Ophthalmic preparation as well as preparation method and application thereof

An ophthalmic preparation and ocular technology, applied in the field of protein engineering, can solve problems such as poor therapeutic effect

Active Publication Date: 2020-11-13
易舟(上海)生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the shortcomings of the preparations used to treat eye diseases in the prior art, such as poor therapeutic effect, the present invention provides an ophthalmic preparation containing transthyretin (TTR), its...

Method used

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  • Ophthalmic preparation as well as preparation method and application thereof
  • Ophthalmic preparation as well as preparation method and application thereof
  • Ophthalmic preparation as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1 Recombinant preparation of human transthyretin (TTR)

[0072] (1) Construction of the recombinant plasmid pETx-rhaPBAD-ttr: transform and restructure the pET-21a plasmid (purchased from the ATCC China Culture Collection Center) (the difference in sequence between the restructured plasmid and the original pET-21a plasmid is About 75%, the specific sequence is shown in SEQ ID NO: 4), and the rhaPBAD promoter (Rhamna Tang inducible) is used to replace the T7 promoter, and the human TTR optimized nucleic acid sequence is connected at the same time (as shown in SEQ ID NO: 2 , the amino acid sequence of TTR is shown in SEQ ID NO:1), and the overall nucleic acid sequence of the resulting plasmid is shown in SEQ ID NO:3. After sequencing verification (the sequencing company is Nanjing GenScript Biotechnology Co., Ltd.), the construction was successful.

[0073] (2) Expression and purification of recombinant human TTR: transform the pETx-rhaPBAD-ttr plasmid constru...

Embodiment 2

[0076] Example 2 Using sodium carboxymethylcellulose as an auxiliary material can promote TTR to pass through the corneal barrier

[0077] (1) Configure the human TTR prepared in Example 1 to be 5-30 μmol / L (containing physiological saline), and apply eye drops to C57BL / 6 mice (8 weeks old) and SD rats (8 weeks old) respectively After 3 hours, they were killed, and the vitreous and fundus samples were taken to extract protein. Using rabbit anti-His-tag antibody as the primary antibody and donkey anti-rabbit antibody as the secondary antibody, the vitreous and SD rats were measured by ELISA. Content of human TTR in fundus samples.

[0078] (2) The human-derived TTR prepared in Example 1 was configured as 10 μmol / L (containing physiological saline and 0-8 mg / mL sodium carboxymethylcellulose (purchased from Sinopharm, with a viscosity of 800-1200CP)), respectively for C57BL / 6 mice (8 weeks old) and SD rats (8 weeks old) were given eye drops, and sacrificed after 3-72 hours, and...

Embodiment 3

[0082] Example 3 Using dextran 70 as an auxiliary material can promote TTR to pass through the corneal barrier

[0083] (1) Configure the human TTR prepared in Example 1 to be 5-30 μmol / L (containing physiological saline), and apply eye drops to C57BL / 6 mice (8 weeks old) and SD rats (8 weeks old) respectively After 3 hours, they were killed, and the vitreous and fundus samples were taken to extract protein. Using rabbit anti-His-tag antibody as the primary antibody and donkey anti-rabbit antibody as the secondary antibody, the vitreous and SD rats were measured by ELISA. Content of human TTR in fundus samples.

[0084] (2) The human TTR prepared in Example 1 was configured to be 10 μmol / L (containing normal saline and 0-0.8 mg / mL dextran 70 (purchased from Sinopharm, with a molecular weight of 64000-76000)), respectively for C57BL / 6 small Rats (8 weeks old) and SD rats (8 weeks old) were given eye drops, and sacrificed after 3-72 hours, and the vitreous and fundus samples we...

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Abstract

The invention provides an ophthalmic preparation, which comprises transthyretin and pharmaceutically acceptable auxiliary materials, and the auxiliary materials are selected from one or more of carboxymethyl cellulose or salt thereof, chondroitin sulfate or salt thereof, and dextran. The invention further provides a preparation method and an application of the ophthalmic preparation. The auxiliarymaterials contained in the ophthalmic preparation and the transthyretin are mutually restrained and synergistically matched to form an organic whole, eye diseases related to ocular angiogenesis and/or ocular retinal leakage and the like can be synergistically treated, and finally, a better treatment effect is obtained.

Description

technical field [0001] The invention belongs to the technical field of protein engineering, and in particular relates to an ophthalmic preparation containing transthyretin and its preparation method and its application in the treatment of ocular diseases related to ocular angiogenesis and / or ocular retinal leakage, etc. . Background technique [0002] The treatment of eye diseases requires the effective delivery of drugs into the eye, especially age-related macular degeneration, retinitis pigmentosa, diabetic retinopathy, retinopathy of prematurity, and neurological changes caused by glaucoma; With more barriers, it is challenging to effectively deliver drugs into the eye. Therefore, ocular drug delivery systems have received extensive attention recently. [0003] According to the anatomical structure of the eye, the eye barrier mainly includes the tear barrier, the corneal / conjunctival barrier and the blood-ocular barrier; while protecting the eye, multiple barriers can e...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K9/00A61K47/38A61K47/36A61P27/02A61P9/10
CPCA61K38/17A61K9/0048A61K47/38A61K47/36A61P27/02A61P9/10
Inventor 辛瑜
Owner 易舟(上海)生物医药有限公司
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