CN109692327A - A kind of application of the nano particle of the application and delivery melittin of melittin - Google Patents
A kind of application of the nano particle of the application and delivery melittin of melittin Download PDFInfo
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- CN109692327A CN109692327A CN201710995177.3A CN201710995177A CN109692327A CN 109692327 A CN109692327 A CN 109692327A CN 201710995177 A CN201710995177 A CN 201710995177A CN 109692327 A CN109692327 A CN 109692327A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1767—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
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- 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
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
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Abstract
The invention discloses a kind of applications of melittin, are related to bioscience and pharmaceutical carrier technical field, and melittin is used to prepare the medicament for the treatment of allergic skin diseases.The invention also discloses a kind of applications of nano particle for delivering melittin, the nano particle of delivery melittin is made of polypeptide and phosphatide, the cholesterol ester of delivery melittin, its medicament for being used to prepare treatment allergic skin diseases, skin lymphatic system can be passed through, it is accumulated in the skin-draining lymph nodes that sensitization is crossed, and the effect of inhibiting T cells with antigenic specificity proliferation and reducing excessive immune reaction is played wherein, and toxic side effect is low.
Description
Technical field
The present invention relates to bioscience and pharmaceutical carrier technical field, and in particular to a kind of application of melittin and delivery bee
The application of the nano particle of phallotoxins.
Background technique
Allergic dermatitis is most commonly seen immunology skin diseases.Mainly swash currently based on the treatment of allergic dermatitis
Plain class drug, however there is a series of secondary works such as induced osteoporosis, hyperglycemia and atrophoderma over the course for the treatment of for hormone
With long-time service can also induce corticosteroid dermatitis.The new drug for being applied to treatment allergic dermatitis of exploitation has very big answer
Use prospect.
For natural bee venom with extensively in traditional medicine, one of them is the immunosuppressive effect of natural bee venom.Currently,
Moxibustion therapy has been efficiently applied to inhibit the immunity diseases such as multiple sclerosis, allergic rhinitis, arthritis.Allergic dermatitis
It is a kind of dermatological disease that immune response mediates.Therefore, have with traditional medicine-bee venom therapy allergic dermatitis huge latent
Power.However, treating the medical evidence of immunity disease and insufficient with bee venom whole components.Specifically, bee venom ingredient
Complexity, the mixture as a kind of intracorporal extraction of honeybee under natural conditions, it is difficult to directly research and applied to clinical spy
Property.Melittin (melittin) is a kind of polypeptide of amphiphilic αhelix, is the main active in bee venom, Zhan Qi
The 40%-60% of dry weight.Melittin has membrane structure very efficient destructive, including plasma membrane and more intracellular
The membrane structure of organelle.Its mechanism of action mainly has to be embedded in phosphatide membrane structure by its amphiphilic structure, then in film
Upper punching, causes membrane structure to rupture, and the variation of film internal and external environment eventually leads to the death of cell；It can also be thin by influencing
Key protein in a plurality of signal path of mediate tumor proliferation intracellular, and then cause Apoptosis.Meanwhile melittin also has
The antibacterial functions of wide spectrum and the function of anti-AIDS.
Simultaneously as the characteristics of melittin toxicity, it is difficult to be directly applied in the disease treatment of living body.Because of bee venom
Peptide can destroy cell membrane rapidly and erythrocyte binding after injecting blood vessel, so that generating strong hemolytic reaction.And directly will
Melittin is treated applied to living body, can be damaged to organism normal cell, be brought great toxic side effect.In addition, melittin
Very short in machine intracorporal half-life period, accretion rate quickly, is unfavorable for clinical disease treatment.
Therefore, a kind of melittin treatment allergic skin diseases are needed and explore its regulatory mechanism in immune system
Technical solution provides new direction for allergic skin diseases healing potion.
Summary of the invention
In view of the deficiencies in the prior art, the purpose of the present invention is to provide a kind of applications of melittin, for making
The medicament of standby treatment allergic skin diseases, can be effectively suppressed T cells with antigenic specificity increment, the cellular inflammation factor is inhibited to release
It puts.
To achieve the above objectives, the technical solution adopted by the present invention is that: a kind of application of melittin, the melittin are used for
The medicament of preparation treatment allergic skin diseases.
Compared with the prior art, the advantages of the present invention are as follows:
Melittin is used to prepare the medicament for the treatment of allergic skin diseases in the present invention, can effectively inhibit inflammation caused by allergy
Disease swelling degree inhibits degree of inflammation to have similar effect with clinical conventional Hormone drug-dexamethasone, can be effectively suppressed anti-
Former specific T-cells increment, inhibits the release of the cellular inflammation factor.
The object of the invention is also to provide a kind of applications of nano particle for delivering melittin, are used to prepare treatment allergy
The medicament of property skin disease, therapeutic effect is good, and toxic side effect is low.
To achieve the above objectives, the technical solution adopted by the present invention is that: it is described delivery melittin nano particle by delivering
Polypeptide and phosphatide, the cholesterol ester composition of melittin；The polypeptide of the delivery melittin is by α helical polypeptide, catenation sequence and bee
Phallotoxins is connected in series in the form of covalent bond；The nano particle of the delivery melittin is used to prepare treatment allergic skin diseases
Medicament.
On the basis of above scheme, the medicament can inhibit T cells with antigenic specificity to rise in value in lymph node, inhibit
The release of the cellular inflammation factor.
On the basis of above scheme, the nano particle of the delivery melittin is also used to prepare pharmaceutical carrier, the medicine
Object carrier is used to load the dye molecule or drug molecule of medical diagnosis on disease.
On the basis of above scheme, the phosphatide is dimyristoyl phosphatidyl choline (DMPC).
On the basis of above scheme, the amino acid sequence of the α helical polypeptide is DWFKAFYDKVAEKFKEAF.
On the basis of above scheme, the amino acid sequence of the melittin is
GIGAVLKVLTTGLPALISWIKRKRQQ。
On the basis of above scheme, the amino acid sequence of the catenation sequence is GSG.
Compared with prior art, it has the advantage that
(1) therapeutic effect is good
The medicament of the treatment allergic skin diseases of the nano particle preparation of the delivery melittin can pass through skin lymph
System is accumulated in the skin-draining lymph nodes that sensitization is crossed, and plays inhibit T cells with antigenic specificity proliferation wherein, reduces
The effect of degree immune response, play good antianaphylactic treatment effect.
(2) toxic side effect is low
The medicament of the treatment allergic skin diseases of the nano particle preparation of the delivery melittin, due to increasing melittin
Interaction between phosphatide is imbedded in melittin in single layer phospholipid layer, so as to avoid melittin and haemocyte and
The direct contact of normal cell, the toxic side effect of intradermal administration are low.
(3) function can be expanded
The nano particle of the delivery melittin can be also used for preparing pharmaceutical carrier, which can load in its core
For disease therapeuticing medicine molecule such as steroids steroid hormone etc., other types of targeted polypeptide can also be loaded simultaneously or is controlled
The property treated polypeptide, the effect of realizing the collaboration targeting or synergistic treatment of disease.
Detailed description of the invention
Fig. 1 is that the melittin of various dose treats the evaluation of Contact hyper sensitization dermatitis；
Fig. 2 is the evaluation that melittin treats Chronic exposure allergic dermatitis；
Fig. 3 is nano particle root of the tail portion's intracutaneous injection using melittin, treats C57BL/6 murine chronic allergic skin
The ear of disease is sliced HE colored graph；
Fig. 4 be deliver melittin nano particle will be compared after living body abdomen intracutaneous injection respectively with free melittin it is small
Mouse skin injury pictorial diagram；
Fig. 5 is that comparison curves is tested in the nano particle for delivering melittin and free melittin living body ear intracutaneous injection swelling
Figure；
Fig. 6 be deliver melittin nano particle and the portion's intracutaneous injection of free melittin root of the tail treat C57BL/6 mouse it is slow
The when m- swelling thickness map of property allergic skin diseases model；
Fig. 7 is the nano particle and free melittin and PBS root of the tail portion's intracutaneous injection treatment C57BL/6 for delivering melittin
The ear of murine chronic allergic skin diseases is sliced HE colored graph；
Fig. 8 is the nano particle core loading fluorescent dye DiR-BOA for delivering melittin, and C57BL/6 mouse is in vivo examined
Nano particle is surveyed in the distribution of skin-draining lymph nodes and histoorgan and the integral fluorescence image of Infiltrating；
Fig. 9 is the lymphocytes in vitro incubation co-cultivation detection cell of the nano particle for delivering melittin and haptens stimulation
The result schematic diagram of proliferation experiment；
Figure 10 is to deliver the nano particle of melittin to co-culture detection carefully with the lymphocytes in vitro incubation of haptens stimulation
The result statistical chart of born of the same parents' proliferation experiment；
Figure 11 is to deliver the nano particle of melittin to be incubated for culture inhibition IFN-γ altogether with the lymphocyte of allergy primary stimuli
Expression；
Figure 12 is that proinflammatory relevant cell factor IFN-γ and IL- in serum can be effectively reduced in the nano particle of delivery melittin
4 expressions.
Specific embodiment
Invention is further described in detail with reference to the accompanying drawings and embodiments.
Embodiment 1: the application of melittin
The embodiment of the invention discloses a kind of application of melittin, melittin is used to prepare treatment allergic skin diseases
Medicament.
Embodiment 2: the application of the nano particle of melittin is delivered
The embodiment of the invention discloses a kind of applications of nano particle for delivering melittin, deliver the nano particle of melittin
It is used to prepare the medicament for the treatment of allergic skin diseases.The medicament can inhibit T cells with antigenic specificity to rise in value in lymph node,
Inhibit the release of the cellular inflammation factor.The nano particle of delivery melittin is also used to prepare pharmaceutical carrier, and the pharmaceutical carrier is for filling
Carry the dye molecule or drug molecule of medical diagnosis on disease.
Wherein, the nano particle for delivering melittin is made of polypeptide and phosphatide, the cholesterol ester of delivery melittin.Phosphatide is
Dimyristoyl phosphatidyl choline (DMPC, 1,2-dimyristoyl-sn-glycero-3-phosphocholine).Deliver bee
The polypeptide of phallotoxins is connected in series in the form of covalent bond by α helical polypeptide, catenation sequence and melittin.The amino of α helical polypeptide
Acid sequence is DWFKAFYDKVAEKFKEAF.The amino acid sequence of α helical polypeptide is shown in SEQ ID NO.1 in sequence table.Bee
The amino acid sequence of phallotoxins is GIGAVLKVLTTGLPALISWIKRKRQQ, and amino acid sequence is SEQ ID in sequence table
Shown in NO.2.The amino acid sequence of catenation sequence is GSG.
Embodiment 3 dissociate melittin apply to living body treatment chronic anaphylaxis skin disease research
Melittin in embodiment 1 applies to living body treatment allergic skin diseases and chronic anaphylaxis skin disease is ground
Study carefully referring to Fig. 1, Fig. 2, Fig. 3.Free melittin corium injection is shown in Fig. 1, is lured in a C57/BL6 mouse single haptens
It in the allergic skin diseases model led, can effectively inhibit inflammation swelling degree caused by allergy, inhibit degree of inflammation and face
Bed conventional Hormone drug-dexamethasone has similar effect.Free melittin corium injection is shown in Fig. 2, in a C57/
In the chronic anaphylaxis skin disease model of the multiple haptens induction of BL6 mouse, it can effectively inhibit inflammation swelling caused by allergy
Degree.Fig. 3 ear skin slice HE coloration result shows that the injection of melittin corium can effectively inhibit allergic skin diseases institute
Caused swelling, inflammatory cell infiltration, epidermal tissue's paraplasm.
The toxicity research that embodiment 4 delivers the nano particle of melittin and free melittin is applied to living body compares
The toxicity that the nano particle of the delivery melittin of embodiment 2 and the free melittin of embodiment 1 are applied to living body is ground
Study carefully comparison referring to fig. 4, Fig. 5.Fig. 4 is the nano particle and free melittin tissue that injection 1mg/mL delivers melittin in skin
It is sliced the comparison of HE coloration result, free melittin will cause more serious inflammatory cell infiltration, tissue and eucaryotic cell structure loss,
And the nano particle for delivering melittin only has a small amount of inflammatory cell infiltration, and without destroying histocyte.Delivery bee is shown in Fig. 5
The nano particle of phallotoxins and free melittin mouse ear intracutaneous injection when the concentration of melittin reaches 1mg/mL
Swelling degree compares after 12h: when delivering injection 1mg/mL concentration dose in the nano particle ear of melittin, will cause 50 μm or so
Swelling；And it will cause 150 μm or so of swelling when injection 1mg/mL concentration dose in free melittin ear.Fig. 4 is delivery
Comparison mouse skin is damaged pictorial diagram after living body abdomen intracutaneous injection respectively by the nano particle of melittin and free melittin.
Free melittin will cause macroscopic skin injury and lasting swelling, and deliver the nano particle of melittin without visible
Skin lesion.
Embodiment 5: research of the nano particle of melittin for living body treatment chronic anaphylaxis skin disease is delivered
The nano particle security application of delivery melittin in embodiment 2 is in living body treatment chronic anaphylaxis skin disease
Research, referring to Fig. 5 and Fig. 6.Fig. 7 ear skin slice HE coloration result shows that the nano particle of melittin can effectively inhibit
Swelling, inflammatory cell infiltration, epidermal tissue's paraplasm caused by quick property skin disease.Figure 12 shows delivery melittin
Proinflammatory relevant cell factor IFN-γ and IL-4 expression in serum can be effectively reduced in nano particle.
Embodiment 6: the nano particle for delivering melittin can be accumulated effectively in skin-draining lymph nodes and inhibit half to resist
The research of former specific C D8+T cell Proliferation and the release for inhibiting the cellular inflammation factor
The nano particle of the delivery melittin of embodiment 2 can be accumulated effectively in skin-draining lymph nodes and inhibit half to resist
The research of former specific C D8+T cell Proliferation and the release for inhibiting the cellular inflammation factor is referring to Fig. 8, Fig. 9, Figure 10 and Figure 11.
As shown in figure 8, the nano particle and core of root of the tail portion two sides intracutaneous injection delivery melittin load fluorescent dye DIR-BOA,
A large amount of fluorescent dye signals are accumulated as the result is shown when 12h, in skin-draining lymph nodes, and are dissected mouse and be found only at mouse skin
Groin and oxter draining lymph node and liver in find stronger fluorescence signal, illustrate delivery melittin nanometer
Particle can enter lymphatic system and adjust immune response, and bear internal organs smaller.Fig. 9 and Figure 10 shows delivery melittin
Nano particle can effectively inhibit the CD8+T cell Proliferation of allergen specificity in lymph node；Figure 11 shows delivery melittin
Nano particle and the lymphocyte of allergy primary stimuli are incubated for culture altogether, detect cell conditioned medium, effectively inhibit inflammation related thin
The expression of intracellular cytokine IFN-γ.
Embodiment 7: the three classes component (phosphatide, cholesteryl ester, polypeptide) of the nano particle of melittin is delivered in clinic
On application
The three classes component (phosphatide, cholesterol acid ester, polypeptide) of the nano particle of the delivery melittin of embodiment 2 is all
Clinically there is application.
Phospholipid substance has been applied to clinical treatment, and liposome is exactly a good example.Doxil is a kind of to use rouge
Plastid package adriamycin is used to treat the drug of Kaposi's sarcoma, and U.S. FDA has been approved by Doxil for oophoroma and multiple
The treatment of property myeloma.
Cholesterol is itself existing substance in human organism, has good biological safety.
Polypeptides matter also there are many being applied to clinical treatment, such as: cervus and cucumis polypeptide is exactly that a kind for the treatment of well is fractured
Drug.Self-bone grafting polypeptide biotic factor as the drug main component, which can effectively facilitate, influences bon e formation and suction in body
The synthesis of the bdgf of receipts.Allium ovalifolium hand-mazz can be obviously promoted the formation of knitting and new bone, adjust bone generation
It thanks, has the effects that anti-inflammatory and antalgic, no obvious adverse reaction, to rheumatoid curative for effect to nonunion osteoporosis
Property arthritis soft tissue injury curative effect is excellent.
The present invention is not limited to the above-described embodiments, for those skilled in the art, is not departing from
Under the premise of the principle of the invention, several improvements and modifications can also be made, these improvements and modifications are also considered as protection of the invention
Within the scope of.The content being not described in detail in this specification belongs to the prior art well known to professional and technical personnel in the field.
Sequence table
<110>Central China University of Science and Technology
Ezhou Industrial Technology Research Institute, the Central China University of Science and Technology
<120>application of the nano particle of the application and delivery melittin of a kind of melittin
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 18
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 1
Asp Trp Phe Lys Ala Phe Tyr Asp Lys Val Ala Glu Lys Phe Lys Glu
1 5 10 15
Ala Phe
<210> 2
<211> 26
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 2
Gly Ile Gly Ala Val Leu Lys Val Leu Thr Thr Gly Leu Pro Ala Leu
1 5 10 15
Ile Ser Trp Ile Lys Arg Lys Arg Gln Gln
20 25
Claims (8)
1. a kind of application of melittin, it is characterised in that: the melittin is used to prepare the medicament for the treatment of allergic skin diseases.
2. a kind of application for the nano particle for delivering melittin, it is characterised in that: the nano particle of the delivery melittin is by transporting
Carry the polypeptide and phosphatide, cholesterol ester composition of melittin；It is described delivery melittin polypeptide by α helical polypeptide, catenation sequence and
Melittin is connected in series in the form of covalent bond；The nano particle of the delivery melittin is used to prepare treatment allergic skin disease
The medicament of disease.
3. a kind of application of nano particle for delivering melittin as claimed in claim 2, it is characterised in that: the medicament can
Inhibit T cells with antigenic specificity increment in lymph node, inhibits the release of the cellular inflammation factor.
4. a kind of application of nano particle for delivering melittin as claimed in claim 2, it is characterised in that: the delivery bee venom
The nano particle of peptide is also used to prepare pharmaceutical carrier, and the pharmaceutical carrier is used to load the dye molecule or drug point of medical diagnosis on disease
Son.
5. a kind of application of nano particle for delivering melittin as claimed in claim 2, it is characterised in that: the phosphatide is two
Dimyristoylphosphatidycholine (DMPC).
6. a kind of application of nano particle for delivering melittin as claimed in claim 2, it is characterised in that: the α spiral is more
The amino acid sequence of peptide is DWFKAFYDKVAEKFKEAF.
7. a kind of application of nano particle for delivering melittin as claimed in claim 2, it is characterised in that: the melittin
Amino acid sequence is GIGAVLKVLTTGLPALISWIKRKRQQ.
8. a kind of application of nano particle for delivering melittin as claimed in claim 2, it is characterised in that: the catenation sequence
Amino acid sequence be GSG.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710995177.3A CN109692327B (en) | 2017-10-23 | 2017-10-23 | Application of nano-particles for carrying melittin |
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| CN201710995177.3A CN109692327B (en) | 2017-10-23 | 2017-10-23 | Application of nano-particles for carrying melittin |
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| CN109692327B CN109692327B (en) | 2022-04-08 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114437193A (en) * | 2022-01-29 | 2022-05-06 | 陕西未来多肽生物科技有限公司 | Gold-melittin nano hybrid and application thereof |
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