A compound containing SKL2001 for the treatment of parasitic infections and proliferative diseases by modulating proteasome activities and for immunomodulatory action

Abstract

The invention relates to a method for treating parasitic infections, immunomodulation, proliferation-related diseases, inflammation and neurodegenerative disorders through inhibition and modulation of the 20S / 26S proteasome by SKL2001.

Description

[0001] A COMPOUND CONTAINING SKL2001 FOR THE TREATMENT OF PARASITIC INFECTIONS AND PROLIFERATIVE DISEASES BY MODULATING PROTEASOME ACTIVITIES AND FOR IMMUNOMODULATORY ACTION

[0002] Technical Field

[0003] The invention relates to a method for treating parasitic infections, immunomodulation, proliferation-related diseases, inflammation and neurodegenerative disorders through inhibition and modulation of the 20S / 26S proteasome by SKL2001.

[0004] Prior Art

[0005] Parasitic worms, also known as helminths (tapeworms, roundworms and flatworms) are large macroparasites; adults are usually visible to the naked eye. Many are intestinal worms that are transmitted through soil and infect the gastrointestinal tract. Other parasitic worms, such as schistosomes, are found in blood vessels. They cause significant health problems both in humans and animals (1). Therefore, many anti helminthic drugs have been developed and used.

[0006] Anthelmintics are a variety of agents and chemical compounds used for a wide range of parasite infections. Anthelmintic agents include benzimidazoles (thiabendazole, mebendazole, albendazole and triclabendazole), ivermectin, nitazoxanide, praziquantel, pyrantel pamoate and niclosamide (2 and 3).

[0007] Their mechanism of action is variable, but often leads to paralysis of the parasitic worm and its release and expulsion. With some exceptions, the agents are parasite category specific. For example, praziquantel is used for the treatment of trematodes and ivermectin for nematodes.

[0008] Albendazole is broadly effective for many nematodes and some trematodes and protozoa. Triclabendazole is the drug of choice for chronic fascioliasis.

[0009] Niclosamide is used for the treatment of tapeworm and intestinal worm infections in general: These include: Taenia saginata (bovine tapeworm), Taenia solium (porcine tapeworm), Diphyllobothrium latum (fish tapeworm), Fasciolopsis buski (large intestinal worm).

[0010] Anthelmintic resistance (AR) is a growing concern for effective parasite control in farmed ruminants globally. Intensified and integrated research is needed to combat AR, develop innovative diagnostic tests to detect helminth infections and AR, and develop sustainable anthelmintic treatment strategies (4).

[0011] Niclosamide acts by killing tapeworms on contact. Adult worms are rapidly killed, probably due to uncoupling of oxidative phosphorylation or stimulation of ATPase activity, although the exact mechanism is not known. As mentioned above, the mechanism of action of niclosamide has not been fully elucidated. However, interestingly, niclosamide has started to be used in many cancer treatments by inhibiting apoptosis and proliferation and enhancing the effect of other anti-cancer agents. In addition, the immunomodulatory effect of niclosamide has emerged as an alternative molecule for many immunologic diseases. Despite this new information, the reason for the anti-cancer effects of niclosamide such as its effects on tapeworms is not known exactly (5).

[0012] As stated in the previous patent we applied for, we have shown that niclosamide dramatically decreases the activities of the proteasome system and accordingly, p21, p27 and p53 proteins increase in the cell. It is known that proteasome activity is directly and indirectly related to oxidative stress, oxidative phosphorylation and ATPase activity (6). Therefore, we suggested that the mechanism of anti-cancer and immunomodulatory effect of niclosamide on tapeworms is due to its inhibition of chemotrypsin, trypsin and caspase-like activities which are proteasome activities.

[0013] In our ongoing studies, we have shown that SKL2001, a chemical that modulates the wnt / b-catenin pathway, dramatically decreased the activities of the proteasome system such as chemotrypsin, trypsin and caspase-like effects, and correspondingly increased p21, p27 and p53 proteins, which are regulated by the proteasome systems (Figure 1 - 4).

[0014] In our next study, we investigated whether SKL2001, which has the same mechanism of action as niclosamide, is directly effective against parasites similar to niclosamide. For this purpose, we tested the effect of SKL2001 on Caenorhabditis elegans (C. elegans), a free-living nematode from roundworms. The results of the study showed that on day 20, 50% of the C. elegans in the medium containing SKL2001 became completely immobile (lifeless) and the amount of swimming C. elegans decreased by 80% compared to the control group (Figure 5).

[0015] Detailed Description of the Invention

[0016] In one embodiment, the invention describes compounds comprising SKL2001, SKL2001-like and a pharmaceutically acceptable carrier which reduce the activity of the 20S and / or 26S proteasome below basal levels.

[0017] The invention inhibits chemotrypsin, trypsin and caspase-like activities of the 20S proteasome and the 26S proteasome.

[0018] The invention provides for inhibition of the proteolytic activity of the immunoproteasome in addition to the 20S proteasome and the 26S proteasome system by the use of SKL2001 and / or salts thereof. The invention also provides compounds for the treatment of parasitic infections, including tapeworms, roundworms and flatworms, by inhibition or modulation of the 20S and / or 26S proteasome.

[0019] This method of reducing proteasome activity is through direct or indirect inhibition of the 20S and / or 26S proteasome.

[0020] It modulates proteasome activity through the ubiquitin-dependent proteasome system (UPS) and / or ubiquitin-independent proteasome system (UIPS) pathways.

[0021] The invention also provides compounds for the treatment of parasitic infections, including tapeworms, roundworms and flatworms that have developed resistance to other anti-parasitic drugs, by inhibition or modulation of the 20S and / or 26S proteasome.

[0022] In some embodiments, parasitic disease includes one of the group of diseases set out in this paragraph; Taenia solium, Taenia saginata, Hymenolepis spp, Echinococcus granulosus, Echinococcus multilocularis, Multiceps multiceps, Schistosoma mansoni, Schistosoma japonicum, Fasciola hepatica, Ascaris spp., Enterobius, Filarioidea, Onchocerca spp., Rhabditis spp., Trichuris spp., Necator americanus, Ancylostoma spp.

[0023] The invention also provides compounds that can be used for the treatment of many proliferative diseases, including cancer by inhibition or modulation of the 20S and / or 26S proteasome.

[0024] The invention also provides compounds for the treatment of proliferative diseases, including cancer, and for the treatment of cell proliferation arrest due to dramatic increase of tumor suppressor proteins such as p21, p27 and p53 by inhibition or modulation of the 20S and / or 26S proteasome. The methods of the present invention may be used to treat a patient in need thereof, who in some embodiments may have cancer, a neurodegenerative disease, an autoimmune disease, an infectious disease, or inflammatory diseases.

[0025] In some embodiments, autoimmune disease includes one of the group of diseases mentioned in this paragraph; an allopecia areata, ankylosing spondylitis, arthritis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune inner ear disease (also known as Meniers disease), autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura, autoimmune hemolytic anemia, autoimmune hepatitis, Bechet's disease, Crohn's disease, diabetes mellitus type 1, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, inflammatory bowel disease, lupus nephritis, multiple sclerosis, myasthenis gravis, pemphigus, pernicious anemia, polyarteritis nodosa, polymyositis, primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, rheumatic fever, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus (SLE), ulcerative colitis, vitiligo and Wegener's granulomatosis.

[0026] In some embodiments, cancer includes one of the types of cancer specified in this paragraph; leukemia; carcinoma of bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid, prostate, head, neck and skin; hematopoietic tumors of lymphoid lineage, acute lymphocytic leukemia; B-cell lymphoma; Burkett's lymphoma; hematopoietic tumors of myeloid lineage, acute and chronic myelogenous leukemias and promyelocytic leukemia; tumors of mesenchymal origin, fibrosarcoma, rhabdomyasarcoma; melanoma; seminoma; teratocarcinoma; osteosarcoma; neuroblastoma and glioma.

[0027] In some embodiments, the inflammation includes one of the disease types mentioned in this paragraph; rheumatoid arthritis, spondyloatopathies, gouty arthritis, osteoarthritis, systemic lupus erythematosus, juvenile arthritis, bronchitis, bursitis, gastritis, inflammatory bowel disease, ulcerative colitis, acne vulgaris, asthma, autoimmune diseases, chronic prostatitis, glomerulonephritis, hypersensitivities, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, sarcoidosis, transplant rejection, vasculitis and interstitial cystitis. Description of the Figures

[0028] Figure 1. Effects of SKL2001 on chemotrypsin activity in HepG2 cells (promega in vivo proteasome activity assay)

[0029] Figure 2. Effects of SKL2001 on trypsin activity in HepG2 cells (promega in vivo proteasome activity assay)

[0030] Figure 3. Effects of SKL2001 on caspase-like activity in HepG2 cells (promega in vivo proteasome activity assay) Figure 4. Effect of SKL2001 on p21, p53 and p27 protein amounts in HepG2 cells

[0031] Figure 5. Effect of SKL2001 on survival and mobility of C.elegans worms.

[0032] REFERENCES

[0033] 1. CDC - Parasites - About Parasites", www.cdc.gov. 20 April 2018

[0034] 2. Taylor-Robinson, David C.; Maayan, Nicola; Donegan, Sarah; Chaplin, Marty; Garner, Paul (11 September 2019). "Public health deworming programmes for soil-transmitted helminths in children living in endemic areas". The Cochrane Database of Systematic Reviews. 9 (11): CD000371.

[0035] 3. "Helminth control in school-age children" (PDF). World Health Organization. 2011. Retrieved 28 July 2015.)

[0036] 4. Aroop Mohanty, T. Shantikumar Singh, Tshering Ongmu Bhutia, Pratima Gupta, Priyanka Gupta A case of albendazole and niclosamide resistant

[0037] Taenia saginata infection, Int J Res Med Sci. 2017 Jun;5(6):2821-2823).

[0038] 5. Haowen Jiang, Albert M. Li, and Jiangbin Ye, The magic bullet: Niclosamide , Front Oncol. 2022; 12: 1004978).

[0039] 6. Min Qiu,J imei Chen,Xiaohong Li, andJian Zhuang , Intersection of the Ubiquitin-Proteasome System with Oxidative Stress in Cardiovascular

[0040] Disease Int. J. Mol. Sci. 2022, 23(20), 12197).

Claims

CLAIMS1. The invention is a composition comprising the SKL2001 molecule for use in reducing proteasome activity and / or regulating the protein homeostasis network and is a pharmaceutically acceptable carrier.

2. A compound according to claim 1, wherein the use of the compound and / or salts thereof inhibits the proteolytic activity of the 20S proteasome, 26S proteasome and immunoproteasome.

3. A compound according to claim 1, wherein SKL2001 causes the activity of the proteasome (20S and / or 26S) to decrease below basal levels.

4. A compound according to claim 1 or 2, wherein SKL2001 inhibits the 20S and / or 26 S proteasome directly or indirectly.

5. A compound according to any one of claims 1 to 4, wherein in particular the chemotrypsin-like activity of the 20S proteasome and the 26S proteasome is inhibited.

6. A compound according to any one of claims 1 to 4, wherein in particular the trypsin-like activity of the 20S proteasome and the 26S proteasome is inhibited.

7. A compound according to any one of claims 1 to 4, wherein in particular the caspase-like activity of the 20 S proteasome and the 26 S proteasome is inhibited.

8. A compound according to any one of claims 1 to 4, wherein the chymotrypsin-like, trypsin-like and caspase-like activities of the 20S proteasome and the 26S proteasome are simultaneously inhibited.

9. A compound according to any one of claims 1 to 4, wherein SKL2001 reduces proteasome activities in a UPS (Ubiquitin-ATP -Dependent Proteolysis) dependent and / or UPS independent manner.

10. A pharmaceutical compound according to any one of claims 1 to 4, characterized in that the compound is present in an amount to effectively inhibit proteasome function in a cell or a mammal.

11. A compound according to any one of claims 1 to 8, wherein SKL2001 provides compounds for the treatment of parasitic infections, including tapeworms, roundworms and flatworms, by inhibition or modulation of the 20S and / or 26S proteasome.

12. A compound according to any one of claims 1 to 8, wherein SKL2001 provides compounds that can be used for the treatment of parasitic infections, including tapeworms, roundworms and flatworms, that have developed resistance to other parasitic drugs, by inhibition or modulation of the 20S and / or 26S proteasome.

13. The compound according to claim 11 or 12, wherein said parasitic infections are, but are not limited to, Taenia solium, Taenia saginata, Hymenolepis spp, Echinococcus granulosus, Echinococcus multilocularis, Multiceps multiceps, Schistosoma mansoni, Schistosoma japonicum, Fasciola hepatica, Ascaris spp., Enterobius, Filarioidea, Onchocerca spp., Rhabditis spp., Trichuris spp., Necator americanus, Ancylostoma spp.

14. A compound according to any one of claims 1 to 8, wherein SKL2001 provides methods of increasing cell proliferation-related proteins p21 p27 and p53 within the cell by reducing proteasome activity15. A compound according to any one of claims 1 to 8, wherein SKL2001 provides compositions and methods of preventing and / or treating cell proliferative diseases, including cancer.

16. A compound according to claim 15, wherein said cell proliferative diseases include, but are not limited to, proteionopathic cell proliferative diseases, disorders and / or various conditions characterized by abnormal cell growth, preferably abnormally increased cell proliferation, cancer, cancermetastasis, immune-mediated reactions and diseases (e.g, transplant rejection, graft vs host disease, immune reaction to gene therapy, autoimmune diseases, pathogen-induced immune dysregulation, etc.), certain circulatory diseases and certain neurodegenerative diseases.

17. A compound according to claim 15 or 16, wherein said cancer is a cancer as set forth in this paragraph, including, but not limited to; carcinoma, leukemias and lymphomas such as cutaneous T-cell lymphomas, peripheral T-cell lymphomas, human T-cell lymphotropic virus-associated lymphomas such as adult T-cell leukemia / lymphoma, B-cell lymphoma, acute lymphocytic leukemia, acute non-lymphocytic leukemias, chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, Hodgkin's disease, non-Hodgkin's lymphomas, multiple myeloma, myelodysplastic syndrome, mesothelioma, head and neck cancers (eg. genitourinary cancers (e.g. prostate, bladder, kidney, uterus, ovary, testis, rectal and colon), lung cancer, breast cancer, liver cancer, colon cancer, small bowel cancer, pancreatic cancer, melanoma and other skin cancers, gastric cancer, central nervous system neoplasms, neuroectodermal cancer, spinal axis tumors, glioma, meningioma and pituitary adenoma, liver cancer and thyroid cancer and / or brain tumors, childhood solid tumors such as neuroblastoma, retinoblastoma, Wilms tumor, bone tumors and soft tissue sarcomas and their metastases.

18. A compound according to claim 17, wherein said autoimmune disease is, but not limited to, the autoimmune disease set forth in this paragraph; ankylosing spondylitis, arthritis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune inner ear disease (also known as Meniers disease), autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura, autoimmune hemolytic anemia, autoimmune hepatitis, Bechet's disease, Crohn's disease, diabetes mellitus type 1, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, inflammatory bowel disease, lupus nephritis, multiple sclerosis, myasthenis gravis, pemphigus, pemic anemia, polyarteritis nodosa, polymyositis, primary biliary cirrhosis, psoriasis, Raynaud's Phenomenon includes rheumatic fever, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemiclupus erythematosus (SLE), ulcerative colitis, vitiligo and Wegener's granulomatosis.

19. A compound according to claim 17, wherein said immune-mediated responses, diseases related to inflammation include, but are not limited to, one of the diseases set forth in this paragraph; rheumatoid arthritis, spondyloathopathies, gouty arthritis, osteoarthritis, systemic lupus erythematosis, juvenile arthritis, bronchitis, bursitis, gastritis, inflammatory bowel disease, ulcerative colitis, acne Vulgaris, asthma, autoimmune dieases, chronic prostatitis, glomerulonephritis, hypersensitivities, inflammatory bowel diseases, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, sarcoidosis, transplant rejection, vasculitis, and interstitial cystitis.