MXPA00003611A - Intra-cancer-cell nuclease activator - Google Patents

Abstract

A drug efficacious for cancer therapy and a novel drug containing a double-stranded RNA such as poly(I).poly(C). Specifically, an intra-cancer-cell nuclease activator containing 2-O-(2-diethylaminoethyl)-carbamoyl-1,3-O-dioleylglycerol and a composite comprising a carrier prepared from a phospholipid as an essential component and poly(I).poly(C) or mismatched poly(I). poly(C).

Description

ACTIVATOR OF THE NUCLEASE INTRA-CANCERIGENA-CELLULAR FIELD OF THE INVENTION («The present invention relates to an activator of cancer cell nuclease The term" activator of cancer cell nuclease "as used in this specification means a drug which activates nucleases 10 in cells cancerigenic and thereby induces apoptosis, and, therefore, death of cancer cells.Also, as used here, the code "I" remains for the inosinic acid, "C" for the cytidic acid, and MA "for the adenylic acid, and 15 WU "for uridyl acid The terms poly (I)" poly (C) unpaired and poly (A) "poly (U) unpaired, mean poly (I) * poly (C) and poly (A) ) "Poly (U), which contain non-complementary nucleic acid bases among those that constitute a double strand as is well known in the art.

BACKGROUND OF THE INVENTION Poly (I) * poly (C) is a double-stranded RNA comprising an acid polyribonucleotide copolymer REF: 119030 polyinosinic and polycytidylic acid, and is known to be a medically active substance having potent activity that induces interferon and unopotent activity. The fact that poly (I) »poly (C) has immunopotent activity suggests that the substance will indirectly inhibit the growth of cancer cells through immune reactions, thus driving many prospectors to explore within their potential utility as a therapeutic drug for malignant tumors. However, the indirect action mediated by the immunity of poly (I) * poly (C) is not potent enough to inhibit the growth of cancer cells ^ and yet, anti-cancer therapy with poly (I) * has not been implemented. poly (C). Therapeutic regimens using poly (I) «poly (C) for other indications based on their activities whether they induce interferon and immunopotent, have not been developed. The poly (A) * poly (U), which is a polyribonucleotide copolymer of polyadenylic acid and polyuridylic acid, the poly (I) * poly (C) unequal, and the poly (A) * poly (U) unequal, are also considered to have similar activities, through variations in degrees.

Meanwhile, as an effective carrier for the intracellular release of drugs, there are known carriers generally called cationic liposomes, such as Lipofectin (trademark), also as a carrier comprising a glycerol derivative such as 2-0- (2). -diethylaminoethyl) carbamoyl-1,3-dioleoylglycerol of the following chemical formula (I) and a phospholipid as essential components [eg, PCT W091 / 17424, PCT W094 / 19314].

CH, -0-CO- (CH. CH = CH (CH,) t CH. -cis I CH -O-C-NHCH, CH, N (CH, CH,), [I] C IH, -0-CO- (CH, CH = CH (CH,) »CH, -cis The cationic liposome is visualized as a small vesicle that has a lipid bilayer structure and assumes a positive charge in aqueous solution. Since such a cationic liposome is positively charged and a double-stranded NRA such as poly (I) «poly (C) is negatively charged in the aqueous solution, the cationic liposome and the poly (I) * poly (C), by For example, they can easily form a complex.

However, they are not known at all, whether complete double-stranded RNAs, for example poly (I) * poly (C), or the complex thereof, with a cationic liposome, will be able to activate the nucleases in the cancer cells to thereby induce apoptosis, and, therefore, the death of cancer cells.

DESCRIPTION OF THE INVENTION The object of the present invention is to provide an effective drug for cancer therapy. The object of the invention is also to provide a novel drug containing a double-stranded RNA, such as poly (I) * poly (C). The inventors of the present invention found after much research that an agent that activates the nuclease of cancer cells is effective in the therapy of malignant tumors and consequently, have developed the present invention. The present invention is therefore directed to a nuclease activator of cancer cells. As long as a substance is an activator of the nuclease of cancer cells, it can be easily determined experimentally, for example by observing the fragmentation of a DNA or RNA as in the Test of Example 2, which is presented later. Commonly, the invention encompasses a composition that *: • activates the cancer cell nuclease, which comprises a complex of an effective carrier for the intracellular release of a drug substance with an element selected from the group consisting of poly (I) «poly (C), poly (I) * poly (C) unpaired, poly (A) * poly (U), and poly (A) «poly (U) unpaired (those double-stranded RNAs, will subsequently be referred to each and collectively as" poly (I) »poly (C) or equivalent, and a composition * that activates the cell nuclease carcinogenic comprising a complex of a cationic liposome with poly (I) * poly (C) or equivalent. The preferred embodiment of the invention includes a composition that activates the nuclease of cancer cells comprising a complex (briefly, the complex) of a carrier (briefly, the carrier), comprising 2-0- (2-diethylaminoethyl) carbamoyl-1,3-O-dioleylglycerol (later referred to as the glycerol derivative) and a phospholipid as components essentials with Poly (I) »poly (C) or equivalent (the composition will be referred to later as the activator of the invention). The preferred exemplary activator of the invention is now described in detail. ("5 The carrier can be generally referred to as a cationic liposome, but does not necessarily need to be strictly in the form of a cationic liposome, so long as it is only functionally qualified to release a drug substance in the cells. The length of the poly (I) poly (C) chain or equivalents, according to the present invention, is not particularly restricted but, taking poly (I) * poly (C) as an example, it is suitable employ one in the range of 50-20,000 base pairs (bp). RNA of 100 to 500 bp is preferred, and one of 200-400 bp is even more preferred. If the length of the chain is less than 50 bp, the RNA will not be effective enough, as long as one is used.

If the chain length exceeds 2,000 bp, a safety problem may occur. The poly (I) * poly (C) within the range of 100-500 bp, is considered to be a balanced RNA in efficacy and safety. Since the poly (I) «poly (C) or Equivalent, usually exist as a given distribution of various chain lengths, the aforementioned chain length of poly (I) * poly (C), is a length of main chain. The phospholipid for use in the present (5) invention, is not particularly restricted, so long as it is a pharmaceutically acceptable phospholipid, including, but not limited to phosphatidy1choline, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, sphingomyelins and lecithin. Hydrogenated phospholipids can also be used. The preferred phospholipid includes egg yolk phosphatidyl choline, egg yolk lecithin, soy lecithin, and egg yolk phosphatide. I also know can use in combination, two or more different phospholipids. Compared with phosphatidylethanolamine, which is commonly used in cationic liposomes, phosphatidylcholine and lecithin are conducive to attenuation significant toxicity without engaging in activity. The proportion of the carrier to poly (I) * poly (C) or equivalent in the complexes, depends on the classes of phospholipids and poly (I) * poly (C) or equivalents, the type of cancer, and other factors but the recommended ratio of poly (I) »poly (C) relative to 10 parts by weight of the carrier is 0.05-10 parts in weight, preferably 0.1-4 parts by weight, and more ("Preferably, 0.5-2 parts by weight.) The ratio of the glycerol derivative to the phospholipid in the carrier depends on the class and amount of the poly (I) poly (C) or equivalent and the phospholipid class, but the recommended proportion of The phospholipid relative to each of the parts by weight of the glycerol derivative is 0.1-10 parts by weight, preferably 0.5-5 parts by weight, and more preferably 1-2 parts by weight. The activator of the invention can by For example, it may be provided in the form of a liquid preparation (an injection or drop infusion), in which the complex is dispersed in the aqueous solution, or a lyophilisate thereof. In the case of a liquid preparation, the concentration The recommended amount of the complex is 0.001-2556 (w / v), preferably 0.01-5% (w / v), and more preferably 0.1-1% (w / v). The activator of the invention may contain pharmaceutically acceptable additives, such as a auxiliary emulsifier, stabilizer, isotonizing agent, and / or pH control agent, in suitable amounts. Specifically, auxiliary emulsifiers such as C6 ~ 22 fatty acids (eg, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, arachidonic acid, docosalhexenoic acid) can be mentioned. its pharmaceutically acceptable salts (eg, sodium salts, potassium salts, calcium salts, etc.), albumin, dextran, etc., stabilizers such as cholesterol, phosphatidine, etc. isotonizing agents such as sodium chloride, glucose, maltose, lactose, sucrose, tr-ehalosa, etc., and pH control agents such as hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, sodium hydroxide, potassium hydroxide , triethanolamine, etc. The activator of the present invention can be produced by the general technology for the production of liposomes. A typical method comprises mixing a predetermined amount of water (eg, water for injection, distilled water for injection, or physiological saline) with a predetermined amount of the glycerol and phospholipid derivative under stirring, dispersing the mixture with a suitable dispersing machine such as a homomixer, ultrasonic disperser, ultrasonic homogenizer, high pressure emulsifier disperser, Microfluidizer (name * >; 5 commercial), Nanomizer (trade name), Ultimizer (trade name), or Manton-Gaulin high-pressure homogenizer, then add a predetermined quantity of poly (I) «poly (C) or equivalent, and redisperse the mixture, to provide the activator of the invention for injection. The optional additives mentioned above can be added to a suitable stage before or after the dispersion. As an alternative, the activator of the invention can be produced by the addition of water to a ternary mixture of the glycerol derivative, phospholipid and poly (I) poly (C) or equivalent, and dispersing the entire mixture. However, a raw dispersion step may be interposed. The activator thus prepared by the above dispersion process can be dry-cooled to provide a lyophilizer activator of the invention. This dry refrigeration operation can be carried out in one way conventional. For example, the activator of the invention, obtained by said dispersion process, is sterilized and distributed in bottles. The filled bottles are subjected to preliminary cooling at about -40 ° -20 ° C, for about 2 hours and, then, to primary drying in vacuo at about 0-10 ° C, and further to secondary drying in vacuo at about 15- 25 ° C. In general, the bottles are flushed with nitrogen gas and stopped to provide the objective lyophilized activator of the invention. The lyophilized stabilizer of the invention may be, in general, reconstituted by the addition of a suitable solvent (for reconstitution) and put to use. The solvent for reconstitution includes water for injection, physiological saline and other ordinary infusions. The volume of the solvent for reconstitution varies with the proposed use and is not particularly restricted but may preferably be 0.5-2 times the volume prior to drying by refrigeration, or not greater than 500 mL. The activator of the invention activates cancer cell nucleases to induce apoptosis and cell death and is only sparingly toxic so that it finds application with dramatic efficacy in cancer therapy, for example hepatocarcinoma, in mammals, including man. Particularly, the activator containing a complex formed between the carrier and the poly (I) «poly (C) is highly effective and is still very low in toxicity. The activator of the invention can be administered intravenously, topically, transmucosally, and by other routes in the treatment of neoplastic diseases. When the activator of the invention is used for the therapy of hepatocarcinoma, it is preferably administered intravenously, within the hepatic artery, or in the portal vein. The dosage of the activator of the invention in cancer therapy depends on the classes of poly (I) «poly (C) or equivalents and phospholipids, the type of cancer, the cancer stage, the age and species of the recipient, the route of administration, and treatment modality among other conditions. In terms of poly (I) * poly (C) or equivalent, the recommended dosage is usually 50 μg-50 mg / man per dose, and preferably 100 μg-2 mg / man per dose. The activator of the invention can be administered in an injection or by drip injection once for 3 times a day, every day, every third day, or on a weekly basis or t biweekly. EXAMPLES The following elaborated examples and test examples illustrate the present invention in further detail. It should be understood that the concentration of the activator of the present invention is variably expressed in the concentration of said poly (I) »poly (C) in the activator.

Example 1 A solution of 40 g of maltose in 100 mL of water for injection was mixed with 2 g of the glycerol derivative and 3 g of purified egg yolk lecithin, and the mixture was treated with a homogenizer for 5 minutes to prepare a dispersion of crude carrier. This crude dispersion was further treated, with a compact emulsifier disperser of upper platform, for 1 hour and adjusted to 250 mL with water for injection. The dispersion of the resulting carrier was recovered. To 25 250 mL of this carrier dispersion was added 150 mL of an aqueous solution containing 500 mg of poly (I) * poly (C) [main chain length: approximately 200 bp] with shaking, and using? a compact emulsifier disperser of upper platform 5, the mixture was further treated for 1 hour to provide the activator of the invention. This activator was then distributed in bottles, 1 L per bottle, and dry-cooled in a conventional manner. 10 Example 2 A solution of 4 kg of sucrose in 10 L of water for injection was mixed with 50 g of the glycerol derivative and 30 mg of egg yolk phosphatide, and the mixture was treated with a Manton-Gaulin high pressure homogenizer. , For 10 minutes. The resulting dispersion was made up to 25 L with water for injection, and recovered. To 20 L of this dispersion m 20 of the carrier, 12 L of an aqueous solution containing 10 g of poly (I) «poly (C) [main chain length: about 200 bp] was added with stirring, and the mixture was stirred. adjusted to pH 5.5 with hydrochloric acid and further treated with a Manton-Gaulin high pressure homogenizer for 30 minutes, to provide the activator of the invention. This activator was then distributed in bottles, 20 mL per bottle, and dry chilled in the conventional manner to provide a lyophilisate. This lyophilized stabilizer was reconstituted by the addition of a commercial infusion of 5% glucose (500 mL).

Example 3 A solution of 20 g of glucose in 100 mL of water for injection was mixed with 2 g of the glycerol derivative and 2 g of soy lecithin, and the mixture was treated with a homogenizer for 5 minutes to prepare a dispersion of raw carrier. This crude dispersion was further treated with a compact emulsifier disperser of upper platform, for 1 hour, and adjusted to 250 mL with water for injection. The resulting carrier dispersion was recovered. To 250 mL of this carrier dispersion, 150 mL of an aqueous solution containing 50 mg of poly (I) «poly (C) [main chain length: about 200 bp] was added with stirring, and using a compact emulsifying disperser of top platform, the mixture was further treated for 1 hour providing the activator of the invention, Example 4 A solution of 40 g of maltose in 100 mL of water for injection was mixed with 1.2 g of the glycerol derivative and 2.0 g of purified egg yolk lecithin, and this crude dispersion was further treated with a compact platform emulsifier disperser. superior for 30 minutes, and it was elaborated up to 250 mL with water for injection. The resulting carrier dispersion was recovered. To 250 mL of this carrier dispersion, 150 mL of a solution / aqueous containing 200 mg of poly (I) «poly (C) [main chain length: approximately 200 bp] was added with stirring, and using an emulsifying disperser compact top platform, the mixture was further treated for 2 hours providing the activator of the invention.

Example 5 A solution of 40 g of maltose in 100 mL of water for injection was mixed with 1.2 g of the glycerol derivative and 2.0 g of purified egg yolk lecithin, and this crude dispersion was further treated with a compact platform emulsifier disperser. superior for 30 minutes, and it was elaborated up to 250 mL with water for injection. He recovered the ("5 dispersion of resulting carrier." To 250 mL of this carrier dispersion, 150 mL of an aqueous solution containing 200 mg of poly (I) "poly (C) [main chain length: approximately 360 base] was added with agitations, and using a compact upper platform emulsifier disperser, the mixture was further treated for 2 hours providing the activator of the invention.

Example 6 A solution of 40 g of maltose in 100 mL of water for injection was mixed with 2 g of the glycerol derivative and 2 g of purified egg yolk lecithin, and the mixture was treated with a homogenizer. for 5 minutes to prepare a crude carrier dispersion. This crude dispersion was further treated with a compact upper platform emulsifying disperser for 1 hour, and worked up to 250 mL with water for injection. He recovered the resulting carrier dispersion. To 250 mL of this carrier dispersion, 150 mL of an aqueous solution containing 250 mg of poly (I) [main chain length: approximately 1419 bp] and 250 mg of poly (C) [chain length] was added. (.main: 1419 base) with shaking, and using a compact upper platform emulsifier disperser, the mixture was further treated for 1 hour providing the activator of the invention.This activator, then, was distributed in flasks, 1 L per bottle, and dry cooled in the conventional manner.

Example 7, 15 A solution of 40 g of maltose in 100 mL of water for injection was mixed with 1.2 g of the glycerol derivative and 2.0 g of purified egg yolk lecithin, and this crude dispersion was further treated with a compact emulsifier disperser. from upper platform for 30 minutes, and made up to 250 mL with water for injection. The resulting carrier dispersion was recovered. To 250 mL of this carrier dispersion, 150 mL of an aqueous solution containing 100 mg of poly (I) was added. [main chain length: approximately 84 base] and poly (C) [main chain length: 76 base] with shaking, and using a compact upper platform emulsifier disperser, the mixture was further treated for 2 hours providing the activator of the invention.

Example 8 The activator of the invention, containing poly (I) «poly (C) [main chain length: about 350 bp] was prepared in the same manner as given in Example 4.

Example 9 The activator of the invention, containing poly (I) * poly (C) [main chain length: about 1450 bp] was prepared in the same manner as given in Example 4.

Example 10 The activator of the invention, containing poly (I) * poly (C) [main chain length: about 80 bp] was prepared in the same manner as given in Example 4 Test Example 1: Growth inhibitory effect in several cell lines (in vitro) A 96-well plate was seeded with cells at a density of 10 4 cells / well. The next day, the activator of Example 4 or adriamycin was added and the cultivation continued. After 3 days, the viable cell count was determined by the MTT method. The results are shown in Tables 1 and 2.

Table 1 (»• The% inhibition was calculated by means of the following equation: O.D value of cells treated with drugs ("5 (1-) x 100% O.D value of cells treated with saline Table 2 % inhibition was calculated by means of the following equation: O.D value of cells treated with drugs ¿2 5 (1-) x 100% O.D value of cells treated with saline It is apparent from Tables 1 and 2, that the activator exhibited strong growth inhibitory effects, in many effects of fibroblasts and epithelial, were completely comparable with the effects of adriamycin, which showed anticancer activity, through the inhibition of nucleic acid synthesis. The activator has effects on the cancer cells of any organ, thus not showing organ specificity. Otherwise, the activator of the invention does not inhibit the growth of 4 lines cells derived from the liver, even at a concentration of 1000 ng / ml. It should be noted that this anticancer effect in vitro was not observed at all with the poly (I) * poly (C) alone, or in the solp carrier and this is an uncountable phenomenon from the simple translocation of poly (I) «poly (C) in cells.

Test Example 2: Observation of apoptosis («5 (1) Fragmentation of DNA and ANR (1) DNA fragmentation To each of the A431 cell lines and KM12-HX cell line, 1 μg / ml of the Activator of Example 4. A431 cells and KM12-HX cells were recovered after 5 hours and 7.5 hours, respectively. Cells were lysed with 5 mM Tris-HCl (pH 8.0) -10 M EDTA-0.5% (v / v) Triton X-100 and centrifuged at 13000 x g for 20 minutes to isolate the fragmented DNA (supernatant) and the chromatin fraction (pellet). Then, 100 μg / ml of Rnase A was allowed to act on the supernatant at 37 ° C for 1 hour, and subsequently, 200 μg / ml of proteinase K and 1% (w / v) of SDS (sodium dodecyl sulfate), were added for the reaction at 50 ° C for 1.5 hours. The fragmented DNA was extracted with phenol-chloroform and subjected to 1.8% agarose gel electrophoresis. As a result, DNA fragmentation was observed in both lines cell phones In the course of time, DNA fragmentation was investigated for the A431 cell line. Thus, a 6-well plate was seeded with A431 cells at a density of 2.8 x 10 5 cells / well and on the next day, the DNA of the cell line was labeled with 2 μCi [3 H] thiamidine. Then, the activator of Example 4 (1 μg / ml) was added and the cells harvested at time intervals. The cells were lysed with 5 mM Tris-HCl (pH 8.0) -10 mM EDTA-0.5% (v / v), Triton X-100 and centrifuged at 13000 xg for 20 minutes to separate the fragmented DNA ( supernatant) of the chromatin fraction (pellet). From the radioactivity measured in the supernatant and in the pellet, respectively, the ratio of fragmented DNA to total DN was calculated. The results are shown in Figure 1. The fragmentation rate was approximately 30% of the total DNA at 3 hours after the addition and not less than 55% at 5 hours, indicating that this fragmentation occurs immediately after intracellular capture of the activator of the invention. (2) Fragmentation of RNA To each of the A431 cell lines, MDA-MB-468 cell line, KB cell line, HeLa S3 cell line, and MCF-7 cell line, 1 μg / ml poly (I) was added. »Poly (C) of the activator of Example 4, and the treated cells recovered after 4 hours. From the recovered cells, the ribosome fraction was separated, and the total RNA was extracted by the ACPC method (Acid-Guanidium-Phenol-chloroform). The RNA was subjected to gel modified with formaldehyde (1.8% agarose gel) electrophoresis and staining of ethidium bromide. As a result, ribosome fragmentation of 28S and 18S RNAs was observed in all cell lines. (2) Effect of a nuclease inhibitor A 96-well plate seeded with HeLaS3 cells at a density of 10 4 cells / well and on the next day, 10 μM of the nuclease inhibitor ATA (aurintricarboxylic acid) and the activator according to Example 4 of the invention, were added simultaneously. The cells were further grown for 3 days and the number of viable cells was determined by the MTT method. The results are shown in Figure 2. It is apparent from Figure 2 that when the intracellular nuclease activity is inhibited by the addition of ATA, the activator of the invention fails to inhibit the growth of cancer cells. In addition, when the addition of ATA was removed from the medium after 8 hours and before the addition of the activator of the invention, the activator fails to express its activity. Therefore, it is thought that the effect of ATA was not that of inhibition of the intracellular take-up of the activator, but that of acting as an inhibitor of the nueclease. (3) The results of the above test indicate that the activator of the invention activates the intracellular nucleases to thereby induce apoptosis of the cancer cells.

Test Example 3: Effect in the murine metastatic hepatocarcinoma model (in vivo).

Using the Balb / c mouse knot, nu / nu / 5 weeks old, male), 106 cells / mouse of the KM12-HX cell line (a human colon cancer cell line which, when transplanted into the lung of the mouse knot, metastasizes to the (L 5 liver, with high efficiency causing neoplastic lesions), was injected into the lung and after 10 minutes, the lung was enucleated. Starting 3 days later, the activator according to Example 4 of the invention was administered twice weekly at substantially constant intervals for 5 consecutive weeks. Two days after the last dose, the liver was isolated, and the number and area of the cancer nodules, formed in the liver 'was determined. The results are shown in Table 3.

Table 3 The figures in the table denote the number and area of the carcinogenic nodes per mouse (mean + S.E). The figure in parentheses, *: significant to p < 0.01 (Dunnett essay). é < 5 Compared with the control group (giving maltose 10%), the inhibition of growth of liver cancer cells was 72% in the group of 30 μg / kg and 91% in the group of 100 μg / kg. 10 In the 100 μg / kg group, 77% inhibition was obtained even when the dosing schedule was once in a week. In addition, liver tissue specimens were prepared and examined biologically. As a result, liver cancer in the control group was low differentiation epithelial adenocarcinoma. The ordinary degree of vascularization feeder was observed. There was no evidence of remarkable cellular immune infiltration. 20 However, the tumor tissues showed local calcification. In the group treated with the activator of the invention, no obvious cancerous cells were detected, but only the calcification remained after the healing.

Thus, the activator of the invention showed significant efficacy in the animal hepatocarcinoma model, within the dose range of 10 μg / kg- 100 μg / kg in a dosing schedule of two times in a week.

Test Example 4: Toxicity study: (1) Expression of hepatoxicity in rats, giving an individual dose (acute toxicity study) Using 8 6-week-old male SD rats, the activator according to Example 4 of the invention, was administered in an individual intravenous dose, and the aminoacyl transferase activity of the serum was determined, after 20 hours. As a result, no death was found, up to 5 mg / kg and a slight elevation of serum aminoacyl transferase in the majority was observed at 5 mg / kg. At 1 mg / kg, the serum aminoacyl transferase level was less elevated. (2) Subacute toxicity study in rats, in two weeks The activator according to Example 4 ^ ~ 4 of the invention, was administered intravenously to 6 A1 5 male SD rats (6 weeks old) daily for 14 consecutive days. As a result, no remarkable signs of toxicity were found at doses up to 1 mg / kg. (3) Study of antigenicity Using male guinea pigs (Hartley strains, 5 weeks old), the antigenicity of the activator was studied according to Example 4 of the invention. As a result, antigenicity was not found at 50 μg / animal. (4) Mutagenicity study File 20 The activator according to Example 4 of the invention was subjected to a reverse mutation dosing test and a chromosomal aberration record. As a result, no mutagenicity was found at 10 μg /. 25 BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing the proportions of DNA fragmentation. The ordinate represents the percentage (%) of DNA fragmentation (* 5 and the abscissa represents time (hr) Figure 2 is a graph showing the effect of the addition of the nuclease inhibitor ATA, where the abscissa represents the concentration ng / ml) of the compliance activator. with Example 4 of the invention, -O- represents the data generated in the free system of ATA, and - • - represents the data generated in the system with ATA.

It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention is that which is clear from the present description of the invention. Having described the invention as above, the content of the following is claimed as property.

Claims (7)

1. A cancer cell nuclease activator, characterized in that it comprises a complex of an effective carrier in the cellular delivery of a drug with poly (I) "poly (C), poly (I)» poly (C) unpaired, poly (A) ) »Poly (U) or poly (A) * poly (U) unpaired.

2. The activator of the cancer cell nuclease, according to claim 1, characterized in that the effective carrier in the intracellular release of a drug is a cationic liposome preparation.

3. An activator of the cancer cell nuclease, characterized in that it comprises a complex of a carrier, which essentially consists of 2-0- (2-diethylaminoethyl) carbamoyl-1,3-dioleolylglycerol and a phospholipid with poly (I) * poly ( C) or poly (I) »poly (C) unequal.

4. The activator of the cancer cell nuclease, according to claim 3 or 4, characterized in that the phospholipid is lecithin. (• 5

5. The activator of the cancer cell nuclease, according to claim 3 or 4, characterized in that the poly (I) »poly (C) has 10 a main chain length within the range of 100 to 500 bp.

6. The activator of the cellular cancer cell nuclease, according to any of claims 3 to 5, characterized in that the cancer is hepatocellular carcinoma.

7. An anticancer composition, characterized in that it comprises the activator of the cancer cell nuclease, in any of claims 1 to 5 and 7. 25