A chlordamine derivative targeting tumor mitochondria, its preparation method and application

Abstract

This invention belongs to the field of pharmaceutical technology, and more specifically, relates to a chlordamine derivative targeting tumor mitochondria, its preparation method, and its application. The chlordamine derivative targeting tumor mitochondria provided by this invention has the structure shown in formula (I): wherein R is phenyl, or a phenyl substituted with one or more substituents selected from halogen, C1-C4 alkyl, nitro, and trifluoromethyl. This type of derivative induces oxidative stress in tumor stem cells by targeting mitochondria, disrupting mitochondrial morphology and function, and can induce apoptosis of tumor stem cells at extremely low concentrations. Compared with chlordamine, the mitochondrial-targeting chlordamine derivative has superior anti-tumor growth and metastasis capabilities and can significantly improve the survival time of tumor-bearing mice.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and more specifically, relates to a chlordamine derivative that targets tumor mitochondria, its preparation method, and its application. Background Technology

[0002] Cancer stem cells are a small population of cells within tumors that possess stem cell-like properties, exhibiting high potential for self-renewal, self-replication, and multi-lineage differentiation. Current research indicates that cancer stem cells are the primary culprits in tumor development, recurrence, metastasis, and drug resistance. Therefore, developing drugs that target cancer stem cells is of significant importance for cancer treatment.

[0003] In recent years, mitochondria have been recognized as a key driving subcellular unit for the origin and development of cancer stem cells. Mitochondria determine the fate of cancer stem cells in areas such as autophagy, energy metabolism, and oxidative stress. Due to the negative potential of the mitochondrial inner membrane, positively charged substances can accumulate efficiently in the mitochondrial matrix against their concentration gradient. Therefore, developing novel small molecule drugs that target mitochondrial function, based on existing or clinically tested small molecule drugs, has become an important strategy for eradicating cancer stem cells.

[0004] Lonidamine (LND) is a mitochondrial metabolic inhibitor that has failed in clinical trials due to its limited anti-tumor efficacy. A key issue to be addressed is how to deliver higher concentrations of lonidamine to mitochondria to enhance its efficacy (especially in its application against cancer stem cells) and thus improve its clinical translation value. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a chlordamine derivative that targets tumor mitochondria, its preparation method and application, so as to solve the technical problems of limited drug concentration delivered to mitochondria by the mitochondrial metabolism inhibitor chlordamine and poor anti-tumor efficacy in the prior art.

[0006] To achieve the above objectives, the present invention provides a chlordamine derivative targeting tumor mitochondria, having a structure as shown in formula (I):

[0007]

[0008] Wherein, R is a phenyl group, or a phenyl group substituted with one or more of the following: halogen, C1-C4 alkyl, nitro, and trifluoromethyl.

[0009] According to another aspect of the present invention, a method for preparing the chlordamine derivative is provided, comprising the following steps:

[0010] (1) 2-hydroxy-5-methyl-isophthalic acid is etherified with a benzyl halide to obtain a bridged intermediate in which the phenolic hydroxyl group is substituted.

[0011] (2) The bridging intermediate described in step (1) is subjected to an esterification reaction with chlordamine to obtain a chlordamine-linker conjugate;

[0012] (3) The conjugate described in step (2) is reacted with 4-carboxytriphenylphosphine bromide to obtain a chlordamine derivative targeting mitochondria.

[0013] Preferably, the benzyl halide is benzyl bromide or benzyl bromide in which one or more of the following are substituted on the benzene ring: C1-C4 alkyl, nitro, or trifluoromethyl.

[0014] According to another aspect of the invention, the use of the described chlordamine derivative in the preparation of a medicament for treating and / or preventing tumors is provided.

[0015] According to another aspect of the invention, the use of the chlordamine derivative described above in the preparation of a drug for killing tumor stem cells is provided.

[0016] Preferably, the tumor is breast cancer, liver cancer, colon cancer, ovarian cancer, melanoma, or pancreatic cancer.

[0017] According to another aspect of the invention, an anticancer drug is provided, comprising the aforementioned mitochondrial-targeting chlordamine derivative and pharmaceutically acceptable additives.

[0018] According to another aspect of the invention, an anticancer drug targeting tumor stem cells is provided, comprising the aforementioned mitochondrial-targeting chlordamine derivative and pharmaceutically acceptable additives.

[0019] Preferably, the dosage form of the anticancer drug is an injection, powder for injection, oral preparation, spray, capsule, or suppository.

[0020] In summary, compared with the prior art, the above-described technical solutions conceived by this invention have the following advantages:

[0021] Beneficial effects:

[0022] (1) The present invention provides a chlordamine derivative based on mitochondria. By conjugating chlordamine with the positively charged triphenylphosphine group, a new type of chlordamine derivative was obtained. Experiments showed that it can accumulate in mitochondria at high concentrations, affect mitochondrial function, and effectively inhibit tumor stem cells.

[0023] (2) This invention provides a class of LND-connector-TPP conjugates, which can target mitochondria, disrupt their morphology and function, cause intracellular redox imbalance, induce tumor apoptosis, and thereby inhibit various tumor stem cells. Compared to LND, the HYL001 synthesized in the preferred embodiment showed a nearly 380-fold increase in the half-maximal inhibitory concentration (IC50) against breast cancer stem cells (BCSCs). This invention fills the gap in the use of chlordamine derivatives to inhibit tumor stem cells, providing a new and effective technology for tumor treatment. Attached Figure Description

[0024] Figure 1 This is a synthetic route diagram for Embodiment 1 (HYL001) of the present invention;

[0025] Figure 2 The proton nuclear magnetic resonance spectrum of Example 1 (HYL001) of this invention;

[0026] Figure 3 The phosphorus nuclear magnetic resonance spectrum of Example 1 (HYL001) of this invention;

[0027] Figure 4 This is the high-resolution mass spectrometry of Example 1 (HYL001) of the present invention;

[0028] Figure 5 The dose-cell survival curves of HYL001 and LND against 4T1 tumor cells and BCSCs are shown.

[0029] Figure 6 Comparison of the inhibitory activities of HYL001 and LND on regenerated breast cancer cells (4T1TRCs) obtained by three-dimensional fibrous gel screening;

[0030] Figure 7 A comparison of the effects of HYL001 and LND on the structure of BCSCs mitochondria;

[0031] Figure 8 A comparison of the effects of HYL001 and LND on the membrane potential of BCSCs mitochondria;

[0032] Figure 9 Comparison of intracellular reduced glutathione concentration changes caused by HYL001 and LND;

[0033] Figure 10 A comparison of the changes in intracellular reactive oxygen species levels caused by HYL001 and LND;

[0034] Figure 11 Comparison of changes in intracellular stem cell marker levels caused by HYL001 and LND;

[0035] Figure 12After treatment with HYL001 and LND respectively, the tumor volume-time curve (content a), tumor weight map (content c), tumor photograph (content c), and the percentage of CD133 positive cells (content d), side group cells (content e), and CD44 positive and CD24 negative cells (content f) in the tumor.

[0036] Figure 13 After treatment with HYL001 and LND respectively, lung metastases were photographed (content a), lung sections were stained with hematoxylin and eosin (content b), the number of lung nodules was quantified (content c), lung wet weight (content d) and survival time (content e) were compared. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0038] This invention provides a chlordamine derivative targeting tumor mitochondria, wherein the derivative has chlordamine as the pharmacophore and triphenylphosphine cation as the target. It has the structure shown in formula (I):

[0039]

[0040] Wherein, R is phenyl, or a phenyl group substituted with one or more substituents selected from halogen, C1-C4 alkyl, nitro, and trifluoromethyl. These derivatives induce oxidative stress in tumor stem cells by targeting mitochondria, disrupting mitochondrial morphology and function, and can induce apoptosis in tumor stem cells at extremely low concentrations. Compared to chlordamine, mitochondrial-targeting chlordamine derivatives exhibit superior anti-tumor growth and metastasis capabilities, and can significantly improve the survival rate of tumor-bearing mice.

[0041] In a preferred embodiment, R is an unsubstituted benzene ring with the structure shown in formula (II), named HYL001.

[0042]

[0043] In some embodiments, the synthesis of HYL001 involves a one-step etherification and a two-step esterification reaction.

[0044] Example 1

[0045] The chlordamine derivative shown in formula (II) was synthesized and named HYL001.

[0046]

[0047] The synthesis route diagram of HYL001 is as follows: Figure 1As shown, its synthesis method includes the following steps:

[0048] (1) 2-Hydroxy-5-methylisophthalic acid (1.7 g, 10 mmol), potassium carbonate (2.1 g, 15 mmol), and benzyl bromide (1.7 g, 10 mol) were added to 10 mL of N',N'-dimethylformamide solvent. The mixture was stirred at 80 °C for 6 hours, and then the reaction was stopped. Post-treatment involved extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, concentration, mixing with silica gel, and purification by column chromatography using petroleum ether:ethyl acetate = 1:1 as the eluent to obtain intermediate 3.

[0049] (2) In a round-bottom flask, intermediate 3 (645.8 mg, 2.5 mmol) was dissolved in 10 mL of dichloromethane, and a mixed solution of chlordamine (642.3 mg, 2 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (479.3 mg, 2.5 mmol), and 4-dimethylaminopyridine (24.3 mg, 0.2 mmol) in 20 mL of dichloromethane was added dropwise. The reaction was stirred at room temperature for 3 hours. After the reaction was completed, the post-treatment was performed by dichloromethane extraction, washing the organic phase with saturated brine, drying with anhydrous sodium sulfate, and concentrating. The concentrate was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain intermediate 4.

[0050] (3) Similar to the synthesis of intermediate 4, HYL001 was obtained through the esterification reaction of intermediate 4 and 4-carboxytriphenylphosphine bromide. The specific steps are as follows: In a round-bottom flask, 4-carboxytriphenylphosphine bromide (443.3 mg, 1.0 mmol) was dissolved in 10 mL of dichloromethane, and intermediate 4 (561.5 mg, 1.0 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (191.7 mg, 1.0 mmol), and 4-dimethylaminopyridine (24.3 mg, 0.2 mmol) in a mixed solution of 20 mL of dichloromethane were added dropwise. The reaction was stirred at room temperature for 4 hours. After the reaction was complete, the post-treatment involved extraction with dichloromethane, washing the organic phase with saturated brine, drying with anhydrous sodium sulfate, and concentrating. The concentrate was purified by silica gel column chromatography (methanol:dichloromethane = 1:20 → 1:10) to obtain a white, foamy solid powder. Figure 2 , Figure 3 and Figure 4 The NMR spectra (1H NMR, 1N P ... and 1N P NMR) and high-resolution mass spectra (HMR) of the solid powder prepared in Example 1 are shown below. NMR and mass spectrometry data: 1H NMR(600MHz, CDCl3)δ8.17(d,J=8.2Hz,1H),7.85–7.72(m,9H),7.68–7.62(m,6H),7.43–7.26(m,10H),7.15(d,J=1.8Hz,1H),7.02–6.98(m,1H), 6.67–6.64(m,1H),5.73(s,2H),5.51(s,2H),5.09(s,2H),4.97(s,2H),2 .44(t,J=6.8Hz,2H),2.29(s,3H),2.04–1.93(m,4H),1.75–1.65(m,2H). 31 P(600MHz,CDCl3)δ24.44.HRMS(ESI):m / z calcd for C 54 H 48 BrCl2N2O5P[M-Br] + 905.2672, found 905.26628. This indicates that HYL001 was successfully prepared.

[0051] Example 2

[0052] Comparison of in vitro antitumor and tumor stem cell activity assays of HYL001 prepared in Example 1 and LND.

[0053] The evaluation of the proliferation activity of 4T1 (mouse breast cancer cells) was conducted using the same method as conventional methods. Specifically, 4T1 tumor cells were seeded in 96-well plates at a seeding density of 5 × 10⁶ cells / well. 3 Cells per well were cultured in 100 μL of medium at 37°C in a 5% CO2 incubator. After 12 hours, the medium was removed, and 100 μL of different concentrations of HYL001 and LND culture medium prepared in Example 1 were added respectively. Cells were incubated for 48 hours. Cell viability was then calculated using the MTT assay.

[0054] The formation of tumor spheres in low-adhesion culture plates is the gold standard for cancer stem cell culture. Mouse breast cancer single cells (4T1) were first cultured in ultra-low adhesion plates using serum-free DMEM / F12 medium supplemented with B-27, 2.05 mM glutamine, 20 ng / mL epidermal growth factor, 10 ng / mL basic fibroblast growth factor, 4 μg / mL insulin, and 0.4% low-endotoxin bovine serum albumin. After 5 days, the spheres were harvested, dissociated into single cells using trypsin, and then recultured in the same medium in 96-well ultra-low adhesion plates to evaluate the efficacy of HYL001 anti-breast cancer stem cells (BCSCs).

[0055] Figure 5IC50 of HYL001 inhibits adhesion of 4T1 tumor cells 50 The value was 26.7 times lower than that of LND, and HYL001 inhibited the IC50 of breast cancer stem cells (BCSCs). 50 The value is 380.5 times lower than that of LND. Therefore, HYL001, which is conjugated with triphenylphosphine bromide, significantly enhances the potency of LND, especially against BCSCs.

[0056] Example 3

[0057] Example 1: Activity assay of HYL001 and LND prepared in vitro against mouse breast cancer tumor regeneration cells (4T1 TRCs).

[0058] 4T1 tumor regeneration cells (4T1 TRCs) were obtained through three-dimensional fibrinogen screening and possessed high tumorigenicity. Specifically, tumor cells were pretreated for 12 hours in conventional rigid plates with 1% DMSO, LND (10-200 μM), and HYL001 (0.1-10 μM), respectively. Then, fibrinogen (2 mg / mL) and the treated cells (4 × 10⁻⁶ cells / mL) were... 4 A mixture of cells / mL was prepared at a 1:1 ratio, and 50 μL of the mixture was inoculated into each well of a 96-well plate, followed by mixing with 1 μL of pre-packed thrombin (0.1 U / μL). After incubating the cell culture plates in a cell culture incubator for 20 minutes, the colloid solidified, and 200 μL of RPMI-1640 medium containing 10% fetal bovine serum was added. Ten areas were randomly selected from each group, and the number and size of colonies were counted or measured.

[0059] Figure 6 The image shows the inhibitory activity of HYL001 and LND on regenerated breast cancer cells (4T1TRCs) obtained by three-dimensional fibrous gel screening. Ctr represents the control group of tumor cells pretreated with 1% DMSO, LND-10μM represents the experimental group of tumor cells pretreated with 10μM LND, HYL001-1μM represents the experimental group of tumor cells pretreated with 1μM HYL001, HYL001-3μM represents the experimental group of tumor cells pretreated with 3μM HYL001, HYL001-5μM represents the experimental group of tumor cells pretreated with 5μM HYL001, and HYL001-10μM represents the experimental group of tumor cells pretreated with 10μM HYL001.

[0060] from Figure 6 It can be seen that HYL001 inhibits the formation and growth of 4T1 TRCs in a dose-dependent manner. In stark contrast, treatment with 10 μM HYL001 showed 100% inhibition of 4T1 TRCs, while LND at the same concentration had almost no inhibitory effect on 4T1 TRCs.

[0061] Example 4

[0062] Example 1: Effects of HYL001 and LND on mitochondrial structure

[0063] Collect BCSCs (approximately 1 × 10⁻⁶) after treatment with 1% DMSO, LND (100 μM), and HYL001 (2 μM) for 12 hours. 8 The mitochondria were washed three times with 2.5% glutaraldehyde at 4°C and PBS, then fixed, dehydrated, sectioned, stained, and observed under a transmission electron microscope.

[0064] Figure 7 In the diagram, Ctr represents the control group treated with 1% DMSO, LND represents the experimental group treated with LND (100 μM), and HYL001 represents the experimental group treated with HYL001 (2 μM). Figure 7 As shown, compared to LND having almost no effect on mitochondrial morphology, HYL001 can disrupt the structure of mitochondria, causing them to become vacuolated.

[0065] Example 5

[0066] Effects of HYL001 and LND on mitochondrial membrane potential of BCSCs in Example 1

[0067] Collect BCSCs (approximately 2 × 10⁻⁶) after treatment with 1% DMSO, LND (100 μM), and HYL001 (2 μM) for 12 hours. 5 The cells were stained with the mitochondrial membrane potential probe JC-1, washed with PBS, and intracellular fluorescence was observed under a confocal microscope.

[0068] like Figure 8 As shown in the confocal laser scanning microscopy images, compared with the untreated or LND-treated groups, the HYL001-treated group had more bright green fluorescence signals (JC-1 monomers) and weaker red fluorescence signals (JC-1 aggregates), indicating that HYL001 treatment caused depolarization of BCSCs mitochondria and loss of membrane potential.

[0069] Example 6

[0070] Effects of HYL001 and LND on intracellular redox balance in BCSCs in Example 1

[0071] BCSCs (approximately 1 × 10⁻⁶) were collected after being treated for 4 hours with 1% DMSO, LND (100 μM), and HYL001 (2 μM), respectively. 6 Intracellular glutathione content was determined using 5,5'-dithiobis(2-nitrobenzoic acid). Figure 9As shown, the intracellular glutathione content in the LND group was comparable to that in the blank group, and HYL001 reduced intracellular glutathione in a dose-dependent manner.

[0072] The level of reactive oxygen species (ROS) in mitochondria was measured using the red mitochondrial ROS probe Mito-SOX. Cell treatment methods were the same as for glutathione assays; after 4 hours of drug treatment, Mito-SOX staining was performed, and flow cytometry was used to quantify the intracellular fluorescence intensity in each group. Figure 10 As shown, the intracellular fluorescence intensity of the LND group was comparable to that of the blank group, and HYL001 increased the content of reactive oxygen species in mitochondria in a dose-dependent manner.

[0073] Therefore, it can be explained that HYL001 targeting mitochondria can cause an intracellular redox imbalance in BCSCs.

[0074] Example 7

[0075] Effects of HYL001 and LND on dryness markers in Example 1

[0076] Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot analysis were used to detect the ribonucleic acid and protein levels of stem cell markers, including SOX2, Nanog, CD133, and OCT4. For qRT-PCR, 4T1 tumor cells treated with 1% DMSO, LND (100 μM), and HYL001 (2 μM) for 12 hours were collected. Total RNA was extracted using the Trizol method, and RNA concentration was measured. cDNA was synthesized by reverse transcription, and then real-time quantitative PCR was performed with corresponding primers. For Western blot analysis, 4T1 tumor cells treated with 1% DMSO, LND (100 μM), and HYL001 (2 μM) for 24 hours were collected. Proteins were extracted, and each sample was separated with 10% sodium dodecyl sulfate-polyacrylamide. The samples were blocked overnight at 4°C with 5% bovine serum albumin, then incubated with the corresponding primary and secondary antibodies. Finally, the protein levels were observed by imaging.

[0077] like Figure 11 As shown, compared to the Ctr and LND groups, the levels of these dry biomarkers were significantly reduced after HYL001 treatment.

[0078] Example 8

[0079] The antitumor efficacy and stemness of HYL001 prepared in Example 1 in mouse breast cancer orthotopic tumors were investigated.

[0080] This invention investigated the antitumor effects of intratumoral injection of HYL001 and LND in mice using a mouse 4T1 mammary carcinoma orthotopic model. The specific steps are as follows:

[0081] Six-week-old, 20g female BALB / c mice were inoculated with 5 × 10⁵ mouse mammary gland 4T1 cell suspensions into the left mammary pad of the fourth pair of mammary glands in their abdomen. 5 A mouse model of 4T1 breast cancer orthotopic tumor was established using [number] cells. When the orthotopic tumor volume was approximately 80 mm², [the tumor was successfully induced]. 3 Mice were randomly divided into three groups of eight each. The groups were administered intratumoral injections of saline, LND, and HYL001, respectively. The dosage of LND and HYL001 was 5 mM / kg during treatment. The first day of administration was designated as Day 1, and subsequent administrations were given at the same dosage on days 1, 4, 7, 10, and 13. From Day 1 onwards, mouse body weight and tumor volume were measured daily, and a tumor volume-time curve was plotted. Mice were sacrificed on day 17, the tumor was dissected, weighed, and photographed.

[0082] Figure 12 Content a shows the tumor volume in mice, indicating that HYL001 significantly inhibits tumor growth, with tumor volumes significantly smaller than those in other groups. Content b shows the weight of the dissected tumors, showing that the tumor weight in the HYL001 group was significantly lower than that in other groups. Content c shows images of the dissected tumors after the experiment, showing that the tumors in the HYL001 group were significantly smaller than those in other groups. This demonstrates that LND-based targeted mitochondrial functional modification can significantly enhance its anti-tumor effect.

[0083] Furthermore, the dissected tumors were ground, filtered into single-cell suspensions, stained with stem cell-related markers (CD133 positive, CD44 positive, CD24 negative, and side population cells), and analyzed by flow cytometry to quantify the proportion of stem cells in each group of tumors.

[0084] Figure 12 Contents d, e, and f show that the proportions of CD133-positive cells, CD44-positive CD24-negative cells, and side population cells in the HYL001 group were all lower than in other treatment groups. This indicates that LND-based targeted mitochondrial function modification can significantly enhance its killing effect on tumor stem cells.

[0085] Example 9

[0086] Evaluation of the anti-metastatic efficacy of HYL001 prepared in Example 1 in a mouse metastatic tumor model

[0087] 2×10 5 Four 4T1 tumor cells were intravenously injected into the tail vein of each mouse. All mice were then randomly divided into three groups, with treatment regimens consistent with the in situ efficacy evaluation experiment. Sixteen days later, the mice were sacrificed, and the number of lung metastatic nodules in each group was counted and hematoxylin-eosin stained.

[0088] Figure 13Content a shows a photograph of the dissected lung, demonstrating that HYL001 significantly inhibits lung metastasis. Content b shows the observation of stained lung sections, indicating that the lung metastasis area treated with HYL001 was smaller than that in other groups. Contents c and d show the number of lung nodules and wet weight measurements; both results show that HYL001 has superior anti-tumor metastasis ability compared to LND. Content e shows the survival time of mice in each group, indicating that HYL001 significantly improves the overall survival of mice compared to LND.

[0089] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A lonidamide derivative targeting tumor mitochondria, characterized in that, It has the structure shown in equation (a): Formula (1) Wherein, R is a phenyl group, or a phenyl group substituted with one or more of the following: halogen, C1-C4 alkyl, nitro, and trifluoromethyl.

2. The method of preparing a lonidamide derivative targeted to tumor mitochondria according to claim 1, wherein, Includes the following steps: (1) 2-hydroxy-5-methylisophthalimide is etherified with a benzyl halide to obtain a bridging intermediate in which the phenolic hydroxyl group is substituted; the benzyl halide is benzyl bromide or benzyl bromide in which one or more of C1-C4 alkyl, nitro or trifluoromethyl groups are substituted on the benzene ring. (2) The bridging intermediate described in step (1) is subjected to an esterification reaction with chlordamine to obtain a chlordamine-linker conjugate; (3) The conjugate described in step (2) is reacted with 4-carboxybutyltriphenylphosphine bromide to obtain a chlordamine derivative targeting mitochondria.

3. The use of the chlordamine derivative targeting tumor mitochondria as described in claim 1 in the preparation of a medicament for treating and / or preventing tumors, wherein the tumor is breast cancer.

4. The use of the chlordamine derivative targeting tumor mitochondria as described in claim 1 in the preparation of a drug for killing tumor stem cells, wherein the tumor is breast cancer.

5. An anticancer drug, characterized by, It contains a mitochondrial-targeting chlordamine derivative as described in claim 1 and pharmaceutically acceptable additives.

6. An anticancer drug targeting tumor stem cells, characterized by, It contains a mitochondrial-targeting chlordamine derivative as described in claim 1 and pharmaceutically acceptable additives.

7. The anticancer drug according to claim 5 or 6, wherein The anticancer drug is available in the form of injection, oral medication, spray, or suppository.

8. The anticancer drug as claimed in claim 5 or 6, wherein The anticancer drug is in the form of powder for injection or capsules.

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