CN1242811C - New use of recombinated hirudin - Google Patents
New use of recombinated hirudin Download PDFInfo
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- CN1242811C CN1242811C CNB031322247A CN03132224A CN1242811C CN 1242811 C CN1242811 C CN 1242811C CN B031322247 A CNB031322247 A CN B031322247A CN 03132224 A CN03132224 A CN 03132224A CN 1242811 C CN1242811 C CN 1242811C
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- ludon
- lepirudin
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- cataract
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Abstract
The present invention discloses a new application of recombinant hirudin, more specifically an application in the preparation of medicines for preventing and curing cataract.
Description
Technical field
The present invention relates to the new purposes of lepirudin 023 ludon, relate in particular to the purposes in pharmaceutical field.
Background technology
Hirudin is 65-66 the amino acid whose albumen that contains from the extraction of Hementaria officianalis salivary gland, be single chain polypeptide, molecular mass is 7000u, contain 3 pairs of disulfide bond in the molecule, formed highly folding, fine and close tertiary structure in the 1-49 position, make this domain firm, and linear C terminal sequence is in order to bind thrombin.As the specific inhibitor of thrombin, it has very strong anticoagulating active, can effectively prevent and treat in the dispersivity blood vessel to condense, and aspect the diseases such as painstaking effort bolt and cerebral thrombosis remarkable effect is being arranged all.Because the natural hirudin source is limited, extensively utilizes biotechnology to prepare lepirudin 023 ludon at present, two class cellular expression systems are respectively escherichia coli (E.coli) and brewer yeast (S.cerevisiae) the most widely.The preparation method of lepirudin 023 ludon is disclosed among the CN1319671A.Clinically, lepirudin 023 ludon be used for the treatment of more unstable angina pectoris (USA), acute myocardial infarction (AMI), angioplasty, postoperative thrombosis, hemodialysis, extracorporeal circulation, disseminated inravascular coagulation (DIC) etc. (MarkwardtF.Hirudin as an inhibitor of thrombin[J] .Methods in Enzymelogy, 1970,19:924) .Clinical experiment shows that lepirudin 023 ludon does not cause allergy, immunoreation, and can not cause the circulatory function obstacle.But do not see pharmacology report and clinical practice that prevention and treatment cataract disease are arranged.
Summary of the invention
The invention discloses a kind of new purposes, the particularly lepirudin 023 ludon application in the medicine of preparation prevention and treatment cataract disease of lepirudin 023 ludon.
In cataractous generating process, all change: quicken as lipid peroxidation with intracrystalline all biochemical metabolisms, aldose reductase activity increases, superoxide dismutase and glutathion reductase isoreactivity reduce, the ATP lowering of concentration, some aminoacid, vitamin and reducing substances reduce, electrolyte balance imbalance and crystalline protein polymerization and structural property change etc.Therefore; it also is many-sided preventing and treating cataractous approach: as utilize antioxidant or antioxidation activator with remove or in and ultraviolet, ionizing radiation, chemical substance and metabolite etc. to crystalline harmful damage; by vitimin supplement, coenzyme (NADP), glutathion and ATP etc.; change to reduce intracrystalline biochemical metabolism; by some reactive groups of closed protein matter, avoid having salt damage chemical substance formation adduct and cause damage with protected protein matter.Whether in pharmacology test, it is useful to improving cataract to detect a medicine, sees usually whether medicine to be measured can make SOD vigor rising in the crystal, the decline of MDA content.
Our test finds that lepirudin 023 ludon is non-stimulated to eye, can significantly postpone the cataract time of origin, alleviates the muddy degree of crystalline lens, reduces MDA content, increases the SOD vigor.Cataract there is significant preventive and therapeutic effect.
In order to bring into play curative effect better, lepirudin 023 ludon is prepared into pharmaceutically common dosage forms, preferred preparation is eye drop, Eye ointments, injection.In the preparation of eye drop, remove adding buffer solution, also can add the non-stimulated short absorbent of eye, thickening agent, isotonic agent, antibacterial etc.The dosage of topical ophthalmic medicine is generally 0.001～0.01g/ day, also can exceed this dosage range according to different situations.
Below be part pharmacology test of the present invention and data, used lepirudin 023 ludon prepares according to the CN1319671A method:
One, eye irritant test repeatedly
Experimental animal: rabbit 1.8-2.3Kg, 4.Nanjing An Limo Science and Technology Ltd., credit number: SCXK (Soviet Union) 20020-0002 test method: undertaken by " preclinical study guideline compilation " bureau of drug administration of Ministry of Health of the People's Republic of China standard, (0.5g/100ml) splashes in the conjunctival sac with the lepirudin 023 ludon eye drop.About 10 seconds of the eyelid of gently sleeping then is to guarantee to be tried thing and partial full contact.An other eye splashes into solvent in the conjunctival sac, in contrast.
Observation period and observation index: 1,24,48,72hr carries out examination of eyes, observed continuously 7 days.Except should observing conjunctiva, cornea, iris, other viewed damages also should write down and report.The each inspection all should be write down the score value of the reaction of eye.
Result of the test: the irritative response score value addition of cornea, iris and the conjunctiva of each animal is got total mark, one group total mark divided by number of animals, is promptly got last score value, see the following form.
Respectively organize the eyes final score each observing time
| The experimental animal group | 1h | 24h | 48h | 72h | 4d | 5d | 6d | 7d |
| Administration one branch hole eyeball contrasts a branch hole eyeball | 0 0 | 0 0 | 0 0 | 0 0 | 0 0 | 0 0 | 0 0 | 0 0 |
Conclusion (of pressure testing): the lepirudin 023 ludon eye drop zest do not occur to administration one branch hole eyeball cornea, iris and conjunctiva, and is the same with contrast one branch hole eyeball; The lepirudin 023 ludon eye drop does not have eye irritation.
Two, control rat cataract test
Experimental animal: 60 of 28 age in days SD rats, body weight 53.5-68g, male and female half and half.Provide the quality certification number by China Medicine University's Experimental Animal Center: SCXK (Soviet Union) 2001-001.
Test method: 60 of 28 age in days SD rats, male and female half and half, be divided into 6 groups at random, every group 10: normal control group (normal saline), model control group (a not administration of modeling), positive controls (catalin Pir:0.4mg/kg), high, medium and low dosage group (the lepirudin 023 ludon 1.68mg/kg of drop in the conjunctival sac, 0.84mg/kg, 0.42mg/kg).Each is organized the administration volume and is 0.25ml/50g.Remove the normal control group, the D-galactose solution of other group lumbar injections 0.08% is equivalent to 20mg/kg, and 1 time/day, lumbar injection is 14 days continuously, and the administration volume is 25ml/kg, and drinks the 10%D-galactose solution.At the 4th day that brings out galactose cataract, put eyes high dose (1.68mg/kg) respectively, middle dosage (0.84mg/kg) and low dosage (0.42mg/kg) lepirudin 023 ludon (5 times/day).Began to observe the turbidity of rat lens on the 3rd day, and write down lenticular variation in the 8th, 14 day, after experiment finishes, get rat lens, measure MDA content and SOD vigor in the crystalline lens with eye-checking instrument.
The crystalline lens observed result is carried out statistical analysis with the Ridit method; In MDA and the SOD measurement result each group all and the cataract model group organize t and check.
Result of the test:
1, lepirudin 023 ludon influence that the rat lens due to the D-galactose is changed.The results are shown in Table 1, table 2.
Table 1, lepirudin 023 ludon are to the influence (the 8th day) of cataract rat lens turbidity (n=10)
| Group | Dosage (mg/kg) | / | + | ++ | +++ | Add up to |
| Dosage r-H low dosage r-H adds up among the negative group model group Pir high dose r-H | ---- ---- 0.4 1.68 0.84 0.42 | 10 0 8 7 8 8 41 | 0 0 2 3 2 1 8 | 0 2 0 0 0 1 3 | 0 ** 8 0 ** 0 ** 0 ** 0 ** 8 | 10 10 10 10 10 10 60 |
/: the nothing muddiness+: slight haze ++: medium muddiness +++: serious muddy
*Compare with model group expression u＞2.58
Table 2, lepirudin 023 ludon are to the influence (the 14th day) of cataract rat lens turbidity (n=10)
| Group | Dosage (mg/kg) | / | + | ++ | +++ | Add up to |
| Dosage r-H low dosage r-H adds up among the negative group model group Pir high dose r-H | ---- ---- 0.4 1.68 0.84 0.42 | 10 0 9 8 7 8 42 | 0 0 1 2 3 1 7 | 0 1 0 0 0 1 2 | 0 ** 9 0 ** 0 ** 0 ** 0 ** 9 | 10 10 10 10 10 10 60 |
/: the nothing muddiness+: slight haze ++: medium muddiness +++: serious muddy
*Compare with model group expression u＞2.58
The result shows: each administration group crystalline lens turbidity all has highly significant difference than model group.
2, respectively organize SOD vigor comparison in the rat lens: the results are shown in Table 3
Table 3 lepirudin 023 ludon is to influence (x ± S) (n=10) of SOD vigor in the cataract rat lens
| Group | Dosage (mg/kg) | SOD(NU/mgprot) |
| Negative group model group pir high dose r-H | ---- ---- 0.4 1.68 | 137.2±13.7 ** 37.5±19.0 133.5±31.3 ** 130.4+24.9 ** |
| Middle dosage r-H low dosage r-H | 0.84 0.42 | 123.6±18.8 ** 122.2±19.2 ** |
*Compare with model group P＜0.01
The result shows: lepirudin 023 ludon high dose group and positive drug catalin be obvious increased SOD vigor all, with model control group utmost point significant difference (p＜0.01) is arranged relatively, and with being proportionate property of dosage.
3, respectively organize MDA content comparison in the rat lens: see Table 4
Table 4 lepirudin 023 ludon is to influence (x ± S) (n=10) of MDA content in the cataract rat lens
| Group | Dosage (mg/kg) | MDA(nmol/mgprot) |
| Dosage r-H low dosage r-H among the negative group model group pir high dose r-H | ---- ---- 0.4 1.68 0.84 0.42 | 6.34±1.19 ** 24.29±4.72 14.81±2.91 ** 15.14±3.32 ** 16.19±3.27 ** 16.87±4.24 ** |
*Compare with model group P＜0.01
The result shows: high, medium and low dosage group of lepirudin 023 ludon and positive drug catalin all can obviously reduce MDA content, with model control group utmost point significant difference (p＜0.01) are arranged relatively, and with being proportionate property of dosage.
Conclusion (of pressure testing): lepirudin 023 ludon can significantly be postponed the cataract time of origin, alleviates the muddy degree of crystalline lens, reduces MDA content, increases the SOD vigor.Double lactose-induced rat cataract of lepirudin 023 ludon has significant preventive and therapeutic effect.
The specific embodiment
Lepirudin 023 ludon eye drop prescription and preparation method thereof:
Lepirudin 023 ludon 0.25g
Boric acid 1.116g
Borax 0.191g
Sodium chloride 0.22g
EDTA 0.5g
Chitosan 0.5g
Ethylparaben 0.03g
Distilled water adds to 100ml
Boric acid, Borax add the buffer solution that water is prepared into pH7.4, dissolve lepirudin 023 ludon, EDTA, chitosan, sodium chloride, the own ester of P-hydroxybenzoic acid with buffer solution, and mixing, filtration, degerming packing promptly get lepirudin 023 ludon eye drop of the present invention.
Claims (1)
1, the application of lepirudin 023 ludon in the medicine of preparation prevention and treatment cataract disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031322247A CN1242811C (en) | 2003-08-04 | 2003-08-04 | New use of recombinated hirudin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031322247A CN1242811C (en) | 2003-08-04 | 2003-08-04 | New use of recombinated hirudin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1483473A CN1483473A (en) | 2004-03-24 |
| CN1242811C true CN1242811C (en) | 2006-02-22 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB031322247A Expired - Fee Related CN1242811C (en) | 2003-08-04 | 2003-08-04 | New use of recombinated hirudin |
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| CN (1) | CN1242811C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102727874B (en) * | 2012-07-18 | 2014-02-19 | 中国药科大学 | Recombinant hirudin eye drops and preparation method thereof |
| CN103071186B (en) * | 2012-12-19 | 2014-05-07 | 华南理工大学 | Surface-modified artificial lens with coating layer carrying drug and preparation method thereof |
| CN104906037A (en) * | 2015-06-23 | 2015-09-16 | 广西复鑫益生物科技有限公司平南分公司 | Recombinant hirudin eye drop and preparation method thereof |
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| CN1483473A (en) | 2004-03-24 |
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Application publication date: 20040324 Assignee: Changzhou Qianhong Biopharma Co., Ltd. Assignor: China Pharmaceutical University Contract record no.: 2010320000308 Denomination of invention: New use of recombinated hirudin Granted publication date: 20060222 License type: Exclusive License Record date: 20100324 |
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Assignee: Changzhou Qianhong Biopharma Co., Ltd. Assignor: China Pharmaceutical University Contract record no.: 2010320000308 Date of cancellation: 20120817 |
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