KR20080016548A - Nutrient compositon for treating sarcoma and prostate cancer - Google Patents

Abstract

The present invention relates to the use of a composition comprising an ascorbic acid compound, a L-lysine compound, a L-proline compound, and a polyphenol compound for the preparation of a pharmaceutical composition for treating fibro-or synovial, sarcoma and prostate cancer. Moreover, the invention further relates to a method of treatment wherein said composition is administered to a subject suffering from fibro-or synovial sarcoma or prostate cancer.

Description

Nutritional composition for treating sarcoma and prostate cancer {NUTRIENT COMPOSITON FOR TREATING SARCOMA AND PROSTATE CANCER}

The present invention relates to the use of a composition comprising an ascorbic acid compound, an L-lysine compound, an L-proline compound, and a polyphenol compound in the manufacture of a pharmaceutical composition for treating fibro- or synovial sarcoma and prostate cancer. The invention also relates to a method of treatment in which said composition is administered to a subject suffering from fibro- or synovial sarcoma, or prostate cancer.

The American Cancer Society estimates that in 2004, approximately 563,700 Americans will die of cancer and that approximately 1,368,030 new cancer patients will be reported (American Cancer Society (2004)). The means of detection have been improved, which has led to early detection of various cancers, thus broadening the range of treatment options available and reducing the associated mortality rates. Haese et al. (2001). In addition, the survival rate is good when diagnosed early. However, the associated prognosis is generally poor in cancers advanced as soft tissue sarcomas and highly metastatic carcinomas. Approximately 90% of all cancer mortality rates occur in patients with metastatic cancer. For example, even when fibrosarcomas are diagnosed early and surgically treated, the probability of cancer spreading to other distant parts of the body is greater than 70%. Papagelopoulos et al. (2002)].

Although surgery is an optional treatment for early stage cancer, its use is generally limited to local tumors and involves numerous possible side effects. Patients who have undergone prostatectomy may have nerve damage, erectile dysfunction, incontinence, or rectal damage. Bishoff et al (1998), Melman et al. (2004), Talcott et al. (2003). Suboptimal methods for treating cancer include hormone therapy and radiation therapy.

Initially, hormone therapy and androgen ablation therapy are advantageous for patients with prostate cancer, but if androgen-independent or sensitive cancer cells appear, this treatment regimen becomes ineffective after up to several years. Isaacs et al. (1999). In addition, patients may consequently experience erectile dysfunction, hot flash, nausea, vomiting, diarrhea, liver complications, bone weakness, breast growth, and / or tenderness [National Cancer Institute (2004), Conde et al. (2003). Chemotherapy and radiation therapy are generally not advantageous for patients with advanced cancer because they focus on cancer cell destruction and do not focus on metastasis. These therapies are not only effective in treatment but also indiscriminately attack all cells, causing cellular damage and destruction of connective tissue in the body and promoting cancer metastasis. Current therapies are clearly limited in the treatment and prevention of metastatic cancer and there is an urgent need for new safe and effective treatment methods.

Several nutrient compounds have been reported to exhibit anticancer activity. Ascorbic acid has been reported to have cytotoxic and antimetastatic activity against malignant cell lines. Koh, Lee et al. (1998), Roomi, House et al. (1998), Naidu, Karl et al. (2003); In addition, low levels of ascorbic acid have been reported in cancer patients. Anthony and Schorah (1982), Nunez, Ortiz de Apodaca et al. (1995), Kurbacher, Wagner et al. (1996)]. ECGC is a potent anticancer agent reported to have a growth inhibitory effect on certain human cancer cell lines. Valcic, Timmerman et. al. (1996), Mukhtar and Ahmed (2000), Yang, Liao et al. (1998). However, the observed effect is rather weak and is not a suitable basis as a treatment method for rapidly growing invasive cancer. Other studies have shown that the synergistic anticancer effect of both ascorbic acid and EGCG and other compounds on several cancer cell lines was greater than that of individual nutrients in tissue culture studies. Netke, Roomi et al. (2003). Cancer cells spread by forming tumors and degrading extracellular matrix (ECM) through various matrix metalloproteinases (MMPs). Nutrients such as lysine and ascorbic acid have been presumed to act as natural inhibitors of ECM proteolysis and as substances with the potential to inhibit tumor growth and expansion (Rath and Pauling (1992)). It has also been suggested that, through inhibition of hyaluronidase, ascorbic acid can prevent metastasis by preventing degradation of the underlying material surrounding the tumor [Cameron et al. (1979)].

There is a clear need for a safe and effective treatment method for controlling the process of cancer metastasis, especially for invasive cancers such as fibro- or synovial sarcoma and prostate cancer.

Accordingly, the technical problem underlying the present invention should be seen as the provision of means and methods for meeting the above-described needs.

This problem is solved by the embodiments characterized by the claims and the specification.

Therefore, the present invention

(a) ascorbic acid compounds,

(b) an L-lysine compound,

(c) an L-proline compound, and

(d) polyphenol compounds

The present invention relates to the use of a composition comprising a fiber- or synovial sarcoma, or a pharmaceutical composition for treating prostate cancer.

The term "composition" includes liquid and solid preparations of the aforementioned nutritional compounds and their gels. Solid preparations may be prepared in suitable forms including tablets, capsules, powders, granules, tea preparations and the like. Methods of preparing the liquid, gel-like and solid formulations referred to herein are well known in the art. The compositions referred to herein may be provided according to the use of the invention as a kit comprising a mixture of compounds or components separately. Can be packaged in the kit in separate vials. The composition is also suitable for human or animal use. Preferably, said animal is a mammal, most preferably a dog, cat or horse.

The amino acids "proline" or "lysine" mentioned in accordance with the invention are preferably L-amino acids. "Proline" or "lysine" also includes hydroxyproline and hydroxylysine and their salts, which are their hydroxyl derivatives.

The term "ascorbic acid" preferably refers to ascorbate, ascorbic acid and salts thereof. Sometimes ascorbate compounds may be referred to as vitamin C.

The term "polyphenol compound" as used according to the present invention refers to the preparation of green tea plants, preferably comprising the polyphenol compounds present in green tea. The polyphenolic compound may be present in up to 30 dry weight percent in green tea. These include bioflavinoids such as flavanols, flavandiols, flavonoids, and phenolic acids. Flavanols represent the most abundant polyphenols in green tea and are commonly known as catechins. Most preferably, the catechin is EGCG, EG, ECG or EC. EGCG refers to (-)-epigallocatechin-3-gallate, EC refers to epicatechin called (-)-epicatechin, and ECG refers to epicatechin-3- called (-)-epicatechin-3-gallate Refers to gallate, and EGC refers to epigallocatechin called (-)-epigallocatechin. Methods of obtaining such formulations are well known in the art. Preferably, the polyphenol is administered in an amount of 200 mg to 5000 mg per person per day.

The compounds mentioned according to the use of the present invention may be admixed in any suitable ratio or amount. Whether such ratios or amounts are appropriate can be determined by one skilled in the art using the assays specified in the accompanying examples, which are mentioned below. Most preferably, the compositions referred to in accordance with the invention comprise 700 mg of vitamin C (as ascorbic acid, Mg, Ca or palmitate ascorbate) in a daily dose; L-lysine 1000 mg; L-proline 750 mg; 500 mg of L-arginine; 200 mg N-acetyl cysteine; 1000 mg of standardized green tea extract (80% polyphenols); Selenium 30 mg; 2 mg of copper; Feed 1 mg of manganese.

Pharmaceutical compositions administered in accordance with the present invention may comprise a pharmaceutically acceptable carrier, diluent, or excipient. The compositions used according to the invention can be prepared by methods known in the art. Each component may be formulated using conventional excipients, diluents, or carriers, and may be prepared in tablets, capsules, suspensions, powders, and the like. Examples of excipients, diluents and carriers include i) fillers and extenders such as starch, sugars, mannitol, and silicic acid derivatives; ii) binders such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl-pyrrolidone; iii) humectants such as glycerol; Disintegrants such as calcium carbonate and sodium bicarbonate; Dissolution retardants such as paraffin; iv) absorption accelerators such as quaternary ammonium compounds; v) surfactants such as acetyl alcohol, and glycerol monostearate; v) adsorptive carriers such as kaolin and bentonite; And vi) lubricants such as talc, calcium and magnesium stearate, and solid polyethyl glycols. The compositions may also be formulated with elixirs or solutions for convenient oral administration, or solutions suitable for parenteral administration, for example by the intramuscular, subcutaneous or intravenous route. Ideally, the formulation is a pill, tablet, capsule, lozenge, liquid or similar dosage form as mentioned above. The composition may be suitable for formulation such as sustained release dosage forms and the like.

As used herein, the term "treatment" is used to mean reducing, inhibiting, attenuating or treating a syndrome accompanying the pathological condition referred to in accordance with the present invention. Treatment is evident by the clinician by monitoring the symptoms accompanying the pathological condition. Symptoms are described in detail in standard textbooks such as Stedman or Pschyrembel. Treatment preferably refers to significant reduction, inhibition, attenuation or treatment. Significance can be determined by standard methods of statistics, such as Student's t-test, chi square test, and the like.

The term “fiber- or synovial sarcoma” refers to a type of cancer of mesenchymal origin that appears in connective tissue and synovial membranes. The term "prostate cancer" relates to all cancer types that originate in the prostate. Preferably, the term refers to prostate carcinoma. Symptoms associated with such diseases or disorders are well known in the art and are described in detail in standard textbooks such as Stedman or Pschyrembel. Thus, the clinician can determine whether the patient suffers from such a disease without further pain. The term "prevention" means that the nutrient composition can be administered to prevent the onset of the aforementioned diseases.

In the underlying studies of the present invention, heterogeneous athymic nude mice of a composition, sometimes referred to as a nutrient mixture (NS) containing lysine, proline, arginine, ascorbic acid, and epigallocatechin gallate The inhibitory effect on the growth of transplanted human cancers was studied. One week after isolation, female mice (group I) 5 to 6 weeks old were inoculated with breast cancer cells MBA-MD-231 3 × 10 ⁶ , male mice were divided into 5 groups and cancer cells 3 × 10 ⁶ were inoculated. Group II, prostate cancer cell PC-3; Group III, colon cancer cell HCT 116; Group IV, melanoma cells A2058; Group V, fibrosarcoma HT 1080; Group VI, synovial sarcoma Hs 701.T. After infusion, each group was randomly divided into two subgroups, fed diets to IA to VIA groups, and fed diets containing 0.5% nutrient mixture to IB to VIB groups (each subgroup was 6 mice). Consisting of). After 4 weeks, mice were euthanized and their tumors excised and weighed and treated for histology. The results showed that nutrient synergism inhibited tumor growth and decreased size in nude mice: while prostate cancer was significantly reduced to 53%, fibrosarcoma to 59%, and synovial sarcoma to 44%. , Breast cancer was less reduced (27%, data not shown). In addition, histological examination showed that the mitotic index was reduced by the nutrient mixture. Nutrient mixtures strongly inhibit tumor growth without any side effects in nude mice, and therefore are proposed as promising anticancer agents.

Thanks to the present invention sarcomas, such as fibrosarcoma, synovial sarcoma and prostate cancer, can be treated without the deleterious side effects of conventional therapies such as surgery and / or radiation therapy.

The definitions and explanations of the above terms are appropriately changed and applied to other embodiments of the present invention described below as necessary.

In a preferred embodiment of the use of the present invention, the ascorbic acid compound is ascorbic acid, a pharmaceutically acceptable ascorbate salt, ascorbate ester, more preferably ascorbyl palmitate, and / or a mixture of these compounds Selected from the group consisting of: More preferably, the pharmaceutically acceptable ascorbate salt is selected from the group consisting of calcium ascorbate salts and magnesium ascorbate salts.

Suitable amounts of the compounds used in the preparation of the compositions mentioned according to the invention are disclosed in detail in document WO 03/057201 A2, which is incorporated herein. Preferably, the ascorbic acid compound is calcium ascorbate and / or magnesium ascorbate and / or ascorbyl palmitate in an amount and / or equivalent amount of 25 mg to 5000 mg of ascorbic acid. All amounts are calculated as the daily dose per individual.

In a further preferred embodiment of the use of the invention said L-lysine compound is selected from the group consisting of L-lysine hydrochloride, L-lysine and pharmaceutically acceptable L-lysine salts.

Suitable amounts of the compounds used in the preparation of the compositions mentioned according to the invention are disclosed in detail in the document WO 03/057201 A2, which is incorporated herein. The L-lysine compound may also preferably be a mixture of two or more compounds of the foregoing group. Preferably, the L-lysine compound is L-lysine in an amount of 50 mg to 5000 mg per person per day.

In another preferred embodiment of the use of the invention said L-proline compound is selected from the group consisting of L-proline hydrochloride, L-proline and pharmaceutically acceptable L-proline salts.

Suitable amounts of the compounds used in the preparation of the compositions mentioned according to the invention are disclosed in detail in the document WO 03/057201 A2, which is incorporated herein. The L-proline compound may also preferably be a mixture of two or more compounds of the foregoing group. Preferably, the L-proline compound is L-proline in an amount of 25 mg to 3000 mg per person per day.

In another preferred embodiment of the use of the invention the composition further comprises a trace element selected from the group consisting of trace elements selenium, manganese, magnesium, calcium and copper.

Suitable amounts of the compounds used in the preparation of the compositions mentioned according to the invention are disclosed in detail in the document WO 03/057201 A2, which is incorporated herein. The composition preferably contains at least two, or at least three trace elements of the aforementioned groups. Preferably, the trace elements can be used together, separately, or in any combination in the following amounts per day per individual: selenium: 1 μg to 200 μg; Copper: 20 μg to 9000 μg; Manganese: 50 μg to 10000 μg; Calcium: 300 mg to 600 mg; Magnesium: 300 mg to 600 mg.

In a further preferred embodiment of the use of the invention, the composition further comprises an L-arginine compound. More preferably, the L-arginine compound is selected from the group consisting of L-arginine hydrochloride, L-arginine and pharmaceutically acceptable L-arginine salts.

Suitable amounts of the compounds used in the preparation of the compositions mentioned according to the invention are disclosed in detail in the document WO 03/057201 A2, which is incorporated herein. The L-arginine compound may also preferably be a mixture of two or more compounds of the foregoing group. Preferably, the L-arginine compound is L-arginine in an amount of 50 mg to 3000 mg per person per day.

In another preferred embodiment of the use of the present invention, the composition may further comprise N-acetyl cysteine.

As used herein, "N-acetyl-cysteine" includes cysteine or cystine (dimer of cysteine) and cysteine salts thereof. Suitable amounts of the compounds used in the preparation of the compositions mentioned according to the invention are disclosed in detail in the document WO 03/057201 A2, which is incorporated herein. Preferably, N-acetyl cysteine is administered in an amount of 10 mg to 1500 mg per person per day.

In a preferred embodiment of the use of the invention the composition is administered in oral, parenteral, subcutaneous, intraarterial or intravenous form.

The present invention also provides a method of treating an individual suffering from fibro- or synovial sarcoma or prostate cancer, comprising administering to a subject suffering from fibro- or synovial sarcoma or prostate cancer in a therapeutically effective amount of a composition as defined herein above. It is about.

Methods of administering such compositions to a subject are well known to those skilled in the art. Particularly preferred techniques are described in detail in document WO 03/057201 A2, which is incorporated herein. In addition, all embodiments of the use of the present invention are suitably modified and applied as necessary to the aforementioned methods for the treatment of fibro- or synovial sarcoma or prostate cancer.

In a preferred embodiment of the method of the invention, the subject is a human.

In an embodiment of a further preferred method of the invention, the composition is administered orally, parenterally, subcutaneously, intraarterally or intravenously.

Complete bibliographic information for the references cited above and below can be found below. The contents disclosed in these references are included herein.

references

Figure 1 shows tumor size and weight for nude mice 4 weeks after treatment with nutrient mixtures (NS) (human breast cancer cells MDA-231 (1A), prostate PC-3 (1B), colon cancer cells HCT 116 ( 1C), melanoma cells A2058 (1D), fibrosarcoma HT 1080 (1E), and synovial sarcoma Hs 701.T (1F)), results are shown as mean ± standard error (SE) for each group.

2 shows the effect of nutrient synergy on tumor presence in male nude mice (2A) and tumor growth in prostate cancer cells PC-3 in nude mice against colon cancer cells HCT 116, melanoma cells A2058 and prostate cancer cell PC-3 Effect of 0.5% nutrient composition (NS) on (tumor value) is shown (2B).

Figure 3 shows human breast cancer cell MDA-231 (3A-control, 3B-0.5% NS), colon cancer cell HCT 116 (3C-control, 3D-) on histological examination 4 weeks after administration of 0.5% nutrient mixture (NS). 0.5% NS), melanoma cells A2058 (3E-control, 3F-0.5% NS), and prostate cancer cell PC-3 (3G-control, 3H-0.5% NS).

The invention is illustrated in the following examples. However, the examples are not intended to limit the scope of the present invention.

Example : Treatment of Different Cancer Types in Nude Mice

All cancer lines used in this study were obtained from the American Type Culture Collection, Rockville, Maryland. This includes: breast cancer (MDA-MB-231), prostate cancer (PC-3), colon cancer (HCT 116), melanoma (A2058), fibrosarcoma (HT-1080) and synovial sarcoma (HTB-93) .

Melanoma and synovial sarcoma cultures are Dulbecco's Modified Eagle Medium, PC-3 is Ham Fs Medium (F12K), Colon Cancer is McCoy medium, HT-1080 is MEM, MBA- MD-231 was maintained in Leibovitz medium. All media was supplemented with 10% fetal calf serum (FBS), 100 U / ml penicillin and 100 μg / ml streptomycin. Medium and serum were obtained from ATCC, and antibiotics (penicillin and streptomycin) were obtained from Gibco BRL (Gibco BRL, Long Island, NY). Near the confluence, cells cultured for inoculation were trypsinized, washed with PBS, 3 × in 0.2 ml of PBS and 0.1 ml of Matrigel (BD Bioscience, Bedford, MD). Diluted to 10 ⁶ cell concentration and emulsified. About six weeks old male and female athymic nude mice (NCr-nu / nu) were purchased from Simonsen Laboratories, Gilroy, Calif., And 12-hour light / 12-hour for a week. Dark schedules were stored in microinsulator cages under pathogen free conditions. All animals were managed according to institutional guidelines on the care and use of experimental animals.

After storage for a week, mice were divided into two groups. Female nude mice were group I, and male nude mice were divided into five group II-VI groups consisting of six mice each. The right flank, the site of injection, was washed with ethanol and inoculated with 3 x 10 ⁶ cancer cells in 0.2 ml of PBS and 0.1 ml of Matrigel as follows: group II, prostate cancer cell PC-3; Group III, colon cancer cell HCT 116; Group IV, melanoma cells A2058; Group V, fibrosarcoma HT 1080; Group VI, synovial sarcoma Hs 701.T. Female mice (Group I) were inoculated with breast cancer cell MBA-MD-231. After infusion, each group was randomly divided into two subgroups, A and B. Six mice were assigned to each subgroup. From the first day, the IA to VIA groups were fed a diet and the IB to VIB groups were fed a diet supplemented with 0.5% nutrient composition. Tumor growth was monitored by measuring the length and width of the tumor with a caliper every three days. Find the tumor volume in mm ³ = length x width ² Calculated using x½. After 4 weeks, mice were euthanized, tumors excised and weighed, fixed in 10% (v / v) buffered formalin and processed for histology.

As a result, it was shown that tumor growth was suppressed and the size was reduced by the nutrient synergy in the IB to VIB groups (nude mice supplemented with 0.5% nutrient mixture). Nutrient synergism had a significant effect on tumor size in all cancer cell lines studied: breast cancer cell MBA-MD-231 27% (p = .0082), prostate cancer cell PC-3 53% (p = .0001 ), Colon cancer HCT 116 63% (p = .0002), melanoma cells A2058 57% (p = .0001), fibrosarcoma HT 1080 59% (p = .0001), and synovial sarcoma Hs 701.T. 44% (p = .0099) (see FIGS. 1A-1E). On biopsy, treatment with nutrient mixtures showed a reduction in mitosis index. Treatment with the nutritional composition resulted in a significant decrease in the average tumor abundance in nude mice xenografted with human cancer cells (prostate, melanoma, and colon) studied over a four week treatment period: 46% of colon (p = .0005) ; Melanoma 32% (p = .0015); Prostate 47% (p = .0001). See FIG. 2A. Treatment over 4 weeks in nude mice supplemented with nutrient composition showed a prostate tumor growth of 36% (p = .002) reduced at 4.5 weeks compared to the control (see FIG. 2B).

All chemicals used were purchased from Sigma (Siga, Missouri, St. Louis) and were of high quality. The stock solution of the nutritional mixture (4.4 g total weight) consists of the following ingredients: 700 mg of vitamin C (as ascorbic acid, Mg, Ca or palmitate ascorbate); L-lysine 1000 mg; L-proline 750 mg; 500 mg of L-arginine; 200 mg N-acetyl cysteine; 1000 mg of standardized green tea extract (80% polyphenols); Selenium 30 mg; 2 mg of copper; Manganese 1 mg. Statistical results are expressed as mean ± standard error for the group. Data was analyzed by an independent sample "t" test.

Claims (15)

(a) ascorbic acid compounds, (b) an L-lysine compound, (c) an L-proline compound, and (d) polyphenol compounds Use of the composition comprising a pharmaceutical composition for treating fibro- or synovial sarcoma or prostate cancer. Use according to claim 1, wherein the ascorbic acid compound is selected from the group consisting of ascorbic acid, pharmaceutically acceptable ascorbate salts, ascorbate esters and / or mixtures of these compounds. 3. Use according to claim 2, wherein said pharmaceutically acceptable ascorbate salt is selected from the group consisting of calcium ascorbate salt and magnesium ascorbate salt. Use according to claim 1, wherein the ascorbate compound is an ascorbate ester. 5. Use according to claim 4, wherein the ascorbate ester is ascorbyl palmitate. The use according to any one of claims 1 to 5, wherein the L-lysine compound is selected from the group consisting of L-lysine hydrochloride, L-lysine and pharmaceutically acceptable L-lysine salts. Use according to any one of claims 1 to 6, wherein the L-proline compound is selected from the group consisting of L-proline hydrochloride, L-proline and a pharmaceutically acceptable L-proline salt. 8. Use according to any of the preceding claims, wherein the composition further comprises a trace element selected from the group consisting of selenium, manganese, magnesium, calcium and copper. The use according to any one of claims 1 to 8, further comprising the composition L-arginine compound. 10. The use of claim 9, wherein the L-arginine compound is selected from the group consisting of L-arginine hydrochloride, L-arginine and pharmaceutically acceptable L-arginine salts. Use according to any of the preceding claims, further comprising the composition N-acetyl cysteine. Use according to any one of the preceding claims, wherein said composition is administered in oral, parenteral, subcutaneous, intraarterial or intravenous form. A method of treating fibro- or synovial sarcoma or prostate cancer, comprising administering to a subject suffering from fibro- or synovial sarcoma or prostate cancer a therapeutically effective amount of a composition as defined in claim 1. Way. The method of claim 13, wherein the subject is a human. The method of claim 13 or 14, wherein the composition is administered orally, parenterally, subcutaneously, intraarterally or intravenously.

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