CN106265737A - A kind of external used medicine for treating skin appendages and itching skin disease and its preparation method and application - Google Patents
A kind of external used medicine for treating skin appendages and itching skin disease and its preparation method and application Download PDFInfo
- Publication number
- CN106265737A CN106265737A CN201610994956.7A CN201610994956A CN106265737A CN 106265737 A CN106265737 A CN 106265737A CN 201610994956 A CN201610994956 A CN 201610994956A CN 106265737 A CN106265737 A CN 106265737A
- Authority
- CN
- China
- Prior art keywords
- cell
- stock solution
- medicine
- preparation
- skin
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Immunology (AREA)
- Cell Biology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Hematology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Virology (AREA)
- Zoology (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a kind of external used medicine for treating skin appendages and itching skin disease and its preparation method and application, described medicine be mainly composed of NK cell stock solution, described NK cell stock solution is carried out amplification cultivation acquisition by NK cell, NK cell stock solution is mixed from different excipient substances, can be made into liquid preparation for external application or external ointment preparation.Compared with prior art, this external used medicine is to utilize the protein of NK emiocytosis and cytokine to be easily absorbed by the skin, and the feature of target cell can be directly acted on, can be directly used for skin appendages and the treatments of itching skin disease such as acne, seborrheic dermatitis, pruritus and prurigo nodularis, curative effect is quick, the immune system of skin can be improved simultaneously, repair impaired skin.Excreted factor in NK cell stock solution also can kill fungus and antibacterial, reaches the purpose of Comprehensive Treatment acne, dermatitis and pruritus, and therapeutic domain is extensive, can prevent repeatedly increasing the weight of of the state of an illness.
Description
Technical field
The present invention relates to field of biological pharmacy, particularly relate to a kind of for treating skin appendages and pruritic skin
Sick external used medicine and its preparation method and application.
Background technology
Acne vulgaris is the most common a kind of dermatosis, has 600,000,000 5 thousand ten thousand patients in the whole world.It is due to hair follicle, skin
Adipose gland blocks, and causes Propiobacterium amount reproduction present in skin and the inflammation that causes.Be mainly in the more face of sebaceous gland,
, often there are blackhead, Whiteheads, red pimple, pustule, cyst etc., and are likely to result in skin in the positions such as cervical region and shirtfront back
Skin pigementation or generation cicatrix.Acne impact is attractive in appearance, and the patient that situation is serious often bears huge mental pressure.Really
The cause of disease cut is the clearest, may relate to the factors such as androgen, sebum generation and Propiobacterium infection, and heredity is also individual important
Aspect.In serious patients with acne blood, androgen precursor substantially increases, and patient with slight symptoms's simply tissue Serum Testosterone Levels raises.
Sebaceous gland hyperfunctioning, generates too much sebum, is also a key factor of acne generation.Propiobacterium infects the immunity caused
React relevant with the generation of struvite skin lesion.
The treatment of acne typically uses medicine for external use and innerlich anwenden: antibiotic, glucocorticoid, tretinoin etc., but this type of medicine
Treatment targeting is single, needs to coordinate other to infect and antimicrobial drug is used in combination, and life-time service exists toxic and side effects.
Seborrheic dermatitis, also known as seborrheic eczema, is to occur at sebaceous gland and enriches the one chronic papulosquamous inflammation at position
Disease dermatoses.Primary disease is more common in adult and neonate, is apt to occur in the sebaceous gland such as head, trunk and enriches district.
Pruritus is a kind of subjective symptoms that many dermatosiss are total, and this cause of disease is because of complexity, and skin exists the sensor of gargalesthesia,
And it is transmitted to brain by teleneuron.The release of affected tissue amine can promote gargalesthesia.Scratch because tenderness has itching-relieving action, but promote
Enter and discharge more histamine, increase the weight of gargalesthesia, form the vicious cycle grabbed of itching, cause even more serious skin injury and inflammation
Disease.
Prurigo nodularis is more common in adult female, is apt to occur in four autopodium sides and brothers back.From the beginning of for indefinite light of number
Red papules, gradually expands Semen Glycines to Semen Viciae fabae size bronzing or taupe hemispherical tuberosity.Rough surface is excipuliform, paroxysm
Property play is itched, and has pigmentation and lichenification because scratching.The chronic course of disease.The cause of disease is not yet clear and definite, patient mostly be allergic constitution with
The sting such as mosquito sand fly are relevant, pathobolism, gastrointestinal dysfunction, endocrine dysfunction, namely may be with this
Sick relevant.
General treatment skin appendages and itching skin disease many employings external and oral medicine comprehensively use, and medicine for external use is main
It is some glucocorticoids, antimicrobial drug, Selsun Blue, vancide ZP lotion, antifungal preparation, sulfur and (or) salicylic acid
Lotion etc., oral medicine use glucocorticoid, tripterygium glycosides, antibiotic, these a few class medicines of vitamin B group, with treatment this type of
Medicine equally exists that treatment is single, and the course for the treatment of is longer, and permanent use exists toxic and side effects, and therefore one comprehensive medicine of research and development makes
Its curative effect is quick, have no side effect in the urgent need to.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, it is provided that one is used for treating skin appendages and pruritus
External used medicine of dermatoses and preparation method thereof, single to solve conventional medication, the course for the treatment of is longer, and permanent use exists poison
The technical problems such as side effect.
The present invention is achieved by the following technical solutions:
The invention provides a kind of external used medicine for treating skin appendages and itching skin disease, described medicine for external use
Thing includes the NK cell stock solution that mass percent is 20%-100%.
Further, described external used medicine is a kind of liquid preparation, named SNK liquid, the component of described SNK liquid and matter thereof
Amount percentage ratio include: NK cell stock solution 70.0-80.0%, glycerol 2.0-5.0%, ethylparaben 0.01-0.03%,
Borax 0.03-0.08%, Oleum menthae 0.5-1.5%, solvent is water.
Further, described external used medicine is a kind of ointment preparation, named SNK cream, the component of described SNK cream and matter thereof
Amount percentage ratio includes: NK cell stock solution 60.0-70.0%, Carbopol 941 are 0.1-0.5%, glycerol 2.0-5.0%, Aloe powder
0.2-0.5%, ethylparaben 0.1-0.5%, hyaluronic acid 0.1-0.3%, Oleum menthae 0.5-1.5%, surplus is
Water.
Present invention also offers the preparation method of the external used medicine of above-mentioned treatment skin appendages and itching skin disease, bag
Include following steps:
(1) take NK cell and carry out amplification cultivation, it is thus achieved that NK cell stock solution;
(2) NK cell stock solution is mixed with external used medicine adjuvant, prepare described external used medicine.
Further, if described medicine is liquid preparation, the mass percent of described NK cell stock solution is 70.0-
80.0%, described adjuvant and mass percent thereof include: glycerol 2.0-5.0%, ethylparaben 0.01-0.03%,
Borax 0.03-0.08%, Oleum menthae 0.5-1.5%, solvent is water;If described medicine is ointment preparation, described NK cell is former
The mass percent of liquid is 60.0-70.0%, and described adjuvant and mass percent thereof include: Carbopol 941 be 0.1-0.5%,
Glycerol 2.0-5.0%, Aloe powder 0.2-0.5%, ethylparaben 0.1-0.5%, hyaluronic acid 0.1-0.3%, thin
Lotus oil 0.5-1.5%, surplus is water.
Further, the method that described NK cell amplification is cultivated, comprise the following steps:
Step one: take fresh blood, extracts the peripheral blood lymphocytes (hereinafter referred to as PBMC) in blood;
Step 2: the celebrating of interleukin I L-2,80U/ml of adding 200U/ml in RPMI-1640 basal medium is the most mould
Element and the hyclone of 10% percentage by weight, it is thus achieved that NK cell culture fluid;
Step 3: take 3.0 × 107-4.0×107Individual PBMC, in a Tissue Culture Flask, adds the NK cell training of 20-30ml
The concentration of nutrient solution and 2ml is 1.0 × 107The NK amplifing reagent of individual/ml, it is thus achieved that initial inoculum;
Step 4: NK cell is inoculated in initial inoculum cultivation 1-2 days, adds according to cell growth state during cultivation
Add NK cell culture fluid;When cultivate to the 3rd day time, by medium centrifugal, it is thus achieved that cell precipitation in add NK cell culture fluid
Continue to cultivate;
Step 5: when NK cell quantity reaches 1.5 × 108Time more than individual/bottle, add 2 NK amplifing reagents and the training of NK cell
Nutrient solution is enlarged cultivating, and increases NK cell culture fluid according to cell growth state and carries out splitting cultivation, make in incubation
Cell concentration is 1.0 × 106Individual/ml, when cultivating to the 12nd day, controls liquid volume added, and making cell concentration is 2.0 × 106Individual/ml;
Step 6: continue to cultivate after the 14th day, collect cell culture fluid, it is thus achieved that NK cell stock solution.
Present invention also offers above-mentioned external used medicine as the application in skin subsidiarity disease therapeuticing medicine.
Further, NK cell stock solution and different external used medicine adjuvants are mixed, prepare liquid preparation for external application and
External ointment preparation, when using above-mentioned external medicine preparation treatment skin appendages and itching skin disease, first uses external use liquid
Body preparation, then carry out mixing treatment with external ointment preparation, to keep drug effect can allow skin absorption well, is used in mixed way number
After it, symptom can alleviate or disappear.
The principle of the present invention is: NK cell (natural killer cell, natural killer cell) is important the exempting from of body
Epidemic disease cell, not only relevant with antitumor, viral infection resisting and immunomodulating, and in some cases participate in allergy and from
The generation of body immune disease.NK cell can kill some antibacterial, fungus etc. in vitro, and this discharges some with NK cell and kills Jie
Matter is relevant.The medium of NK release mainly has perforin, NK cytotoxic factor and TNF etc..And in NK cell cultivation process, can close
Becoming and secrete cytokine profiles, play immunoregulation effect, these cytokines include MIP-1 α, MIP-1 β, RANTES, list
Monocyte chemoattractant protein, interleukin, colony stimulating factor, tumor necrosis factor, interferon and perforin etc..This biological preparation
Use NK cell stock solution treatment skin appendages and itching skin disease, utilize the egg of NK emiocytosis in NK cell stock solution just
White matter and cytokine directly can act on the feature of target cell, meanwhile, containing abundant inflammation in NK cell stock solution by skin permeation
Inflammation factor, after this inflammatory factor is absorbed by the skin, induction body produces humoral immunization and cell immune response, improves skin certainly
The resistivity of body, and accelerate to repair impaired tissue and cell, reach the effect for the treatment of. simultaneously.
Compared with prior art, the invention provides a kind of external for treating skin appendages and itching skin disease
Medicine and its preparation method and application, this external used medicine utilizes the protein of NK emiocytosis and cytokine easily to be inhaled by skin
Receive, and the feature of target cell can be directly acted on, can be directly used for acne, seborrheic dermatitis, pruritus and prurigo nodularis
Deng skin appendages and the treatment of the skin subsidiarity disease such as pruritic, curative effect is quick, can improve the siberian crabapple of skin simultaneously
System, repairs impaired skin.Excreted factor in NK cell stock solution also can kill fungus and antibacterial, reach Comprehensive Treatment acne,
Dermatitis and the purpose of pruritus, therapeutic domain is extensive, can prevent repeatedly increasing the weight of of the state of an illness.
Detailed description of the invention
Embodiment 1
1, material:
The conventional method that the present embodiment method therefor is the most all known to those skilled in the art dawn, used
The material such as reagent, if no special instructions, be commercially available purchase product.
2, method
2.1 preparation NK cell stock solution
Step one: PBMC extracts:
Fresh blood is centrifuged 10 minutes in 740g, it is thus achieved that the faint yellow blood plasma in upper strata and lower floor's hemocyte, by thin for lower floor's blood
Born of the same parents mix according to volume ratio 1:1 with normal saline, take 50ml centrifuge tube, and the blood cell suspension of dilution is layered on 15ml lymphocyte
Separating on liquid, 400g is centrifuged 30 minutes.Collecting the membranaceous cellular layer of middle white, add brine, 400g is centrifuged 10 points
Clock, abandons supernatant, and repeated washing is once.Finally precipitate with appropriate culture fluid re-suspended cell, i.e. obtain PBMC.
Step 2: the preparation of NK cell culture fluid:
200U/ml is added in Gibco RPMI Medium 1640basic culture fluid (buying in GIBCO company of the U.S.)
The gentamycin of interleukin I L-2,80U/ml and the Gibco FBS of 10% percentage by weight (buy in U.S. GIBCO public
Department), it is thus achieved that NK cell culture fluid.
Step 3: the preparation of NK cell initial inoculum:
Take 3.0 × 107-4.0×107Individual PBMC, in a T75 culture bottle, adds the NK cell culture fluid and 1 of 20-30ml
Propping up NK amplifing reagent, wherein, described NK amplifing reagent (sees Chinese patent for a kind of CD8 α-interleukin-22 1-CD137 complex
Document CN102154207A), can directly obtain by the way of market is bought, the product rule of the NK amplifing reagent that the present invention uses
Lattice are that 2ml/ props up, and concentration is 1 × 107Individual/ml.
Step 4: NK cell is initial to be cultivated:
NK cell is inoculated in above-mentioned T75 culture bottle cultivation, wherein:
Within 1st, 2 days, note observation of cell state.If cell state well can not process.If culture fluid cell density is obvious
Strengthen, NK cell culture fluid 10ml can be added.
3rd day by medium centrifugal, it is thus achieved that cell precipitation be transferred in new T75 culture bottle, and add the NK of 30ml
Cell culture fluid continues to cultivate.
4th day observation of cell, if cell state is good, can not process, if cell density is excessive, can take the circumstances into consideration to add NK thin
Born of the same parents' culture fluid.
5th day, if the 4th day untreated, and cell state was good, if cell density is big, then added NK cell culture fluid
20ml。
6th day observation of cell, adds culture fluid 30-50ml according to cell density.
Step 5: NK cell expansion is cultivated:
7th day counting, if cell quantity is more than 1.5 × 108Individual/bottle, then add 2 NK amplifing reagents, if cell quantity
At 30-60 million, then length added NK amplifing reagent again to the 8th day.Then add NK cell culture fluid, make total cell concentration exist
1.0×106Individual/ml.Incrementally increasing with volume of culture, can proceed to T175 and cultivate the cultivation of square vase extended volume, culture fluid volume is big
It is more than 2.0 × 10 in 120ml or cell quantity8Individual/bottle time, then can be transferred to culture bag cultivate.
8-11 days every days observation of cell so that cell concentration is 1.0 × 106Individual/about ml, is added by counting and cultivates
Liquid so that cell concentration is 1.0 × 106Individual/ml.
12-14 days every days observed, when cell quantity is more than 3.0 × 109Individual/bag, and when volume is more than 1200ml, will
Cell is divided into 2 bags.
Step 6: collection NK cell stock solution:
Can be collected when cell grows to requirement using or frozen.Generally, within 14 or 15 days, collect
Cell, because now cell is in exponential phase, in the best state, the slip of cell state meeting the most backward, now cell can
To 3 × 109More than individual, NK cell (CD3-CD56+) purity can reach more than 80%.
2.2 preparation SNK liquid:
By weight, NK cell stock solution 80%, glycerol 2.0%, ethylparaben 0.01%, Borax are taken
0.03%, Oleum menthae 0.5%, is substantially soluble in sterilized water, sterilizing subpackage after mixing, it is thus achieved that described SNK liquid.
2.3 preparation SNK cream:
By weight, take NK cell stock solution 70%, Carbopol 941 be 0.1%, glycerol 2.0%, Aloe powder 0.2%, to hydroxyl
Yl benzoic acid ethyl ester 0.1%, hyaluronic acid 0.1%, Oleum menthae 0.5%, surplus is sterilized water, sterilizing subpackage after mixing, it is thus achieved that
Described SNK cream.
3, clinical verification
Clinical experiment is divided into 3 groups carry out, particularly as follows:
Experimental group 1: select 8 examples to suffer from the patient of acne vulgaris, 6 examples suffer from the patient of seborrheic dermatitis, pruritus 5 example patient
With prurigo nodularis patient 4 example, carry out smearing or hydrojet treatment with SNK liquid, use 2-3 time every day, add up clinical treatment outcome.
Experimental group 2: select 8 examples to suffer from the patient of acne vulgaris, 6 examples suffer from the patient of seborrheic dermatitis, pruritus 5 example patient
With prurigo nodularis patient 4 example, carry out smearing treatment with SNK cream, use 2-3 time every day, add up clinical treatment outcome.
Experimental group 3: select 8 examples to suffer from the patient of acne vulgaris, 6 examples suffer from the patient of seborrheic dermatitis, pruritus 5 example patient
With prurigo nodularis patient 4 example, smear with SNK liquid and cream or hydrojet treatment, use 2-3 time every day, add up clinical treatment outcome.
Result is as shown in table 1 below:
Table 1:SNK liquid and SNK cream clinical trial results statistical table
Result shows, the NK cell stock solution of the present invention, SNK liquid and SNK cream are to acne, seborrheic dermatitis, pruritus and knot
Joint property prurigo all has certain therapeutic effect, especially notable to the therapeutic effect of acne vulgaris, can thoroughly cure acne, alleviate fat
The symptom of property dermatitis of overflowing, and need not long-term prescription and take innerlich anwenden, greatly reduce toxic and side effects.Wherein: first group and
Second component not Wei SNK liquid and SNK cream, SNK liquid is different due to the content of NK cell stock solution with in SNK cream, and curative effect somewhat has difference
Not, SNK liquor ratio cream is effective.(first it is coated with SNK liquid, repaste SNK cream) when SNK liquid and SNK cream are used together, both can ensure NK liquid
In active factors efficiently and be persistently less readily available for absorption by the skin, curative effect can be improved again, both are used in combination best results.
Embodiment 2
Present embodiments providing a kind of SNK liquid, by weight, described SNK liquid is by NK cell stock solution 70.0%, glycerol
5.0%, ethylparaben 0.03%, Borax 0.08%, Oleum menthae 1.5% and appropriate amounts of sterilized water composition, wherein, described
The preparation method of NK cell stock solution is with embodiment 1.
Choosing suffers from patients with acne vulgaris 3 example, pruritus patient 2 example, is directly sprayed on affected part with above-mentioned SNK liquid and controls
Treat, use 2-3 time every day, add up clinical treatment outcome.
It was found that above-mentioned SNK liquid is notable to the therapeutic effect of acne vulgaris, 7 days symptoms of severe patient substantially alleviate, even
Continuous use can be fully recovered after 2 weeks;Have certain therapeutic effect to pruritus, patients with mild can be fully recovered in 2 weeks, and severe patient is after 2 weeks
Symptom is clearly better.
Embodiment 3
Present embodiments providing a kind of SNK cream, by weight, described SNK cream is by NK cell stock solution 60.0%, carbomer
941 be 0.5%, glycerol 5.0%, Aloe powder 0.5%, ethylparaben 0.5%, hyaluronic acid 0.3%, Oleum menthae
1.5% and suitable quantity of water composition.
Choosing suffers from seborrheic dermatitis patient 2 example, utilizes above-mentioned SNK cream to carry out smearing treatment, uses 2-3 time every day, statistics
Clinical treatment outcome.
It was found that after using 2 weeks, the symptom of 2 example patients with seborrheic dermatitis all takes an evident turn for the better, and this SNK cream pair is described
Seborrheic dermatitis has certain therapeutic effect.
Embodiment 4
Present embodiments providing a kind of SNK liquid, by weight, described SNK liquid is by NK cell stock solution 20%, glycerol 5% He
Appropriate amounts of sterilized water forms, and utilizes described SNK liquid to treat acne vulgaris, necessarily controls it was found that acne vulgaris is had by this SNK liquid
Therapeutic effect, after patient uses 10 days continuously, symptom all has certain improvement.
Embodiment 5
Present embodiments providing a kind of SNK liquid, by weight, described SNK liquid is by NK cell stock solution 50%, glycerol 5% He
Appropriate amounts of sterilized water forms, and utilizes described SNK liquid treatment prurigo nodularis patient after 30 days, and symptom disappears substantially.
Embodiment 6
Present embodiments providing a kind of SNK liquid, described SNK liquid is the NK cell stock solution of 100%, utilizes described SNK liquid to control
Treat prurigo nodularis can fully recover after 25 days.
In theory, the content of the NK cell stock solution in medicine is the highest, and active factors content is the highest, and effect is the best, but considers
In the case of ease of use and increase percutaneous permeability and protectiveness, need to the addition of on the basis of NK cell stock solution
Auxiliary element, uses the NK cell stock solution of 100% to treat, its clinical trial results and the SNK of embodiment 1 in embodiment 6
Liquid and cream are compared, and there is no very big difference on therapeutic effect.
It is more than a kind of detailed embodiment of the present invention and concrete operating process, is to be front with technical solution of the present invention
Put and implement, but protection scope of the present invention is not limited to the above embodiments.
Claims (8)
1. the external used medicine being used for treating skin appendages and itching skin disease, it is characterised in that described external used medicine
Including the NK cell stock solution that mass percent is 20%-100%.
A kind of external used medicine for treating skin appendages and itching skin disease the most according to claim 1, it is special
Levying and be, described medicine is a kind of liquid preparation, and its component and mass percent thereof include: NK cell stock solution 70.0-80.0%,
Glycerol 2.0-5.0%, ethylparaben 0.01-0.03%, Borax 0.03-0.08%, Oleum menthae 0.5-1.5%, molten
Agent is water.
A kind of external used medicine for treating skin appendages and itching skin disease the most according to claim 1, it is special
Levying and be, described medicine is a kind of ointment preparation, and its component and mass percent thereof include: NK cell stock solution 60.0-70.0%,
Carbopol 941 is 0.1-0.5%, glycerol 2.0-5.0%, Aloe powder 0.2-0.5%, ethylparaben 0.1-0.5%,
Hyaluronic acid 0.1-0.3%, Oleum menthae 0.5-1.5%, surplus is water.
4. the preparation of the external used medicine for treating skin appendages and itching skin disease as described in claim 1-3
Method, it is characterised in that comprise the following steps:
(1) take NK cell and carry out amplification cultivation, it is thus achieved that NK cell stock solution;
(2) NK cell stock solution is mixed with external used medicine adjuvant, prepare described external used medicine.
The preparation of a kind of external used medicine for treating skin appendages and itching skin disease the most according to claim 4
Method, it is characterised in that if described medicine is liquid preparation, the mass percent of described NK cell stock solution is 70.0-
80.0%, described adjuvant and mass percent thereof include: glycerol 2.0-5.0%, ethylparaben 0.01-0.03%,
Borax 0.03-0.08%, Oleum menthae 0.5-1.5%, solvent is water;If described medicine is ointment preparation, described NK cell is former
The mass percent of liquid is 60.0-70.0%, and described adjuvant and mass percent thereof include: Carbopol 941 be 0.1-0.5%,
Glycerol 2.0-5.0%, Aloe powder 0.2-0.5%, ethylparaben 0.1-0.5%, hyaluronic acid 0.1-0.3%, thin
Lotus oil 0.5-1.5%, surplus is water.
The preparation of a kind of external used medicine for treating skin appendages and itching skin disease the most according to claim 4
Method, it is characterised in that the method that described NK cell amplification is cultivated, comprises the following steps:
Step one: take fresh blood, extracts peripheral blood lymphocytes PBMC in blood;
Step 2: add in RPMI-1640 basal medium interleukin I L-2,80U/ml of 200U/ml gentamycin and
The hyclone of 10% percentage by weight, it is thus achieved that NK cell culture fluid;
Step 3: take 3.0 × 107-4.0×107Individual PBMC, in a Tissue Culture Flask, adds the NK cell culture fluid of 20-30ml
It is 1.0 × 10 with the concentration of 2ml7The NK amplifing reagent of individual/ml, it is thus achieved that initial inoculum;
Step 4: NK cell is inoculated in initial inoculum cultivation 1-2 days, adds NK according to cell growth state during cultivation
Cell culture fluid;When cultivate to the 3rd day time, by medium centrifugal, it is thus achieved that cell precipitation in add NK cell culture fluid continue
Cultivate;
Step 5: when NK cell quantity reaches 1.5 × 108Time more than individual/bottle, add 2 NK amplifing reagents and NK cell culture fluid enters
Row amplification culture, increases NK cell culture fluid according to cell growth state in incubation and carries out splitting cultivation, making cell dense
Degree is 1.0 × 106Individual/ml, when cultivating to the 12nd day, controls liquid volume added, and making cell concentration is 2.0 × 106Individual/ml;
Step 6: continue to cultivate after the 14th day, collect cell culture fluid, it is thus achieved that NK cell stock solution.
7. the external used medicine as described in claim 1-3 is as the application in skin subsidiarity disease therapeuticing medicine.
A kind of external used medicine the most according to claim 7 as the application in skin subsidiarity disease therapeuticing medicine, its
It is characterised by, NK cell stock solution is mixed with external used medicine adjuvant, prepare liquid preparation for external application and external ointment preparation, first
With liquid preparation for external application, then using external ointment preparation, immixture is in affected part.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610994956.7A CN106265737A (en) | 2016-11-11 | 2016-11-11 | A kind of external used medicine for treating skin appendages and itching skin disease and its preparation method and application |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610994956.7A CN106265737A (en) | 2016-11-11 | 2016-11-11 | A kind of external used medicine for treating skin appendages and itching skin disease and its preparation method and application |
Publications (1)
Publication Number | Publication Date |
---|---|
CN106265737A true CN106265737A (en) | 2017-01-04 |
Family
ID=57721150
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610994956.7A Pending CN106265737A (en) | 2016-11-11 | 2016-11-11 | A kind of external used medicine for treating skin appendages and itching skin disease and its preparation method and application |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106265737A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110200996A (en) * | 2019-05-16 | 2019-09-06 | 安徽瑞达健康产业有限公司 | A kind of application of cell composition on skin ulcer |
CN111712227A (en) * | 2017-12-14 | 2020-09-25 | 绿十字生命健康有限公司 | Cosmetic and pharmaceutical compositions for relieving atopic dermatitis, alopecia and trauma or reducing skin wrinkles |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102256608B (en) * | 2008-12-18 | 2013-12-04 | 阿波科学有限公司 | Pharmaceutical preparation comprising supernatant of blood mononuclear cell culture |
CN105754941A (en) * | 2016-04-18 | 2016-07-13 | 广州复大医疗股份有限公司 | In-vitro induction and proliferation culture method for peripheral blood NK (natural killer) cells |
CN105925527A (en) * | 2016-05-19 | 2016-09-07 | 成都百赛泰科生物科技有限公司 | Kit for preparing NK cells and application method thereof |
-
2016
- 2016-11-11 CN CN201610994956.7A patent/CN106265737A/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102256608B (en) * | 2008-12-18 | 2013-12-04 | 阿波科学有限公司 | Pharmaceutical preparation comprising supernatant of blood mononuclear cell culture |
CN105754941A (en) * | 2016-04-18 | 2016-07-13 | 广州复大医疗股份有限公司 | In-vitro induction and proliferation culture method for peripheral blood NK (natural killer) cells |
CN105925527A (en) * | 2016-05-19 | 2016-09-07 | 成都百赛泰科生物科技有限公司 | Kit for preparing NK cells and application method thereof |
Non-Patent Citations (1)
Title |
---|
宗金宝等: "NK细胞与皮肤病", 《中国麻风皮肤病杂志》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111712227A (en) * | 2017-12-14 | 2020-09-25 | 绿十字生命健康有限公司 | Cosmetic and pharmaceutical compositions for relieving atopic dermatitis, alopecia and trauma or reducing skin wrinkles |
EP3725296A4 (en) * | 2017-12-14 | 2021-09-29 | Green Cross WellBeing Corporation | Cosmetic composition and pharmaceutical composition for alleviating atopic dermatitis, hair loss, and wounds or reducing skin wrinkles |
CN110200996A (en) * | 2019-05-16 | 2019-09-06 | 安徽瑞达健康产业有限公司 | A kind of application of cell composition on skin ulcer |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101328219B (en) | Nano liposome anti-HPV, gynecological inflammation pathogen specific compound IgY and combined preparation thereof | |
CN106727649A (en) | A kind of multicomponent parenteral solution | |
CN103861079A (en) | Traditional Chinese medicine composition with effects of clearing heat, relieving pain and eliminating tumors | |
CN103405577B (en) | The Pharmaceutical composition of a kind of enhancing immunity, fatigue alleviating | |
CN106265737A (en) | A kind of external used medicine for treating skin appendages and itching skin disease and its preparation method and application | |
CN101011559B (en) | Traditional Chinese medicine preparation for treating malignant tumor and its preparation method | |
CN103330924A (en) | Biological extraction method of traditional Chinese medicine activin | |
CN106266908A (en) | A kind of medicine treating persistence low grade fever | |
CN102416144B (en) | Chinese medicine for treating infantile convulsions | |
CN105664140A (en) | Glycopeptide composition as well as preparation method and application thereof | |
CN106389467A (en) | External use medicine for treating dermatitis and eczema type skin diseases, and preparation method and application thereof | |
CN106344613A (en) | Application of NK cell stock solution in preparation of medicine for treating skin diseases | |
CN102764294B (en) | Cough relieving and sputum eliminating combination and preparation method thereof | |
CN111558031A (en) | Pure plant multifunctional eye special nutrient solution and preparation method thereof | |
CN105943651A (en) | Traditional Chinese medicine composition for treating premature ovarian failure, and application thereof | |
CN105943820A (en) | Traditional Chinese medicine compound for resisting swine influenza and porcine reproductive and respiratory syndrome and extraction method thereof | |
CN106344612A (en) | External medicine for treating autoimmune dermatosis and preparation method and application thereof | |
CN105233144A (en) | Traditional Chinese medicine preparation for treating dermatomycoses of animals and preparation method of traditional Chinese medicine preparation | |
CN103638471A (en) | Immune stimulant used for improving immunity function of piglets | |
CN100382837C (en) | A medicine for treating children's excessive blink, hyperkinetic syndrome and Tottre's syndrome and method for preparing same | |
CN103721147B (en) | For treating the Chinese medicine composition of chicken coccidiasis | |
CN106491645A (en) | One kind is used for treating physical property dermatosiss and the dermopathic external used medicine of animality and its preparation method and application | |
CN104645213A (en) | External traditional Chinese medicine for treating warts | |
CN103479714B (en) | The novelty teabag of YUPINGFENG KELI | |
CN106173409A (en) | A kind of ablactational baby pig feed additive and preparation method thereof |
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: 20170104 |
|
RJ01 | Rejection of invention patent application after publication |