CN1739665A - Method of preparing active medicinal matter of lignin from broad-leaved tree and its use - Google Patents
Method of preparing active medicinal matter of lignin from broad-leaved tree and its use Download PDFInfo
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Abstract
The present invention relates to medicine industry, and is especially method of preparing lignin as active medicinal matter from broad-leaved tree and its use in treating acute soft tissue damage and skin damage. The present invention prepares lignin with poplar, birch and other broad-leaved tree material, and through the steps of water vapor extraction in alkaline environment to obtain fraction solution, naturally degrading at normal temperature in the presence of polysaccharidase for 2-18 months to obtain degraded liquid with pH lowered from 14 to 6, separating the degraded liquid to obtain active matter of molecular weight less than 4000, and sterilizing to obtain the medicine liquid for treating acute soft tissue damage and skin damage. The method liquid is applied to skin wound to reach astringency and inflammation-diminishing effects.
Description
Technical field
The present invention relates to medical medicine industry, refer in particular to a kind of method that from broad leaf tree, prepares active medicinal lignin matter and the treatment acute soft tissue sprain and skin injury aspect purposes.
Background technology
Lignin is that a class is a skeleton with phenylpropyl alcohol alkane monomer, has cancellated high molecular polymer, is one of key component of plant cell wall.Since the kind difference of plant, the geographic difference of growing environment, and the extracting method difference of using, and the molecular weight of the lignin that obtains is also different with molecular structure, has caused the purposes difference of lignin.Domestic and international at present application and the research to lignin mainly is: 1, lignin directly concentrates and can be used as the fuel utilization in the papermaking wastewater.2, lignin product is mainly used in: mud water reducer, oil drilling mud pour-point depressant, rubber stiffener, feed additive, the report of aspects such as adhesive and synthetic resin.
Literature survey and data-searching show, external comparatively active in the research aspect medical to lignin, seen about carrying out the research of AIDS resisting poison, cancer, non-cancer, hormone dependence disease and cardiovascular disease have been prevented the report of aspect with lignin.But as stop blooding, invigorate blood circulation, the research of traumatic injury medicine do not appear in the newspapers, as US Patent Application 20040048804.US Patent Application 20020061854 has narrated by giving the cancer of Hydroxymatatairesinoin (hydroxymatairesinol) to the people, some non-cancer, hormone relies on the prevention method of disease and cardiovascular disease, relates to a method that increases metabolite (metabolite) level of enteral fat (enterolactone) in the human serum or another kind of Hydroxymatatairesinoin (hydroxymatairesinol) simultaneously.The metabolite Hydroxymatatairesinoin of wherein using in this invention (hydroxymatairesinol) can be the main lignin of loose Hydroxymatatairesinoin fruit timber/coniferous timber, separate in the compression wood of its fragment (containing branch, legendary small dragon with horns branch and burl) and obtain from bulk.
Summary of the invention
The purpose of this invention is to provide a kind of method that from broad leaf tree, prepares active medicinal lignin matter and the treatment acute soft tissue sprain and skin injury aspect purposes, it can play anti-inflammatory analgetic, blood circulation promoting and blood stasis dispelling.
Preparation process of the present invention is:
1. be the feedstock production lignin active substance with broad leaf trees such as poplar, birch, in alkaline environment, extract with water vapour, because lignin and hemicellulose all are the ingredients of plant cell, hemicellulose-be acidic polysaccharose, vegetable polysaccharides is followed outflow in extracting the lignin process, also rising thereupon of minimizing PH along with vegetable polysaccharides, effluent is elevated to PH14 gradually by PH5, one batch of material continuous extraction in 12 hours ending PH14 finishes, and cuts different fractions according to the variation of PH in the different time is corresponding.
With each fraction solution of above-mentioned preparation in the presence of polyase, room temperature is placed down and was carried out natural degradation lignin unit in 2-18 month, degradation solution is ended between the PH6 by degraded by PH14.With degradation solution process dialysis, contrary impregnating method, methods such as ultrafiltration are carried out initial gross separation to degradation solution, the molecular weight of inside and outside dialysis solution is respectively＜and 300,＜4000, remove＜300 low-molecular-weight composition, the part material of residue＜4000 molecular weight is stable performance, does not have toxic blood circulation promoting and blood stasis dispelling material substantially.
3. get the above-mentioned molecular weight for preparing＜4000, pH value neutrality is extracted under agitation dripping nitric acid during the degradation solution between the subacidity adds reactor, isolate water-insoluble solid, it is lignin-polysaccharide complex active substance, be dissolved in the inorganic salts such as potassium salt, sodium salt, ammonium salt, dissolubility is 1-8%.
4. with the above-mentioned lignin-polysaccharide complex active substance that obtains, be mixed with 0.01--0.05% with above-mentioned inorganic salt, at 100 ℃ of following boiling reaction 2-4 hour sterilizings or with the aperture is that the Mycoderma that removes of 0.22 μ m carries out degerming, obtains can be used for treating that acute soft tissue is sprained and the medicinal liquid of skin injury.
5. get the part active substance of molecular weight＜4000 that above-mentioned steps 3 obtains, further separate with Pu polysaccharide gel G15, acute soft tissue is sprained and skin injury aspect disease for the 300-1500 active medicinal lignin matter is used for the treatment of to get molecular weight.
Because the chemical constitution of the different lignins of lignin raw material sources Different Extraction Method is also different, and the present invention obtains containing in the active medicinal lignin matter structures such as magnolol, Syringin, ferulic acid, salicoside, functions such as that these structures have respectively is antibiotic, antiulcer, hemostasis are invigorated blood circulation, anti-inflammatory analgetic, be to be skeleton because of lignin again with phenylpropyl alcohol alkane monomer, has the network structure amorphous high polymer, this structure of lignin has objectively also formed a physical barriers, stop the intrusion of antibacterial effectively, accelerate the healing of wound.Sprain and during skin injury, use the present invention to rub wiping immediately at acute soft tissue, hemostasis, pain relieving within 3-20 minute.As long as be coated in during skin injury on the wound surface, can play convergence, antiinflammation.And the blood that simultaneously stasis of blood is amassed in the blood vessel outside spreads, and makes it by skin absorbs, and medicine of the present invention can be realized taking stopgap measures effecting a permanent cure.
The specific embodiment
1, preparation active medicinal lignin matter
Getting poplar is the feedstock production lignin active substance, extracts with water vapour in alkaline environment, and batch of material continuous extraction in 12 hours ending PH14 finishes, and cuts different fractions according to the variation of PH in the different time is corresponding.In the presence of polyase, room temperature is placed down and was carried out natural degradation lignin unit in 2-18 month with each fraction solution of above-mentioned preparation, and degradation solution is ended between the PH6 by degraded by PH14.With degradation solution process dialysis, contrary impregnating method, methods such as ultrafiltration are carried out initial gross separation to degradation solution, under agitation dripping nitric acid in the part material of the molecular weight＜4000 adding reactor in the dialysis solution inside and outside extracting extracts, isolate water-insoluble solid, it is lignin-polysaccharide complex active substance, be dissolved in potassium salt, sodium salt, in the inorganic salts such as ammonium salt, dissolubility is 1-8%, being mixed with 0.01% medicinal liquid is that the Mycoderma that removes of 0.22 μ m carries out degerming at 100 ℃ of following boiling reaction 2-4 hour sterilizings or with the aperture, obtain the medicinal liquid that is used to treat, carry out animal experiment.
2. animal experiment
1) animal skin acute toxicity test
50 of extracting waste Cavia porcelluss are divided 5 groups at random, and 10 every group, depilatory is at Cavia porcellus spinal column both sides depilation 40cm
2, being used for experiment after 24 hours, the damaged skin group is scratched skin (degree of being with the oozing of blood) with syringe needle.Intact skin and damaged skin, high dose group only is coated with liquid medicine 2ml/, low dose group 1ml/ only, the blank group is coated with a water 2ml/ water (till being coated with several times), be coated with the liquid medicine group after 24 hours, with warm water flush away liquid medicine, observe and to go behind the medicine in 1,24,48,72 hour and 7 days skin to have or not situations such as ANOMALOUS VARIATIONS and fur, breathing, four skins.
Experimental result
Observe in the flush away liquid medicine 1 hour to 7 days and respectively organize guinea pig skin and all do not have phenomenons such as the erythema of appearance, edema, meanwhile, animal diet followed is normal, fur gloss, breathe, activity freely, do not see ANOMALOUS VARIATIONS such as the watery nasal discharge of shedding tears.Show that this medicine does not have acute toxic reaction to skin.
2) irritant test of the skin of animal
Get 36 Cavia porcelluss, divide 6 groups at random, 6 every group with depilatory at Cavia porcellus spinal column both sides depilations 20Cm
2, being used for experiment after 24 hours, damaged skin scratches skin (degree of being with the oozing of blood) with syringe needle.High dose 2ml/, low dosage 1ml/, a matched group 2ml/ water.
Table one medicinal liquid is to the irritant test of the skin of animal
| Irritant reaction | High dose | Low dosage | Matched group | ||||
| Complete | Damaged | Complete | Damaged | Complete | Damaged | ||
| Erythema | 1, no erythema 2, reluctantly visible 3, obviously visible 4, moderate is to serious 5, Hol and incrustations | - - - - - | 5/6 1/6 - - - | - - - - - | 5/6 1/6 - - - | - - - - - | - - - - - |
| Edema | 1, no edema 2, strong visible 3, visible 4, show as seen | - - - | - - - | - - - | - - - | - - - | - - - |
Transdermal stimulation reaction standards of grading judge that mean scores is 0.16, the basic nonirritant of decidable.
3) skin anaphylactic test of animal
Get 30 Cavia porcelluss, divide 3 groups at random, each 5 first group of every group of male and female are for being subjected to the reagent group, second group of positive medicine matched group, and the 3rd group is the blank group.Spinal column both sides depilation 40Cm
2, be used for experiment after 24 hours, at first sensitization contact.Be subjected to reagent thing group only to be applied to depilation district, left side, continue 6 hours, in the 7th day and the 14th day, with quadrat method coating thing with 0.2ml/.Blank group and positive drug control group are with above-mentioned same method sensitization.(positive drug control group is with 1%2.4 dinitro-chloro-benzene sensitization) each group is all after last sensitization 14 days, and coating removed medicine after 6 hours, reached 12,24,48,72 hours at once and observed the skin allergy situation.
Table two medicinal liquid is to the skin anaphylactic test of animal
| The anaphylaxis situation | Be subjected to the reagent thing | Rigidity medicine matched group | The blank group | |
| Erythema | 1. there are not erythema 2. slight 3. moderates, 4. severes, 5. purplish red eschars | - - - - - | 2/10 7/10 1/10 - - | - - - - - |
| Edema | 1. anhydrous tumor 2. slight 3. moderates, 4. severes 5. are serious | - - - - - | - - - - - | - - - - - |
Exciting 24,48,72 hours skin not see that erythema appears in 1 example, so decidable FA1 does not have irritated reaction substantially.
4) influence of xylol induced mice auricle inflammation
Get 50 of male mice in kunming, body weight 22 ± 2g, divide 5 groups at random, every group 10, irritating stomach respectively gives, dosage get little in big by (respectively 0.2,0.4,0.5ml/10g/ inferior), aspirin 0.1g/Kg/ time and 0.5ml/10g normal saline, administration every day 2 times, for three days on end, after the last administration 1 hour, only evenly be applied in two sides before and after the ear of a mice left side with dimethylbenzene 0.02ml/, auris dextra is in contrast, after 1 hour mice being taken off cervical vertebra puts to death, laying class auricle at the same position of two ears and weigh with the card punch of the straight warp of 9mm, is auricle swelling degree with the difference of two auricle weight, the results are shown in Table:
The influence of table three medicinal liquid xylol induced mice auricle inflammation (X ± SD, n=10)
| Group | Auricle swelling degree (mg) | Auricle edema rate (%) | Suppression ratio (%) |
| The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | 10.8±1.7 9.7±2.6 9.0±2.0 * 8.8±2.2 * 8.4±2.3 ** | 164.6±33.4 140.0±42.1 134.5±39.4 128.8±42.2 120.7±34.1 | 14.9％ 18.3％ 21.7％ 26..7％ |
Compare with matched group,
*P＜0.05,
*P＜0.01
In the medicinal liquid, heavy dose of group all can significantly suppress the Mice Auricle inflammation due to the dimethylbenzene, the suppression ratio maximum can be 21.7%, illustrates to have certain antiinflammatory action.
5) influence of Dichlorodiphenyl Acetate induced mice writhing response
Get 50 of male mice in kunming, body weight 22 ± 2g divides 5 groups at random, 10 every group, irritating stomach respectively gives, dosage get little in big (respectively 0.2,0.4,0.5ml/10g/ time), aspirin reaches 0.5ml/10g normal saline, administration every day 2 times 0.1g/Kg/ time, for three days on end, 1 hour lumbar injection 0.6% acetum 0.1ml/10g after the last administration observes each treated animal in 15 minutes and turns round the body number of times by what acetic acid brought out, the results are shown in Table.
Table four medicinal liquid Dichlorodiphenyl Acetate induced mice writhing response influence X ± SD, n=10
| Group | Turn round the body number of times | Suppression ratio (%) |
| The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | 47.5±12.4 36.6±10.1 * 34.0±9.3 * 32.6±10.5 ** 31.5±12.2 ** | 22.9 28.4 31.4 33.7 |
Compare with matched group,
*P＜0.05,
*P＜0.01
With the normal saline matched group relatively, the mouse writhing reaction due to each dosage Dichlorodiphenyl Acetate of medicinal liquid has obvious inhibitory action, 22.9%, 28.4%, 31.4%, illustrate that it has certain analgesic activity suppression ratio is respectively:.
6) to the rheol influence of stasis syndrome rat blood flow
Get 48 of SD rats, body weight 300-350g male and female half and half are divided 6 groups at random by body weight, 8 every group, irritate stomach respectively and award medicinal dose and be (0.4,0.8,1.0ml/100g) and 1ml/Kg Radix Salviae Miltiorrhizae Injection, blank group and model control group award the normal saline of 1ml/100g.Each is organized and is administered once every day, and administration is 7 days altogether.After administration in the 6th day 1 hour, except that the blank group, the equal subcutaneous injection 0.1% epinephrine 0.2ml of all the other 5 treated animals, inject the same epinephrine of measuring again 1 time after 2 hours, inject once more and rat was put into frozen water in back 2 hours and soaked 5 minutes, water is can't help in fasting, preparation stasis syndrome model.After the administration in the 7th day 1 hour, pentobarbital sodium 40mg/Kg penetrates anesthesia in the abdominal cavity, the blood-letting of carotid artery intubate, the EDTA anticoagulant detects whole blood low shear rate (10mPa.s), middle shear rate (50mPa.s), high shear rate viscosity (120mPa.s), erythrocyte sedimentation rate and packed cell volume and aggregate index.Result of the test sees Table
The table five couple rheol X ± SD that influences of stasis syndrome rat blood flow, n=8
| Group | Blank group | Model group | Small dose group | Middle dosage group | High dose group | The Radix Salviae Miltiorrhizae group |
| Low cutting cut the 50mPa.s height and cut 120mPa.s erythrocyte sedimentation rate mm/h hematocrit % aggregate index among the 10mPa.s | 12.7±4.4 66.6±1.52 5.31±0.93 2.13±1.13 43.8±6.0 2.34±0.44 | 20.9±2.9 ** 9.42±1.0 ** 6.95±0.6 ** 3.23±0.89 * 49.6±3.9 * 3.14±0.29 ** | 20.1±3.9 9.17±1.33 6.77±0.80 3.13±0.83 49.25±2.55 3.09±0.38 | 16.53±3.62 * 7.86±1.15 * 6.03±0.8 * 3.00±0.53 48.3±2.7 2.76±0.47 | 15.7±3.88 ** 7.55±1.43 ** 5.83±0.81 ** 2.25±0.71 * 45.5±4.0 2.61±0.48 * | 15.8±4.8 * 7.64±1.66 * 5.82±1.02 * 2.13±0.83 14.4±5.6 * 2.57±0.50 * |
Compare with the blank group,
*P＜0.05,
*P＜0.01
This medicinal liquid has some improvement to stasis syndrome model hemorheology, can reduce whole blood low cut, in cut, high shear rate viscosity, reduce erythrocyte sedimentation rate, reduce erythrocyte aggregation index, illustrate that this medicinal liquid has certain function of promoting blood circulation to disperse blood clots.
3. clinical experiment
Be used for zooperal medicinal liquid with above-mentioned, further separate, get molecular weight and be used for clinical experiment for the 300-1500 active medicinal lignin matter with Pu polysaccharide gel G15.
Use this medicine as follows to the embodiment that treats the 40 routine patients of soft tissue contusion and carry out observation of curative effect:
(1) 40 people of treatment, male 30 examples, women 10 examples, minimum 6 years old of age, maximum 70 years old, acute stage receive treatment 35 routine non-acute stages 5 example.
(2) clinical manifestation: damaged soft tissue swelling, blood stasis, local pain has skin surface breakage out of the ordinary, and concurrent infection betides the joint, the contusion around the carpal joint, around the articulations digitorum manus, around the joints of foot, limitation of activity.Rub with this medicine and to put 2-10 minute on the skin, hematoma obviously disappears.
(3) 35 routine acute contusion patients, single rubbing in the part with this medicinal liquid put 2-10 minute on the skin, every day 2-5 time, other any medicines of no use, 5 examples are once with after the other drug treatment, and the person uses this medicinal liquid instead because of skin injury infects not more, smeared inferior to the skin ulcer face in one day 4, smeared repeatedly 2-3 minute at every turn.Injured wound surface is because of infecting obstinate, and available this medicinal liquid is smeared, and its inflammation disappears gradually, and wound surface oozes out minimizing, and can grow granulation tissue rapidly with wound repairing, and the wound surface after the reparation is smooth, cures the back wound and does not stay scar substantially.Therapeutic outcome sees Table
Table is observed in the treatment of 40 routine soft tissue contusions
| Time | Soft tissue contusion's (number) | Account for injured ratio (%) |
| Recovery from illness on the 3rd | 25 | 62.5 |
| Recovery from illness on the 5th | 12 | 30 |
| Recovery from illness on the 10th | 3 | 7.5 |
Claims (3)
1. from broad leaf tree, prepare the method for active medicinal lignin matter, it is characterized in that:
(1) be the feedstock production lignin active substance with broad leaf tree poplar, birch, in alkaline environment, extract with water vapour, because lignin and hemicellulose all are the ingredients of plant cell, hemicellulose-be acidic polysaccharose, vegetable polysaccharides is followed outflow in extracting the lignin process, also rising thereupon of minimizing PH along with vegetable polysaccharides, effluent is elevated to PH14 gradually by PH5, one batch of material continuous extraction in 12 hours ending PH14 finishes, and cuts different fractions according to the variation of PH in the different time is corresponding;
(2) with each fraction solution of above-mentioned preparation in the presence of polyase, room temperature is placed down and was carried out natural degradation lignin unit in 2-18 month, degradation solution is ended between the PH6 by degraded by PH14; With degradation solution through dialysis, contrary impregnating method, methods such as ultrafiltration are carried out initial gross separation to degradation solution, the molecular weight of inside and outside dialysis solution is respectively＜300,＜4000, removes＜300 low-molecular-weight composition the part material of residue＜4000 molecular weight;
(3) get the above-mentioned molecular weight for preparing＜4000, pH value neutrality is extracted under agitation dripping nitric acid during the degradation solution between the subacidity adds reactor, isolate water-insoluble solid, it is lignin-polysaccharide complex active substance, be dissolved in the inorganic salts such as potassium salt, sodium salt, ammonium salt, dissolubility is 1-8%;
(4) with the above-mentioned lignin-polysaccharide complex active substance that obtains, be mixed with 0.01-0.05% with above-mentioned inorganic salt, at 100 ℃ of following boiling reaction 2-4 hour sterilizings or with the aperture is that the Mycoderma that removes of 0.22 μ m carries out degerming, obtains can be used for treating that acute soft tissue is sprained and the medicinal liquid of skin injury.
2. the method that from broad leaf tree, prepares active medicinal lignin matter according to claim 1, it is characterized in that: the part active substance of getting molecular weight＜4000 that above-mentioned steps 3 obtains, further separate with Pu polysaccharide gel G15, getting molecular weight is the 300-1500 active medicinal lignin matter.
3. the purposes of preparation active medicinal lignin matter from broad leaf tree is characterized in that getting the above-mentioned molecular weight for preparing and sprains and skin injury aspect disease for 300-1500 active medicinal lignin matter treatment acute soft tissue.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111803402A (en) * | 2020-08-21 | 2020-10-23 | 长春科技学院 | Preparation method of lignin-based sunscreen cream |
| WO2023128777A1 (en) | 2021-12-30 | 2023-07-06 | University Of Belgrade | Synthesized lignin oligomers, method for obtaining and use thereof |
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2005
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111803402A (en) * | 2020-08-21 | 2020-10-23 | 长春科技学院 | Preparation method of lignin-based sunscreen cream |
| WO2023128777A1 (en) | 2021-12-30 | 2023-07-06 | University Of Belgrade | Synthesized lignin oligomers, method for obtaining and use thereof |
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