CN119454681A

CN119454681A - Use of renal glutaminase inhibitors and combined use thereof

Info

Publication number: CN119454681A
Application number: CN202411590552.2A
Authority: CN
Inventors: 陈锦泽; 马亮
Original assignee: Beijing Runzhou Biotechnology Co ltd
Current assignee: Beijing Runzhou Biotechnology Co ltd
Priority date: 2024-11-08
Filing date: 2024-11-08
Publication date: 2025-02-18

Abstract

The invention relates to a renal glutaminase inhibitor and application of the renal glutaminase inhibitor and the combined drug. The invention obtains the specific KGA inhibitor compound by analyzing, screening and identifying in the known compound library through the molecular docking technology, and the compound has better KGA inhibition property and can obviously improve the effect of treating fibrosarcoma tumor after being combined with the specific genistein. After the two are prepared into the medicine box, the medicine box has excellent application prospect.

Description

Kidney type glutaminase inhibitor and application of combination thereof

Technical Field

The application relates to the field of biopharmaceuticals, in particular to a renal glutaminase inhibitor and application of the renal glutaminase inhibitor in combination.

Background

The kidney-shaped glutaminase GLS is encoded by a GLS gene located on human chromosome 2, with different splice variants. The longest one, known as renal glutaminase (KGA), the human KGA sequence was deduced from sequencing of a large number of isolated cDNAs, and more targeted studies have been made on the already found KGA analogues of murine and porcine. The GLS gene was studied and found to consist of 19 exons, exceeding 82kb in length, splicing of exons 1-15 to GAC and splicing of exons 1-14 and 16-19 to KGA. The mRNA of GAM contains the 2-3 intron, which is distinguished from GAC/KGA starting at residue 162. The C-terminal sequence of GAM is "VSFYIFLS" encoded by introns 2-3. Introns are very rare as alternative splicing forms in mammalian cells, and therefore it cannot be determined whether GAM is the result of a gene defect in the isolated cell due to cDNA.

The role of renal glutaminase in cancer has been the focus of research. Cancer genomic profile (TCGA) pan-oncogene expression data shows that GLS is highly expressed in acute myelogenous leukemia, adrenocortical carcinoma, triple negative breast cancer, colorectal cancer, renal clear or papillary cell carcinoma, lung adenocarcinoma, melanoma, mesothelioma, pancreatic cancer, sarcoma and thyroid cancer. Among them, GLS was found to be critical for the growth of acute myelogenous leukemia, breast cancer, colorectal cancer, renal cancer, lung cancer, melanoma, and pancreatic cancer cells through small molecule inhibitors and gene silencing experiments. Although TCGA data showed relatively low GLS expression in glioblastoma cell lines, they were still very sensitive to glutaminase inhibitors in vitro. These results indicate that GLS is critical for the growth of various cancer cell lines, and inhibition of this enzyme is a potentially valuable therapeutic strategy.

During the 80 years after glutaminase was found, the only chemical inhibitor of interest was DON (6-diazo-5-oxo-1-isoleucine). DON is a glutamine mimetic that irreversibly binds to catalytic serine of two glutaminase isozymes. DON has been the most widely used inhibitor in the last decade, although other small molecules have also been used to inhibit GLS (particularly membrane impermeable molecules for determining subcellular localization). DON, however, has limited its use due to its target's broad nature and severe toxicity. In 2007, another inhibitor BPTES of GLS was found. BPTES is a long, highly flexible and C2 symmetrical molecule that binds GLS in a 1:2 ratio and is located at the binding interface of the dimer. BPTES has approximately 1000 times higher affinity for GLS than LGA. However, to date, the only GLS enzyme used in the study was a recombinant mutant GLS enzyme, BPTES similar to its binding site and LGA. Since BPTES and GLS crystal structure complexes have been reported, BPTES has been the focus of research, focusing mainly on increasing inhibitor affinity and improving its pharmaceutical properties.

Furthermore, in a new study, researchers from university of tokyo, university of gentamicin Ying Yi, university of ninety, etc. have found that inhibition of mouse kidney-type glutaminase (kidney-type glutaminase, KGA) -dependent glutaminolysis can eliminate senescent cells. RNA interference (RNAi) is specifically used to find enzymes required for the survival of senescent cells, which are subsequently induced to die. This study involved the use of RNA interference to find enzymes required for the survival of senescent cells. This prompted them to carefully study glutamine metabolism, particularly glutaminase 1 (glutaminase, GLS 1). Tests have shown that it is critical for the survival of senescent cells. These researchers then inhibited the glutaminase 1 pathway in the mice tested. After allowing these changes time to play a role, inhibiting this pathway can lead to death of senescent cells. In the long term this also reduces age-related organ problems and also reduces obesity-related health problems.

However, at present, there are few studies on inhibitors of the kidney-type glutaminase GLS and there are few alternatives available.

Disclosure of Invention

In one aspect, the invention provides a specific KGA inhibitor.

The KGA inhibitor is a compound.

The KGA inhibitor compounds of the invention have the structural formula:

the compounds can also be synthesized according to prior art CN 102030700B.

Furthermore, the invention provides a pharmaceutical composition for treating fibrosarcoma tumor, which comprises the KGA inhibitor compound and pharmaceutical excipients.

Furthermore, the pharmaceutical excipients can be the excipients widely used in the field of pharmaceutical production. Adjuvants are used primarily to provide a safe, stable and functional pharmaceutical composition, and may also provide means for allowing the subject to dissolve at a desired rate after administration, or for promoting effective absorption of the active ingredient after administration of the composition. The pharmaceutical excipients may be inert fillers or provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients can include one or more of binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, binders, disintegrants, lubricants, anti-adherent agents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, reinforcing agents, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

Still further, materials that may be pharmaceutically acceptable excipients include, but are not limited to, ion exchangers, aluminum stearate, lecithin, serum proteins such as human serum proteins, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, lanolin, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch, celluloses and their derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate, gum powders, malt, gelatin, talc, adjuvants such as cocoa butter and suppository waxes, oils such as peanut oil, cotton seed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil, glycol compounds such as propylene glycol and polyethylene glycol, esters such as ethyl oleate and ethyl laurate, agar, buffers such as magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogen-free water, isotonic salts, ringer's solution, ethanol, phosphate buffer solutions, and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate, colorants, release agents, coating materials, sweeteners, flavoring agents and fragrances, preservatives and antioxidants.

Substances that may be pharmaceutically acceptable excipients include, but are not limited to, ion exchangers, aluminum stearate, lecithin, serum proteins, such as human serum proteins, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, lanolin, sugars, such as lactose, glucose and sucrose; starches such as corn starch and potato starch, celluloses and their derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate, gum powders, malt, gelatin, talc, adjuvants such as cocoa butter and suppository waxes, oils such as peanut oil, cotton seed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil, glycol compounds such as propylene glycol and polyethylene glycol, esters such as ethyl oleate and ethyl laurate, agar, buffers such as magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogen-free water, isotonic salts, ringer's solution, ethanol, phosphate buffer solutions, and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate, colorants, release agents, coating materials, sweeteners, flavoring agents and fragrances, preservatives and antioxidants.

The pharmaceutical compositions of the present invention may be prepared in accordance with the disclosure using any method known to those of skill in the art. For example, conventional mixing, dissolving, granulating, emulsifying, levigating, encapsulating, entrapping or lyophilizing processes.

The therapeutically effective amount of the compositions of the present invention can vary depending on a variety of factors, such as the method of administration, the target site, the condition of the patient, and the like. Therefore, when the composition of the present invention is used in a human body, the amount to be administered should be determined to be an appropriate amount that is compatible with safety and effectiveness. The amount for the human body can also be estimated from the effective dose determined by animal experiments. Matters to be considered in determining an effective amount are described in, for example, hardman and Limbird, goodman AND GILMAN's The Pharmacological Basis of Therapeutics, 10 th edition (2001), pergamon Press, and E.W. Martin, remington's Pharmaceutical Sciences, 18 th edition (1990), mack publishing company.

The compositions of the present invention may comprise carriers, diluents, excipients conventionally used in biological agents, and mixtures of two or more of the foregoing. The pharmaceutically acceptable carrier may be any carrier suitable for in vivo delivery of the composition, without particular limitation. For example, the compounds described in Merck Index (13 th edition, merck & co. Inc.) may be used, saline solution, sterile water, ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of the above, and other conventional additives (e.g., antioxidants, buffered solutions, bacteriostats (bacteristats), etc.) may be added as necessary. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to prepare injections (e.g., aqueous solutions, suspensions, emulsions), pills, capsules, granules or tablets. In addition, the compositions may be formulated according to the respective diseases or ingredients by a suitable method in the art or a method disclosed in Remington's Pharmaceutical Science (Mack publishing company, easton PA, 18 th edition, 1990).

The compositions of the present invention may additionally comprise one or more active ingredients having the same or similar function. The compositions of the present invention may comprise from about 0.0001wt% to about 10wt%, preferably from about 0.001wt% to about 1wt%, relative to the total weight of the composition.

The compositions of the present invention may be administered parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) or orally, depending on the method of interest. The dosage administered may vary depending on body weight, age, sex, health condition, diet of the particular patient, period of administration, method of administration, clearance rate, severity of disease, etc. The daily dosage of the composition of the present invention is from about 0.0001mg/mL to about 10mg/mL, preferably from about 0.0001mg/mL to about 5mg/mL, and more preferably the daily dosage of the composition is administered once a day or several times a day in divided fashion.

Furthermore, the present invention provides a kit for the treatment of fibrosarcoma tumor, said kit comprising the KGA inhibitor compound of the invention and genistein.

Further, the dosage ratio of the two medicines is 1:1-1:5, and more preferably 1:2.

Furthermore, the present invention also provides the use of a KGA inhibitor compound and genistein for the preparation of a kit for the treatment of fibrosarcoma tumors.

Further, the kit is used in an amount that is pharmaceutically effective.

The pharmaceutically effective amount may be a total amount of, but is not limited to, about 0.00001mg/kg to about 10mg/kg, preferably about 0.0001mg/kg to about 1mg/kg. The dosage administered may vary depending on body weight, age, sex, health condition, diet of the particular patient, period of administration, method of administration, clearance rate, severity of disease, etc.

The individual may be a vertebrate, preferably a mammal, more preferably a laboratory animal (e.g., rat, rabbit, guinea pig, hamster, dog, and cat), and most preferably a ape (e.g., chimpanzee and gorilla). The method of administration may be oral or parenteral. For parenteral administration, intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, epidural injection, cerebrovascular injection, or intrathoracic injection may be selected.

Further, based on the mechanism of the present invention, the kit can also be used for the treatment of other cancers.

The term "cancer" as used herein refers to solid tumors and hematological tumors selected from the group consisting of breast cancer, prostate cancer, cervical cancer, ovarian cancer, gastric cancer, colorectal cancer (i.e., including colon and rectal cancers), pancreatic cancer, liver cancer, brain cancer, neuroendocrine cancer, lung cancer, kidney cancer, hematological malignancies (e.g., leukemia), melanoma, and sarcoma. More particularly preferred cancers are selected from the group consisting of hematological malignancies, prostate cancer, breast cancer, cervical cancer, ovarian cancer, colorectal cancer, melanoma, and lung cancer. In a particularly preferred embodiment, the cancer is Acute Myelogenous Leukemia (AML) or prostate cancer.

Further, the medicine in the medicine box is solid dispersion. Solid dispersions, particularly MBP and/or spray-dried products obtainable according to the provided methods, can be used in various forms for administration of poorly water soluble drugs (e.g. compound a), particularly for oral dosage forms. Exemplary dosage forms include powders or granules, tablets, capsules or pills that may be dried or reconstituted by the addition of water to form a paste, slurry, suspension or solution for oral ingestion. Various additives may be mixed, milled or granulated with the solid dispersions as described herein to form materials suitable for the above dosage forms. Potentially beneficial additives may generally fall into the group of other matrix materials or diluents, surfactants, drug complexing or solubilizing agents, fillers, disintegrants, binders, lubricants and pH modifying agents (e.g., acids, bases or buffers). Examples of other matrix materials, fillers or diluents include lactose, mannitol, xylitol, microcrystalline cellulose, calcium pyrophosphate and starch. Examples of surfactants include sodium lauryl sulfate and polysorbate 80. Examples of drug complexing or solubilizing agents include polyethylene glycol, caffeine, xanthene, gentisic acid and cyclodextrin. Examples of disintegrants include sodium carboxymethyl starch, sodium alginate, sodium carboxymethyl cellulose, methyl cellulose, and croscarmellose sodium. Examples of binders include methylcellulose, microcrystalline cellulose, starches, and gums such as guar gum and gum tragacanth. Examples of lubricants include magnesium stearate and calcium stearate. Examples of the pH modifier include acids such as citric acid, acetic acid, ascorbic acid, lactic acid, aspartic acid, succinic acid, phosphoric acid, etc., bases such as sodium acetate, potassium acetate, calcium oxide, magnesium oxide, trisodium phosphate, sodium hydroxide, calcium hydroxide, aluminum hydroxide, etc., and buffers generally comprising a mixture of an acid and a salt of the acid. At least one function of including such a pH modifier is to control the dissolution rate of the drug, matrix polymer, or both, thereby controlling the local drug concentration during dissolution.

Advantageous effects

The invention obtains the specific KGA inhibitor compound by analyzing, screening and identifying in the known compound library through the molecular docking technology, and the compound has better KGA inhibition property.

Since combination therapy is a common and effective form of treatment for tumor therapy, it is not easy to choose with what drug combination,

The inventor can obviously improve the effect of treating fibrosarcoma tumor after the compound is used in combination with specific genistein by identifying dozens of drug combinations. The synergistic effect is not significant with other common drugs such as paclitaxel.

The compounds of the invention have better safety and the preparation methods are also mature and known in the art.

Drawings

FIG. 1 is a graph showing the tumor-inhibiting effect of each treatment group

Detailed Description

Specific embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. While specific embodiments of the invention are shown in the drawings, it should be understood that the invention may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

EXAMPLE 1 screening for inhibitors of human KGA recombinant proteins

The amino acid sequence of the human KGA functional active region is as follows

IPDFMSFTSH IDELYESAKK QSGGKVADYIPQLAKFSPDL WGVSVCTVDG QRHSTGDTKV

PFCLQSCVKP LKYAIAVNDL GTEYVHRYVG KEPSGLRFNK LFLNEDDKPH NPMVNAGAIV

VTSLIKQGVN NAEKFDYVMQ FLNKMAGNEY VGFSNATFQS ERESGDRNFA IGYYLKEKKC

FPEGTDMVGI LDFYFQLCSI EVTCESASVM AATLANGGFC PITGERVLSP EAVRNTLSLM

HSCGMYDFSG QFAFHVGLPA KSGVAGGILL VVPNVMGMMC WSPPLDKMGN SVKGIHFCHD

LVSLCNFHNY DNL recombinant expression of Pichia pastoris by Detai organism to obtain functional active protein KGA, and after SDS-PAGE and human kidney type glutaminase ELISA kit (product number 2H-KMLJh315182, brand carvedilol camilo) are used for identifying active recombinant protein, regulating the protein concentration to 1mg/mL for standby.

And (3) regulating the final concentration of the KGA inhibitor compound (formula 1) obtained by screening the molecular docking technology to 0-100 mu M in an enzyme reaction system. Specifically, the preparation method comprises the steps of K ₂PO₄ M and 0.15M,

Tris-ACETATE PH.650.00 mM, bovine serum albumin BSA 0.10mg/mL, EDTA 0.25mM,DTT 1.00mM,NAD 4.00mM, glutamine 0-20mM, glutamate dehydrogenase GDH 1unit, KGA

1.00. Mu.M, compound 0-100. Mu.M.

A reaction system was prepared at the above concentration without adding the enzyme KGA and the compound. The reaction group without adding the compound is taken as a blank group, the protein KGA is added in the last step, the change of OD value of the reaction system at 340nm in 10min is measured at 37 ℃, the final volume of the enzyme reaction system is 100.00 mu L,3 compound holes are reacted each, the reaction system without adding the protease KGA and the compound is prepared according to the same steps, the compound and the enzyme KGA are incubated for 10min at 37 ℃, then the enzyme reaction system is added to start the reaction, the final volume is 100.00 mu L, the change of OD value in 10min is measured, 3 compound holes are used for each reaction condition, the inhibition OD value of the compound on the enzyme reaction system is measured, BPTES is taken as a positive control compound, the inhibition activity of the compound on the KGA is measured by the same steps, and the IC50 value of the compound on the KGA is calculated. The results are shown in Table 1.

IC50 values of the compounds of Table 1 for KGA

Type of compound	IC50(μM)
		KGA inhibitor compounds (formula 1)	2.03±0.12
BPTES positive control	8.67±0.56

As can be seen from Table 1, the KGA inhibitor compounds of the invention (formula 1) have a better inhibitory activity against KGA.

EXAMPLE 2 cytotoxicity assay of KGA inhibitor Compound (formula 1)

Human fibrosarcoma cell HT1080 was purchased from cloud clone (Beijing) Biotechnology Inc., model YKLHT-1080. Taking out frozen cells in a liquid nitrogen tank, quickly thawing in a 37 ℃ water bath, transferring to a 10cm cell culture dish, adding 10mL of a culture medium containing 10% FBS, culturing for 4h at 37 ℃ by 5% CO ₂, sucking the supernatant and the cells which are not adhered to the wall after 4h, discarding, washing once by using an isotonic FBS phosphate buffer solution, discarding, and adding 10mL of a cell culture medium containing 10% FBS; when the cell density reached about 80%, pancreatin was digested, counted, diluted to a density of 5 ten thousand/mL with a medium containing 10% FBS, added to a 96-well plate with a lance at 100.00. Mu.L/well (5000 pieces/well) and cultured in a cell culture box for 12 hours with 100.00. Mu.L of FBS added to the outermost round of the 96-well plate, 5) diluted the compound with a cell culture medium (containing 10% FBS) to a concentration gradient of 200.00. Mu.M, 100.00. Mu.M, 50.00. Mu.M, 25.00. Mu.M, 12.50. Mu.M, 6.25. Mu.M, 3.12. Mu.M, 1.56. Mu.M, 0.78. Mu.M, and aspirated with a lance at 96-well plate (as thoroughly aspirated as possible), 3 compound-containing medium was added to each of 3 multiplex wells at 37℃in a cell culture box, CCK-8 was added to a cell culture medium without FBS for 12 hours so that the final concentration of CCK-8 was 10%, and the cell proliferation was inhibited at 96% by measuring cell culture medium at 96% IC at 37℃of 300.37.37 nm, 0.5.5 nm, respectively. BPTES was used as a control. The results are shown in Table 2.

IC50 values of compounds of Table 2 for tumor cells HT1080

Type of compound	IC50(μM)
		KGA inhibitor compounds (formula 1)	5.13±0.34
BPTES positive control	48.17±0.76

As can be seen from Table 2, the KGA-inhibitor compounds of the invention (formula 1) have significantly higher proliferation-inhibiting activity on tumor cells than the positive control compounds BPTES which have been reported. Has better activity of inhibiting tumor cells.

Example 3 verification of therapeutic Effect of Compounds and combinations thereof

Balb/C nude mice were supplied by Beijing Vietnam laboratory animals Inc.

Taking human fibrosarcoma HT1080 cell strain in logarithmic growth phase, digesting with 0.25% trypsin at room temperature, centrifuging to remove supernatant, centrifuging and washing with serum-free corresponding culture solution for 2 times, counting cells, and regulating cell density to obtain cell suspension of about 5×10 ¹⁰/L cells. 0.2ml of the prepared cell suspension was extracted and inoculated under the skin of the right axilla of Balb/C nude mice. When the tumor volume reaches about 100mm ³, animals are grouped for administration.

The successfully vaccinated human fibrosarcoma mice were randomly divided into model control group, compound high, medium, low dose group and positive control group and combination dosing group, and genistein monotherapy group. The compound administration dose is 2mg/kg/2d, 1mg/kg/2d, 0.5mg/kg/2d. 2 times per week, 3wk continuous intraperitoneal injections. The control group was given the same compound with cyclophosphamide dosing of 2mg/kg/2d 2 times per week, 3wk continuous intraperitoneal injection. The combination group was a compound in combination with genistein (purchased from sigma) at a dose of 1mg/kg/2d and genistein at 2mg/kg/2d, administered 2 times per week, followed by 3wk intraperitoneal injections. The Genistein monotherapy group was Genistein 2mg/kg/2d, and was given 2 times per week for continuous 3wk intraperitoneal injection. The model group was injected with physiological saline. Animals of each group were sacrificed 24h after the last dose, the tumor peeled off and the tumor weight was measured, and the average tumor weight and tumor suppression rate of each group were calculated. Tumor inhibition rate (%) = (average tumor weight of control group-average tumor weight of administration group)/average tumor weight of control group 100%. The results are shown in FIG. 1.

As can be seen from fig. 1, each treatment group had a significant tumor-inhibiting effect (P < 0.01) relative to the model group. And from the dosage of the compound, the tumor inhibition rate of the tumor is gradually improved along with the increase of the dosage. And the therapeutic effect of the compound is better than that of the positive control group. The compound of the invention and the genein medicine obtained by screening are combined to have obvious synergistic treatment effect, the tumor inhibition rate reaches (98.14 +/-2.92)%, the effect is far better than that of the medicines in the prior art, and the compound has better treatment effect.

Before mice of each treatment group were sacrificed, each mouse was tested for no obvious symptoms of toxic side effects. After the mice are sacrificed, livers are taken to detect corresponding enzyme activity indexes, and obvious liver poisoning symptoms are avoided. The compound is proved to have better safety.

It should be noted that the above embodiments are merely for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that the technical solution described in the above embodiments may be modified or some or all of the technical features may be equivalently replaced, and these modifications or substitutions do not deviate from the essence of the corresponding technical solution from the scope of the technical solution of the embodiments of the present invention.

In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the spirit and scope of the invention as defined by the appended claims and their equivalents.

Claims

1. Use of a KGA inhibitor compound and genistein in the manufacture of a kit for the treatment of fibrosarcoma tumors, wherein the KGA inhibitor compound has the structural formula:

2. The use according to claim 1, wherein the ratio of KGA inhibitor compound to genistein in the kit is 1:2.

3. The use according to claim 1, wherein the fibrosarcoma tumor is caused by HT 1080.

4. Use of a KGA inhibitor compound for the manufacture of a medicament for the treatment of fibrosarcoma tumors, wherein the KGA inhibitor compound has the structural formula: