CN116549459B - Application of pregnane alkaloid derivative in preparation of drugs for treating fundus diseases - Google Patents
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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
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
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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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0048—Eye, e.g. artificial tears
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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Abstract
The invention belongs to the technical field of medicines, and discloses application of pregnane alkaloid derivatives in preparation of medicines for treating fundus diseases. The invention overcomes the defect of clinical application of anti-VEGF antibody medicines in the prior art, discovers and proves the application of pregnane alkaloid derivative in medicines for treating fundus diseases, can effectively treat AMD, PDR and ROP, is hopeful to become a new medicine for treating fundus diseases, further improves patient compliance by eye surface eye drop administration, and avoids side effects caused by intravitreal injection.
Description
Technical Field
The invention belongs to the technical field of medicines, and in particular relates to application of pregnane alkaloid derivatives in preparation of medicines for treating fundus diseases.
Background
Age-related macular degeneration (Age-related Macular Degeneration, AMD), proliferative diabetic retinopathy (Proliferative Diabetic Retinopathy, PDR), retinopathy of prematurity (Retinopathy of Prematurity, ROP) are common irreversible blinding fundus diseases, which are commonly characterized by fundus pathology, proliferative neovascular, and are also major factors in blinding. Currently, the main clinical therapeutic drugs are anti-VEGF antibody drugs, including ranibizumab, bevacizumab and the like. However, such antibody drugs are required to be repeatedly injected through the vitreous cavity, patient compliance is poor, risks of complications such as endophthalmitis are increased, and treatment cost is high. In addition, the effective rate of the antibody medicines is about 40% -60% clinically, and a large number of patients are insensitive to VEGF medicines or develop drug resistance. Therefore, the clinical application of the antibody medicament is greatly limited, and medicaments which can be replaced, have good curative effect and are convenient to administer are still required to be developed for treating the fundus neovascularization diseases.
Compared with macromolecular biological medicines, the micromolecular medicines have the advantages of high stability, easy administration, low cost and the like. At present, in preclinical research, small molecular angiogenesis inhibitors applied to anti-tumor neovascular treatment or natural products and derivatives with anti-angiogenesis activity are applied to research or patent report on treating fundus neovascular diseases. In addition, small molecule inhibitors applied to the treatment of ocular fundus neovascular diseases have not been marketed until now.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides application of pregnane alkaloid derivatives in preparing medicaments for treating fundus diseases.
The technical scheme adopted for solving the technical problems is as follows:
an application of pregnane alkaloid derivative in preparing a medicament for treating fundus diseases, wherein the pregnane alkaloid derivative has a structural formula as follows:
。
further, the ocular fundus disease includes wet age-related macular degeneration and/or proliferative diabetic retinopathy and/or retinopathy of prematurity.
Further, the application also comprises application in medicines for inhibiting ocular choroidal neovascularization.
Further, the use also includes use in inhibiting the non-perfused area of the retina of the eye and/or neovascular medicament.
Further, the administration mode of the pregnane alkaloid derivative comprises the following steps: intravitreal injection, subretinal, subconjunctival, ocular surface, postbulbar, and/or peribulbar. Optimal intravitreal injection administration.
Further, the solvent of the medicine is physiological saline solution containing dimethyl sulfoxide with the volume concentration of 0.2-0.05%. Preferably 0.1% dimethyl sulfoxide (volume fraction).
Further, the drug was administered at a concentration of 2.5 mg/ml, 5 mg/ml and 10 mg/ml. And most preferably 10 mg/ml.
Further, the administration volume of the medicine is 0.5-2 microliters. Preferably 1 microliter.
Further, the administration frequency of the medicine is 1-5 times per week for one week. Preferably 1 time.
The invention has the advantages and positive effects that:
1. the invention overcomes the defect of clinical application of anti-VEGF antibody medicines in the prior art, discovers and proves the application of pregnane alkaloid derivative in medicines for treating fundus diseases, can effectively treat AMD, PDR and ROP, is hopeful to become a new medicine for treating fundus diseases, further improves patient compliance by eye surface eye drop administration, and avoids side effects caused by intravitreal injection.
2. The invention discloses an application of pregnane alkaloid derivative in fundus diseases, and the pregnane alkaloid derivative can inhibit proliferation and migration of retinal endothelial cells and the capability of forming tubules on a cell model.
3. The invention discloses application of pregnane alkaloid derivative in fundus diseases, which can effectively inhibit leakage and proliferation of pathological new blood vessels of choroid in an animal model of wet AMD.
4. The invention discloses application of pregnane alkaloid derivative in fundus diseases, which can effectively inhibit formation of a retinal perfusion-free region and proliferation of retinal pathological new blood vessels in an animal model of PDR.
5. Compared with the commercial drug ranibizumab, the pregnane alkaloid derivative is high in stability, less influenced by environmental factors such as temperature and humidity and easy to store for a long time; simple synthesis equipment, simple synthesis process and low cost of synthesis raw materials.
6. The use of pregnane alkaloid derivatives of the present invention in ocular fundus diseases includes, but is not limited to, intravitreal injection, subretinal, subconjunctival, ocular surface, postbulbar or peribulbar administration. Minimally invasive or non-invasive modes of administration can promote patient compliance.
7. The in-vitro and in-vivo experiments prove that the pregnane alkaloid derivative inhibits proliferation of choroidal neovascularization on one hand and retinal non-perfusion areas and neovascularization on the other hand. The pregnane alkaloid derivative can effectively treat AMD, PDR and ROP, is expected to replace the existing anti-VEGF antibody medicament, and has important clinical application value.
8. The pregnane alkaloid derivative is a novel pregnane alkaloid angiogenesis inhibitor (CN 201310749522.7) which is obtained by taking pregnane alkaloid extracted from a Tujia folk medicine, namely, lycopodium clavatum as a lead compound for derivative synthesis. Early researches found that the pregnane alkaloid derivative can regulate and control HIF-1 alpha/VEGF/VEGFR 2 signal channels by targeting and combining HSP90 alpha, thereby remarkably inhibiting tumor angiogenesis. In view of the excellent anti-angiogenic activity and the characteristic of local administration of eyes, the pregnane alkaloid derivative provided by the invention is expected to exert an excellent curative effect in treating ocular fundus neovascular diseases.
Drawings
FIG. 1 is a diagram showing the cytotoxicity of pregnane alkaloid derivatives against monkey retinal endothelial cells;
FIG. 2 is a graph showing the ability of pregnane alkaloid derivatives of the present invention to inhibit proliferation of monkey retinal endothelial cells;
FIG. 3 is a graph showing the ability of pregnane alkaloid derivatives of the present invention to inhibit migration of retinal endothelial cells in monkeys: wherein, (A) migration condition diagram, (B) statistical analysis diagram;
fig. 4 is a graph showing the ability of pregnane alkaloid derivatives of the present invention to inhibit the formation of tubules in monkey retinal endothelial cells: wherein, (A) a tubule formation map, (B) a statistical analysis map;
FIG. 5 is a graph showing leakage of pregnane alkaloid derivatives of the present invention for inhibiting choroidal neovascularization in a wet AMD animal model (laser induced choroidal neovascularization model): wherein, (a) fundus photography and fluorescein sodium fundus angiography, (B) fluorescence intensity map of the leak area, (C) Choroidal Neovascularization (CNV) grade statistics;
FIG. 6 is a graph showing inhibition of proliferation of choroidal neovascularization in wet AMD animal models (laser induced choroidal neovascularization models) by pregnane alkaloid derivatives of the present invention: wherein, (a) a vascular specific marker IB4 fluorescent staining profile, (B) a Choroidal Neovascularization (CNV) area profile, (C) an OCT imaging profile, (D) a Choroidal Neovascularization (CNV) thickness profile;
fig. 7 is a graph showing inhibition of retinal no-perfusion region formation and proliferation of new blood vessels in PDR animal models (oxygen-induced retinal neovascular models) with pregnane alkaloid derivatives according to the present invention: wherein, (A) a vascular specific marker IB4 fluorescent staining chart, (B) a retinal perfusion-free area statistical chart, and (C) a retinal neovascularization area statistical chart.
Detailed Description
The following describes the embodiments of the present invention in detail, but the present embodiments are illustrative and not limitative, and are not intended to limit the scope of the present invention.
The raw materials used in the invention are conventional commercial products unless specified; the methods used in the present invention are conventional in the art unless otherwise specified.
An application of pregnane alkaloid derivative in preparing a medicament for treating fundus diseases, wherein the pregnane alkaloid derivative has a structural formula as follows:
。
preferably, the ocular fundus disease comprises wet age-related macular degeneration and/or proliferative diabetic retinopathy and/or retinopathy of prematurity.
Preferably, the use further comprises use in a medicament for inhibiting ocular choroidal neovascularisation.
Preferably, the use further comprises use in inhibiting the non-perfused area of the retina of the eye and/or neovascular medicament.
Preferably, the pregnane alkaloid derivative is administered by a method comprising: intravitreal injection, subretinal, subconjunctival, ocular surface, postbulbar, and/or peribulbar. Optimal intravitreal injection administration.
Preferably, the solvent of the medicine is physiological saline solution containing dimethyl sulfoxide with the volume concentration of 0.2-0.05%. Preferably 0.1% dimethyl sulfoxide (volume fraction).
Preferably, the drug is administered at a concentration of 2.5 mg/ml, 5 mg/ml and 10 mg/ml. And most preferably 10 mg/ml.
Preferably, the administration volume of the drug is 0.5-2 microliters. Preferably 1 microliter.
Preferably, the administration frequency of the medicine is 1-5 times per week for one week. Preferably 1 time.
Specifically, the relevant preparation and detection examples are as follows:
example 1: preparation of pregnane alkaloid derivative dosing solution
Embodiments are described below: precisely weighing pregnane alkaloid derivative with certain mass, dissolving in dimethyl sulfoxide solution with certain volume, and fully vortex to prepare stock solution with concentration of 2.5 g/ml, 5 g/ml and 10 g/ml. And (3) absorbing a certain volume of the stock solution, adding the stock solution into 1000 times of physiological saline, and fully swirling to obtain pregnane alkaloid administration solution with the concentration of 2.5, 5 and 10 mg/ml.
Example 2: cytotoxicity of pregnane alkaloid derivative on monkey retinal endothelial cells
Embodiments are described below: culturing monkey retinal endothelial cells (RF/6A), and inoculating 96-well plates with cell number of 5×10 per well 4 After cell attachment proliferation to 70% of the plate bottom area, gradient concentration pregnane alkaloid derivatives (0, 0.01, 0.05, 0.1, 0.5,1,2.5, 5, 10, 25, 50, 100. Mu.M) were added, incubated for 24 hours, the medium was discarded, 100. Mu.L of phenol red-free medium was replaced, and 10. Mu.L of 0.5 mg/mL MTT (3- (4, 5-dimethylthiazole-2) -2, 5-diphenyltetrazolium bromide) solution was added and incubated in an incubator for 4 hours. The absorbance of each well was measured at the wavelength of enzyme-linked immunosorbent assay 450 nm.
Implementation results: as shown in FIG. 1, the pregnane alkaloid derivative of the present invention has an IC50 of 7.229. Mu.M for RF/6A cytotoxicity, and has no effect on cell viability at drug concentrations of 2.5. Mu.M or less.
Example 3: pregnane alkaloid derivative capable of inhibiting proliferation of monkey retinal endothelial cells
Embodiments are described below: culturing monkey retinal endothelial cells RF/6A, taking 96-well plate, and inoculating 5×10 cells per well 4 After cell adhesion proliferation to 70% of the plate bottom area, pregnane alkaloid derivatives (0, 0.5,1, 2.5. Mu.M) containing VEGF (vascular endothelial growth factor, 20 ng/mL) were added at gradient concentration, incubated for 24 hours, the medium was discarded, 100. Mu.L of phenol red-free medium was replaced, 10. Mu.L of 0.5 mg/mL MTT solution was added, and incubated in an incubator for 4 hours. The absorbance of each well was measured at the wavelength of enzyme-linked immunosorbent assay 450 nm.
Implementation results: as shown in fig. 2, the pregnane alkaloid derivative of the present invention can inhibit VEGF-induced proliferation activity of RF/6A in a dose-dependent manner at a non-cytotoxic concentration (0.5,1,2.5 μm), and has significant differences (P <0.05, P <0.01, P < 0.001) through statistical analysis.
Example 4: pregnane alkaloid derivative capable of inhibiting migration of monkey retinal endothelial cells
Embodiments are described below: inoculating 5X 10 per well in 12-well plate 5 After the cells were grown to the bottom of the plate and incubated with pregnane alkaloid derivatives (0, 0.5,1,2.5. Mu.M) for 24 hours, a 10. Mu.L gun head was used to gently draw a straight line at the bottom of each well plate to cause a flaw in the cells at the bottom of the plate. PBS was washed, serum-free medium was changed, photographed 0, 6, 8, 12, 24 hours after scratch, and cell mobility was calculated.
Implementation results: as shown in fig. 3, the pregnane alkaloid derivatives of the present invention are capable of dose-dependently inhibiting VEGF-induced RF/6A migration distance at non-cytotoxic concentration (0.5,1,2.5 μm) (panel a), and have significant differences (P <0.05, P <0.01, P < 0.001) through statistical analysis (panel B).
Example 5: pregnane alkaloid derivative capable of inhibiting tubule formation of monkey retina endothelial cells
Embodiments are described below: RF/6A after 24 hours of incubation with pregnane alkaloid derivatives (0, 0.5,1,2.5. Mu.M) was inoculated into Matrigel-coated 48 well plates, and after 6, 8, 10, 12 hours of cell inoculation, the cells were observed for tube formation, and the tube lengths of the tubes were analyzed by photographing.
Implementation results: as shown in fig. 4, the pregnane alkaloid derivatives of the present invention are capable of dose-dependently inhibiting VEGF-induced RF/6A tubule length at non-cytotoxic concentration (0.5,1,2.5 μm), and have significant differences (P <0.05, P <0.01, P < 0.001) as determined by statistical analysis.
In summary, the pregnane alkaloid derivative disclosed by the invention can inhibit proliferation, migration and tubule formation of VEGF-induced RF/6A in a dose-dependent manner at a non-cytotoxic concentration.
Example 6: pregnane alkaloid derivatives for inhibiting leakage and proliferation of choroidal neovascularization in wet AMD animal model (laser induced choroidal neovascularization model)
Embodiments are described below: a laser-induced choroidal neovascularization model was used as an animal model of wet AMD. C57BL/6 mice of 8 weeks old were selected, 4 spots were photocoagulated between blood vessels with a spot diameter of 50. Mu.M and an exposure time of 0.05 s, power 400 mW, focused on Bruch membranes, and the occurrence of air bubbles or slight bleeding accompanied by breaking of Bruch membranes was marked as an effective photocoagulation point by using 532 nm multiple laser to surround the distance of 2-3 discs. The case of Bruch's membranes was shown using Optical Coherence Tomography (OCT). After laser irradiation, the medicine is administered in groups, and the specific groups are as follows: blank (negative) control, positive control, 2.5. 2.5 mg/mL, 5. 5 mg/mL and 10. 10 mg/mL groups for a total of 5 groups. Wherein the blank control group is prepared by injecting 2 μl of physiological saline into vitreous cavity after laser injury, the positive control group is injected with 2 μl of 10 mg/mL of ranibizumab (Norhua pharmaceutical Co., ltd., north adapted), the pregnane alkaloid derivatives are dosed with 10 mg/mL group, and the experiment groups are respectively injected with 2 μl of 2.5, 5, 10 mg/mL pregnane alkaloid derivatives. And (3) after the model is induced, the 7 th gastrodia elata drunk mouse is subjected to mydriasis, 20 mg/mL FITC-dextran is injected intravenously, a fundus fluorescence contrast image is observed and shot by using a fundus imaging instrument of the small animal, and the change condition of each layer of retina is detected by using OCT. After imaging, the mice were removed quickly from their eyes, fixed overnight in 4% pfa, and fluorescent staining with choroidal-scleral complex fixed patches and vascular endothelial cell antibody (IB 4) was performed. The staining procedure is specifically performed by washing the tissue with a physiological saline solution containing 0.3% Triton x100 followed by co-incubation with IB4 at 4 ℃ for 12 hours. The tissue was further washed with physiological saline, then observed and photographed using a confocal laser scanning microscope, and proliferation of the new blood vessel was analyzed.
Implementation results: as shown in fig. 5, the pregnane alkaloid derivatives of the present invention inhibit leakage of choroidal neovascularization in a wet AMD animal model (laser induced choroidal neovascularization model). Wherein, the A graph is the condition of fundus angiography by fundus photography and fluorescein sodium fundus angiography, and the B graph and the C graph are the statistics of fluorescence intensity of a leakage area (a cluster fog-like part is a leakage area) and Choroidal Neovascularization (CNV) grade according to the condition of fundus angiography. As shown in figure a, very pronounced vascular leakage (diffuse, haze-like, high fluorescence areas) was observed in the blank control, and the case of vascular leakage in the negative positive control and the dosing group was significantly improved. As the concentration of pregnane alkaloid derivative increased, the fluorescence intensity in the vascular leak area (panel B) decreased while the CNV grade (panel C) decreased, and each of the dosing groups had significant differences (P <0.05, P < 0.01) compared to the placebo group.
Fig. 6 shows that a pregnane alkaloid derivative according to the present invention inhibits proliferation of choroidal neovascularization in a wet AMD animal model (laser induced choroidal neovascularization model). Wherein, the A graph shows proliferation of the suprachoroidal neovascularization marked by fluorescent-labeled vascular specific marker IB4, and the B graph shows statistics of areas according to each proliferation point in the A graph. As shown, in panel a, proliferation of pathologic neovasculature (bolus-like highlighting) was significantly reduced in the positive control group and the dosing group compared to the blank control group. As the concentration of pregnane alkaloid derivatives increased, the area of new blood vessels (panel B) decreased, with a significant difference (P < 0.001) in the 10 mg/mL dosing group compared to the placebo group. In panel C, the choroid thickness of the ocular fundus of each group of mice under OCT imaging is shown to be significantly reduced compared to the placebo group, and the results of statistical analysis (panel D) show that as the concentration of pregnane alkaloid derivative increases, the choroid thickness is continuously reduced, and there is a significant difference between the 5 mg/mL and 10 mg/mL groups compared to the placebo group (P <0.01, P < 0.001).
In conclusion, the pregnane alkaloid derivative disclosed by the invention can inhibit leakage and proliferation of choroidal neovascularization in a dose-dependent manner in a wet AMD animal model (laser-induced choroidal neovascularization model).
Example 7: pregnane alkaloid derivative for inhibiting formation of retinal perfusion-free region and proliferation of new blood vessel in PDR animal model (oxygen-induced retinal neovascular model)
Embodiments are described below: an oxygen-induced retinopathy model was used as an animal model of PDR. Will be newly outThe 7-day-old C57BL/6 rats and the female rats were placed in an oxygen chamber (75% O) 2 ) After 5 days of feeding, the mixture was taken out on the 12 th day after birth and returned to the normal environment (21% O) 2 ) The cultivation was continued for 5 days. During this period, the 2 nd day after the intake (the 9 th day after the birth) is the peak period of no perfusion region formation, and the 5 th day after the discharge (the 17 th day after the birth) is the peak period of retinal neovascularization proliferation. The non-perfused region refers to a region where the retina of a young mouse naturally degenerates to form a blood vessel-free region under the condition of high oxygen in an oxygen cabin, and forms the non-perfused region. After the young mice are subjected to oxygen cabin, in normal environment, the non-perfused area is lack of oxygen due to no blood vessels, so that a great amount of proliferation of new blood vessels is induced. Delivery (12 th day after birth) starts group administration, and the specific group is: blank (negative) control, positive control, 2.5. 2.5 mg/mL, 5. 5 mg/mL and 10. 10 mg/mL groups for a total of 5 groups. Wherein, after taking out, 1 mu L of physiological saline is injected into the vitreous cavity, 1 mu L of 4 mg/mL of ranibizumab is injected into the positive control group, and 1 mu L of 2.5, 5 and 10 mg/mL of pregnane alkaloid derivatives are respectively injected into the experimental group. On postnatal day 17, the mice' eyeballs were removed, fixed overnight in 4% PFA, and fixed patches of retina and fluorescent staining of vascular endothelial cell antibody (IB 4) were performed by washing the tissues with saline containing 0.3% Triton x100 followed by co-incubation with IB4 at 4 ℃ for 12 hours. The tissue was washed with physiological saline, then observed and photographed using a confocal laser scanning microscope, and the areas of the perfused areas and proliferation of new blood vessels were analyzed.
Implementation results: fig. 7 shows that a pregnane alkaloid derivative according to the present invention inhibits the formation of a retinal perfusion-free region and proliferation of new blood vessels in a PDR animal model (oxygen-induced retinal neovascular model). Wherein, the A diagram is the proliferation condition of the neovascular on the retina marked by the fluorescent marked vascular specific marker IB4 after the retina is tiled and the partial enlarged diagram thereof. In addition, marking a non-perfusion area in the A graph, and carrying out statistics on the areas of the non-perfusion areas of the blank control group, the positive control group and each administration group, wherein the statistical result is shown in the B graph; the neonatal vascular area was marked in panel a and the areas of the non-perfused areas were counted for the blank control group, the positive control group and each of the dosing groups, with the statistics shown in panel C. The results show that in panel a, the perfused areas and the neovascularization of the positive control group and of the respective dosing groups are significantly reduced compared to the blank control group. Statistical analysis showed that as pregnane alkaloid derivative concentration increased, the areas of non-perfused areas and areas of new blood vessels decreased, and that each of the dosing groups had significant differences (P <0.05, P <0.01, P < 0.001) compared to the placebo group. Furthermore, the non-perfused area of the 10 mg/mL dosing group was significantly reduced compared to the positive control group (P < 0.001).
In summary, the pregnane alkaloid derivatives of the invention are capable of dose-dependently inhibiting the formation of retinal perfusion-free areas and proliferation of new blood vessels in PDR animal models (oxygen-induced retinal neovascular models). And administration of 10 mg/mL has a stronger effect of inhibiting formation of a retinal perfusion-free region than ranibizumab.
Although embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the disclosure of the embodiments.
Claims (6)
1. The application of pregnane alkaloid derivative in preparing medicament for treating fundus diseases is characterized in that: the pregnane alkaloid derivative has the structural formula as follows:
；
the fundus disease is wet age-related macular degeneration or proliferative diabetic retinopathy.
2. The use according to claim 1, characterized in that: the administration mode of the pregnane alkaloid derivative comprises the following steps: intravitreal injection, subretinal, subconjunctival, ocular surface, postbulbar, and/or peribulbar.
3. Use according to claim 1 or 2, characterized in that: the solvent of the medicine is physiological saline solution containing dimethyl sulfoxide with the volume concentration of 0.05-0.2%.
4. Use according to claim 1 or 2, characterized in that: the drug was administered at concentrations of 2.5 mg/ml, 5 mg/ml and 10 mg/ml.
5. Use according to claim 1 or 2, characterized in that: the administration volume of the medicine is 0.5-2 microlitres.
6. Use according to claim 1 or 2, characterized in that: the administration frequency of the medicine is 1-5 times per week, and the medicine is used for one week.
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| 热休克蛋白90抑制剂治疗新生血管性眼病的研究现状;袁冬青 刘庆淮;中华眼底病杂志;第28卷(第5期);摘要，第546段左栏第1段 * |
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