CN104873562A - Preparation method of water-soluble preparation containing isatan and application thereof for treating high altitude alopecia - Google Patents
Preparation method of water-soluble preparation containing isatan and application thereof for treating high altitude alopecia Download PDFInfo
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- CN104873562A CN104873562A CN201510314722.9A CN201510314722A CN104873562A CN 104873562 A CN104873562 A CN 104873562A CN 201510314722 A CN201510314722 A CN 201510314722A CN 104873562 A CN104873562 A CN 104873562A
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Abstract
The invention relates to a preparation method of a water-soluble medicine preparation containing isatan, and an application thereof for treating high altitude alopecia. The preparation method of the medicine comprises the following steps of adopting the nanometer film (model: JCM-2012-100N) and macroporous adsorbing resin (name: styrene type non-polar copolymer; model: D101) technique to extract the isatis indigotica fortune containing 1% of isatan, enriching and extracting the effective ingredient of isatan, and preparing the crystallizing powder; re-dissolving the crystallizing powder by 10% of distilled water (g/L), and applying to the clinics to obviously treat a high altitude alopecia patient. The prepared medicine has the advantages that the effect of treating the high altitude alopecia is obvious, and is better than the treating effect of 5% minoxidil and normal saline; any allergic reaction on human body is avoided, the side effect on the human body caused by the existing hormone treating the high altitude alopecia is overcome, and the physical pressure on the high altitude alopecia patient is reduced.
Description
Technical field
The present invention relates to the water soluble preparation containing Isatis indigotica Fort. glycosides of the high altitude alopecia for the treatment of, particularly relate to a kind of clinical practice for the treatment of the water soluble preparation containing Isatis indigotica Fort. glycosides of high altitude alopecia.
Background technology
High altitude alopecia is a kind of disease common in highlands, and according to Epidemiological study, in more than height above sea level 3000m area, its sickness rate is up to about 8%.Along with building up of Qinghai-Tibet Railway, this disease of high altitude alopecia will be further outstanding.The treatment of current high altitude alopecia, mainly with whole body or topical application containing hormone medicine, but life-time service hormone can produce a series of obvious side effect, as: Ku Xinshi levies, thinning of skin etc.The stimulating hair growth method such as employing Rhizoma Zingiberis Recens, hot pepper paste of folklore, curative effect is only transient.In view of the side effect that the high altitude alopecia of hormone therapy brings, how to reduce or be current problem demanding prompt solution without the high altitude alopecia of hormone therapy.
Summary of the invention
High altitude alopecia is because High aititude causes in-vivo tissue cell hypoxia, the atrophy of hair follicle anoxia, thus the regeneration metabolism affecting hair, the anagen phase is shortened, and catagen extends, and hair is stagnated and caused alopecia in the early stage state of growth.Wu Sima grass is rich in the active ingredient Isatis indigotica Fort. glycosides and mustard glycosides that promote hair follicle activation, Isatis indigotica Fort. glycosides has the effects such as antiviral, antiinflammatory, Anti-endotoxin Activity, antitumor and enhancing immunity, raw property or lymphatic temperament disease is cured mainly in the traditional Chinese medical science, as weak in capillary root, alopecia, prematured white hair, raw property headache, anus disease, hemorrhoid etc.The visible following ancient books and records of effectiveness of Isatis indigotica Fort. glycosides are recorded: 1. " note doctor allusion quotation ": " restrain astringent or styptic treatment for spontaneous sweating, logical impaction brightness etc.For black etc. of having hair dyed." 2. " white palace ": " astringing to arrest bleeding, the effectiveness such as speckle dispelling blazes, and more wound cures the wound, dye hair color hair.Control various hemorrhage, various speckle (black speck, white macula, purpura), nascent wound, hair white etc." Xinjiang Uygur woman just wipes eyebrow with Wu Sima (European mutation Isatis indigotica Fort.) leaf juice since the childhood, thus allow eyebrows, black, bright, close.For analyzing the effect of the high altitude alopecia of Wu Sima grass treatment further, our specific enrichment extracts its effective ingredient Isatis indigotica Fort. glycosides, is prepared into crystalline powder, and Preliminary Applications, in high altitude hair loss patient, finds that it has definite therapeutical effect.
The object of the present invention is to provide a kind of Isatis indigotica Fort. glycosides aqueous solubility pharmaceutical formulations for the treatment of high altitude alopecia.
The Isatis indigotica Fort. glycosides aqueous solubility pharmaceutical formulations that another object of the present invention is to prepare treats the application of high altitude alopecia in clinical.
The object of the invention is to complete like this:
Treat a medicine Isatis indigotica Fort. glycosides aqueous solubility pharmaceutical formulations for high altitude alopecia, its preparation process is:
A. cleaned by Wu Sima grass, put into 40 DEG C of baking ovens and dry, take out the Wu Sima grass 13.2g of drying, add the water extraction 3 times of 8 times amount, each 0.5 hour, merge extractive liquid, hold over night, gets supernatant;
B. under room temperature environment, supernatant prepared by a step is put into IK16 type centrifuge 3000rpm/min, centrifugal 10min, after centrifugal, by the micropore nanofiltration membrane of supernatant with 0.22 μm of aperture, repeat 3 times, merge ultrafiltrate, then concentrate by NF membrane;
C. concentrated solution rotary evaporator b step obtained continues concentrated under 70 DEG C of environment, concentrated solution is added in macroporous resin, gradient elution is carried out with 30%, 50%, 70%, 90% ethanol water, collect ethanol elution, reclaim ethanol elution and be concentrated into fluid extract, with after distilled water diluting with the extracted with diethyl ether 3 times of equivalent, isolate water layer;
D. in the isolated water layer of step c, add the ethyl acetate of 2 times of water layer amounts, hot reflux 5-6 hour, collect ethyl acetate layer, at normal atmospheric pressure, ethyl acetate layer is concentrated into 1/10th volumes, left at room temperature crystallize, filter, obtain loose blue glycosides crude product;
E. heat d. step water dissolution pine blue glycosides crude product 30 ~ 45 DEG C, filter while hot, place cooling crystallization, 5 times repeatedly, can obtain loose blue glycosides fine work, be canescence imperfect crystal formation powder;
F. press the blue glycosidal crystalline powder of concentration (g/L) distilled water redissolution e. step gained pine of 10%, with the filtering with microporous membrane in 0.22 μm of aperture, obtain the water miscible preparation of loose blue glycosides, by 10ml/ bottle, aseptic subpackaged, for subsequent use.
Physicochemical property containing the water miscible preparation of loose blue glycosides:
Color: colourless;
Form: liquid;
PH value: 7.75;
Dissolubility: with the loose blue glycosidal crystalline powder of distilled water redissolution, can dissolve instantaneously.
The beneficial effect of advantage of the present invention and generation is:
The reason of high altitude alopecia is that High aititude causes in-vivo tissue cell hypoxia, and the atrophy of hair follicle anoxia, thus the regeneration metabolism affecting hair, make the anagen phase shorten, and catagen extends, and hair is stagnated and caused alopecia in the early stage state of growth.Wu Sima grass is rich in the active ingredient Isatis indigotica Fort. glycosides and mustard glycosides that promote hair follicle activation, Isatis indigotica Fort. glycosides has the effects such as antiviral, antiinflammatory, Anti-endotoxin Activity antitumor and enhancing immunity, raw property or lymphatic temperament disease is cured mainly in the traditional Chinese medical science, as weak in capillary root, alopecia, prematured white hair, raw property headache, anus disease, hemorrhoid etc.The present invention collects effective ingredient Isatis indigotica Fort. glycosides in Wu Sima grass, for the high altitude alopecia of clinical treatment, observes its therapeutic effect, finds that there is definite therapeutical effect, achieve optimum economic benefit and social benefit.
Medicine prepared by application the present invention has obvious curative effect in the high altitude alopecia for the treatment of, obviously be better than other medicines (as hormone) to the therapeutic effect of high altitude alopecia, and the extraction of this medicine have employed " NF membrane and macroporous adsorbent resin " technology, leaching process is without additional additive, the water miscible preparation of the blue glycosides of pure pine is used for human body scalp without any anaphylaxis, this just lays a good foundation in clinical for this medicinal application, and this is also the core place of key technology of the present invention.
Accompanying drawing explanation
Photo before Fig. 1 alopecia
Photo before the high altitude alopecia agent treatment of Fig. 2
Photo after the high altitude alopecia agent treatment of Fig. 3
Detailed description of the invention
Wu Sima grass is a kind of Isatis indigotica Fort. specific to Xinjiang, China, grows north latitude about 37 degree.The nearly kind more than 30 in the Isatis indigotica Fort. platymiscium whole world, it is mainly distributed in Central Europe, Mediterranean Region, West Asia and Central Asia.Isatis indigotica Fort. in domestic 6 kinds and 1 mutation, 5 kinds and 1 mutation (namely what is often called Wu Sima grass) are produced in Xinjiang.China is one of NATURAL DISTRIBUTION center of Isatis indigotica Fort. platymiscium, and Xinjiang is Chinese Isatis indigotica Fort. plant the most intensively NATURAL DISTRIBUTION center.Wu Sima grass in the present invention derives from Xinjiang Urumqi.
In view of the compositions such as Isatis indigotica Fort. glycosides in Wu Sima grass have effects such as promoting hair follicle activation, antiviral, antiinflammatory, Anti-endotoxin Activity antitumor and enhancing immunity, the present invention is intended to utilize Isatis indigotica Fort. glycosides function in Wu Sima grass to prepare a kind of water soluble preparation containing Isatis indigotica Fort. glycosides and is applied to the treatment of high altitude alopecia.
One prepares Isatis indigotica Fort. glycosides aqueous solubility pharmaceutical formulations method, the steps include:
1. cleaned by 50g Wu Sima grass, put into 40 DEG C of baking ovens and dry, take out the Wu Sima grass 13.2g of drying, add the water extraction 3 times of 8 times amount, each 0.5 hour, merge extractive liquid, 100ml, hold over night, gets supernatant 98ml;
2. under room temperature environment, supernatant is put into IK16 type centrifuge (SIGMA company of the U.S.) the centrifugal 10min of 3000rpm/min, after centrifugal, the supernatant 92ml micropore NF membrane (model: NF membrane JCM-2012-100N) in 0.22 μm of aperture is filtered, repeat 3 times, merge ultrafiltrate 86ml, then concentrate by NF membrane;
3. concentrated solution 30ml rotary evaporator is continued under 70 DEG C of environment concentrated, concentrated solution is added in macroporous resin (the nonpolar copolymer of styrene type, model: D101 type), gradient elution is carried out with 30%, 50%, 70%, 90% ethanol water, collect ethanol elution 100ml, reclaim ethanol elution and be concentrated into fluid extract, with after 50 ml distilled waters dilutions with the extracted with diethyl ether 3 times of equivalent, isolate water layer 44ml;
4. in isolated water layer, add the ethyl acetate hot reflux 5-6 hour of 2 times, collect ethyl acetate layer, be concentrated into 1/10th volumes at normal atmospheric pressure, left at room temperature crystallize, filter, obtain loose blue glycosides crude product 0.142g;
5. heating water dissolves loose blue glycosides crude product to 35 DEG C, and filtering while hot, let cool crystallize, 5 times repeatedly, can obtain loose blue glycosides fine work 0.113g, is canescence imperfect crystal formation powder.
6. the loose blue glycosidal crystalline powder of concentration (g/L) distilled water redissolution of 10% is pressed, with the filtering with microporous membrane in 0.22 μm of aperture, containing the water miscible preparation of loose blue glycosides, by 10ml/ bottle, aseptic subpackaged, for subsequent use.
the clinical practice of medicine
(1) the present invention is applied to affected part when patient's informed consent.
Dosage: 1ml/cm
2
Store method: white plastic bottle encapsulates
(2) in order to therapeutic effect of the present invention is described, the present invention is divided into three groups plateau type hair loss patient: i.e. medication therapy groups, 5% minoxidil treatment group, blank group.Treat respectively, statistical analysis carried out to result:
1. medication therapy groups (Isatis indigotica Fort. glycosides treatment group): selected for 35 example ages at the high altitude hair loss patient of 12 ~ 64 years old, during treatment, with cotton swab, the water soluble preparation containing Isatis indigotica Fort. glycosides is embrocated in affected part, every day 2 times, each 1ml/cm
2, treat 4 weeks, treated rear observation March.
2. 5% minoxidil treatment group: patient 26 example selecting the different alopecia district of same patients head in medication therapy groups, external 5% minoxidil embrocates affected part, every day 2 times, 1ml/cm
2, treat 4 weeks, treated rear observation March.
3. blank group: select high altitude hair loss patient 10 example in medication therapy groups, embrocate affected part with normal saline external, every day 2 times, 1ml/cm
2, treat 4 weeks, treated rear observation March.
4. therapeutic effect criterion: disease damage portion skin revitalization terminal hair is greater than 20/cm
2for curing, be less than 20/cm
2for effectively, invalid without changing into after treatment.
5. the cure rate of the high altitude alopecia of Drug therapy and effective percentage compare (see table 1) with 5% minoxidil group and blank group, medication therapy groups and 5% minoxidil group and the blank effectiveness comparison (see table 2) organizing three middle of the month after the treatment.
Table 1 is situation after group high altitude alopeciaing therapeutic February respectively
* compare with blank group, P<0.01,
* compare with 5% minoxidil group, P<0.01
Contrast on effect in 3 months after the treatment of table 2 each group completes
Table 1 shows: in medication therapy groups of the present invention, 35 routine high altitude hair loss patients, and except 1 routine patient's disease damage portion skin is without change, all the other have 26 routine patient's disease damage portion skins to occur that newborn terminal hair is greater than 20/cm
2, 8 examples occur that newborn terminal hair is less than 20/cm
2, effective percentage is 97%, and this compares with 5% minoxidil treatment group the curative effect achieving affirmative.In 35 examples of medication therapy groups, there are 15 examples to be the high altitude hair loss patient of recurrent and repeatedly clinical treatment is invalid, after Drug therapy, except 1 example is invalid, all the other all have certain curative effect to this 15 routine patient, and medicine same determined curative effect in the high altitude alopecia for the treatment of recurrent is described.In 35 examples of medication therapy groups, the present invention selects 26 examples, at alopecia district drug application---the water miscible preparation of loose blue glycosides and 5% minoxidil respectively that same patients head is different, 7 routine patient's disease damage portion skins are without change, and all the other have 14 routine patient's disease damage portion skins to occur that newborn terminal hair is greater than 20/cm
2, 5 examples occur that newborn terminal hair is less than 20/cm
2, 5% minoxidil treated effect is 73%.Compare from 5% minoxidil treatment group and medication therapy groups and find that medicine is when treating high altitude alopecia, it promotes that the speed of natural on-off cycles of hair growth is faster than 5% minoxidil treatment group, and new piliation is also obviously better than 5% minoxidil treatment group in glossiness.Except this, the present invention also makes blank with normal saline, finds that there is 1 example and newly sends out hair loss patient disease damage portion skin high altitude and occur that newborn terminal hair is greater than 20/cm
2, 2 routine neopathy damage portion skins occur that newborn terminal hair is less than 20/cm
2effective percentage is 10%, and none example of the high altitude alopecia of recurrent effectively, illustrate for the new high altitude alopecia sent out, there is certain stimulation and have certain curative effect, then effect is poor with common stimulation for the high altitude alopecia of recurrent, and this also used the external curing alopecia effects such as Rhizoma Zingiberis Recens, hot pepper paste, carbolic acid consistent clinically with the past.
Table 2 is medication therapy groups, 5% minoxidil group and blank group therapeutic effect compare.By in drugs compared treatment group, 5% minoxidil group and blank group therapeutic process, the indexs such as the skin color of patient, the length of terminal hair/fine hair on birds or animals gross appreciation, newly piliation, can intuitively to the difference of three groups of therapeutic effect.The value of the growth terminal hair/fine hair on birds or animals hair of normal person's hair is about 7:1, this ratio of high altitude hair loss patient obviously reduces, through three groups of treatments, patient's alopecia district terminal hair/fine hair on birds or animals gross appreciation all has increase in various degree, and this ratio of medication therapy groups is pushed the speed and Hair growth rate is obviously better than 5% minoxidil group and blank group.
fig. 1-3 ispatient XX, male, 27 years old, head adopted cotton swab to embrocate in high altitude alopecia place by the water soluble preparation containing Isatis indigotica Fort. glycosides after there is high altitude alopecia February, every day 2 times, each 1ml/cm
2, treat 4 weeks, treated rear observation March.After 3 months, visible patient's alopecia district has newborn terminal hair to grow, and in unit are, number of hairs increases.Fig. 1 is before alopecia, and Fig. 2 is before high altitude alopeciaing therapeutic, and Fig. 3 is for after use Isatis indigotica Fort. glycosides Solution In The Treatment, and Fig. 3 can find out: the alopecia position hair of Drug therapy is grown.
From clinical practice, we recognize: high altitude alopecia is a kind of disease common in highlands, according to Epidemiological study, in more than height above sea level 3000m area, its sickness rate is up to about 8%, its reason is that High aititude causes in-vivo tissue cell hypoxia, the atrophy of hair follicle anoxia, thus the regeneration metabolism affecting hair, make the anagen phase shorten, catagen extends, and hair is stagnated and caused alopecia in the early stage state of growth.In addition high altitude alopecia also affects by the irradiation of plateau intense UV rays and plateau psychologic factors etc.; High aititude makes quality of human sleeping poor, too increases the sickness rate of high altitude alopecia, on dietary habit simultaneously, High aititude air pressure is lower, major part food could will eat after pressure cooker processing, thus causes the disappearance of a large amount of vitamin, and this is also another major incentive of high altitude alopecia.Wu Sima grass is rich in the active ingredient Isatis indigotica Fort. glycosides and mustard glycosides that promote hair follicle activation, and Isatis indigotica Fort. glycosides has the effects such as antiviral, antiinflammatory, Anti-endotoxin Activity antitumor and enhancing immunity.The present invention collects effective ingredient Isatis indigotica Fort. glycosides in Wu Sima grass and is prepared into Isatis indigotica Fort. glycosides water soluble preparation, determining that its biological safety is guaranteed, and under meeting the prerequisite of ethics, be used for the treatment of high altitude hair loss patient, obtain tangible result, thus further disclose the effect that the careless active ingredient Isatis indigotica Fort. glycosides be rich in of Wu Sima has promotion hair follicle growth.
Claims (2)
1. prepare a method for Isatis indigotica Fort. glycosides aqueous solubility pharmaceutical formulations, the steps include:
A. cleaned by Wu Sima grass, put into 40 DEG C of baking ovens and dry, take out the Wu Sima grass of drying, add the water extraction 3 times of 8 times amount, each 0.5 hour, merge extractive liquid, hold over night, gets supernatant;
B. under room temperature environment, supernatant prepared by a step is put into IK16 type centrifuge 3000rpm/min, centrifugal 10min, after centrifugal, by the micropore nanofiltration membrane of supernatant with 0.22 μm of aperture, repeat 3 times, merge ultrafiltrate, then concentrate by NF membrane;
C. concentrated solution rotary evaporator b step obtained continues concentrated under 70 DEG C of environment, concentrated solution is added in macroporous resin, gradient elution is carried out with 30%, 50%, 70%, 90% ethanol water, collect ethanol elution, reclaim ethanol elution and be concentrated into fluid extract, with after distilled water diluting with the extracted with diethyl ether 3 times of equivalent, isolate water layer;
D. in the isolated water layer of step c, add the ethyl acetate of 2 times of water layer amounts, hot reflux 5-6 hour, collect ethyl acetate layer, at normal atmospheric pressure, ethyl acetate layer is concentrated into 1/10th volumes, left at room temperature crystallize, filter, obtain loose blue glycosides crude product;
E. heat blue glycosides crude product to 30 ~ 45 DEG C of Step d water dissolution pine, filter while hot, place cooling crystallization, 5 times repeatedly, can obtain loose blue glycosides fine work, be canescence imperfect crystal formation powder;
F. press the blue glycosidal crystalline powder of concentration g/L distilled water redissolution step e gained pine of 10%, with the filtering with microporous membrane in 0.22 μm of aperture, obtain the water miscible preparation of loose blue glycosides, by 10ml/ bottle, aseptic subpackaged, for subsequent use.
2. use claim 1 to prepare the application of Isatis indigotica Fort. glycosides aqueous solubility pharmaceutical formulations in the high altitude alopecia for the treatment of.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111035668A (en) * | 2019-12-11 | 2020-04-21 | 中国科学院新疆理化技术研究所 | Preparation method and application of instant isatis tinctoria leaf extract |
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| CN101701028A (en) * | 2009-10-26 | 2010-05-05 | 新疆奥斯曼生物科技有限公司 | Preparation method of indole glycoside |
| CN102133166A (en) * | 2011-03-18 | 2011-07-27 | 乌鲁木齐一枝好生物科技有限公司 | Plant hair reviving, blacking and strengthening nutrient solution and preparation method thereof |
Non-Patent Citations (1)
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111035668A (en) * | 2019-12-11 | 2020-04-21 | 中国科学院新疆理化技术研究所 | Preparation method and application of instant isatis tinctoria leaf extract |
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