A glucosamine compound and a pharmaceutically acceptable salt thereof suitable for use in the treatment of breast cancer and relief of symptoms resulting from such diseases

Abstract

The invention relates to a glucosamine compound or a pharmaceutically acceptable salt thereof for use in a method for the treatment of breast cancer disease and symptoms resulting from such diseases in humans in need thereof.

Description

[0001] A GLUCOSAMINE COMPOUND AND A PHARMACEUTICALLY ACCEPTABLE SALT THEREOF SUITABLE FOR USE IN THE TREATMENT OF BREAST CANCER AND RELIEF OF SYMPTOMS RESULTING FROM SUCH DISEASES

[0002] TECHNICAL FIELD

[0003] The invention relates to a glucosamine compound or a pharmaceutically acceptable salt thereof for use in a method for the treatment of breast cancer disease and symptoms resulting from such diseases in humans in need thereof.

[0004] The invention also relates to in vitro studies to demonstrate the effect of the glucosamine compound or a pharmaceutically acceptable salt thereof in breast cancer disease.

[0005] PRIOR ART

[0006] Breast cancer is defined as a type of cancer which occurs as a result of the uncontrolled proliferation of cells in the breast tissue. If these cells tend to spread, they are called malignant, and if they continue to stay where they are, they are called benign tumors. Breast cancer is the most common type of cancer in women.

[0007] Breast cancer can often be detected during physical examination of women. Masses smaller than 1 cm are usually detected by radiological examination during medical examination. The cancerous mass is usually hard, immobile, rough and irregular edged.

[0008] As a result of the diagnosis of breast cancer in the patient, there are various treatment methods applied in the technique. The first stage of breast cancer treatment is surgical treatment. In the surgical procedure, cancerous tissue is removed from the breast. If deemed necessary, the axillary lymph nodes are also cleaned. The surgical procedure may be aimed at preserving the breast without removing it or removing the entire breast completely.

[0009] Another method is radiotherapy or chemotherapy methods. All of these treatment methods cause more or less damage to the human body and there is a possibility that the cancer will recur in the following processes. This situation creates problems in terms of forming a definitive treatment opinion. In the next period, it is expected to be kept under constant observation and monitored with routine examination.

[0010] Experts carry out research and development activities in the relevant technical field in order to perform breast cancer treatment with non-tiring, long and non-damaging drug treatment methods for the patient. For this reason, it has become a necessity to obtain pharmaceutical components and compounds which are new drug candidates for the relief of breast cancer and symptoms resulting from such diseases in the relevant technical field.

[0011] BRIEF DESCRIPTION OF THE INVENTION

[0012] Breast cancer, as it is known in the art, is one of the most common types of cancer in women. In cases where the said cancer treatment is not performed, it has rapid metastasis feature and causes the cancer to spread to other organs. Radiotherapy treatments, surgical operations and / or chemotherapy treatments are mostly performed for the treatment of breast cancer in the relevant technical field. These treatment methods are very tiring, damaging, long and costly for patients. In addition, the duration of treatment is quite long and recurrence is possible. Therefore, it has been determined that studies should be carried out on new drug candidates for the treatment of breast cancer and the elimination of symptoms caused by breast cancer for the relevant technical field. The present inventors present a compound and a pharmaceutical salt thereof with pharmaceutical effect which can be an alternative to known technical solutions in the art for the treatment of breast cancer and the relief of symptoms caused by such diseases.

[0013] BRIEF DESCRIPTION OF DRAWINGS

[0014] Fig.1-a) represents flow cytometry histogram analysis of MCF-7 and 4T1 cells in the presence and absence of D-GIcNAc.

[0015] Fig. 1-b) represents statistical analysis of MCF-7 and 4T1 cells in the presence and absence of D-GIcNAc. Fig. 2-a) represents flow cytometry histogram analysis of MCF-7 and 4T1 cells in the presence and absence of D-GIcNAc.

[0016] Fig. 2-b) represents flow cytometry of MCF-7 and 4T 1 cells in the presence and absence of GIcNAc. In statistical analysis, the significance degree of the differences is shown in a bar graph.

[0017] Fig. 3 represents flow cytometry point-plot analysis of MCF-7 and 4T1 cells. Early apoptosis (lower right quadrant) was increased in MCF-7 and 4T1 cells in the presence of 2mM GIcNAc. The quadrants are as follows: Lower left: Living cells, Lower right: early apoptosis, Upper right: Late apoptosis, Upper left: Necrosis.

[0018] Fig. 4 represents a statistical analysis of apoptosis of MCF-7 and 4T 1 cells.

[0019] Fig. 5 represents tumor pathology and statistical analysis of breast tissues in the Murine 4T1 model.

[0020] Fig. 6 represents the interaction of GIcNAc on the Her2 protein.

[0021] DETAILED DESCRIPTION OF THE INVENTION

[0022] In this detailed description, the subject of the invention relates to a glucosamine compound or a pharmaceutically acceptable salt thereof for use in a method for the treatment of breast cancer and its symptoms in humans in need thereof, and is explained only by examples which do not create any limiting effect for a better understanding of the subject.

[0023] The term "treatment' used within the scope of the present invention refers to the recovery of people in the treatment of breast cancer defined within the scope of the present invention.

[0024] The expression "relief of symptoms caused by breast cancer" used within the scope of the present invention refers to the elimination of at least one of the symptoms and / or disorders caused by the said diseases. The term "breast cancer" used within the scope of the present invention refers to the presence of abnormally differentiated cells in the breast tissue. Within the scope of the invention, breast cancer can be ductal carcinoma in situ (DCIS), lobular carcinoma in situ (LCIS), invasive breast cancer, invasive (or infiltrative) ductal carcinoma, invasive (or infiltrative) lobular carcinoma, inflammatory breast cancer, Paget's disease, medullary breast cancer, mucinous breast cancer, tubular breast cancer, adenoid cystic breast cancer, metaplastic breast cancer, angiosarcoma of the breast, basal-like breast cancer, phyllodes or cystocarcinoma phyllodes and papillary breast cancer. The types of cancer mentioned can occur at any stage, for example, at stages I, II, III or IV or in their subgroups.

[0025] The inventors propose the molecule N-acetyl-D-glucosamine (abbreviated as D-GIcNAc) for the treatment of breast cancer and the relief of symptoms caused by these diseases in a human in need thereof. The chemical name of the compound N-acetyl-D- glucosamine is p-D-(Acetylamino)-2-deoxy-glucopyranose, and its chemical structure is indicated by formula 1 .

[0026] Formula 1 .

[0027] The present inventors investigated the immunomodulatory effect of D-GIcNAc on breast cancer as a candidate compound by applying it to the xenograft murine model in vitro to MCF-7 and 4T 1 cell lines, which are the most commonly studied breast cancer cells, with an effective dose determined using the machine learning method. As a result of the researches, it has been shown that D-GIcNAc inhibits cell proliferation by promoting apoptosis in MCF-7 and 4T1 cell lines in a concentration-dependent manner. In another study, increased levels of D-GIcNAc suppressed the migration ability in the body, cell proliferation and clonal formation of MDA-MB-231 cells.

[0028] Other studies by the present inventors have shown that Fas expression increases in MCF-7 and 4T1 cell lines depending on D-GIcNAc concentrations and accordingly suppresses proliferation in cancer cells.

[0029] As it is known in the art, sialylation usually increases in tumor cells and combines with N-glycans and O-glycans. N-glycan sialylation may act as a regulatory mechanism for various receptor tyrosine kinases shown following activation of MET and RON receptors by a2-3Sia. In contrast to the action of apoptosis-inducing galectins, death receptor ligands and chemotherapeutic drugs, a2-6Sia increased on N-glycans due to ST6GAL1 up-regulation in cancer cells increases integrin-mediated cell motility and protects cells against apoptosis. In this case, exogenously administered D-GIcNAc has been found to have a reversible effect which may prevent cells from being protected against apoptosis. In fact, the sialic acid-binding Ig-like lectin mechanism appears to be a pathway for cancer cells to hide from immune cells by receptors mimicking immunoglobulin (Ig)-type lectins with an N terminal V-set Ig-like domain containing sialic acid-binding sites which activate or inhibit the immune response on the sialic acid-binding transmembrane cell surface which are members of the immunoglobulin superfamily which act as immune regulators. In the light of the findings of previous studies on sialic acid-binding Ig-like lectin mechanisms, it has been determined that D-GIcNAc can replace lectin found in the sialic acid-binding Ig lectin mechanism, where cancer cells can be deceived and lead to apoptosis. As parameters related to the inhibition of carcinogenic cells, significant reductions in metastasis and increases in apoptosis and Fas expression were observed in D-GIcNAc-treated cells and these are shown in Figs. 1 , 2, 3, 4 and 5. Accordingly, the ability of D-GIcNAc to bind on HER2, proven by molecular docking and dynamics studies, has been revealed to have a HER2-GlcNAc interaction, which may be the cause of Fas expression and apoptosis induction. A decrease in vascularity and regression in metastasis have been observed in mouse murine model studies, which may be related to the anti-tumoral effect of D-GIcNAc.

[0030] O-GIcNAcylation has an effect on cancer cell metabolism due to changes in the stability or activity of transcription factors and kinases which can be modulated by the administration of D-GIcNAc. D-GIcNAc can be evaluated for larger-scale therapeutic purposes in future clinical trials due to its potential synergistic effect estimated with licensed cancer drugs, as well as other proposed methods developed to block tumor cells through the administration of sialic acid mimics or targeted delivery of sialidases to tumor cell surfaces.

[0031] HER2, a ligandless tyrosine kinase receptor of the HER family, is often overexpressed in breast cancer due to its proto-oncogene properties. HER2 is known to suppress cancerous cell apoptosis leading to uncontrolled proliferation and tumor growth. In the studies carried out within the scope of the invention, it was determined that the binding of D-GIcNAc on HER2 directed the cell to apoptosis.

[0032] As a result of the researches, the present inventors have shown that D-GIcNAc has the potential to reduce the proliferation of mammary gland tumor cells by increasing Fas expression and inducing early apoptosis in tumor cells. In addition, it has been determined that systemic administration of D-GIcNAc can reduce mitosis and angiogenesis and therefore reduce the tumor size of breast cancer cells.

[0033] When Fig. 1 is examined, it has been found that 1 mM, 2 mM and 4 mM GIcNAc significantly reduced the proliferation rate in MCF-7 cells and that 2 mM and 4 mM D- GIcNAc significantly reduced the proliferation rate in 4T 1 cells (p<0.05).

[0034] When Figs. 2 a) and b) are examined, it has been found that 1 mM, 2 mM and 4 mM D- GIcNAc significantly increased Fas expression in MCF-7 and 4T1 cells (p<0.05).

[0035] In Figure 3, the quadrants are as follows: Lower left: Living cells, Lower right: early apoptosis, Upper right: Late apoptosis, Upper left: Necrosis. Total apoptotic cells were calculated by subtracting the percentage of living cells from the total cells.

[0036] In Fig. 4, living cells in MCF-7 and 4T1 cells in the presence of 1 mM, 2 mM and 4 mM D-GIcNAc were found to be significantly reduced with increasing percentage of early apoptosis.

[0037] In Fig. 5, it has been found that a) Angiogenesis was significantly reduced by D-GIcNAc in the post-treatment group, b) Mitosis was significantly reduced by D-GIcNAc in both the simultaneous and post-treatment groups, c) Necrosis tended to decrease in the posttreatment group, but the difference did not occur significantly compared to the breast cancer group d) Pleomorphism was not significantly altered by D-GIcNAc administration in the simultaneous or post-treatment groups e) Tumor-infiltrating cells were significantly increased in the simultaneously GIcNAc administered group compared to the breast cancer group, f) the tumor size decreased significantly in the post-treatment group compared to the breast cancer group.

[0038] In Fig. 5, g) the use of D-GIcNAc after tumor development; marked necrosis and inflammation, low pleomorphism and low angiogenesis are indicated by red arrows (10X magnification).

[0039] In Fig. 5, h) Simultaneous administration of D-GIcNAc during tumor development; significant necrosis, angiogenesis and inflammation are indicated by red arrows (10X magnification).

[0040] Fig. 5 shows i) Untreated breast cancer; Marked pleomorphism and high mitosis in breast tumor tissue.

[0041] Fig. 6 shows the interaction of GIcNAc on the Her2 protein.

[0042] In an embodiment of the invention, D-GIcNac, which is represented by formula 1 suitable for the invention, is used in the treatment of breast cancer.

[0043] In an embodiment of the invention, D-GIcNac, represented by formula 1 , is used in the production of a drug suitable for use in the treatment of breast cancer.

[0044] The invention relates to a pharmaceutical composition containing D-GIcNac, also referred to as formula 1 .

[0045] These pharmaceutical compounds may contain D-GIcNac, referred to as formula 1 , as well as at least one additional active compound.

[0046] Said pharmaceutical compound is combined with D-GIcNac, referred to as formula 1 , as well as at least one active compound with an antineoplastic effect. Said pharmaceutical composition includes D-GIcNAc, referred to as formula 1 , and the active compounds as antineoplastic compounds can be selected from a group of or from a double or triple combinations of cyclophosphamide, ifosfamide, temozolomide, capecitabine, 5-fluorouracil, methotrexate, gemcitabine, pemetrexet, mitomycin, blemycin, epirubicin, doxorubicin, etoposide, paclitaxel, irinotecan, docetaxel, vincristine, carboplatin, cisplatin, oxaliplatin, bevacizumab, cetuximab, gefitinib, imatinib, trastuzumab, denosumab, rituximab, sunitinib, zoledronate, abirateron, anastrozole, bicalutamide, exemestane, goserelin, medroxyprogesterone, octreotide, tamoxifen, dendamustine, carmustine, chlorambucil, lomustin, melphalan, procarbazine, streptozosin, fludarabine, raltitrexed, actinomycin D, dactinomycin, doxorubicin, mitoxantrone, eribulin, topotecan, vinblastine, vinorelbin, afatinib, aflibersept, crizotinib, dabrafenim, ibandronic acid, pamidronate, bexarotan, buserelin, cyproterone, degarelix, folinic acid, fulvestrant, lanreotide, lenalidomide, letrozole, leuprorelin, megestrol, mesna and thalidomide.

[0047] D-GIcNac, which is referred to as Formula 1 suitable for the invention, and the dose range in which it can be used can be determined according to the needs of the patient and the stage of the disease.

[0048] The protection scope of the invention is specified in the appended claims and cannot be limited to what is described in this detailed description for illustrative purposes. Further, it is clear that a person skilled in the art can put forward similar structures in the light of the above, without leaving the main theme of the invention.

Claims

CLAIMS1. A glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use in a method for the treatment of breast cancer disease and symptoms resulting from such diseases in humans in need thereof,Formula 1 .

2. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to Claim 1 , wherein it is administered with a pharmaceutically acceptable carrier or excipient.

3. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to Claim 1 or 2, wherein said compound [3-D- (Acetylamino)-2-deoxy-glucopyranose4. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to one of the preceding claims, wherein it is administered orally.

5. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to one of the preceding claims, wherein it is administered after the person exhibits a symptom of breast cancer.

6. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to one of the preceding claims, wherein the breastcancer is ductal carcinoma in situ (DCIS), lobular carcinoma in situ (LCIS), invasive breast cancer, invasive (or infiltrative) ductal carcinoma, invasive (or infiltrative) lobular carcinoma, inflammatory breast cancer, Paget's disease, medullary breast cancer, mucinous breast cancer, tubular breast cancer, adenoid cystic breast cancer, metaplastic breast cancer, angiosarcoma of the breast, basal-like breast cancer, phyllodes or cystocarcinoma phyllodes and papillary breast cancer.

7. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to one of the preceding claims, wherein it is administered in conjunction with at least one other therapeutic agent.

8. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to one of the preceding claims, wherein it is administered in conjunction with at least one other antineoplastic agent.

9. The glucosamine compound of Formula 1 or a pharmaceutically acceptable salt thereof for use according to Claim 8, wherein the active compounds as antineoplastic compounds can be selected from a group of or from a double or triple combinations of cyclophosphamide, ifosfamide, temozolomide, capecitabine, 5-fluoro uracil, methotrexate, gemcitabine, pemetrexet, mitomycin, blemycin, epirubicin, doxorubicin, etoposide, paclitaxel, irinotecan, docetaxel, vincristine, carboplatin, cisplatin, oxaliplatin, bevacizumab, cetuximab, gefitinib, imatinib, trastuzumab, denosumab, rituximab, sunitinib, zoledronate, abirateron, anastrozole, bicalutamide, exemestane, goserelin, medroxyprogesterone, octreotide, tamoxifen, dendamustine, carmustine, chlorambucil, lomustin, melphalan, procarbazine, streptozosin, fludarabine, raltitrexed, actinomycin D, dactinomycin, doxorubicin, mitoxantrone, eribulin, topotecan, vinblastine, vinorelbin, afatinib, aflibersept, crizotinib, dabrafenim, ibandronic acid, pamidronate, bexarotan, buserelin, cyproterone, degarelix, folinic acid, fulvestrant, lanreotide, lenalidomide, letrozole, leuprorelin, megestrol, mesna and thalidomide.

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