Application of asparagine synthetase activator or asparagine

Abstract

The invention belongs to the technical field of medicines, and provides application of an asparagine synthetase activator or asparagine, and particularly, the asparagine synthetase activator or asparagine has an obvious treatment effect on cataract as a medicine. By using the small molecular asparagine synthetase activator, the occurrence process of cataract is effectively slowed down, lenticular opacification is avoided, and the pharmaceutical composition is expected to become a new medicine for treating age-related cataract.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to an asparagine synthetase (Recombinant Asparagine Syn thetase, ASNS) activator or the application of asparagine (ASN). Background Art

[0002] Cataract, or clouding of the lens of the eye, is the leading cause of blindness worldwide. Cataracts involve a multifactorial disease process and may be caused by various toxic factors, environmental stress, aging, diabetes, genetic factors, etc., which can lead to the occurrence of many types of cataracts, such as age-related cataracts, diabetic cataracts, and congenital cataracts. Diabetic cataracts are one of the most common complications of diabetes and one of the main causes of visual impairment in diabetic patients. Patients with diabetic cataracts account for a large proportion of all cataract patients. Age-related cataracts are common in people over the age of 40, and with age, the lens becomes more cloudy and hardens, eventually blocking light from reaching the retina.

[0003] The cause of cataracts remains elusive, and many factors, including genetics, metabolism, nutrition, and environment, are involved in its formation, but it is known that damage or death of lens epithelial cells is associated with cataracts in many animal models and human case studies. In the development of diabetic cataracts, due to cortical thickening, the lens thickness of diabetic patients is thicker than that of normal people, causing lens metabolism disorders, resulting in lens protein denaturation and turbidity. At this time, the lens thickness, refractive power, light scattering, etc. change, and the light is blocked by the turbid lens and scattered, so it cannot be projected on the retina, which eventually leads to blindness. The development of diabetic cataracts involves multiple mechanisms, which are still unclear. There are three main hypotheses: non-enzymatic glycosylation of lens proteins, osmotic pressure theory, and oxidative damage mechanism.

[0004] Currently, there is no specific drug for cataracts, and surgical treatment is still the main treatment. The most common drugs for treating cataracts are antioxidants (glutathione, vitamins, etc.), aldose reductase inhibitors (sorbinir, epalrestat, torestat, bonacrestat, etc.), glycosylation inhibitors (aminoguanidine, pyruvate, taurine, etc.), and Chinese herbal preparations (Cornus polysaccharides, Cuscuta seeds, lanosterol, etc.). However, there are still some patients in clinical practice who cannot achieve the expected results of the above treatment measures, and there are postoperative complications, surgical costs, and the large number of people who need to undergo surgery, which still imposes a burden on society and many patients.

[0005] Therefore, it is imperative to find more suitable small molecule drugs to effectively intervene and treat cataracts. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides an asparagine synthetase activator or the use of asparagine.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] The present invention provides an asparagine synthetase activator or use of asparagine in preparing medicines for preventing and treating cataract and / or related diseases.

[0009] As a preferred embodiment of the present invention, the asparagine synthetase activator is specifically cabozantinib.

[0010] As another preferred embodiment of the present invention, the cabozantinib increases the production of PPi.

[0011] As another preferred embodiment of the present invention, the cabozantinib has the effect of preventing 1,2-DHN-induced lens opacity.

[0012] As another preferred embodiment of the present invention, the cabozantinib is used to improve naphthalene cataracts.

[0013] As another preferred embodiment of the present invention, the drug is a lyophilized preparation or a solution preparation.

[0014] As another preferred embodiment of the present invention, the administration method of the drug includes at least one of oral administration, intravenous injection, intravitreal injection, subretinal injection, subconjunctival injection, eye drop treatment and gene therapy.

[0015] As another preferred embodiment of the present invention, the cataract-related diseases include at least one of other ophthalmic diseases, post-cataract surgery diseases and central nervous system diseases.

[0016] Specifically, the cataract-related diseases include, but are not limited to, ophthalmic diseases such as diabetic retinopathy, central and branch retinal vein occlusion, choroidal neovascularization, uveitis, age-related macular degeneration, central serous chorioretinopathy, retinitis pigmentosa, neuromyelitis optica, eye tumors, retinal vasculitis, macular epiretinal membrane, hypertension, ocular trauma and other ophthalmic diseases, as well as post-cataract surgery (such as Irvine-Gass syndrome) diseases, and central nervous system diseases such as multiple sclerosis (MS), Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), Huntington's disease (HD), etc.

[0017] Among them, in different types of cataract models, including age-related cataract-naphthalene cataract model (Naphthalene) and diabetic cataract (Diabetic cataract), although the causes of the disease vary from disease to disease, it was found that asparagine synthetase showed a trend of first high and then low.

[0018] Compared with the prior art, the technical solution provided by the present invention has at least the following advantages:

[0019] The present invention proposes to enhance the activity of ASNS or supplement the ASNS production product asparagine by exogenous supplementation of asparagine synthetase activator (ASNS activator) to alleviate and delay the development of cataracts. This strategy is expected to become a new method for treating eye-related diseases such as diabetic cataracts and age-related cataracts, thereby providing a potential drug intervention for cataract treatment.

[0020] In summary, the use of an ASNS activator and its product asparagine in the preparation of drugs for preventing and treating cataracts and / or related diseases provided in the embodiments of the present invention clarifies that the activation of ASNS has a significant therapeutic effect on cataracts. The present invention can effectively slow down the progression of cataracts and avoid lens opacity by using an ASNS activator or asparagine, and is expected to become a new target for the treatment of age-related cataracts, providing a reference for the development of cataract drugs.

[0021] The present invention focuses on ASNS as the treatment target, broadening the idea of ​​cataract prevention and treatment. In addition, the present invention's research found that ASNS may be involved in the occurrence and development of cataracts, and can be used as a new target for the treatment of neuroinflammatory diseases. The study of ASNS further supplements the pathogenesis of neuroinflammatory diseases and provides a theoretical basis for a better understanding and treatment of cataracts. Through the in vivo model study of ASNS agonists, the present invention proposes to enhance ASNS activity and thus reduce and delay the development of cataracts by exogenous supplementation of ASNS agonists, thereby applying it to the treatment of cataracts, which is expected to become a new method for the prevention and treatment of cataracts. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Reflects the activation effect of LCVS101 (Cabozantinib) on ASNS;

[0023] Figure 2 Reflect the effect of LCVS101 in delaying cataract progression in vitro;

[0024] Figure 3 Reflect the anti-cataract effect of LCVS101 in animal cataract models;

[0025] Figure 4 This indicates that asparagine improves the lens opacity induced by 1,2-DHN, a metabolite of naphthalene, in in vitro crystal culture;

[0026] Figure 5 Reflect the anti-cataract effect of asparagine in animal cataract models. DETAILED DESCRIPTION

[0027] The inventors have found that asparagine synthetase converts aspartic acid and glutamine into asparagine and glutamate through an adenosine triphosphate (ATP)-dependent reaction, and its activation can delay the occurrence and development of cataracts.

[0028] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0029] It should be noted that the experimental methods not specifically described in the following embodiments are all performed in accordance with conventional biochemical experimental methods described in "Biochemical Experimental Techniques" or "Cold Spring Harbor Protocols" or the experimental operations recommended by commercial reagent suppliers.

[0030] Example 1

[0031] This example is used to verify that LCVS101 activates ASNS enzyme activity, and specifically includes:

[0032] To detect the activity of ASNS in each reaction, the activity was monitored by measuring the concentration of pyrophosphoric acid (PPi) produced by the enzymatic reaction. The amide transfer reaction catalyzed by ASNS requires aspartate as a substrate, glutamine as a nitrogen source, and is dependent on ATP. To detect the activity of rhASNS protein, the ability of ASNS to catalyze the production of PPi at different concentrations was tested in the presence of sufficient substrate, nitrogen source, and ATP. Figure 1 This reflects the activation effect of LCVS101 on ASNS. The drug with a final concentration of 1 μM was added to the reaction system (0.4 μg rhASNS, 2 mM ATP, 8 mM L-Aspartic acid and 40 mM L-Glutamine), and the fluorescence value (λex = 316 / λem = 456 nm) was detected after incubation at room temperature for 30 minutes. The PPi concentration in each sample solution was calculated using the four-parameter logistic regression equation fitted by the AAT Bioquest online program to detect the effects of different drugs on ASNS enzyme activity. Figure 1 As can be seen from the figure, the drug LCVS101 significantly increased PPi production, which indicates that LCVS101 can activate ASNS activity.

[0033] Example 2

[0034] This example is used to verify that LCVS101 can improve lens opacity caused by naphthalene cataract during in vitro lens culture, and specifically includes:

[0035] The intact lens was first incubated in a culture medium containing 50 μM 1,2-DHN for 24 h, and then incubated in a culture medium containing 1 μM CVS101 for another 24 h. After the incubation, the lens was transferred to a PBS solution for morphological examination and photographed. Figure 2It reflects the effect of LCVS101 in delaying the progression of cataract under in vitro culture. NC / - is the normal group, and the lens was cultured in DMEM culture medium for 48 hours. NC+LCVS101 is the drug control group, and the lens was cultured in DMEM culture medium for 24 hours, and then continued to be cultured in culture medium containing 1μM drug for 24 hours. DHN / - is the cataract modeling group, and the lens was first cultured in culture medium containing 50μM 1,2-dihydroxynaphthalene (1,2-DHN) for 24 hours, and then continued to be cultured in DMEM culture medium for 24 hours. DHN+LCVS101 is the experimental drug group, and the lens was first cultured in culture medium containing 50μM 1,2-DHN for 24 hours, and then continued to be cultured in culture medium containing 1μM drug for 24 hours. From Figure 2 It can be seen that the lens of the normal group remained transparent after 48 hours of incubation, the lens of the drug control group remained transparent, and the lens of the 1,2-DHN control group was obviously turbid. Compared with the 1,2-DHN control group, the lens turbidity of the LCVS101-treated group was relatively low, showing moderate turbidity. These observations indicate that LCVS101 can prevent 1,2-DHN-induced lens turbidity and play a certain therapeutic role.

[0036] Example 3

[0037] This example is used to verify that LCVS101 can improve lens opacity caused by naphthalene cataract in a rat naphthalene cataract model, and specifically includes:

[0038] Cataract model was established in cataract model group by intragastric gavage of naphthalene, and LCVS101 was injected intraocularly in treatment group by gavage of naphthalene. Figure 3 This reflects the anti-cataract effect of LCVS101 in the animal cataract model. CTL is the normal control group, Model+LCVS101 is the rats treated with naphthalene via gastric gavage and intraocular injection of cabozantinib, and Model is the rats cataract modeling group via gastric gavage of naphthalene. From the lens photography, it can be seen that cabozantinib has improved the opacity of the lens to a certain extent. Figure 3 It can be seen that compared with the normal group, the lens of the naphthalene cataract group has been significantly cloudy. Compared with the naphthalene cataract group, LCVS101 can prevent the lens opacity of rats and play a certain therapeutic role.

[0039] In summary, LCVS101 can be used as an ASNS agonist, has a certain protective effect on naphthalene cataracts, and is expected to become a new drug for the treatment of age-related cataracts and related eye diseases.

[0040] Example 4

[0041] This example is used to verify that asparagine can improve lens opacity caused by naphthalene cataract during in vitro lens culture, and specifically includes:

[0042] In the model group, the intact lens was incubated in a culture medium containing 100 μM 1,2-DHN for 48 h. In the treatment group, asparagine was added to the 100 μM 1,2-DHN solution for treatment. After the incubation, the lens was transferred to a PBS solution for morphological examination and photographed. Figure 4 It reflects that asparagine improves lens opacity induced by naphthalene metabolite 1,2-DHN in in vitro lens culture. After rat lenses were isolated from the body and cultured in culture medium, the control group was treated with different concentrations of asparagine, the model group was treated with 100μM 1,2-DHN, and the treatment group was treated with 100μM 1,2-DHN and different concentrations of Asn. From the lens photography results, it can be seen that asparagine has improved the opacity of the lens to a certain extent. Figure 4 It can be seen that the lens of the normal group remained transparent after 48 hours of incubation, the lens of the asparagine control group remained transparent, and the lens of the 1,2-DHN model group was obviously turbid. Compared with the 1,2-DHN model group, the lens turbidity of the asparagine treatment group was relatively low, showing mild turbidity. These observations indicate that asparagine can prevent 1,2-DHN-induced lens turbidity and play a certain therapeutic role.

[0043] Example 5

[0044] This example is used to verify that asparagine can improve lens opacity caused by naphthalene cataract in a rat naphthalene cataract model, and specifically includes:

[0045] Cataract model was established in cataract model group by intragastric administration of naphthalene, and that in treatment group was treated by intragastric administration of naphthalene and asparagine at the same time. Figure 5 This reflects the anti-cataract effect of asparagine in animal cataract models. N is the normal control group, NC+Asn is a rat treated with naphthalene and asparagine by intragastric gavage, and the model is a rat cataract modeling group by intragastric gavage of naphthalene. From the lens photography, it can be seen that asparagine also improves the opacity of the lens in the animal cataract model. Figure 5 It can be seen that compared with the normal group, the lens of the naphthalene cataract group has been significantly cloudy. Compared with the naphthalene cataract group, asparagine can prevent the lens opacity of rats and play a certain therapeutic role.

[0046] In summary, the ASNS agonist LCVS101 and the ASNS product asparagine have a certain protective effect on naphthalene cataracts and are expected to become new drugs for the treatment of age-related cataracts and related eye diseases.

[0047] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. Use of an asparagine synthetase activator or asparagine in the preparation of a drug for preventing and treating cataract and / or related diseases.

2. The use according to claim 1, characterized in that: The asparagine synthetase activator is specifically cabozantinib.

3. The use according to claim 2, characterized in that: The cabozantinib acts to increase the production of PPi.

4. The use according to claim 2, characterized in that: The cabozantinib acts to prevent 1,2-DHN-induced lens opacity.

5. The use according to claim 2, characterized in that: The cabozantinib is used to improve naphthalene cataract.

6. The use according to claim 1, characterized in that: The drug is a lyophilized preparation or a solution preparation.

7. The use according to claim 6, characterized in that: The administration method of the drug includes at least one of oral administration, intravenous injection, intravitreal injection, subretinal injection, subconjunctival injection, eye drop treatment and gene therapy.

8. The use according to claim 1, characterized in that: The cataract-related diseases include at least one of other ophthalmic diseases, post-cataract surgery diseases and central nervous system diseases.

Images