yellow, it should change the fresh culture of drug containing during processing, ensure to fill
The nutrient of foot, consumed for cell metabolism)；
2.3.4 sample collection (whole process operates on ice)：
Each treatment group is put on ice for after 48h, sucks nutrient solution, is cleaned 2 times, filled with the phosphate buffer of precooling
The cell pyrolysis liquid for adding 100 μ L precoolings after residual liquid per hole point is blotted, rocking 6 orifice plates, lysate is fully covered adherent thin
Born of the same parents, scraped with cell and 1.5mL EP pipes (import) are transferred to after quickly cell is scraped, 4 DEG C of 1000rpm, abandoned after centrifuging 5min
Clearly；The PBS liquid of 100 μ L precoolings is added, vortex oscillation instrument mixes, and 1000 turns/min of temperature 4 DEG C, supernatant is abandoned after centrifuging 5min；
2.3.5 melanin is collected and dissolved：
After EP pipes are placed on the blending instrument of 4 DEG C of temperature fully mixing 40min, 4 DEG C of 12000rpm of temperature, 20min is centrifuged,
Supernatant is shifted into clean EP pipes, precipitation adds 190 μ L 1mol/L NaOH, temperature 60 C water-bath 2h, or 80 DEG C of temperature
Water-bath 1h, mixing is turned upside down during waters for several times, the melanin granule for being deposited in ttom of pipe is fully dissolved；
2.3.6 sample protein assay：
Supernatant protein content is determined, using Bo Maide Bradford protein quantification kits, is operated according to kit
Step is carried out, and calculates the protein concentration C in each hole；
2.3.7 melanin light absorption value determines：
Water-bath takes 150 μ L to determine measure OD values at 405nm into 96 hole elisa Plates after terminating；
2.3.8 statistical analysis：
Calculate melanocyte relative amount, melanocyte relative amount=melanocyte OD405/ protein concentration C, melanogenesis activity ratio=reality
Test a group melanocyte relative amount/solvent control group melanocyte relative amount × 100%；
3. experimental result：
The given formulation of embodiment 1-7 leucoderma dimension medicine compound is weighed, prepares the sample solution of various concentrations, detection
Its influence to B16 cell melanin contents, the results are shown in Table 3；
Influence of the 3 different extracts of table to B16 cell melanin contents
Conclusion：The dimension medicine compound different dosage forms of leucoderma just have preferably raising melanin thin in 5 μ g/mL of low concentration
The bioactivity of born of the same parents' melanin content, and bioactivity has preferable concentration dependent.Result of the test shows that different dosage forms are equal
B16 melanoma cells rise B16 cell amount can be influenceed, there is good anti-leucoderma activity.
Claims (2)
1. the dimension medicine compound of a kind of leucoderma, it is characterised in that it by bulk drug is Carum carvi 100-5 parts that the compound, which is, psoralea corylifolia 5-
50 parts, puncture vine 5-50 parts, ajowan-caraway seed 5-50 parts, turpeth 5-50 parts, auxiliary material is Steviosin 10-100 parts, sorbic acid 10-
100 parts, sucrose 10-100 parts, lactose 10-100 parts, microcrystalline cellulose 10-100 parts, mannitol 10-100 parts, starch 10-100
Part, dextrin 10-100 parts, magnesium stearate 1-10 parts, sodium carboxymethyl starch 5-40 parts or low-substituted hydroxypropyl methylcellulose 5-40 parts are pressed
Oral liquid, syrup, granule, hard shell capsules or tablet is made in conventional method.
2. the dimension medicine compound preparation method of leucoderma according to claim 1, it is characterised in that following step is pressed in concrete operations
It is rapid to carry out：
A, the Carum carvi 100-5 parts screened are crushed, crosses 10-80 mesh sieves, the water immersion 4-12 for adding 4-20 times of mass ratio to measure is small
When, volatile oil 2-8 hours are extracted with volatile oil extractor, collect volatile oil and aqueous extract respectively；
B, the volatile oil in step a is included according to a conventional method with beta-schardinger dextrin, obtained inclusion compound is dried, standby；
C, the psoralea corylifolia 5-50 parts screened, puncture vine 5-50 parts, ajowan-caraway seed 5-50 parts, turpeth 5-50 parts are mixed broken
Or crush, 10-40 mesh sieves are crossed, the alcohol reflux extraction for being 10-95% with the 4-20 times of water measured or concentration or Soakage extraction 1-4 times,
Each 1-4 hours, filtration, merging filtrate, respectively obtain water or ethanol extract；
D, Aqueous extracts in step c are merged with Aqueous extracts in step a, is concentrated under reduced pressure, dries, crushes, obtain extract powder；
Or will merge when alcohol extract is concentrated under reduced pressure into no ethanol flavor in step c with Aqueous extracts in step a, it is concentrated under reduced pressure, dries, powder
It is broken, obtain extract powder；
E, inclusion compound in step b is merged with extract powder in step d, adds Steviosin 10-100 parts, sorbic acid 10-100 parts, pure
Water, after being uniformly dissolved, plate-frame filtering, dispense, sterilizing, oral liquid is made according to a conventional method；
Or merge inclusion compound in step b with extract powder in step d, add sucrose 10-100 parts, pure water, after being uniformly dissolved, mistake
Filter, dispense, sterilizing, syrup is made according to a conventional method；
Or merge inclusion compound in step b with extract powder in step d, add lactose 10-100 parts, mannitol 10-100 parts, sucrose
10-100 parts or dextrin 10-100 parts, fully mix, granule is made according to a conventional method；
Or merge inclusion compound in step b with extract powder in step d, add lactose 10-100 parts, microcrystalline cellulose 10-100 parts, carboxylic
Methyl starch sodium 5-40 parts, low-substituted hydroxypropyl methylcellulose 5-40 parts, magnesium stearate 1-10 parts, fully mix, according to a conventional method
Hard shell capsules or tablet is made.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1264590A (en) * | 1999-02-26 | 2000-08-30 | 魏鹏 | Medicine for treating leukoderma and preparing process thereof |
| CN101224241A (en) * | 2007-01-20 | 2008-07-23 | 成爱华 | Melanin regeneration pill |
| CN101766779A (en) * | 2010-02-09 | 2010-07-07 | 闫明 | Compound medicine for vitligo treatment and preparation method thereof |
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2017
- 2017-09-13 CN CN201710821860.5A patent/CN107569532A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1264590A (en) * | 1999-02-26 | 2000-08-30 | 魏鹏 | Medicine for treating leukoderma and preparing process thereof |
| CN101224241A (en) * | 2007-01-20 | 2008-07-23 | 成爱华 | Melanin regeneration pill |
| CN101766779A (en) * | 2010-02-09 | 2010-07-07 | 闫明 | Compound medicine for vitligo treatment and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 国家中医药管理局: "《中华本草 维吾尔药卷》", 31 December 2005, 上海科技出版社 * |
| 梁启成等: "《中国壮药学》", 31 May 2005, 广西民族出版社 * |
| 陈琼等: "《中药制剂技术》", 30 November 2014, 中国农业大学出版社 * |
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