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CN-121983326-A - Target protein inhibitor for preventing and treating proliferative diabetic retinopathy, screening method and application thereof

CN121983326ACN 121983326 ACN121983326 ACN 121983326ACN-121983326-A

Abstract

The invention relates to the field of biological medicine and ophthalmic diseases, in particular to a target protein inhibitor for preventing and treating proliferative diabetic retinopathy, a screening method and application thereof. The screening method comprises the steps of 1) obtaining a three-dimensional crystal structure of CTSH, 2) constructing a binding pocket at an active site of the CTSH, 3) carrying out molecular butt joint and scoring on a small molecular library, screening candidate compounds with binding energy lower than a preset threshold value, and 4) carrying out molecular dynamics simulation on the candidate compounds and CTSH complexes, and selecting compounds with stable structure and lower binding free energy as inhibitors. The inhibitor obtained by screening comprises eriocitrin or polygala sucrose ester B or AP-III-a4 inhibitor. The invention confirms that CTSH has causal driving effect on PDR through genetic causal inference, and can obviously inhibit inflammation and pathological angiogenesis by regulating CTSH expression under high sugar condition, and finally effective inhibitor components are obtained through screening.

Inventors

  • ZHAO QIUCHEN
  • CUI XUEHAO
  • HAN QUANHONG
  • Hui Jingwen

Assignees

  • 剑康生物科技(嘉兴)有限责任公司

Dates

Publication Date
20260505
Application Date
20260126

Claims (9)

  1. 1. A screening method of a target protein inhibitor for preventing and treating proliferative diabetic retinopathy is characterized in that, The screening method comprises the following steps: 1) Obtaining a three-dimensional crystal structure of CTSH; 2) Constructing a binding pocket at its active site; 3) Molecular docking and scoring are carried out on the small molecule library, and candidate compounds with binding energy lower than a preset threshold value are screened; 4) Molecular dynamics simulation is carried out on candidate compounds and CTSH complexes, and compounds with stable structure and low binding free energy are selected as inhibitors.
  2. 2. The method according to claim 1, wherein, The small molecule library of step 3) comprises a database of natural products, synthetic compounds, or known drugs; In the step 3), the molecular docking uses Glide software, and the preset binding energy threshold value is less than or equal to-10 kcal/mol; in the step 4), the molecular dynamics simulation time is 50-200 ns, the simulation environment is aqueous solution, the temperature is 300-310K, and the pressure is 0.9-1.1 atm.
  3. 3. The protein inhibitor of interest for preventing and treating proliferative diabetic retinopathy obtained by the screening method according to claim 1 or 2, The inhibitor is a CTSH inhibitor; The CTSH inhibitor is a small molecule compound, polypeptide, antibody, siRNA, shRNA, antisense oligonucleotide or CRISPR gene editing vector.
  4. 4. The protein inhibitor of interest for use in the prevention and treatment of proliferative diabetic retinopathy of claim 3, The inhibitor comprises eriocitrin or polygala sucrose ester B or AP-III-a4 inhibitor.
  5. 5. The use of an inhibitor according to claim 3 or 4, The inhibitor is used for preventing and treating proliferative diabetic retinopathy.
  6. 6. The use of an inhibitor according to claim 5, wherein, The application method comprises the following steps: 1) Detecting the expression level of CTSH in a biological sample; 2) Comparing the detection result with a reference threshold value, judging the risk of the individuals for generating PDR, and judging the individuals with high risk or the diagnosed patients when the detection result exceeds the reference threshold value; 3) Administration of CTSH inhibitors to high risk individuals or diagnosed patients.
  7. 7. The use of an inhibitor according to claim 6, wherein, The biological sample in the step 1) is peripheral blood, serum, plasma, vitreous humor or aqueous humor; The detection method comprises immunological detection or mass spectrum detection, wherein the detection environment temperature is 4-37 ℃, and the reaction duration is 0.5-4 h.
  8. 8. The use of an inhibitor according to claim 6, wherein, The reference threshold in the step 2) is a CTSH concentration range established based on healthy people or diabetic retinopathy-free people, and the threshold is set to be the mean value of the healthy people or the diabetic retinopathy-free people plus or minus 2 standard deviations.
  9. 9. The use of an inhibitor according to claim 6, wherein, Step 3) when administering the CTSH inhibitor to a high risk individual or to a diagnosed patient: The inhibitors may be used alone or in combination with anti-VEGF agents; routes of administration include intravitreal injection, systemic administration or topical administration; the administration dosage is effective amount, specifically 0.1-100 mg/kg per dose, and the administration frequency is weekly to monthly.

Description

Target protein inhibitor for preventing and treating proliferative diabetic retinopathy, screening method and application thereof Technical Field The invention relates to the field of biological medicine and ophthalmic diseases, in particular to a target protein inhibitor for preventing and treating proliferative diabetic retinopathy, a screening method and application thereof. Background Diabetic retinopathy (Diabetic Retinopathy, DR) is one of the leading causes of blindness in people of global working age, with Proliferative Diabetic Retinopathy (PDR) being its most severe stage, characterized by retinal ischemia, abnormal neovascularization, and fibrovascular hyperplasia. Existing treatment means include glycemic control, laser photocoagulation, and intravitreal anti-Vascular Endothelial Growth Factor (VEGF) drug injection. However, clinical practice shows that anti-VEGF treatment has the problems of incomplete response, frequent recurrence, heavy injection burden and the like, and the pathogenesis of PDR involves multi-channel disorders such as inflammation, immune activation, metabolic reprogramming and the like outside VEGF channels. Therefore, there is an urgent need to find upstream regulatory factors and novel therapeutic targets located at the junction of inflammation, metabolism and angiogenesis. In recent years, multiple sets of studies suggest that lysosomal cysteine protease family members play a key role in the regulation of immune metabolism. Among them, cathepsin H (CTSH) is a candidate target of interest due to its high expression in myeloid cells and its potential function in inflammation, oxidative stress and angiogenesis. Disclosure of Invention The invention aims to solve the following problems in the prior art: 1. The lack of reliable biomarkers useful for early prediction and risk stratification of PDR; 2. the existing treatment mainly aims at VEGF (vascular endothelial growth factor) pathway, and key pathological processes such as inflammation, immunity, metabolic reprogramming and the like cannot be effectively considered; 3. Lack of drug targets and corresponding inhibitor/diagnostic agent compositions directed against upstream key signaling nodes. Based on multi-mathematic causal deduction, single-cell transcriptome and in-vitro function experiments, the invention confirms that CTSH plays causal and upstream regulation roles in PDR generation and development for the first time, and accordingly provides the following technical scheme. A screening method of target protein inhibitor for preventing and treating proliferative diabetic retinopathy, The screening method comprises the following steps: 1) Obtaining a three-dimensional crystal structure of CTSH; 2) Constructing a binding pocket at its active site; 3) Molecular docking and scoring are carried out on the small molecule library, and candidate compounds with binding energy lower than a preset threshold value are screened; 4) Molecular dynamics simulation is carried out on candidate compounds and CTSH complexes, and compounds with stable structure and low binding free energy are selected as inhibitors. As a preferred alternative to this, The CTSH crystal structure source in step 1) is a public database (such as PDB database); The small molecule library of step 3) comprises a database of natural products, synthetic compounds, or known drugs; In the step 3), the molecular docking uses Glide software, and the preset binding energy threshold value is less than or equal to-10 kcal/mol; in the step 4), the molecular dynamics simulation time is 50-200 ns, the simulation environment is aqueous solution, the temperature is 300-310K, and the pressure is 0.9-1.1 atm. A target protein inhibitor for preventing and treating proliferative diabetic retinopathy, The inhibitor is a CTSH inhibitor; The CTSH inhibitor is a small molecule compound, polypeptide, antibody, siRNA, shRNA, antisense oligonucleotide or CRISPR gene editing vector. As a preferred alternative to this, The inhibitor comprises eriocitrin or polygala sucrose ester B or AP-III-a4 inhibitor. The application of the target protein inhibitor for preventing and treating proliferative diabetic retinopathy, The inhibitor is used for preventing and treating proliferative diabetic retinopathy. As a preferred alternative to this, The application method comprises the following steps: 1) Detecting the expression level of CTSH in a biological sample; 2) Comparing the detection result with a reference threshold value, judging the risk of the individuals for generating PDR, and judging the individuals with high risk or the diagnosed patients when the detection result exceeds the reference threshold value; 3) Administration of CTSH inhibitors to high risk individuals or diagnosed patients. As a preferred alternative to this, The biological sample in the step 1) is peripheral blood, serum, plasma, vitreous humor or aqueous humor; The detection method comprises immunological detection (such as