CN111109198A - Method for constructing uveitis animal model - Google Patents
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- CN111109198A CN111109198A CN202010068340.3A CN202010068340A CN111109198A CN 111109198 A CN111109198 A CN 111109198A CN 202010068340 A CN202010068340 A CN 202010068340A CN 111109198 A CN111109198 A CN 111109198A
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- 206010046851 Uveitis Diseases 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000010171 animal model Methods 0.000 title claims abstract description 16
- 238000010276 construction Methods 0.000 claims abstract description 14
- 241000124008 Mammalia Species 0.000 claims abstract description 8
- 241000282414 Homo sapiens Species 0.000 claims description 14
- 238000002347 injection Methods 0.000 claims description 10
- 239000007924 injection Substances 0.000 claims description 10
- 241000282567 Macaca fascicularis Species 0.000 claims description 8
- 241000282560 Macaca mulatta Species 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 7
- 238000012216 screening Methods 0.000 claims description 2
- 238000007920 subcutaneous administration Methods 0.000 claims 1
- 241000282693 Cercopithecidae Species 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 2
- 238000011156 evaluation Methods 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 abstract description 2
- 230000000857 drug effect Effects 0.000 abstract 1
- 239000002547 new drug Substances 0.000 abstract 1
- 210000001508 eye Anatomy 0.000 description 9
- 206010051625 Conjunctival hyperaemia Diseases 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 238000002583 angiography Methods 0.000 description 5
- 238000012014 optical coherence tomography Methods 0.000 description 5
- 208000006069 Corneal Opacity Diseases 0.000 description 4
- 206010030113 Oedema Diseases 0.000 description 4
- 210000003161 choroid Anatomy 0.000 description 4
- 210000004087 cornea Anatomy 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000004410 intraocular pressure Effects 0.000 description 4
- 238000011809 primate model Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 210000003484 anatomy Anatomy 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 210000004240 ciliary body Anatomy 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 230000001575 pathological effect Effects 0.000 description 3
- 230000007170 pathology Effects 0.000 description 3
- 230000035479 physiological effects, processes and functions Effects 0.000 description 3
- 210000001525 retina Anatomy 0.000 description 3
- 230000002207 retinal effect Effects 0.000 description 3
- 210000001210 retinal vessel Anatomy 0.000 description 3
- 238000010254 subcutaneous injection Methods 0.000 description 3
- 239000007929 subcutaneous injection Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 241000700198 Cavia Species 0.000 description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 201000010183 Papilledema Diseases 0.000 description 2
- 206010038886 Retinal oedema Diseases 0.000 description 2
- 208000034700 Vitreous opacities Diseases 0.000 description 2
- 210000002159 anterior chamber Anatomy 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 230000001886 ciliary effect Effects 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 210000005081 epithelial layer Anatomy 0.000 description 2
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 201000011195 retinal edema Diseases 0.000 description 2
- 208000002306 Eye Manifestations Diseases 0.000 description 1
- 208000003098 Ganglion Cysts Diseases 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- LJUIOEFZFQRWJG-KKIBDXJDSA-N S-[2,3-bis(palmitoyloxy)propyl]-Cys-Ser-Lys-Lys-Lys-Lys Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCC)CSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O LJUIOEFZFQRWJG-KKIBDXJDSA-N 0.000 description 1
- 208000005400 Synovial Cyst Diseases 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 210000000795 conjunctiva Anatomy 0.000 description 1
- 201000001891 corneal deposit Diseases 0.000 description 1
- 231100000269 corneal opacity Toxicity 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000010339 dilation Effects 0.000 description 1
- 230000002497 edematous effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000009540 indirect ophthalmoscopy Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 201000004614 iritis Diseases 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004126 nerve fiber Anatomy 0.000 description 1
- 230000008397 ocular pathology Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 108091008695 photoreceptors Proteins 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 210000003583 retinal pigment epithelium Anatomy 0.000 description 1
- 210000001957 retinal vein Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/20—Animals treated with compounds which are neither proteins nor nucleic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/106—Primate
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a construction method of a uveitis monkey model. The method is used for treating uveitis of a mammal by subcutaneously injecting HS6101 into the mammal for medical experiment. The uveitis animal model constructed by the invention provides a new clinical pre-drug effect evaluation tool for the development of new drugs for uveitis.
Description
Technical Field
The invention belongs to the field of disease models, and particularly relates to a method for constructing a uveitis animal model.
Background
Uveitis, also known as uveitis, is a general term for inflammation of iris, ciliary body, and choroid tissues. The disease is a common disease in ophthalmology, can cause some serious complications and sequelae, and is one of the main blinding causes. In order to develop a drug for uveitis, a corresponding experimental animal model is essential.
At present, the animal model of uveitis is constructed by a method of inducing autoimmunity and immune mediation by using a plurality of photorefractive tissues including photoreceptor protein, melanin components and the like. Animals mostly use big mice, guinea pigs and rabbits, and have larger differences with human fundus anatomical structures, physiology and pathology; the anatomical structure, physiology and pathology of the eyeground of the non-human primate are relatively similar, but the non-human primate uveitis model is not reported at present.
HS6101, also known as CBLB612, is a novel derivative of a lead medicament CBLB-601. Research shows that HS6101 can inhibit the apoptosis of normal cells induced by stress, and the research shows that the effect of the HS6101 can be used for protecting healthy tissue cells in radiation and chemotherapy. To date, no clinical patent medicine of the compound is available. Nor was any information available on the use of this compound to construct monkey uveitis.
Disclosure of Invention
The invention aims to provide a monkey uveitis animal model and a construction method thereof.
Firstly, the invention provides a method for constructing a uveitis animal model, which is a method for injecting HS6101 into a mammal to make the mammal suffer from uveitis.
In the aforementioned model construction method, the mammal is a non-human primate. The model construction method as described above, wherein the non-human primate is a cynomolgus monkey or a rhesus monkey.
The model construction method as described above, wherein the HS6101 single injection dose is: 250-1000 μ g/kg.
The model building method comprises the following steps of: each injection was 250. mu.g/kg, 2 times per week for 1-4 weeks.
The model building method comprises the following steps of: 500 μ g/kg 2 times a week for 4 weeks.
In the aforementioned model building method, the injection is subcutaneous injection.
The invention further provides application of HS6101 in constructing a uveitis animal model.
The invention also provides application of the uveitis animal model constructed by the method in screening medicines for treating uveitis.
The inventor accidentally finds in research work that HS6101 can induce animals to generate acute uveitis, so a brand-new method for constructing a uveitis animal model is provided from a design idea different from the prior art.
Firstly, the invention firstly proposes that HS6101 is used for constructing a uveitis model, and expands the medical scientific research application of HS 6101.
In addition, the invention firstly constructs a non-human primate model of uveitis, compared with models of large mice, guinea pigs, rabbits and the like, the ocular fundus anatomical structure, physiology and pathology of the model are closer to those of human beings, and the model is more suitable for the development of related treatment methods and medicines of human uveitis.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The foregoing aspects of the present invention are explained in further detail below with reference to specific embodiments. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1: uveitis model-cynomolgus monkey eye performance.
FIG. 2: uveitis model-rhesus eye manifestations.
FIG. 3: intra-ocular examination of cynomolgus monkeys-OCT examination; a is before molding, B is after molding; the ordinate Thickness (Thickness) is in μm.
FIG. 4: cynomolgus monkey intraocular examination-fundus fluorography.
Detailed Description
Example 1 construction of a non-human primate model of uveitis
Cynomolgus monkeys were taken and injected subcutaneously with 250 ug/kg of HS6101 at a frequency of 2 times per week for 1-4 weeks.
Example 2 construction of non-human primate model of uveitis
Rhesus monkeys were injected subcutaneously with 1000. mu.g/kg of HS6101 at a frequency of 2 injections per week for 1-4 weeks.
Example 3 construction of non-human primate model of uveitis
Rhesus monkeys were injected subcutaneously with 500. mu.g/kg HS6101 at a frequency of 2 times per week for 1-4 weeks.
The invention is further described below in the form of experimental cases.
Experimental example 1 construction of uveitis cynomolgus monkey model
1. Experimental animals: a cynomolgus monkey.
2. Method of producing a composite material
HS6101 was given subcutaneously 2 times a week for 4 weeks at a dose of 500. mu.g/kg per administration with 4 cynomolgus monkeys.
Slit-lamp biomicroscopy, indirect ophthalmoscopy and intraocular pressure examinations were performed on days 8, 12, 18 and 25, respectively. Fundus fluorescence angiography and OCT (optical coherence tomography) examination on day 18. After the experiment, the eyeball is subjected to pathological examination.
3. Results
Monkeys injected within 1 week all developed ocular and conjunctival congestion (conjunctival congestion and ciliary congestion) with edema and pigmentation, and opaque cornea (fig. 1).
Eye examination on day 8 revealed decreased intraocular pressure, mild corneal haze, conjunctival congestion (1/4 ratio), anterior chamber exudation (2/4 ratio), large limbal pigment deposition (2/4 ratio), mild vitreous haze (1/4 ratio), disc edema (3/4 ratio), and the like.
Eye examination on day 12 revealed increased anterior chamber exudation (3/4 rate), decreased intraocular pressure (3/4 rate), mild corneal opacity (2/4 rate), mild vitreous opacity (3/4 rate), and retinal edema on and around the optic disc (3/4 rate). The degree of conjunctival congestion (1/4 ratio) was reduced compared to the former.
Eye examination on day 18 revealed a reduction in the symptoms of conjunctival congestion, corneal clouding, etc., as compared to the earlier stage. The OCT examination of the intraocular examination proves that the retina around the blood vessel of the fundus is obviously edematous and thickened, the reflection of the blood vessel of the retina is enhanced, and the definition of the boundary of each layer of the structure of the retina is reduced; fundus fluorescence angiography examination does not show retinal vessel changes, but fluorescein appears in the later period of angiography, indicating that the inner eye has fluorescence leakage.
Histopathological examination at day 25 revealed inflammatory foci in the ciliary body, choroid, retinal blood vessels, cornea (limbus), and conjunctival epithelial layer.
And judging that the acute uveitis model is successfully constructed by combining the clinical pathological characteristics and the histopathological changes of the ophthalmic examination.
4. Conclusion
The cynomolgus monkey can successfully induce an acute uveitis disease model by subcutaneous injection of HS 6101.
Experimental example 2 construction of uveitis rhesus monkey model
1. Experimental animals: rhesus monkey.
2. Method of producing a composite material
HS6101 was given subcutaneously 2 times a week for 4 weeks at a dose of 500. mu.g/kg per administration of 4 rhesus monkeys.
3. Results
Animals receiving injections within 1 week developed ocular bulbar conjunctival congestion (conjunctival congestion and ciliary congestion) with nebulous clouding of the cornea (fig. 2).
On day 8, eye examination revealed mixed redness of the conjunctiva of the eyes (3/4 ratio), increased or decreased intraocular pressure (2/4 ratio), corneal haze edema (1/4 ratio), glary flare (2/4 ratio), post-corneal deposits (2/4 ratio), disc edema (1/4 ratio), mild vitreous opacity (2/4 ratio), and varicose dilation of retinal veins (1/4 ratio).
On day 12, except for increased disc and peripheral retinal edema (3/4 rate), the severity of ocular pathology generally decreased;
OCT examination at day 18 confirmed that the retinal thickness increased and the structures of the retinal nerve fiber layer, ganglion cell layer, retinal pigment epithelium layer, and choroid were sharply demarcated (FIG. 3). Fundus fluorescence angiography examination showed no retinal vascular changes, but in the late phase of angiography fluorescein appeared in part of the eye indicating a fluorescence leak in the inner eye (fig. 4).
On day 25, histopathological examination revealed inflammatory foci in the ciliary body, choroid, retinal blood vessels, cornea (limbus), and conjunctival epithelial layer.
And judging that the acute uveitis model is successfully constructed by combining the clinical pathological characteristics and the histopathological changes of the ophthalmic examination.
4. Conclusion
Subcutaneous injection of HS6101 into rhesus monkeys successfully induced an acute uveitis disease model.
In conclusion, HS6101 is used for constructing a primate uveitis disease model, and a clinical prodrug efficacy evaluation tool closer to human is provided for developing new uveitis drugs.
Claims (9)
1. A method for constructing a uveitis animal model, which is a method for injecting HS6101 into a mammal to make the mammal suffering from uveitis; the structural formula of HS6101 is as follows:
2. the method of construction of claim 1, wherein the mammal is a non-human primate.
3. The method of claim 2, wherein the non-human primate is a cynomolgus monkey or a rhesus monkey.
4. The method of claim 3, wherein the single injection dose is: 250-1000 μ g/kg.
5. The construction method according to claim 4, wherein the injection method of HS6101 is: each injection was 250. mu.g/kg, 2 times per week for 1-4 weeks.
6. The construction method according to claim 5, wherein the injection method of HS6101 is: 500 μ g/kg 2 times a week for 4 weeks.
7. The method of claim 1, wherein the injection is subcutaneous.
Use of HS6101 in the construction of an animal model of uveitis.
9. Use of an animal model of uveitis constructed according to the method of any one of claims 1 to 6 in screening for a medicament for the treatment of uveitis.
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| CN2019102565979 | 2019-03-29 | ||
| CN201910256597 | 2019-03-29 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113057142A (en) * | 2021-03-30 | 2021-07-02 | 四川大学华西医院 | Method for constructing intraretinal and/or subretinal fibrosis animal model |
| CN113252912A (en) * | 2021-07-16 | 2021-08-13 | 中山大学中山眼科中心 | Auxiliary prevention and treatment system for uveitis |
| CN114568343A (en) * | 2022-03-11 | 2022-06-03 | 成都合拓创展生物科技有限公司 | Headwear for preparing eye disease model, application of headwear and preparation method of eye disease model |
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| CN114568343B (en) * | 2022-03-11 | 2023-04-07 | 成都合拓创展生物科技有限公司 | Headwear for preparing eye disease model and application thereof |
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