CN109568322A - Purposes of the mycophenolate in the drug of preparation treatment leucoderma - Google Patents
Purposes of the mycophenolate in the drug of preparation treatment leucoderma Download PDFInfo
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- CN109568322A CN109568322A CN201811465719.7A CN201811465719A CN109568322A CN 109568322 A CN109568322 A CN 109568322A CN 201811465719 A CN201811465719 A CN 201811465719A CN 109568322 A CN109568322 A CN 109568322A
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- Prior art keywords
- mycophenolate
- quinhydrones
- purposes
- group
- leucoderma
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
Abstract
The present invention relates to medical applications fields, and in particular to a kind of new application of mycophenolate is more particularly to purposes of the mycophenolate in the drug of preparation treatment leucoderma.The depigmentation model that the present invention is induced by establishing quinhydrones, smears mycophenolate external preparation on the animal of melanocyte function missing, promotes the local efficacy of secondary color at leucoderma skin lesion, has good pigment recovery effects.By the related skin lesion melanocyte GAP-associated protein GAP and genetic enrichment identified after confirming pigment recovery, the application range of mycophenolate is expanded, the selection of patients with vitiligo medication is expanded.
Description
Technical field
The present invention relates to medical applications fields, and in particular to a kind of new application of mycophenolate is more particularly to mycophenolic acid
Purposes of the ester in the drug of preparation treatment leucoderma.
Background technique
Leucoderma is a kind of depigmentation disease, is characterized at disease damage being symmetrical, the white patch of bilateral, lesion master
It is distributed in back, wrist, forearm, face, neck, multiple positions such as surrounding genital.Currently, the specific morbidity machine of leucoderma disease
System is not clear that existing etiology includes theory Of heredity, theory of autoimmunity, spirit and neurochemistry theory, melanocyte
Cell autoclasia theory, microelement deficiencies theory and other factors.The current treatment means of leucoderma have drug therapy,
The methods of physiotherapy, surgical operation therapy need safer effective drug.
Mycophenolate (MMF) is a kind of neotype immunosuppressant, is 2 one morpholinylethyl ester products of mycophenolic acid (MPA),
Now common name examines phenol for wheat.Can selectively inhibit T, the carnine acidohydrogenase in B cell and play immunosupress
Effect, adverse reaction is lighter, it is still lacked in existing report and whether has medicative research to leucoderma.
Summary of the invention
The purpose of the present invention is to provide a kind of external preparation of mycophenolate and its in the new of preparation treatment leucoderma
Medical usage.For the present invention by building leucoderma animal model, the pigment that verifying mycophenolate external preparation induces quinhydrones is de-
Lose the pigment restitution of model, it was demonstrated that mycophenolate external preparation can be as new the answering of preparation treatment leucoderma medicament
With.
The present invention provides purposes of the mycophenolate in the drug of preparation treatment leucoderma.Wherein, the drug is
The pharmaceutical composition being made of mycophenolate and excipient is added pharmaceutically acceptable auxiliary using mycophenolate as effective component
The preparation that material or complementary ingredient are prepared.Wherein, the pharmaceutical composition is local topical formulation.Further preferably
Ground, the local topical formulation are liniment, ointment, gelling agent, aerosol, spray or transdermal patch.Mycophenolate is in institute
The weight percentage in local topical formulation stated is 6% ~ 24%.
Mycophenolate is prepared into local topical formulation, by Transdermal absorption, increases the local organization free drug of skin
Concentration, can effectively improve the effect of immunosupress pathogenic T lymphocyte, preferably plays it and promotes to answer at leucoderma skin lesion
The local efficacy of color reduces adverse reaction.It of particular concern is, it is in conjunction with the embodiments middle to find mycophenolate external preparation pair
The depigmentation model of quinhydrones induction has quite ideal therapy of vitiligo effect, and in drug systemic administration route, do not have still
Sign can prompt mycophenolate to have such pharmacological activity.
According to the physicochemical property of mycophenolate, local topical formulation of the present invention is cream;The cream is
Mycophenolate powder is dissolved in alcohol, object is mixed until completely dissolved and adds in excellent color woods (Eucerin) emulsifiable paste, stirring is equal
Even to be made mycophenolate emulsifiable paste, the mass ratio of the Mycophenolate mofctil and excellent color woods emulsifiable paste is 1 ~ 4:15.
The present invention also provides prepare the liniment of mycophenolate, ointment, cream, gelling agent, aerosol, spray,
The preparation method of the external preparations such as transdermal patch: ethyl alcohol, propylene glycol and distilled water, phosphate-buffered have been selected in the prescription of liniment
Basis of the liquid as liniment;Carboxymethyl cellulose, glycerol, ethylparaben, urea, card have been selected in the preparation of ointment
Bo Mu -940, Tween 80, triethanolamine and distilled water are used to prepare.In view of the Transdermal absorption of mycophenolate is less desirable,
In order to preferably promote to absorb, it can choose suitable penetrating agent in the preparation and be added preparation prescription, in the embodiment of the present invention
The penetrating agents such as dimethyl sulfoxide, laruyl alcohol, propylene glycol, azone, urea, oleic acid have been selected to increase and absorb.
Compared with prior art, the medicine have the advantages that
The principal pathogenetic inducement of leucoderma is the missing of melanocyte function in its skin, and the present invention is by establishing quinhydrones induction
Depigmentation model smears mycophenolate emulsifiable paste, the results showed that pigment restores to 80% on the animal of melanocyte function missing
More than, there are good pigment recovery effects.It is related by the skin lesion melanocyte after related the identification also recovery of susceptible of proof pigment
Albumen and genetic enrichment.In addition, peripheral blood compares conscience kidney index of correlation before and after the medication of cavy depigmentation model, have no significant
Difference, it was demonstrated that drug is to the harmless effect of the functions such as conscience kidney.The drugs such as hormone used in more current therapy of vitiligo are more pacified
Entirely.The present invention uses the outer painting dosage form of mycophenolate to treat leucoderma depigmentation model for the first time, and therapeutic effect is significant, has bright
Aobvious technical effect;The application range for expanding mycophenolate expands the selection of patients with vitiligo medication.
Detailed description of the invention
The leucoderma illustraton of model of cavy when Fig. 1 is the 18th day, wherein figure (a) be own control side, figure (b) for cavy take off
Hair-fields, figure (c) are quinhydrones decoloration side;
The histological stain figure of the leucoderma model of cavy when Fig. 2 is the 18th day;Wherein, scheme (a), figure (b) is that itself is right respectively
According to the H&E colored graph of side and quinhydrones decoloration side;With Masson-Fontana colored graph;Scheme (c), figure (d) is control sides respectively
With the Masson-Fontana colored graph of quinhydrones decoloration side;
Fig. 3 is quinhydrones group and quinhydrones+mycophenolate group depigmentation spot distribution situation figure;
Fig. 4 is that Image-Pro Plus software calculates medication side pigment on the right side of control group, quinhydrones group and quinhydrones+mycophenolate group
Depigmentation spot accounts for the percentage of the total depilation area in right side;
Fig. 5 is H&E dyeing, Masson-Fontana colored graph, and in figure, a, c, e are control group, quinhydrones group and quinhydrones+mould respectively
The H&E colored graph of phenols acids group;In figure, b, d, f are control group, quinhydrones group and quinhydrones+mycophenolate group Masson- respectively
Fontana colored graph;
Fig. 6 is the quinhydrones decoloration side skin lesion Western bloting lab diagram of cavy;Wherein, figure a is the β-of phosphorylation
The expression of catenin and internal reference (GAPDH) in control group, quinhydrones group and quinhydrones+mycophenolate group;Scheming b is that GSK3 β is being compareed
Expression in group, quinhydrones group and quinhydrones+mycophenolate group;Figure c is the quantization of figure a;Figure d is the quantization to figure b;
Fig. 7 is the mRNA expression comparative diagram of Wnt access proinflammatory gene;Wherein, figure a is to Axin2, Dkk and Dact2 gene
Measurement, figure b is measurement to TGF-β and IFN-γ gene;
Fig. 8 is guinea pig serum darling renal function correlation factor expression figure.
Specific embodiment
With reference to the accompanying drawing, the present invention is further illustrated, these embodiments are merely to illustrate the present invention rather than limitation
The scope of the invention;Test method without specific conditions in embodiment, according to normal conditions;The reagent and biological material
Material, unless otherwise specified, commercially obtains.
Embodiment 1: leucoderma animal model preparation
It establishes leucoderma animal model: choosing healthy ater cavy 60, weight 300-350g is male, is raised in river
Soviet Union university animal experimental center.Guinea pig back hair is cut short, rosin and the mixing of paraffin one to one are placed in water-bath completely
Melt, pours before the rosin and paraffin after thawing in guinea pig back, tear after rosin and paraffin, cavy hair is complete
It tears and drags, depilation in cavy every 7 days is primary, and every cavy sets up own control, is own control side at the depilation of left side, smears 5% physiology
Salt water 1ml;It decolourizes side, to smear 6% hydroquinone (quinhydrones) 1ml for quinhydrones at the depilation of right side;Twice daily;It sees for the first time within 18th day
It observes depigmentation spot at the depilation of right side to occur, the continuous coating of skin 50 days at cavy depilation.Cavy is white when Fig. 1 is the 18th day
Purplish or white patches on the skin wind illustraton of model;Wherein, (a) is own control side, is (b) quinhydrones decoloration side;As shown in Figure 1, at the 18th day, quinhydrones decoloration
There is patch shape depigmentation spot in side, in distribution is dispersed in, is similar to leucoderma.For black hair follicle and substrate at identification depigmentation spot
The layer whether complete depigmentation of melanocyte has carried out hematoxylin eosin staining method (H&E) dyeing and the black of histotomy at skin lesion
Plain masson-fontana silver dyeing (Masson-Fontana) dyeing identification, the histology dye of the leucoderma model of cavy when Fig. 2 is the 18th day
Chromatic graph;From Figure 2 it can be seen that the H&E dyeing display black hair follicle of own control side is normal in figure (a), and scheme quinhydrones decoloration side in (b)
Black hair follicle disappear, equally, relative to figure (c) own control side, scheme the Masson-Fontana dyeing of (d) quinhydrones decoloration side
Display decoloration spot basal layer melanocyte missing, therefore, it was demonstrated that the success of leucoderma model foundation.
Cavy is randomly divided into quinhydrones group, quinhydrones+mycophenolate group and negative control group;Quinhydrones is only smeared in quinhydrones group,
2 times a day;18% mycophenolate emulsifiable paste is smeared after smearing quinhydrones 1 hour in quinhydrones+mycophenolate group, it is each daily to apply 2 times;Yin
Property control group only smears 5% physiological saline 1ml, 2 times a day.
Quinhydrones group and quinhydrones+mycophenolate group depigmentation spot distribution situation when observing 30 days, 40 days and 50 days respectively.
Fig. 3 is quinhydrones group and quinhydrones+mycophenolate group depigmentation spot distribution situation figure;As seen from Figure 3, in quinhydrones group, 30 days
When the right neck smeared of quinhydrones when there is plaquelike depigmentation spot, 40 days depigmentation spot extend to the small of the back and face
The fusion of depigmentation spot is in flakes when product expansion, 50 days and area becomes larger;As it can be seen that over time, quinhydrones group pigment
Depigmentation spot area is gradually increased;And the appearance of the substantially colorless plain depigmentation spot of quinhydrones+mycophenolate group.To more precisely compute pigment
The area of depigmentation spot, the present embodiment calculate the area percentage of hickie using Image-Pro Plus software using color range method, by
Shown in Fig. 4, when being 75%, 50 day when depigmentation spot accounts for 54%, 40 day of the quinhydrones decoloration side depilation gross area at quinhydrones group 30 days
It is 89%;And it is when depigmentation spot accounts for 19%, 40 day of the quinhydrones decoloration side depilation gross area when quinhydrones+mycophenolate group 30 days
It 16%, is 12% at 50 days;Control group occurs without decoloration spot, it is known that, mycophenolate has resisted the depigmentation effect of quinhydrones.
Embodiment 2: histological characterization of the mycophenolate to the pneumatic object model function and effect of leucoderma
Control group, quinhydrones group and quinhydrones+mycophenolate group quinhydrones decoloration side skin lesion tissue is taken to cook frozen section respectively, to slice
Do H&E dyeing and Masson-Fontana dyeing.Fig. 5 is H&E colored graph and Masson-Fontana dyeing in embodiment 2
Figure;In figure, a, c, e are control group, quinhydrones group and quinhydrones+mycophenolate group H&E colored graph respectively;In figure, b, d, f are respectively
Control group, quinhydrones group and quinhydrones+mycophenolate group Masson-Fontana colored graph;As seen from Figure 5, mould compared with quinhydrones group
Phenols acids restore melanin hair follicle and basal layer melanocyte, and melanin hair follicle restores the horizontal melanin hair follicle with control group
It is close, reach 80% or more.
Embodiment 3: mechanism of action of the mycophenolate to the pneumatic object model of leucoderma
At 50 days, control group, quinhydrones group and the decoloration side skin lesion of quinhydrones+mycophenolate group cavy quinhydrones are extracted by rules of organization
The Western bloting experiment that histone is completed;Fig. 6 is that the quinhydrones decoloration side skin lesion Western bloting of cavy is real
Test figure;Wherein scheme a, the beta-catenin (β-catenin) and internal reference (GAPDH) for being phosphorylation are in control group, quinhydrones group and hydrogen
Expression in quinone+mycophenolate group;Scheming b is glycogen synthase kinase-3 (GSK3 β) in control group, quinhydrones group and quinhydrones+mould phenol
Expression in acid esters group;Figure c is the quantization of figure a;Figure d is the quantization to figure b;As seen from Figure 6, Western blot shows
Mycophenolate can improve the Wnt access hair follicle stem cells growth associated protein beta-catenin (β-catenin) of quinhydrones induction
Reduction effect;And reduce leucoderma sample Wnt access inflammatory factor glycogen synthase kinase-3 (GSK3 β) caused by quinhydrones
It increases.
Embodiment 4: mycophenolate plays therapeutic effect to the pneumatic object model of leucoderma by Wnt access
RNA at the decoloration side skin lesion of control group, quinhydrones group and quinhydrones+mycophenolate group cavy quinhydrones is taken at 50 days, passes through reverse
The programs such as record, PCR are completed to Wnt access proinflammatory gene: Axin2, Dkk, Dact2, transforming growth factor-β (TGF-β),
The measurement of interferon-γ (IFN-γ);Fig. 7 is the mRNA expression comparative diagram of Wnt access proinflammatory gene;Wherein, figure a is pair
The measurement of Axin2, Dkk and Dact2 gene, figure b are the measurements to TGF-β and IFN-γ gene;As seen from Figure 7, mycophenolate
The Wnt access related inflammatory gene that quinhydrones can be made to excite: Axin2, Dkk, Dact2, transforming growth factor-β (TGF-β),
Interferon-γ (IFN-γ) reduces, and further illustrates that mycophenolate has the function of inhibiting or treat to leucoderma.
Embodiment 5: the drug safety identification of mycophenolate
Control group, quinhydrones group and quinhydrones+mycophenolate group cavy Peripheral Blood is taken to carry out darling renal function at 50 days confirmatory
Test, Fig. 8 guinea pig serum darling renal function correlation factor expression figure;As seen from Figure 8, glutamic-pyruvic transaminase (ALT), millet straw turn
Adnosine deaminase (AST), urea nitrogen (BUN), creatinine (CREA), creatine kinase isozyme (CK-MB) are in quinhydrones group, quinhydrones+mycophenolate
Significant difference is had no between group and control group, and mycophenolate is demonstrated to the darling renal function of leucoderma model not by hepatic and renal function
It causes to damage, prompts mycophenolate on darling renal function without influence, mycophenolate has reliable safety.
Embodiment 6: the preparation of Mycophenolate mofctil cream
3g, 6g, 9g, 12g mycophenolate powder are dissolved in the alcohol of 5g95% respectively, after mycophenolate powder is completely dissolved
Mixture is added in excellent color woods (Eucerin) emulsifiable paste of 45g, it is respectively 6%, 12%, 18%, 24% that concentration is made after mixing evenly
Mycophenolate emulsifiable paste.
Embodiment 7: the preparation of Mycophenolate mofctil liniment
Mycophenolate mofctil 60g
Ethyl alcohol 35ml
Propylene glycol 180ml
Tween 80 2g
Phosphate buffer (PBS) adds to 1000ml
Mycophenolate mofctil: being first dissolved in ethyl alcohol, propylene glycol and the Tween 80 of recipe quantity by preparation process, and the phosphate of recipe quantity is added
Buffer (PBS).
Embodiment 8: the preparation of Mycophenolate mofctil liniment
Mycophenolate mofctil 90g
Ethyl alcohol 200ml
Propylene glycol 500ml
Distilled water adds to 1000ml
Mycophenolate mofctil: being first dissolved in the ethyl alcohol and propylene glycol of recipe quantity by preparation process, then adds to 1000ml with distilled water.
Liniment prepared by embodiment 7 and embodiment 8 is packed into specific container (sprayer or atomizer), can be used as
Spray uses.
Embodiment 9: the ointment of Mycophenolate mofctil
Mycophenolate mofctil 240g
Carboxymethyl cellulose 50g
Azone 10ml
Glycerol 400g
Ethylparaben 2g
Distilled water adds to 1000g
Preparation process: carboxymethyl cellulose and glycerol and azone are mixed, and are then added and are dissolved with recipe quantity Mycophenolate mofctil
Hot distilled water, place after being swelled into gel, add the aqueous solution containing ethylparaben, add water to enough.
Embodiment 10: the ointment of Mycophenolate mofctil
Mycophenolate mofctil 120g
Carbomer-940 1g
Glycerol 65g
Tween 80 2g
Ethylparaben 0.5g
Triethanolamine 4g
Distilled water adds to 1000g
Preparation process: the Carbomer-940 of recipe quantity is added in the aqueous solution containing Mycophenolate mofctil, forms the acid of low viscosity
Property solution, stirring be completely dissolved Mycophenolate mofctil, after the glycerol, Tween 80 and ethylparaben of recipe quantity is added, with three ethyl alcohol
Amine adjusts pH value to 7.0.
Embodiment 11: the cream of Mycophenolate mofctil
Mycophenolate mofctil 180g
Carboxymethyl cellulose 50g
Glycerol 200g
Ethylparaben 5g
Urea 12g
Distilled water adds to 1000g
Preparation process: carboxymethyl cellulose and glycerol are mixed, the hot distilled water containing recipe quantity Mycophenolate mofctil is then added, puts
It sets after being swelled into gel, adds the aqueous solution containing urea and ethylparaben, add water to enough.
Embodiment 12: Mycophenolate mofctil aerosol
Mycophenolate mofctil 100g
Ethyl alcohol 50ml
Dicholorodifluoromethane 150g
Vitamin C 7.5g
Sterile distilled water adds to 1000ml
Preparation process: the Mycophenolate mofctil of recipe quantity, vitamin C being dissolved in the mixed liquor of sterile distilled water and ethyl alcohol, filtered, and used
Sterile distilled water adds to 1000ml, and divided dose is filling, sealing-in dose valve system, and repressurization injects dicholorodifluoromethane respectively,
To obtain the final product.
Embodiment 13: Mycophenolate mofctil spray
Mycophenolate mofctil 60g
Ethyl alcohol 30ml
Dimethyl sulfoxide 15ml
Vitamin C 8g
Sterile distilled water adds to 1000ml
Preparation process: the Mycophenolate mofctil, dimethyl sulfoxide, vitamin C of recipe quantity are dissolved in sterile distilled water, after ethyl alcohol is added
Filtered through sand core funnel, add to 1000ml with sterile distilled water, divided dose encapsulating in spray manual pump sys to get.
Embodiment 14: the transdermal patch of Mycophenolate mofctil
Mycophenolate mofctil 25g
Ethyl alcohol 30ml
Laruyl alcohol 15g
Vitamin C 7.5g
Appropriate acrylate pressure-sensitive adhesive
Preparation process: the heating of left Mycophenolate mofctil, vitamin C and laruyl alcohol is dissolved in ethyl alcohol, pours into acrylate under stiring
It is stirred until homogeneous, is put to room temperature, film in pressure sensitive adhesive, then heat volatile organic solvent, cover polyolefin film by lining, cut
Cut component to get.
Embodiment 15: the gelling agent of Mycophenolate mofctil
Mycophenolate mofctil 100g
Glycerol 30ml
Azone 14g
Carbomer-940 20g
Appropriate triethanolamine
Distilled water adds to 1000g
Preparation process: taking glycerol, Carbomer-940, azone to set in mortar and grind well, and adds appropriate distilled water, is slowly added dropwise three
Ethanol amine adjusts PH to 7.0 or so, stirring while adding, makes into gel, and Mycophenolate mofctil is added and is uniformly mixed, adds distilled water extremely
1000g is stirred evenly, and is dispensed to obtain the final product.
Claims (8)
1. purposes of the mycophenolate in the drug of preparation treatment leucoderma.
2. the purposes of the mycophenolate according to claim 1, which is characterized in that the drug be by mycophenolate and
The pharmaceutical composition of excipient composition, using mycophenolate as effective component.
3. the purposes of mycophenolate according to claim 2, which is characterized in that the pharmaceutical composition is local topical
Preparation.
4. the purposes of mycophenolate according to claim 3, which is characterized in that the local topical formulation be liniment,
Ointment, cream, gelling agent, aerosol, spray or transdermal patch.
5. the purposes of mycophenolate according to claim 3, which is characterized in that mycophenolate is in the local topical system
Weight percentage in agent is 6% ~ 24%.
6. the purposes of mycophenolate according to claim 4 or 5, which is characterized in that the local topical formulation is cream
Paste.
7. the purposes of mycophenolate according to claim 6, which is characterized in that the cream is by mycophenolate powder
It is dissolved in alcohol, will be added in excellent color woods emulsifiable paste until completely dissolved, and be stirred evenly and mycophenolate emulsifiable paste is made.
8. the purposes of mycophenolate according to claim 6, which is characterized in that the Mycophenolate mofctil and excellent color woods emulsifiable paste
Mass ratio is 1 ~ 4:15.
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CN201811465719.7A CN109568322A (en) | 2018-12-03 | 2018-12-03 | Purposes of the mycophenolate in the drug of preparation treatment leucoderma |
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN111973600A (en) * | 2020-08-20 | 2020-11-24 | 华中农业大学 | Application of mycophenolic acid or derivative thereof in preparation of canine distemper virus inhibitor |
Citations (1)
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AU2003204344A1 (en) * | 2002-05-23 | 2003-12-11 | Milder, Dan George | Compositions and methods for the treatment of autoimmune diseases and neurological disorders |
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2018
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Patent Citations (1)
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AU2003204344A1 (en) * | 2002-05-23 | 2003-12-11 | Milder, Dan George | Compositions and methods for the treatment of autoimmune diseases and neurological disorders |
Non-Patent Citations (2)
Title |
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FARHAD HANDJANI等: "Topical mycophenolate mofetil in the treatment of vitiligo: a pilot study", 《DERMATOLOGY PRACTICAL & CONCEPTUAL》 * |
杨坷等: "白癜风免疫发病机制研究进展", 《中国麻风皮肤病杂志》 * |
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CN111973600A (en) * | 2020-08-20 | 2020-11-24 | 华中农业大学 | Application of mycophenolic acid or derivative thereof in preparation of canine distemper virus inhibitor |
CN111973600B (en) * | 2020-08-20 | 2021-11-16 | 华中农业大学 | Application of mycophenolic acid or derivative thereof in preparation of canine distemper virus inhibitor |
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