Application of glutaminase inhibitor in preparation of medicine for treating psoriasis

Abstract

The invention discloses application of a glutaminase inhibitor in preparation of a medicine for treating psoriasis. In-vitro cell experiments prove that the glutaminase inhibitor can inhibit keratinocyte metabolism glutamine, inhibit miR-31 induced up-regulated keratinocyte mitochondrial respiration, inhibit miR-31 induced mTOR pathway, reduce cell activity and promote cell apoptosis, and has an anti-inflammatory effect; animal experiments prove that the glutaminase inhibitor can be used for effectively treating imiquimod-induced mouse psoriasis-like pathology change, including remarkably reducing the Baker score of skin lesion, effectively reducing the epidermal hypertrophy degree, remarkably reducing the number of subepidermal capillaries, remarkably reducing the number of epidermal Ki67positive cells and reducing the expression of epidermal GLS. Therefore, the glutaminase inhibitor can be used for preparing the medicine for treating psoriasis.

Description

technical field

[0001] The invention belongs to the technical field of medicine and relates to the use of glutaminase inhibitors, in particular to the application of glutaminase inhibitors in the preparation of medicines for treating psoriasis. Background technique

[0002] Psoriasis is a chronic inflammatory skin disease with skin erythema, papules, and scales as the main clinical manifestations, and the global incidence is about 1-3%. The main pathological features of psoriasis are excessive epidermis proliferation and local inflammatory cell infiltration, and the metabolic spectrum of local skin lesions changes. The pathogenesis is complex, involving genetics, immune-inflammation, vascular abnormalities, etc. Procrastination is hard to heal. Drugs currently used to treat psoriasis mainly include glucocorticoids, retinoic acid drugs, vitamin D3 derivatives, anthranol, cytotoxic drugs, etc. Among them, tazarotene, moderate and strong glucocorticoids Hormone and calcipotri...

Examples

Embodiment 1CB-839

[0044] Example 1 CB-839 inhibits the uptake and metabolism of keratinocyte glutamine

[0045] Add 10 μM CB-839 (CB-839 group) and equal volume of DMSO (Ctrl group) to the culture medium of keratinocyte HaCaT cells respectively, after 12 hours of intervention, collect the cell culture medium, and use the method of targeted metabolomics Detect the contents of glutamine (glutamine) and glutamic acid (glutamate) in the collected cell culture fluid, and by comparing with fresh culture fluid, calculate the amount of glutamine (ie glutamine) taken up by cells under different intervention conditions. consumption of amide) and the amount of glutamic acid secreted into the cell culture medium (i.e. the secretion of glutamic acid), the results are as follows Figure 1-2 shown. Depend on Figure 1-2 The results showed that CB-839 can significantly inhibit the uptake of glutamine by keratinocytes (P<0.001), and reduce the amount of glutamic acid secreted by keratinocytes into the culture...

Embodiment 2

[0048] Example 2 CB-839 inhibits miR-31-induced mitochondrial respiration of keratinocytes

[0049] Oxygen consumption rate (OCR) is the oxygen consumption rate of cells, which is an indicator of mitochondrial respiration capacity. Oxygen consumption detection principle: The value displayed before adding oligo represents the basic oxygen consumption of cells, including mitochondrial oxidative phosphorylation and proton leakage oxygen consumption, that is, after protons pass through the mitochondrial membrane to form potential energy through the respiratory chain, a part Proton reflux can form ATP by ATP synthase, which converts potential energy into energy in ATP. A portion passes through the mitochondrial membrane but is only oxidized, and the potential energy is converted into heat, but not used to synthesize ATP. Oligo is an ATP synthase inhibitor. The reduced oxygen consumption after adding this drug represents the oxygen consumption used by the body for ATP synthesis, an...

Embodiment 3

[0052] Example 3 Western Blotting Experiment of CB-839 Inhibiting the mTOR Pathway Induced by miR-31

[0053] Western Blotting experiments can detect the expression of the target protein. 24 hours after HaCaT cells were transfected with control miRNAmimic or miR-31mimic, they were intervened with DMSO and 10 μM CB-839; among them, those who were given DMSO intervention 24 hours after transfection with control miRNAmimic were recorded as the Ctrl-mimic group, and those transfected with miR-31mimic for 24 hours The miR-31mimic group was given after DMSO intervention, the CB839 group was given 10 μM CB-839 intervention 24 hours after transfection of control miRNA mimic, and the miR-31mimic group was given 10 μM CB-839 intervention 24 hours after miR-31mimic transfection. -31+CB839 group. In addition, HaCaT cells were transfected with control siRNA (ThermoFisher Scientific, AM4635) and GS-siRNA (ThermoFisher Scientific, AM16708) 24 hours after each intervention with DMSO as a con...

technical field

[0001] The invention belongs to the technical field of medicine and relates to the use of glutaminase inhibitors, in particular to the application of glutaminase inhibitors in the preparation of medicines for treating psoriasis. Background technique

[0002] Psoriasis is a chronic inflammatory skin disease with skin erythema, papules, and scales as the main clinical manifestations, and the global incidence is about 1-3%. The main pathological features of psoriasis are excessive epidermis proliferation and local inflammatory cell infiltration, and the metabolic spectrum of local skin lesions changes. The pathogenesis is complex, involving genetics, immune-inflammation, vascular abnormalities, etc. Procrastination is hard to heal. Drugs currently used to treat psoriasis mainly include glucocorticoids, retinoic acid drugs, vitamin D3 derivatives, anthranol, cytotoxic drugs, etc. Among them, tazarotene, moderate and strong glucocorticoids Hormone and calcipotri...

Examples

Embodiment 1CB-839

[0044] Example 1 CB-839 inhibits the uptake and metabolism of keratinocyte glutamine

[0045] Add 10 μM CB-839 (CB-839 group) and equal volume of DMSO (Ctrl group) to the culture medium of keratinocyte HaCaT cells respectively, after 12 hours of intervention, collect the cell culture medium, and use the method of targeted metabolomics Detect the contents of glutamine (glutamine) and glutamic acid (glutamate) in the collected cell culture fluid, and by comparing with fresh culture fluid, calculate the amount of glutamine (ie glutamine) taken up by cells under different intervention conditions. consumption of amide) and the amount of glutamic acid secreted into the cell culture medium (i.e. the secretion of glutamic acid), the results are as follows Figure 1-2 shown. Depend on Figure 1-2 The results showed that CB-839 can significantly inhibit the uptake of glutamine by keratinocytes (P<0.001), and reduce the amount of glutamic acid secreted by keratinocytes into the culture...

Embodiment 2

[0048] Example 2 CB-839 inhibits miR-31-induced mitochondrial respiration of keratinocytes

[0049] Oxygen consumption rate (OCR) is the oxygen consumption rate of cells, which is an indicator of mitochondrial respiration capacity. Oxygen consumption detection principle: The value displayed before adding oligo represents the basic oxygen consumption of cells, including mitochondrial oxidative phosphorylation and proton leakage oxygen consumption, that is, after protons pass through the mitochondrial membrane to form potential energy through the respiratory chain, a part Proton reflux can form ATP by ATP synthase, which converts potential energy into energy in ATP. A portion passes through the mitochondrial membrane but is only oxidized, and the potential energy is converted into heat, but not used to synthesize ATP. Oligo is an ATP synthase inhibitor. The reduced oxygen consumption after adding this drug represents the oxygen consumption used by the body for ATP synthesis, an...

Embodiment 3

[0052] Example 3 Western Blotting Experiment of CB-839 Inhibiting the mTOR Pathway Induced by miR-31

[0053] Western Blotting experiments can detect the expression of the target protein. 24 hours after HaCaT cells were transfected with control miRNAmimic or miR-31mimic, they were intervened with DMSO and 10 μM CB-839; among them, those who were given DMSO intervention 24 hours after transfection with control miRNAmimic were recorded as the Ctrl-mimic group, and those transfected with miR-31mimic for 24 hours The miR-31mimic group was given after DMSO intervention, the CB839 group was given 10 μM CB-839 intervention 24 hours after transfection of control miRNA mimic, and the miR-31mimic group was given 10 μM CB-839 intervention 24 hours after miR-31mimic transfection. -31+CB839 group. In addition, HaCaT cells were transfected with control siRNA (ThermoFisher Scientific, AM4635) and GS-siRNA (ThermoFisher Scientific, AM16708) 24 hours after each intervention with DMSO as a con...

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