CN104367992A - New application of type I collagen in preparation of medicament for treating and/or preventing psoriasis - Google Patents
New application of type I collagen in preparation of medicament for treating and/or preventing psoriasis Download PDFInfo
- Publication number
- CN104367992A CN104367992A CN201410556819.6A CN201410556819A CN104367992A CN 104367992 A CN104367992 A CN 104367992A CN 201410556819 A CN201410556819 A CN 201410556819A CN 104367992 A CN104367992 A CN 104367992A
- Authority
- CN
- China
- Prior art keywords
- ntx
- purposes according
- animal tissue
- protein
- albumen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to a new application of type I collagen, namely a new application of the type I collagen in preparation of a medicament for treating and/or preventing psoriasis, and in particular relates to the new application of the type I collagen in preparation of the medicament for treating and/or preventing psoriasis.
Description
Technical field
The present invention relates to the novelty teabag of NTx albumen, the novelty teabag, particularly NTx protein peptide that namely treat and/or prevent in psoriasis in preparation are preparing the novelty teabag treated and/or prevented in psoriasis.
Background technology
Psoriasis is the inflammatory dermatoses of chronic recurrent of a kind of induce by multifactor, polygenic inheritance, and it has a strong impact on the quality of life of patient.It is introduced, its Therapeutic Principle is symptom management, delay of progression, mitigation symptoms, reduces recurrence.Clinically, psoriatic medicine and method are divided into topical therapeutic and systematic treating.Topical therapeutic comprises the use medicine such as glucocorticoid, dithranol or photochemotherapy; Systematic treating comprises the oral retinoic acid of use, methotrexate, ciclosporin and biological preparation.The position that the selection factor affecting Therapeutic Method comprises the age, psoriatic type, skin lesion involve and degree, the treatment in past and the disease of merging.Biological preparation is different with the principle of traditional therapy, and both can synergism.
Psoriasis diagnosis basis:
1. primary lesion be foxtail millet grain to Semen phaseoli radiati size pale red pimple, above cover multilamellar silvery white squama, strike off rear visible thin film and petechial hemorrhage phenomenon.In the course of disease, skin lesion form can have a drop-wise to arrive the differentiation of map shape again to coin shape.Border is clear, the pruritus of normal companion's varying degree.
2. skin lesion is apt to occur in scalp and extremity stretch side.Hair damages and often causes hair and become tufted outward appearance, but do not accompany alopecia.
3. minority case can involve margo palpebrae, lip, buccal mucosa, glans penis and foreskin.
4. deck is often in pitting, also can turn yellow, thicken and fingernail stripping.
5. be generally the heavy summer in winter light, normal recurrent exerbation.
6. histopathology has additive diagnostic value, and show that keratinization of epidermis is complete, visible microabscess is formed, acanthosis, and nail is prominent to be stretched down regularly, and dermal papilla is bar-shaped, inside has and bends and the blood capillary of expansion.
7. the classification of the order of severity: before working out rational therapeutic scheme to psoriatic, clinicist needs to assess the psoriatic order of severity.A method simply defining severity of psoriasis is called very regular: namely body surface area (BSA) >10% (areas of 10 palms) or psoriatic lesion area >10% that gets involved is severe psoriasis.BSA<3% is slight, and 3% ~ 10% is moderate.
Medicine and Scheme Choice:
1. local topical medicine: select the various external preparation such as emollient, keratoplastics, keatolytics, vitamin D 3-derivatives, glucocorticoid, tretinoin preparation, dithranol, tar class according to the state of an illness.Selection medication and administration time length should depending on the state of an illness.In/potent glucocorticoid, calcipotriol, tazarotene can be used as the first-line drug of topical therapeutic.
2. naturopathy: the naturopathy means such as long wave ultraviolet (UVA), photochemotherapy (PUVA), wide range UVB, narrow spectrum UVB, laser, photodynamics can be selected.
3. Systemic administration: tretinoin medicines, immunosuppressant, biological preparation, anti-infectives, immunomodulator, determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs.
4. conjoint therapy: conjoint therapy refers to two or more method coupling, often and phototherapy or systematic treating coupling, thus it is minimum that the untoward reaction of various treatment is down to for topical therapeutic.Phototherapy also can with multiple biological preparation therapeutic alliance psoriasis, improve treatment effective percentage.
5. sequential therapy refers to and first uses a kind of potent medicine to remove skin lesion, then uses a kind of medicine that is safer, weak effect instead and carrys out maintaining treatment.
Above Therapeutic Method can be used alone or use in conjunction, has different use taboos during application for the psoriasis of various Clinical types and dissimilar psoriasis.
Collagen protein is a kind of structural protein of extracellular matrix, and English name " collagen ", is developed by Greek, GL-PP, in white, containing a small amount of galactose and glucose, be the main component of extracellular matrix (ECM), account for 85% of collagen fiber solids.Collagen protein is ubiquitous a kind of high molecular weight protein in animal body, is mainly present in the connective tissue (bone, cartilage, skin, tendon, tough etc.) of animal, accounts for 25% ~ 30% of total protein in mammalian body, be equivalent to 6% of body weight.Many marine organisms, as the skin of Fish, protein content is even up to more than 80%.To body and internal organs play support, protection, combine and formed boundary every etc. effect.Except biomechanics aspect, also there is the functions such as the transport of such as signal transduction, somatomedin and cytokine.
Containing a large amount of glycine, proline, hydroxyproline and hydroxylysine, alanine and glutamic acid in collagen protein, be insoluble to diluted acid dilute alkaline soln, not easily by general protease hydrolysis, not easily digested.Hydroxyproline and hydroxylysine are that other protein are unexistent, hydroxylysine residues is glycosylation necessary in tropocollagen molecule, it is closely related with the formation that is crosslinked and fiber of lysine and tropocollagen molecule, and the hydroxyl of hydroxyproline (being formed after the hydroxylation of proline hydroxylase) is all absolutely necessary, so the stable in properties of collagen protein for the formation of hydrogen bond and the stable of triple helix structure.And the essential amino acids content such as tryptophan, tyrosine and methionine are low, therefore collagen protein belongs to incomplete protein.
Collagen peptide is from collagen protein, extract the product obtained, the mean molecule quantity of collagen peptide is at about 1.000Da, three grades of superhelixes of relative collagen protein complexity, the linear structure of collagen peptide makes its digestibility up to 80% (collagen protein is <=1%).Collagen peptide is the hydrolyzate of collagen protein.Being the minimum unit of collagen protein, is a kind of high-purity peptide having glycine in N end band, and compared with the molecular weight of common collagen protein 2.000 ~ 3.000Da, collagen peptide can be absorbed and utilized by the body without the need to decomposing.
Up to now, found the collagen molecules that more than 19 kinds aminoacid sequences are different, these collagen molecules are very different on aminoacid sequence, in morphosis, physicochemical properties and physiological function.
Protein is topmost component in organism, and the skin, the hair that comprise covering human body are all made up of protein.Therefore, protein almost dominates the whole vital movement of biosphere, as cell division, metabolism, transmission of information, and the resistivity etc. to disease, be all that dependent protein matter has been come.Therefore, in cosmetics, using protein as active substance.The natural materials that U.S. CTFA cosmetic material handbook is employed, with in " functional cosmetics raw material ", has collagen protein or its hydrolyzate (hydroxyproline, proline, serine) in the natural materials selected.Collagen protein has whitening function, this is due to melanin mainly quinoid polymer, containing tyrosine residue in collagen protein, due to the similarity of chemical constitution, the tyrosine of tyrosine residue in skin of the collagen protein added in cosmetics is competed and is combined with the active center of tryrosinase, thus restraint of tyrosinase catalysis propylhomoserin is converted into DOPA quinone, stops melanic formation in skin, reach whitening function.
In mammal, it has been found that the collagen protein of 27 types, wherein I, II, III, IV, V, Ⅺ, X X IV and X X VII Collagen Type VI be fibril collagen protein, all the other all belong to non-protofibre collagen protein.In collagen protein, glycine accounts for 1/3 of total amino acid content, and hydroxyproline accounts for 10% of total amino acid content.Skeleton is made up of 1/3 Organic substance and 2/3 inorganic matter, wherein the 80%-90% of organic component is NTx albumen, the macromole that the subunit of NTx albumen is made up of 3 polypeptide chains, form many covalent bonds between its polypeptide chain, interlinkage is tridimensional network.In NTx, hydroxyproline is about 0.7 ~ 0.8 with the ratio of proline.
Understanding at present due to psoriasis is a kind of disease throughout one's life, there is no effective medicine at present.And clinical treatment mainly uses immunosuppressant, but these immunosuppressant curative effects are scarcely desirable and toxic and side effects is large.In existing open source literature, collagen protein is common to be brightened for beauty treatment, the report that so far there are no uses NTx albumen or NTx protein peptide to treat psoriasis.
Summary of the invention
The object of this invention is to provide NTx albumen and prepare the novelty teabag treated and/or prevented in psoriasis;
Another object of the present invention is to provide NTx protein peptide and is preparing the novelty teabag treated and/or prevented in psoriasis;
Another object of the present invention is to provide the preparation method of NTx albumen;
Another object of the present invention is to provide the preparation method of NTx protein peptide.
The object of the invention is by following technical scheme realize:
NTx albumen of the present invention, NTx protein peptide come from animal tissue, comprise skeleton, fish scale, fin etc.
Inventor finds in the psoriatic therapeutic process of research, and NTx albumen, NTx protein peptide can obviously improve guinea pig experimental psoriasis.NTx albumen, NTx protein peptide treat psoriatic clinical research and the display of pharmacodynamic study result: NTx albumen, NTx protein peptide not only can treat psoriasis; with put on the skin compared with glucocorticoid calcipotriol outside existing Western medicine therapy; discovery NTx albumen, NTx protein peptide are not only treated psoriatic effect and are better than prior art; the skin smooth of Cavia porcellus can also be made; moist, play effect of protection skin.
On this basis, the present inventor, by after on purpose controlling NTx albumen, NTx protein peptide use amount, makes it be used for the treatment of and/or prevents psoriatic more remarkable effect.
NTx albumen, NTx protein peptide are for the preparation for the treatment of and/or preventing psoriatic medicine, and when during Clinical practice, every day, oral dose amounted to protein 1600-10000mg, effect is better;
Preferably, best results when every day, oral dose amounted to protein 3000-6000mg.
Onset of psoriasis reason is complicated, present study general is thought, the principal element that psoriatic morbidity relates to has the change of keratinocyte (KC cell), dendritic cell (DC cell), mastocyte (MC cell), macrophage and T cell function.These cells and relevant cell factor thereof, chemotactic factor interphase interaction and then mediate a series of inflammatory reaction, epidermal proliferation, parakeratosis, acanthosis and Blood vessel pattern etc.
[8].Deepen continuously along with to the study on mechanism of the cytokine related in psoriatic generation and time-continuing process and ceS signal molecules, these important intermolecular interactions and cytokine provide many therapy target, and the curing psoriasis medicine much researched and developed for these target spots has gone through go on the market or carrying out clinical trial.
The significant curative effect that NTx albumen and NTx protein peptide obtain when treating psoriasis, according to a preliminary estimate with following mechanism about the protein content ratio in: NTx albumen and NTx protein peptide and normal human skin is comparatively close, DNA and keratin synthesis in specified protein contained by it or some hormone energy regulating cell, antiproliferative effect, inducing cell normal differentiation, immunity moderation intracellular cytokine discharges, induction KC produces the medium with antiinflammatory or immunoregulation effect, the generation of inflammation-inhibiting cytokine, thus produce the psoriatic antiinflammatory anti-proliferative effect for the treatment of.
Can say so, NTx albumen and NTx protein peptide, the most close with the protein ratio in normal human skin, NTx albumen and NTx protein peptide over the course for the treatment of its effect are balance adjustment, regulate DNA and keratin synthesis in cell, not not merely Competitive assays, so effect of the existing whitening of ability, in treatment psoriasis, demonstrate pleasantly surprised effect again.
The safety of patent clinical practice of the present invention is higher, and to can yet be regarded as a kind of good medicine in the psoriatic Drug therapy for the treatment of, it has widened the psoriatic new method for the treatment of, treats left untoward reaction after psoriasis for patient solves prior art simultaneously.
Therefore, the novelty teabag that applicant provides NTx albumen, NTx protein peptide can be used in treating psoriasis aspect.
The safety of patent clinical practice of the present invention is higher, and to can yet be regarded as a kind of good medicine in the psoriatic Drug therapy for the treatment of, it has widened the psoriatic new method for the treatment of, treats left untoward reaction after psoriasis for patient solves prior art simultaneously.
Therefore, the novelty teabag that applicant provides NTx albumen, NTx protein peptide can be used in treating psoriasis aspect.
The invention also discloses the preparation method of NTx albumen, comprise the following steps: choose animal tissue, pulverize with after clear water rinsing, add animal tissue weight 3-10 water heating extraction 1-8 hour doubly, extract 1-3 time, filter, merging filtrate, dry, obtain NTx albumen.
The invention also discloses the preparation method of NTx albumen, comprise the following steps: choose animal tissue, cleaning Hou Jia animal tissue weight 5-10 acid soak 5-120 minute doubly, filter, animal tissue pulverizes with after clear water rinsing, add animal tissue weight 3-10 water heating extraction 1-8 hour doubly, extract 1-3 time, filter, merging filtrate, drying, obtains NTx albumen.
The invention also discloses the preparation method of NTx protein peptide, comprise the following steps: get NTx albumen, add protease hydrolysis 20-60 minute when 50-60 DEG C after, heat inactivation, gets supernatant, concentrated, dry, obtains NTx protein peptide.
NTx protein peptide disclosed by the invention can also be prepared by following steps and obtain: choose animal tissue, clean, drain, and pulverizes, and adds animal tissue weight 5-10 acid soak 5-120 minute doubly, filters, and is 5-6 with clear water rinsing to pH; Add the NaCl solution immersion 1-10 hour that concentration is 1-5%, added NaCl solution is 8-12 times of animal tissue's weight; Be chilled to 3-10 DEG C, add protease, hydrolysis 2-6 hour, after hydrolysis terminates, heats deactivation in 20-40 minute; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide.
Described in preparation method of the present invention, animal tissue comprises skeleton, fish scale, fin.
When adding acid soak described in preparation method of the present invention, select any one among the hydrochloric acid of 0.5-2%, glacial acetic acid, lactic acid, acetic acid, citric acid or malic acid.
Protease described in preparation method of the present invention is any one among neutral protease, acid protease or alkaline protease.
Protease described in preparation method of the present invention is any one or two or more combinations among pepsin, trypsin, cathepsin, papain or subtilisin.
Preparation described in preparation method of the present invention treats and/or prevents leucoderma medicament, is to be prepared from by NTx albumen or NTx protein peptide and pharmaceutically acceptable carrier combination.
The novelty teabag that NTx albumen provided by the invention or NTx protein peptide can be used in treating psoriasis aspect has the following advantages:
1, NTx albumen provided by the invention, NTx protein peptide are native protein, and safety non-toxic, can long-term taking.
2, the present invention is after treatment guinea pig experimental psoriasis, and bilateral skin of pinna cosmetic variation, skin color transfer to normally, smooth surface, and alopecia phenomenon makes moderate progress, and the blood capillary of expansion is repaired, and various symptom obviously alleviates or disappears; Histology Baker marks and inflammatory cell infiltration scoring obvious rising compared with blank group, and difference has statistical significance (P < 0.01), proves that the psoriatic model of Cavia porcellus is successfully established.And treatment group histology Baker marks and inflammatory cell infiltration scoring obvious reduction compared with observation group after modeling, difference has statistical significance (P < 0.01), and the every Score index for the treatment of group is all better than matched group, there is significant difference (P<0.05).Overall treatment effect shows: the present invention can improve Cavia porcellus ear Pigs with Psoriasis symptom and pathological characters thereof, and therapeutic effect optimum is NTx protein peptide, and NTx albumen takes second place, but both equal significances are better than matched group.
3, NTx albumen, the NTx protein peptide prepared of the present invention is simple for process, can be used for producing popularization greatly.
Detailed description of the invention
The present invention is further illustrated below by embodiment.It should be understood that embodiments of the invention are for illustration of the present invention instead of limitation of the present invention.Essence according to the present invention all belongs to the scope of protection of present invention to the simple modifications that the present invention carries out.Except as otherwise noted, the percent of the amount of alcohol in the present invention is percentage by volume, and v/v represents the volume ratio of solution.
Embodiment 1:
Choose Os Gallus domesticus, after cleaning, add the soak with hydrochloric acid 5 minutes of 0.5% of Os Gallus domesticus weight 5 times, filter, pulverize after the rinsing of Os Gallus domesticus clear water, add the water heating extraction 1 hour of Os Gallus domesticus weight 10 times, extract 3 times, filter, merging filtrate, drying, obtains NTx albumen, and purity is 30%.
Embodiment 2:
Choose Os Sus domestica, clean, drain, pulverize, the citric acid adding 2% of Os Sus domestica weight 5 times soaks 5 minutes, filters, and is 6 with clear water rinsing to pH; Add concentration be 1% NaCl solution soak 10 hours, added NaCl solution is 12 times of Os Sus domestica weight; Be chilled to 10 DEG C, add trypsin, be hydrolyzed 6 hours, after hydrolysis terminates, heat deactivation in 40 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 78%.
Embodiment 3:
Choose fishbone, the lactic acid adding 2% of fishbone weight 10 times after cleaning soaks 120 minutes, filters, pulverizes after the rinsing of fishbone clear water, adds the water heating extraction 8 hours of fishbone weight 3 times, extracts 1 time, filters, filtrate, and drying, obtain NTx albumen, purity is 45%.
Embodiment 4:
Choose fishbone, clean, drain, pulverize, the glacialacetic acid adding 1.5% of fishbone weight 10 times soaks 120 minutes, filters, and is 5 with clear water rinsing to pH; Add concentration be 5% NaCl solution soak 1 hour, added NaCl solution is 8 times of fishbone weight; Be chilled to 3 DEG C, add cathepsin, be hydrolyzed 2 hours, after hydrolysis terminates, heat deactivation in 20 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 60%.
Embodiment 5:
Choose Os Bovis seu Bubali, clean, drain, pulverize, with Os Bovis seu Bubali weight 8 times 2% malic acid soak 20 minutes, filtering, is 5 with clear water rinsing to pH; Add concentration be 2% NaCl solution soak 8 hours, added NaCl solution is 10 times of Os Bovis seu Bubali weight; Be chilled to 5 DEG C, add subtilisin, be hydrolyzed 4 hours, after hydrolysis terminates, heat deactivation in 30 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 50%.
Embodiment 6:
Choose fin, clean, drain, the acetic acid adding 1% of fin weight 10 times soaks 100 minutes, filters, and is 6 with clear water rinsing to pH; Add concentration be 4% NaCl solution soak 3 hours, added NaCl solution is 12 times of fin weight; Be chilled to 8 DEG C, add pepsin, be hydrolyzed 5 hours, after hydrolysis terminates, heat deactivation in 40 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 90%.
Embodiment 7:
Choose fish scale, after cleaning, add the water of fish scale weight 4 times amount, extract 4 times, Extracting temperature is 120 DEG C, merges 4 extracting solution, filters, filtrate concentrates, and obtains fish scale extract, adds 0.3% neutral protease insulation hydrolysis 30 minutes, after hydrolysis terminates, heat deactivation in 30 minutes, obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 85%.
Embodiment 8:
Choose fish scale, after cleaning, add the soak with hydrochloric acid 5 minutes of 0.5% of fish scale weight 5 times, filter, pulverize after the rinsing of fish scale clear water, add the water heating extraction 1 hour of fish scale weight 10 times, extract 3 times, filter, merging filtrate, drying, obtains NTx albumen, and purity is 80%.
Embodiment 9:
Choose fish scale, clean, drain, the citric acid adding 2% of fish scale weight 5 times soaks 5 minutes, filters, and is 6 with clear water rinsing to pH; Add concentration be 1% NaCl solution soak 10 hours, added NaCl solution is 12 times of fish scale weight; Be chilled to 10 DEG C, add trypsin, be hydrolyzed 6 hours, after hydrolysis terminates, heat deactivation in 40 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 97%.
Embodiment 10:
Choose fish scale, the lactic acid adding 2% of fish scale weight 10 times after cleaning soaks 120 minutes, filters, pulverizes after the rinsing of fish scale clear water, adds the water heating extraction 8 hours of fish scale weight 3 times, extracts 1 time, filters, filtrate, and drying, obtain NTx albumen, purity is 75%.
Embodiment 11:
Choose fish scale, clean, drain, the glacialacetic acid adding 0.5% of fish scale weight 10 times soaks 120 minutes, filters, and is 5 with clear water rinsing to pH; Add concentration be 5% NaCl solution soak 1 hour, added NaCl solution is 8 times of fish scale weight; Be chilled to 3 DEG C, add cathepsin, be hydrolyzed 2 hours, after hydrolysis terminates, heat deactivation in 20 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 83%.
Embodiment 12:
Choose fish scale, after cleaning, add the soak with hydrochloric acid 5 minutes of 0.5% of fish scale weight 5 times, filter, pulverize after the rinsing of fish scale clear water, add the water heating extraction 1 hour of fish scale weight 10 times, extract 3 times, filter, merging filtrate, dry, obtain NTx albumen, 50 DEG C time, add papain hydrolysis after 60 minutes, heat inactivation, get supernatant, concentrated, dry, obtain NTx protein peptide, purity is 65%.
Embodiment 13:
Choose fish scale, clean, drain, the malic acid adding 2% of fish scale weight 8 times soaks 20 minutes, filters, and is 5 with clear water rinsing to pH; Add concentration be 2% NaCl solution soak 8 hours, added NaCl solution is 10 times of fish scale weight; Be chilled to 5 DEG C, add subtilisin, be hydrolyzed 4 hours, after hydrolysis terminates, heat deactivation in 30 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 90%.
Embodiment 14:
Choose fish scale, clean, drain, the acetic acid adding 1% of fish scale weight 10 times soaks 100 minutes, filters, and is 6 with clear water rinsing to pH; Add concentration be 4% NaCl solution soak 3 hours, added NaCl solution is 12 times of fish scale weight; Be chilled to 8 DEG C, add pepsin, be hydrolyzed 5 hours, after hydrolysis terminates, heat deactivation in 40 minutes; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide, purity is 60%.
Embodiment 15:
Example 1 gained NTx albumen, add protease hydrolysis when 50-60 DEG C after 60 minutes, heat inactivation, gets supernatant, concentrated, and dry, obtain NTx albumen, purity is 65%.
Effect experiment
The effect of experimental example 1 pair of guinea pig experimental curing psoriasis
1 experiment material
1.1 laboratory animals: healthy guinea pig 200, initial weight 250-300g, is provided by Guangxi University of Chinese Medicine's animal experimental center.
1.2 Experimental agents:
NTx albumen: the embodiment of the present invention 1, embodiment 8 prepare gained sample;
NTx protein peptide: the embodiment of the present invention 2, embodiment 9 prepare gained sample;
Formalin: be made into 10% concentration for subsequent use;
Ethanol: be made into 60%, 70%, 80%, 90%, 100% concentration for subsequent use;
Dimethylbenzene; Hematoxylin; Yihong; Calcipotriol;
5% Propranolol ointment: get propranolol hydrochloride 5g and be dissolved in the ethanol of appropriate 50%, add people Azone-propylene glycol 5mL as compound accelerant, adding ketone K (PVPK) 305g is filmogen, and the ethanol finally adding 50% makes into 100mL, stirs evenly and get final product.
2 experimental techniques
2.1 animal grouping and modelings
After Cavia porcellus adaptability raises 3 weeks, random take out 20 and do blank group, remaininging 180, to be evenly applied to Cavia porcellus with 5% Propranolol ointment hard of hearing, 1cm
2medication 0.3g, smoothens with Glass rod, every day 3 times, thickness 1.0mm.To the 3rd weekend, the random 2O that puts to death only does ear tissue pathological examination, if any Psoriasis-like Pathological Changes, then points out modeling success, and is classified to model group.All the other 160 model mouses are divided into 8 groups at random: observation group, matched group, treatment 1 group after modeling, treat 2 groups, treat 3 groups, treat 4 groups, treat 5 groups, treat 6 groups, often organize 20.
2.2 administration
Blank group, model group do not do any process.
Administration from the 10th day of modeling type, every day 2 times, totally 4 weeks.
Matched group: put calcipotriol outward on the skin appropriate,
Treat 1 group: oral embodiment 1 NTx albumen, one time oral dose amounts to protein 5mg/100g;
Treat 2 groups: oral embodiment 8 NTx albumen, one time oral dose amounts to protein 5mg/100g;
Treat 3 groups: oral embodiment 2 NTx protein peptide, one time oral dose amounts to protein 5mg/100g;
Treat 4 groups: oral embodiment 9 NTx protein peptide, one time oral dose amounts to protein 5mg/100g;
Treat 5 groups: oral embodiment 2 NTx protein peptide, one time oral dose amounts to protein 5mg/100g, and it is appropriate to put calcipotriol outward on the skin.
Treat 6 groups: oral embodiment 9 NTx protein peptide, one time oral dose amounts to protein 5mg/100g, and it is appropriate to put calcipotriol outward on the skin.
Matched group and treatment 1-6 group irradiate weekly NB-UVB 3 times, and predose is 70%MED, increases progressively 20% at every turn, 3 times weekly, totally 4 weeks.
2.3 specimen preparations: the skin of pinna specimen that the 4th week is taken off, skin histology fixes 2 weeks with 10% formalin immediately respectively, running water 24h will be organized, the up dehydration of 70%, 80%, 90%, 95%, 100% ethanol is used respectively to high concentration by low concentration, dimethylbenzene is transparent, paraffin embedding, section, incubator 45 DEG C of roasting 24h.Dimethylbenzene dewaxing 10min × 3 time.By high concentration to low-concentration ethanol descending enter water: 100%, each 1min of the ethanol of 95%, 90%, 85%, 80%, 70%, 50%.Tap water 3min.Brazilwood extract dyeing 5min.Tap water 5min.L% acidic alcohol differentiation 20s.Tap water 1min.1% ammonia returns blue 20s.Tap water 1min.Eosin stains lmin.By low concentration to the up dehydration of high concentration ethanol, dimethylbenzene is transparent, neutral gum mounting, and every Cavia porcellus is got both sides ear specimen and make 3 slice, thin pieces.
3 experimental results
3.1 perusals: the change of gross examination of skeletal muscle bilateral skin of pinna cosmetic variation, skin color, blood capillary, squama dropping situations etc.The results are shown in Table 1.
The each treated animal visual results of table 1
Blank group Cavia porcellus is in high spirits, and reaction is quick, and bellicose, hair is dense glossy, is close to skin, and diet, two is just normal.All occur after all modeling animal l weeks that spirit is poor, bradykinesia, happiness is sleeping, and feed reduces, weight loss, and hair is matt, easily comes off.After 3 weeks, modeling animal above-mentioned symptom increases the weight of and occurs ear's phenomenon of scratching.
Modeling is after 2 weeks, and coating place, model group ear of animal back hair part comes off without Rhizoma Imperatae, and the hard of hearing local skin of Some Animals is red, and telangiectasis is obvious.Blank group Cavia porcellus ears hair is normal without coming off.
During medication, after modeling, observation group's symptom is without significant change; After treating 4 weeks, treatment 1-6 group and matched group Cavia porcellus above-mentioned symptom obviously alleviate; And treatment 1-6 group above-mentioned symptom disappears substantially, has compared notable difference with matched group.
4 period-luminosity Microscopic observation E coloration results after 3.2 medications
Light Microscopic observation every Cavia porcellus both sides ear makes 3 sections, under light microscopic, random selecting 6 visuals field, utilize computer image analysis software, carry out graphical analysis to choosing the visual field, carry out histological score assessment with reference to Baker method, and inflammatory cell is counted.The results are shown in Table 2, table 3.
The each treated animal histomorphometric outcomes of table 2
Table 3 Cavia porcellus ear Psoriasis-like Pathological Changes histological score and the scoring of inflammatory cell infiltration situation
Note: after modeling, observation group compares with blank group, △ P<0.01; Compare with observation group after modeling, * P<0.01; Compare with matched group, #P<0.05.
Result of study shows, skin lesion and the histopathology performance of observation group after modeling, matched group and treatment 1-6 group meet psoriatic tissue pathological manifestations, and histology Baker marks and inflammatory cell infiltration scoring obvious rising (P<0.01) compared with blank group, proves that psoriatic guinea pig model is successfully established.Matched group and the histology Baker for the treatment of 1-6 group mark and inflammatory cell infiltration scoring obvious reduction (P<0.01) compared with observation group after modeling, and each main organs of blank group Cavia porcellus does not find tissue pathologies change, prove that matched group and treatment 1-6 group all can improve the pathological change of Cavia porcellus ear Psoriasis-like Model.Treatment 1-6 group is compared with matched group, and the skin lesion performance improvement degree such as erythema, pimple, speckle are higher, have significant difference (P<0.05), and display treatment 1-6 group clinical efficacy is obviously better than matched group.
Treating 5 groups is treatment 3 groups and matched group drug combination, and curative effect is better than treatment 2 groups, the independent medication treatment of matched group respectively; Treating 6 groups is treatment 4 groups and matched group drug combination, and curative effect is better than treatment 4 groups, the independent medication treatment of matched group respectively.Oral more desirable with external drug combination effect during prompting Clinical practice.
Conclusion: NTx albumen and NTx protein peptide are after treatment guinea pig experimental psoriasis, bilateral skin of pinna cosmetic variation, skin color transfer to normally, smooth surface, and alopecia phenomenon makes moderate progress, the blood capillary of expansion is repaired, and various symptom obviously alleviates or disappears; Histology Baker marks and inflammatory cell infiltration scoring obvious rising compared with blank group, and difference has statistical significance (P < 0.01), proves that the psoriatic model of Cavia porcellus is successfully established.And the histology Baker treating 1-6 group marks and inflammatory cell infiltration scoring obvious reduction compared with observation group after modeling, difference has statistical significance (P < 0.01), and the every Score index for the treatment of 1-6 group is all better than matched group, has significant difference (P<0.05).
Overall experiment effect display: the present invention can improve Cavia porcellus ear Pigs with Psoriasis symptom and pathological characters thereof, and therapeutic effect optimum is NTx protein peptide, and NTx albumen takes second place, but both equal significances are better than matched group.
Although above with general explanation, detailed description of the invention and test, the present invention is described in detail, and on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (14)
1. NTx albumen is preparing the novelty teabag treated and/or prevented in psoriasis.
2. purposes as claimed in claim 1, is characterized in that: described NTx albumen is NTx protein peptide.
3. purposes according to claim 1 and 2, treats and/or prevents psoriatic medicine described in it is characterized in that, every day, oral dose amounted to protein 1600-10000mg.
4. purposes according to claim 1 and 2, treats and/or prevents psoriatic medicine described in it is characterized in that, every day, oral dose amounted to protein 3000-6000mg.
5. purposes according to claim 1, it is characterized in that described NTx albumen is prepared by following steps and obtains: choose animal tissue, pulverize with after clear water rinsing, add animal tissue weight 3-10 water heating extraction 1-8 hour doubly, extract 1-3 time, filter, merging filtrate, drying, obtains NTx albumen.
6. purposes according to claim 1, it is characterized in that described NTx albumen is prepared by following steps and obtains: choose animal tissue, cleaning Hou Jia animal tissue weight 5-10 acid soak 5-120 minute doubly, filter, animal tissue pulverizes with after clear water rinsing, add animal tissue weight 3-10 water heating extraction 1-8 hour doubly, extract 1-3 time, filter, merging filtrate, drying, obtains NTx albumen.
7. purposes according to claim 2, is characterized in that described NTx protein peptide is prepared by following steps and obtains: get NTx albumen, add protease hydrolysis 20-60 minute when 50-60 DEG C after, heat inactivation, gets supernatant, concentrated, drying, obtains NTx protein peptide.
8. purposes according to claim 2, is characterized in that described NTx protein peptide is prepared by following steps and obtains: choose animal tissue, clean, drain, pulverize, adding animal tissue weight 5-10 acid soak 5-120 minute doubly, filter, is 5-6 with clear water rinsing to pH; Add the NaCl solution immersion 1-10 hour that concentration is 1-5%, added NaCl solution is 8-12 times of animal tissue's weight; Be chilled to 3-10 DEG C, add protease, hydrolysis 2-6 hour, after hydrolysis terminates, heats deactivation in 20-40 minute; Obtain hydrolyzed solution, dry after hydrolyzed solution is concentrated, obtain NTx protein peptide.
9. the purposes according to any one of claim 5,6 or 8, is characterized in that described animal tissue comprises skeleton, fish scale, fin.
10. the purposes according to claim 6 or 8, when adding acid soak described in it is characterized in that, selects any one among the hydrochloric acid of 0.5-2%, glacial acetic acid, lactic acid, acetic acid, citric acid or malic acid.
11. purposes according to claim 7 or 8, is characterized in that described protease is any one among neutral protease, acid protease or alkaline protease.
12. purposes according to claim 7 or 8, is characterized in that described protease is any one or two or more combinations among pepsin, trypsin, cathepsin, papain or subtilisin.
13. purposes according to claim 1, is characterized in that being prepared from by NTx albumen according to claim 5 and pharmaceutically acceptable carrier combination.
14. purposes according to claim 2, is characterized in that being prepared from by the NTx protein peptide described in claim 6 or 7 and pharmaceutically acceptable carrier combination.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410556819.6A CN104367992A (en) | 2014-10-20 | 2014-10-20 | New application of type I collagen in preparation of medicament for treating and/or preventing psoriasis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410556819.6A CN104367992A (en) | 2014-10-20 | 2014-10-20 | New application of type I collagen in preparation of medicament for treating and/or preventing psoriasis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104367992A true CN104367992A (en) | 2015-02-25 |
Family
ID=52547351
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410556819.6A Pending CN104367992A (en) | 2014-10-20 | 2014-10-20 | New application of type I collagen in preparation of medicament for treating and/or preventing psoriasis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104367992A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107488696A (en) * | 2017-09-19 | 2017-12-19 | 广东雅道生物科技有限公司 | One breeder bone double enzymolysis method |
| CN107670026A (en) * | 2017-10-25 | 2018-02-09 | 界首市菁华科技信息咨询服务有限公司 | One kind is used to prevent antipsoriatic spray and preparation method thereof |
| CN108355130A (en) * | 2018-05-22 | 2018-08-03 | 界首市菁华科技信息咨询服务有限公司 | It is a kind of to treat psoriasic temperature sensitive type collagen composite antiphlogistic gel spray and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101489541A (en) * | 2006-03-14 | 2009-07-22 | 比奥福什克宁制药科学院 | Use of polyamines in the treatment of psoriasis |
| US20120141575A1 (en) * | 2010-12-02 | 2012-06-07 | Dr. Suwelack Skin & Health Care Ag | Collagen For Use In The Treatment Of Skin Diseases |
| CN102688483A (en) * | 2011-03-22 | 2012-09-26 | 王成德 | Composition for treating dermatosis, preparation containing composition and preparation method thereof |
-
2014
- 2014-10-20 CN CN201410556819.6A patent/CN104367992A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101489541A (en) * | 2006-03-14 | 2009-07-22 | 比奥福什克宁制药科学院 | Use of polyamines in the treatment of psoriasis |
| US20120141575A1 (en) * | 2010-12-02 | 2012-06-07 | Dr. Suwelack Skin & Health Care Ag | Collagen For Use In The Treatment Of Skin Diseases |
| CN102688483A (en) * | 2011-03-22 | 2012-09-26 | 王成德 | Composition for treating dermatosis, preparation containing composition and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 胡建平: "《鱼胶原蛋白的开发与应用》", 31 March 2014, 四川大学出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107488696A (en) * | 2017-09-19 | 2017-12-19 | 广东雅道生物科技有限公司 | One breeder bone double enzymolysis method |
| CN107670026A (en) * | 2017-10-25 | 2018-02-09 | 界首市菁华科技信息咨询服务有限公司 | One kind is used to prevent antipsoriatic spray and preparation method thereof |
| CN108355130A (en) * | 2018-05-22 | 2018-08-03 | 界首市菁华科技信息咨询服务有限公司 | It is a kind of to treat psoriasic temperature sensitive type collagen composite antiphlogistic gel spray and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8383100B2 (en) | Method to treat and prevent skin diseases with Porifera-based therapeutic compositions treating and preventing skin diseases | |
| Yoon et al. | Supplementing with dietary astaxanthin combined with collagen hydrolysate improves facial elasticity and decreases matrix metalloproteinase-1 and-12 expression: A comparative study with placebo | |
| US20190358270A1 (en) | Porifera-based therapeutic compositions for treating and preventing skin diseases | |
| Nordlund et al. | The normal color of human skin | |
| Yoo et al. | Effects of egg shell membrane hydrolysates on UVB-radiation-induced wrinkle formation in SKH-1 hairless mice | |
| CN104367992A (en) | New application of type I collagen in preparation of medicament for treating and/or preventing psoriasis | |
| Kim et al. | Thread Embedding Acupuncture Inhibits Ultraviolet B Irradiation‐Induced Skin Photoaging in Hairless Mice | |
| CN104491003B (en) | Add taste Yupingfeng gelling agent and its preparation method and application | |
| Madireddy et al. | Hypopigmented Macules | |
| KR20120029346A (en) | Beauty drink containing apple extract and collagen tripeptide | |
| JP2008063315A (en) | Compounding ingredient for makeup product and makeup product improving texture | |
| CN104382943A (en) | Novel purpose of scale extract in aspect of preparation of drug for treating and/or preventing psoriasis | |
| CN104323966A (en) | Golden hydrotherapy composition and using method thereof | |
| Van Abbe et al. | Pharmaceutical and Cosmetic Products for Topical Administration: Pharmaceutical Monographs | |
| CN105473186A (en) | Topical composition for stimulating epidermis and dermis layers of the skin | |
| CN101791282B (en) | Beauty lotion and preparation method and application thereof | |
| Jadach et al. | Use of Collagen in Cosmetic Products | |
| Bucay et al. | Low molecular weight heparan sulfate containing facial skin care for reducing inflammation and restoring aged‐skin homeostasis | |
| MXPA01001014A (en) | Composition for stimulating the synthesis of the melanic pigment and process for obtaining it. | |
| Camacho-Martinez | Actinic keratosis | |
| CN105287496B (en) | A kind of pharmaceutical composition and preparation method thereof for treating ichthyosis | |
| Lübbe et al. | Propolis, beeswax, and the sensitization potential of topical calcineurin inhibitors | |
| Singh | Can we prevent skin aging? | |
| CN104324054A (en) | Novel application of I-type collagen in preparation of medicine for treating and/or preventing leukoderma | |
| Pandey et al. | Nanocosmetics and Skin Health: A Comprehensive Review of Nanomaterials in Cosmetic Formulations |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150225 |
|
| RJ01 | Rejection of invention patent application after publication |