EA010263B1 - A method for chemoprevention of prostate cancer - Google Patents

Abstract

This invention relates to the chemoprevention of prostate cancer and, more particularly, to the use of raloxifene or tamovifene or a derivative thereof, isomer, or pharmaceutically acceptable salts, esters, or N-oxides, and mixtures thereof for the preparation a pharmaceutical composition for the treatment of precancerous precursors of prostate adenocarcinomaiesions, for the preparation a pharmaceutical composition for reducing the amount of precancerous precursors of prostate adenocarcinomaiesions, and for the preparation a pharmaceutical composition for inhibiting or suppressing precancerous precursors of prostate adenocarcinomaiesions.

Description

The invention relates to chemoprophylaxis of prostate cancer and, more specifically, to a method of suppressing or inhibiting latent prostate cancer, comprising administering a chemoprophylactic agent and its analogues and metabolites to a mammal. A chemoprophylactic agent prevents recurrence or suppresses or inhibits prostate carcinogenesis and treats prostate cancer.

Background of the invention

Prostate cancer is one of the most common cancers among men in the United States, where hundreds of thousands of new cases are diagnosed each year. Unfortunately, it is found that in more than 60% of newly diagnosed cases, prostate cancer progresses pathologically and is not treated, and the prognosis is disappointing. One approach to this problem is to detect prostate cancer at an earlier stage using screening programs, thus reducing the number of patients with advanced prostate cancer. However, another strategy is to develop drugs to prevent prostate cancer. Every third man over the age of 50 has a latent form of prostate cancer, which can be activated before the life-threatening clinical form of prostate cancer. It is shown that the incidence of latent tumors of the prostate gland increases significantly with each decade over a period of life from 50 years (5.314%) to 90 years (40-80%). The number of people with latent prostate cancer is the same for all cultures, ethnic groups and races, but the incidence of clinically aggressive cancer differs markedly. This suggests that environmental factors may play a role in the activation of latent prostate cancer. Thus, developing a strategy for chemoprophylaxis against prostate cancer can be a powerful overall blow in both medical and economic terms for prostate cancer.

Toremifene is an example of the triphenylakenes compound described in US Pat. Nos. 4,696,949 and 5,491,173, Τοίνο1η е1 а1., Included in this description as references. Parenteral and topical administration of toremifene-containing compositions to mammals is described in US Pat. No. 5,571,534, 1st Alen et al., And in US Pat. No. 5,605,700, Geoscadopo et A1., Which are included in this description by reference.

US Pat. No. 5,595,985, LaFe, incorporated herein by reference, also describes a method for treating benign prostatic hyperplasia using a combination of 5a-reductase inhibitor and a compound that binds androgen receptors and blocks access to them. One example of compounds that block androgen receptors is flutamide.

In US patent No. 4329364 and 4474813, No. E1 A1, included in this description as a reference, describes pharmaceutical preparations containing flutamide, to delay and / or prevent the onset of prostate cancer. The drug may be in the form of a capsule, tablet, suppository, or elixir. Despite these developments, there remains a need for tools and methods that are effective in preventing prostate cancer.

Because of the high incidence of disease and death from prostate cancer, it is necessary to develop strategies for chemoprophylaxis against this devastating disease. Understanding the factors contributing to prostate carcinogenesis, including initiation, promotion and development of prostate cancer, will provide molecular mechanistic criteria for appropriate intervention points to prevent or stop the carcinogenic process. New approaches are urgently needed both at the scientific and clinical levels to reduce the incidence of prostate cancer, as well as to stop or cause regression of latent prostate cancer. As the incidence of prostate cancer rises dramatically at an age when men confront other competing causes of mortality, simply slowing the development of prostate adenocarcinoma can be both a more appropriate and cost-effective health strategy. The present invention is directed to meeting such a need.

In addition, since the prostate gland intraepithelial neoplasia is on the direct causal pathway to prostate cancer, and its presence specifically indicates an increased risk of prostate cancer, in men diagnosed with prostate gland intraepithelial neoplasia, dramatic changes occur in their quality of life. The only way to establish a diagnosis of prostate intraepithelial neoplasia is prostate biopsy. However, when the diagnosis of intraepithelial neoplasia of the prostate gland is established, the usual medical observation is that the patient should undergo a biopsy more often and visit a doctor. In addition, the patient and the doctor are very worried because the diagnosis of prostate cancer is imminent. Currently, there is no treatment available for patients who have prostate intraepithelial neoplasia.

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Brief description of the invention

The invention relates to a method of preventing prostate carcinogenesis in a subject, comprising administering to a mammalian subject a pharmaceutical preparation containing raloxifene, or tamoxifen, or an analogue thereof, a derivative, an isomer, and their pharmaceutically acceptable salts, esters or Ν-oxides, and mixtures thereof.

This invention relates to a method of suppressing or inhibiting latent prostate cancer in a subject, comprising administering to a mammalian subject a pharmaceutical preparation containing raloxifene or tamoxifen, or an analog, derivative, isomer, and their pharmaceutically acceptable salts, esters or-oxides, and their mixes.

This invention relates to a method of reducing the risk of developing prostate cancer in a subject, comprising administering to a mammalian subject a pharmaceutical preparation containing raloxifene, or tamoxifen, or an analogue thereof, a derivative, an isomer, and their pharmaceutically acceptable salts, esters or Ν-oxides, and mixtures thereof .

This invention relates to a method for increasing the survival rate of a subject having prostate cancer, comprising administering to a mammalian subject a pharmaceutical preparation containing raloxifene or tamoxifen, or an analogue thereof, a derivative, an isomer, and their pharmaceutically acceptable salts, esters or оксид-oxides, and mixes.

This invention relates to a method of treating a subject having prostate cancer, comprising administering to a mammalian subject a pharmaceutical preparation containing raloxifene or tamoxifen, or an analogue thereof, a derivative, an isomer, and their pharmaceutically acceptable salts, esters or Ν-oxides, and mixtures thereof.

This invention relates to a method of reducing the amount of pre-cancerous precursors of pathological changes in prostate adenocarcinoma in a subject, comprising administering to a mammalian subject a pharmaceutical preparation containing raloxifene or tamoxifen, or an analog, derivative thereof, isomer, as well as their pharmaceutically acceptable salts, esters or Ν -oxides and mixtures thereof.

In one embodiment, the precancerous precursor of adenocarcinoma of the prostate gland is intraepithelial neoplasia of the prostate gland (). In one embodiment, the precancerous precursor of adenocarcinoma of the prostate gland is a high degree intraepithelial neoplasia of the prostate gland (ΡΙΝ).

The present invention relates to a safe and effective method of preventing prostate carcinogenesis, suppressing or inhibiting latent prostate cancer and is particularly useful for treating subjects with an increased risk of developing prostate cancer, for example, subjects with benign prostatic hyperplasia, intraepithelial neoplasia of the prostate gland () either with abnormally high levels of circulating prostate-specific antibodies (Ρ8Α), or su Objects with a family history of prostate cancer.

Brief Description of the Drawings

FIG. 1. A graph illustrating the chemopreventive effect of torenifen on the model ΤΚΑΜΡ.

FIG. 2A-C. H & E slices showing cells of the ventral lobe of the prostate in healthy mice and prostate cancer in ΤΚΑΜΡ-mice participating in the study.

FIG. 3. Effect of torenifen on the development of the ventral lobe of the prostate gland in the ΤΚΑΜΡmouse.

FIG. 4. The effect of toreniphene on the appearance of a tumor in-mice.

FIG. 5. Effect of toreniphene on tumor development on model ΤΚΑΜΡ.

FIG. 6A-B. Comparison of the effect of torenifen relative to placebo on tumor growth.

Detailed Description of the Invention

This invention relates to the following methods:

1) a method of preventing prostate carcinogenesis;

2) a method of suppressing or inhibiting prostate cancer;

3) a way to reduce the risk of developing prostate cancer;

4) a way to increase the degree of survival of the subject;

5) a method of treating prostate cancer;

6) the method of regression of the intraepithelial neoplasia of the prostate gland and ways to reduce the number of pathological changes in the intraepithelial neoplasia of the prostate gland of a high degree by introducing a chemopreventive agent described in this description.

In one embodiment of the invention, the chemoprophylactic agent is raloxifene or tamoxifen, its analog, derivative, isomer, and their metabolites.

As shown in this specification, tamoxifen, faslodex, and raloxifene, which are examples of anti-estrogens, are chemoprophylactic agents for the prostate gland and anti-prostate intraepithelial neoplasia agents.

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Intermediate target biomarkers are measurable biological changes in tissue that occur between the initiation and the development of obvious neoplasia. The biomarker states if the end goal — the incidence of cancer — is also reduced by the intended chemopreventive agent. Intermediate biomarkers for cancer can be classified into the following groups: histological markers, proliferation and differentiation markers, and biochemical markers. With any chemoprophylaxis strategy, the availability of histologically recognizable and recognized precancerous pathological changes constitutes an important starting point. For the prostate, the histological marker is a precancerous precursor of adenocarcinoma of the prostate, an example of which is intraepithelial neoplasia of the prostate gland (ΡΙΝ). ΡΙΝ appears as an abnormal proliferation in the prostate grooves of the precancerous focus of cell dysplasia and η διι carcinoma without stromal invasion. ΡΙΝ and histological prostate cancer morphometrically and phenotype are similar. Thus, development ΡΙΝ of a high degree is an important stage in the development path through which ΡΙΝ, histological prostate cancer, invasive clinical prostate cancer and metastasis develop in a normal prostate gland.

It has been shown that intraepithelial neoplasia of the prostate gland is a precancerous pathological change or a precursor of adenocarcinoma of the prostate gland. Intraepithelial neoplasia of the prostate gland is an abnormal proliferation in the prostate grooves of the pre-malignant foci of cell dysplasia and ίη δίΐιι carcinoma without stromal invasion. Intraepithelial neoplasia of the prostate gland is the most accurate and reliable marker of prostate carcinogenesis and can be used as a suitable target when testing chemoprophylactic agents for the prostate gland. Intraepithelial neoplasia of the prostate gland as a marker is important for the prediction of adenocarcinoma, and its identification serves as a basis for a repeated biopsy for simultaneous or subsequent invasive carcinoma. Most studies suggest that most patients with prostate intraepithelial neoplasia will develop cancer within 10 years. Of interest is the fact that the intraepithelial neoplasia of the prostate gland does not contribute to the content of "8" in serum, which is not surprising, since unlike prostate cancer, the intraepithelial neoplasia of the prostate gland does not encroach on the vasculature of the prostate gland to pass the "8" into the bloodstream. Thus, the intraepithelial neoplasia of the prostate gland may be even more important than prostate cancer with respect to an increase in serum Ρ8Ά.

This invention relates to the use of a composition and a pharmaceutical composition for preventing prostate carcinogenesis, suppressing or inhibiting prostate cancer, reducing the risk of developing prostate cancer, increasing the survival rate of a subject, treating prostate cancer, regressing prostate intraepithelial neoplasia and reducing the intraepithelial changes high prostate neoplasia by introducing chemoprotein ilakticheskih means mentioned above and a carrier or diluent, and / or pharmaceutically acceptable carrier for them, diluents, salts, esters or Ν-oxides, and mixtures thereof.

The present invention relates to a safe and effective method of preventing carcinogenesis, suppressing or inhibiting latent prostate cancer and is particularly useful for treating subjects with an increased risk of developing prostate cancer, for example, subjects with benign prostatic hyperplasia, intraepithelial neoplasia of the prostate gland () or abnormal high levels of circulating antibodies specific for the prostate gland or having a family history of cancer edstatelnoy gland. In one embodiment of the invention, the subject is a mammal. In another embodiment, the subject is human. The invention covers pure (() - and (E) -isomers of the compounds and their mixtures, as well as a pair of pure (CK.88) - and (B8.8B) -enantiomers and their mixtures.

The invention includes pharmaceutically acceptable salts of amino-substituted compounds with organic and inorganic acids, for example, citric acid and hydrochloric acid. The invention also includes α-oxides of amino-substituted compounds of formula (I). Pharmaceutically acceptable salts can also be obtained from phenolic compounds by treatment with inorganic bases, for example, sodium hydroxide. You can also get the esters of phenolic compounds with aliphatic and aromatic carboxylic acids, for example, esters of acetic acid and benzoic acid.

The term “pharmaceutical composition” as used herein means a therapeutically effective amount of an agent together with suitable diluents, preservatives, solubilizers, emulsifiers, adjuvant and / or carriers. As used herein, the term “therapeutically effective amount” refers to an amount that provides a therapeutic effect for a given condition and mode of administration. Such compositions are liquids or lyophilized or otherwise dried formulations and include diluents with different contents.

- 3 010263 buffer substances (for example, Tris-HC1, acetate, phosphate), pH and ionic strength, additives such as albumin or gelatin to prevent adhesion to surfaces, detergents (for example, Tween 20 or Tween 80, Pluronic E68, salts bile acids), solubilizers (for example, glycerin, polyethylene glycol), antioxidants (for example, ascorbic acid, sodium metabisulfite), preservatives (for example, thimerosal, benzyl alcohol, parabens), fillers or tonicity modifiers (for example, lactose, mannitol, polymers, such as polyethylene glycol, covalently attached to a protein, complexes with metal ions or substances included in or attached to polymer particles, such as polyacrylic acid, polyglycolic acid, hydrogels, etc., or included in liposomes, microemulsions, micelles, monolamellar or multilamellar vesicles, hemolized red blood cells or spheroplasts. Such compositions will affect the physical state, solubility, stability, degree of release of η νίνο, and degree of purification of ίη νίνο. Compositions with controlled or delayed release include the composition in lipophilic depot (for example, fatty acids, waxes, oils). The invention also encompasses compositions in the form of particles with a polymer coating (for example, poloxamers and poloxamines). Other embodiments of the compositions of the invention include particle forms with protective coatings, protease inhibitors or penetration enhancers for various modes of administration, including parenteral, pulmonary, nasal, and oral. In one embodiment, the pharmaceutical composition is administered parenterally, paracanerally, transmucosally, transdermally, intramuscularly, intravenously, intradermally, subcutaneously, intraperitoneally, intraventricularly, intracranially and into the tumor. The dosage may be in the range of 580 mg / day. In another embodiment, the dosage is in the range of 35-66 mg / day. In another embodiment, the dosage is in the range of 40-60 mg / day. In another embodiment, the dosage is in the range of 4560 mg / day. In another embodiment, the dosage is in the range of 15-25 mg / day. In another embodiment, the dosage is in the range of 55-65 mg / day. The dosage may be 60 mg / day. The dosage may be 20 mg / day. The dosage may be 45 mg / day.

A tablet for oral administration contains 88.5 mg of a chemopreventive agent, which is equivalent to 60 mg of a chemopreventive agent.

In addition, the pharmaceutically acceptable carrier used in this case is well known to those skilled in the art and includes, but is not limited to, 0.01-0.1 M, preferably 0.05 M, phosphate buffer or 0.8% saline. In addition, such pharmaceutically acceptable carriers can be aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous carriers are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil, and organic esters for injection, such as ethyl oleate. Water carriers include water, alcoholic-aqueous solutions, emulsions or suspensions, including saline and buffered media. Carriers for parenteral administration include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, Ringer's lactate or non-volatile oils. Intravenous carriers include fluid and nutritional supplements, electrolytes such as those based on Ringer's dextrose, and the like. Preservatives and other additives may also be present, such as, for example, antimicrobial agents, antioxidants, cobalt inert gases, and the like.

The term adjuvant refers to a compound or mixture that enhances the immune response to an antigen. An adjuvant can serve as a tissue depot, gradually releasing an antigen, and also as an activator of the lymphoid system that nonspecificly enhances the immune response (Noob A1 A1., 1shipo160, 8esop EB, 1984, Wei) Atsch / Sittd8: Meplo Rugk, CaPGog and MecPo, CpGog and MecPi, CaPGog and MecPG, CuPGog and MecPG, CaPGog and MecPG. , R. 384). Often, the primary infection with an antigen alone in the absence of an adjuvant is not enough to detect a humoral or cellular immune response. Adjuvants include Freund's complete adjuvant, Freund's incomplete adjuvant, saponin, mineral gels such as aluminum hydroxide, surfactants such as lysolecithin, pluronic polyols, polyanions, peptides, oily or hydrocarbon emulsions, hemocyanins of snail lymph, dinitrophenol and hemocyanins. an adjuvant useful for humans, such as BCG (Laste Sa1teTse-Siepp) and Capsuleus. and other substances. Preferably, the adjuvant is pharmaceutically acceptable.

Compositions with controlled or delayed release include a composition in a lipophilic depot (eg, fatty acids, waxes, oils). The invention also encompasses compositions in the form of particles with a polymer coating (for example, poloxamers and poloxamines) and the compound in combination with antibodies directed against tissue-specific receptors, ligands or antigens, or in combination with tissue-specific receptor ligands. Other embodiments of the compositions of the invention include particle forms with protective coatings, protease inhibitors or penetration enhancers for various modes of administration, including parenteral, pulmonary, nasal, and oral. It is known that compounds modified by covalent addition of water-soluble polymers, such as polyethylene glycol, that is the main part of a non-alcohol and a polyethylene glycol, polyvinyl alcohol, polyvinyl pyrolidone, and polyvinylpyrrolidone, polyvinyl pyrrolidone, polyvinyl pyrrolidone, polyproline, polyvinyl pyrolidone, polyvinyl glycol, polyvinyl pyrolidone, polyvinyl alcohol, polyvinyl pyrrolidone or polypropylene glycol. e1 al., 1981, Heyagan e1 al., 1982 and Kaie e1 al., 1987). Such modifications can also increase solubility.

- 4 010263 compounds in aqueous solutions, eliminate aggregation, enhance the physical and chemical stability of the compound and significantly reduce the immunogenicity and reactivity of the compound. As a result, the desired biological activity can be achieved by less frequent administration of such adducts of polymers and compounds or by administering smaller doses than in the case of an unmodified compound.

In yet another embodiment, the pharmaceutical composition may be delivered in a controlled release system. For example, the agent can be administered using intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other means. In one of the variants, a pump can be used (see Kapdet, cited above; 8Neyop, SNF Stj. Ke £. Wuscheb., 14: 201 (1987); Visayteb e! A1., 8dedegu, 88: 507 (1980); 8aibek f! a1., N. Epd1. 1. Meb., 321: 574 (1989)). In another embodiment, polymeric materials may be used. In another embodiment, a controlled release system may be placed near the treatment target, i.e. in this case, only a part of the systemic dose is required (see, for example, Co-op. in Metablis1 Arrysiopes of the Sopteine K. Ketease, cited above, νο1. 2, p. 115-138 (1984)). Preferably, the controlled release device is administered to a subject in the vicinity of a location of inappropriate immune activation or a tumor. Other controlled release systems are described in a review (8c1fse, 249: 1527-1533 (1990)).

The method of the present invention for preventing prostate carcinogenesis includes administering to a mammal a pharmaceutical preparation containing a chemopreventive agent, or a salt or metabolite thereof. The pharmaceutical preparation may contain a single chemopreventive agent or may also contain a pharmaceutically acceptable carrier, and may be in solid or liquid form, such as tablets, powders, capsules, pills, solutions, suspensions, elixirs, emulsions, gels, creams or suppositories, including including rectal and urethral suppositories. Pharmaceutically acceptable carriers are resins, starches, sugars, cellulosic materials, and mixtures thereof. A pharmaceutical preparation containing a chemopreventive agent can be administered to a subject, for example, by subcutaneous implantation of a granule; in another embodiment, the granule provides controlled release of the chemopreventive agent over time. The drug can also be administered by intravenous, intraarterial, or intramuscular injection of a liquid preparation, by oral administration of a liquid or solid preparation, or by topical administration. Administration can also be done using a rectal suppository or a urethral suppository. The pharmaceutical preparation may also be a composition for parenteral administration; in one embodiment, the composition contains liposomes comprising a complex of a chemopreventive agent, such as, for example, toremifene, and cyclodextrin, as described in previously cited US Pat. No. 5,571,534,11,1pepe e! a1.

The pharmaceutical preparations of the invention can be obtained by known methods of dissolving, mixing, granulating or tableting. For oral administration, chemoprophylactic agents or their physiologically tolerable derivatives, such as salts, esters, Ν-oxides, etc., are mixed with additives common for this purpose, such as excipients, stabilizers, or inert diluents, and are converted by conventional means into a suitable form for administration, such as tablets, coated tablets, hard or soft gelatin capsules, water, alcohol or oil solutions. Examples of suitable inert fillers are the usual bases for tablets, such as lactose, sucrose, or corn starch, in combination with binders like gum arabic, corn starch, gelatin, or disintegrating agents such as corn starch, potato starch, alginic acid, or with lubricants like stearic acid or magnesium stearate. Examples of suitable oily excipients or solvents are vegetable or animal oils, such as sunflower oil or fish liver oil. Drugs can be effective in the form of dry and wet granules. For parenteral administration (subcutaneous, intravenous, intraarterial or intramuscular injection), chemoprophylactic agents or their physiologically tolerable derivatives, such as salts, esters,-oxides, etc., are converted into a solution, suspension or emulsion, if necessary, with substances conventional and suitable for this purpose, for example, solubilizers or other auxiliary substances. Examples are sterile liquids, such as water and oils, with or without the addition of a surfactant and other pharmaceutically acceptable adjuvants. Examples of oils are those derived from petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, or mineral oil. In general, water, saline, aqueous dextrose, and solutions of related sugars, and glycols, such as propylene glycols or polyethylene glycols, are preferred liquid carriers, in particular, for injection solutions.

The preparation of pharmaceutical compositions containing the active ingredient is well known in the art. Typically, such compositions are prepared as an aerosol of a polypeptide delivered to the nasal sinuses, or as injectables, or as liquid solutions or suspensions, but it is also possible to obtain solid forms suitable for preparing a solution or suspension before injection. The drug can also be emulsified. The active medicinal ingredient is often mixed with ex

- 5 010263 patients, who are pharmaceutically acceptable and compatible with the active ingredient. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol or the like, and combinations thereof. In addition, if necessary, the composition may contain small amounts of auxiliary substances, such as wetting agents or emulsifiers, buffer substances to stabilize the pH, enhancing the effectiveness of the active ingredient.

The active ingredient can be introduced into the composition in neutralized forms of pharmaceutically acceptable salts. Pharmaceutically acceptable salts include additive salts (formed by free amino groups of a polypeptide molecule or antibody) formed with inorganic acids, such as, for example, hydrochloric or phosphoric acids, or organic acids such as acetic, oxalic, tartaric, almond and the like Salts formed by free hydroxyl groups can also be prepared with inorganic bases, such as, for example, sodium, potassium, ammonium, calcium or iron hydroxides, and organic bases such as isopropylamine, trimethylamine, 2-ethylaminoethanol, histidine, procaine, and so on. P.

For topical administration on the surface of the body using, for example, creams, gels, drops, etc. get chemoprophylactic agents or their physiologically tolerable derivatives, such as salts, esters, оксид-oxides, etc., and used in the form of solutions, suspensions or emulsions in a physiologically acceptable diluent with or without a pharmaceutical carrier.

In another embodiment, the active compound can be delivered in a vesicle, in particular, a liposome (see Laideg, Scepter, 249: 1527–1533 (1990); Taga1 a1., In Hockecke Thiegaru οί 1пGesboi8 Egea ^ e apb Sapseg, ёί Gesboi8 Egea ^ e apb Sapseg, ёί Gesboi8 Egea ^ e apb Sapseg, iore Vegeyet Plebner (éb§.) .188, No. \\ Wogk, p. 353-365 (1989); Loreh-Vegeyet, ibid, pp. 317-327; see ibid., In general).

The pharmaceutical compositions of the present invention are particularly useful for treating a subject with an increased risk of developing prostate cancer. High-risk subjects include, for example, people with benign prostatic hyperplasia, prostate intraepithelial neoplasia (ΡΙΝ) or abnormally high levels of circulating antibodies specific for the prostate gland (P8A), or a family member with prostate cancer.

In addition, a prostate chemoprophylaxis agent can be administered in combination with other cytokines or growth factors, including ΙΡΝ-γ or ΙΡΝ-α, ΙΡΝ-β; interleukin (1b) 1, 1b-2, 1b-4, 1b-6, 1b-7, 1b-12, tumor necrosis factor ΤΝΡ-α, ΤΝΡ-β, granulocytic colony-stimulating factor (C-C8P), C8P soil-macrophage ( CM-C8P) and others; additional molecules, including members of the superfamily of integrins and members of the superfamily 1d, such as LPA-1, LPA-3, SE22 and B7-1, B7-2, and others, and T-cell stimulation cc-1CAM-1.

The chemopreventive agent may precede or follow the treatment with a DNA damaging agent at intervals from minutes to weeks. Applications and methods are known to those skilled in the art. The means or factors that damage DNA are known to those skilled in the art and denote any chemical compound or treatment method that induces DNA damage when applied to a cell. Such means and factors include radiation and waves that induce DNA damage, such as gamma radiation, x-rays, UV radiation, microwaves, electron emission, and the like. Various chemical compounds, also described as chemoprophylactic agents, function as inducing DNA damage, and it is assumed that they are all used in the combined treatment methods described in this specification. Chemoprophylactic agents intended for use include, for example, adriamycin, 5-fluorouracil (5RI), etoposide (ΥΡ-16), camptothecin, actinomycin-, mitomycin C, cisplatin (COOP), and even hydrogen peroxide. The invention also encompasses the use of a combination of one or more DNA damaging agents, whether they are based on radiation or on actual compounds, such as applying X-rays with cisplatin or using cisplatin with etoposide.

In another embodiment of the invention, a localized tumor site can be irradiated with radiation that damages DNA, such as X-rays, UV light, gamma rays, or even microwaves. Alternatively, tumor cells can be brought into contact with a DNA damaging agent by administering to the subject a therapeutically effective amount of a pharmaceutical composition containing a DNA damaging compound, such as adrimycin, 5-fluorouracil, etoposide, camptothecin, actinomycin-Ό, mitomycin C or preferably cisplatin. DNA damaging agents also include compounds that affect DNA replication, mitosis and chromosomal segregation. Such chemotherapeutic agents include adrimycin, also known as doxorubicin, etoposide, verapamil, podophyllotoxin, etc.

Other factors that cause DNA damage and are widely used include well-known factors such as gamma radiation, x-rays and / or targeted delivery of radioactive isotopes to tumor cells. Other forms of DNA damaging factors are also contemplated, such as microwaves and UV radiation. Most likely all of these factors

- 6 010263 act in a wide range of DNA damage on DNA precursors, DNA replication and repair and on the assembly and preservation of chromosomes.

As an ordinary person skilled in the art can easily imagine, the methods and pharmaceutical compositions of the present invention are particularly suitable for administration to a mammal, preferably a human.

Transgenic Mouse Prostate Adenocarcinoma

The study of chemoprophylaxis of prostate cancer was hampered by the absence of suitable animal models. The recent development of transgenic mouse prostate adenocarcinoma (TCAMR) makes it possible to investigate chemoprophylaxis. In the model TCAMR described in Stiepetd e! A1., Progressor of the World Trade Union, Progress. №11. Asab. 8c1. I8A, 1995, νο1.92, p. 3439-3443, the RV-8U40 large T-antigen (RV-Tad) transgene is specifically expressed in the mouse prostate gland epithelial cells. As a result, this model has several advantages over the models that currently exist:

1) mice develop early progressive forms of epithelial hyperplasia of the prostate gland - at the age of 10 weeks and invasive carcinoma at about 18 weeks;

2) the picture of metastatic dissemination of prostate cancer resembles prostate cancer in humans, and the lymph node, lung, kidney, adrenal gland and bone are common metastasis areas;

3) development, as well as promotion of prostate cancer, may follow within a relatively short period of 10-30 weeks;

4) tumors appear with a frequency of 100% and

5) animals can be screened for the presence of a prostate cancer transgene prior to the onset of clinical prostate cancer in a direct test with a toremifene treatment, which can change the carcinogenesis of the prostate gland.

The transgenic mouse model of TCAMR is an excellent model of ίη νίνο for determining the mechanisms for initiating and promoting prostate cancer and for testing the effectiveness of potential toremifene. In these mice, epithelial prostatic hyperplasia, RHY and then prostate cancer develop progressively in a short period (<17 weeks).

Chemoprophylactic treatment of the hybrid mouse TCAMR is started postnatally after 30 days using toremifene at a level of about 0.5–50 mg per 1 kg of the subject’s mass per day, preferably about 6–30 mg per 1 kg of the subject’s mass per day. Toremifene is conveniently produced in 21-day and 90-day granules (manufactured by Kekeags on Ates, Sarasota, Florida) and delivered as subcutaneous implants. Control animals receive placebo implants. In each drug-treated group, animals were sacrificed at the age of 5, 7, 10, 15, 20, 25, 30, 40, and 50 weeks before the palpable tumor developed. Blood is collected by treatment time to assess changes in serum testosterone and estradiol. Prostate tissues are collected for morphometric, histological and molecular studies.

Use the following test procedures:

1) periodically analyze the prostate gland to detect changes in the morphology of the prostatic grooves with time with and without treatment; Examples are given in FIG. 2. Tissue sections were assessed histologically using the H & E method and standard Ma55op-1ps1ute staining. The occurrence of RHL is estimated and distributed by degrees (I - moderate, III - severe);

2) measure the content of estradiol in serum and total testosterone (VA) for each age interval and evaluate any changes in the content of these hormones as a result of the action of toremifene.

Results of immuno-histochemical analysis

Images in the microscope of each tissue section are evaluated using quantitative image assessment using a computer (computer and monitor Mac 9500M 32), using a Kobak'.'8 460 camera on a microscope # which is M1goryo! -EX, and quantification is carried out using a quantitative system image analysis using color (SEB 8 reps titer 3.1, 8 papa1, Bc., AU), which distinguishes differences in staining of tissue sections. Establish thresholds for the identification of various components of prostate tissue. Photographic densities of areas corresponding to each of these tissue components are calculated for each full screen of a color monitor. In general, data for sections of the prostate gland are averaged over 5 screens. Immunohistochemical images can be ditalized and quantified in order to be able to be statistically evaluated by determining sample correlation and probability coefficients (2-sequential).

Chemoprophylactic Activity Study

Undertake research to test the effectiveness of toremifene on transgenic animals TCAMR (RVTad X ECM) (provided by Dr. Yottapp Steepetd, Journal Coedé about Mebouche, TX). In these mice, preliminary signs of cancer appear early - at 10 weeks. The males in the progeny of transgenic TCAMR are screened for the large T-antigen transgene, and positive males are used in the study. Antiestrogen toremifene, which is tested for its possible chemoprofys

- 7 010263 lactic effect, injected into the prepared granules (ΙηποναΙίνο Veseags οί Athens, Sarasota, Florida), and begin chemoprophylactic treatment of mice postnatally on day 30 (average mouse weight 14 g). Mice in each of the four groups of 10–12 animals are given subcutaneous implantation of toremifene-containing pellets with a 90-day release. The dosage of the diffusing drug fitted for growth associated with weight changes is created for delivery or low dose (6 mg / kg) or high dose (30 mg / kg) toremifene. Control animals (η = 10) receive placebo implants. Treatment efficacy is assessed by the absence of palpable tumor formation. Tumors of the prostate glands of mice are collected and evaluated by molecular and histological methods.

Using a transgenic model of TUMP prostate cancer, in which each animal inheriting a prostate cancer gene develops prostate cancer, it has been shown that toremifene both increases the latent period and reduces the incidence of prostate cancer.

As seen in FIG. 1, the effect of toremifene is effective both at low and at high dose. The formation of a tumor of the ventral lobe of the prostate gland in a mouse TAMPA is observed at week 17 in the placebo group (η = 10), at week 19 in the group treated with a high dose of toremifene (η = 12), and at week 28 in the group treated with a low dose of toremifene (η = 12). Thus, in the case of 5 treatments with toremifene, the period of latency for the development of cancer of the ventral lobe of the prostate gland in TMAH mice is increased by essentially 11 weeks.

Since the tumor development in 50% of the animals treated with toremifene was not achieved during the study period, the comparison between the groups is carried out over the time during which 25% of the animals had tumors. Tumors are palpated in 25% of 10 animals at week 23 in the placebo group and at 30–31 weeks in the high and low dose groups of toremifene, with a delay of 7–8 weeks. Both low and high doses of toremifene, versus placebo, are significant according to the 1-rank analysis and Wilcoxon statistical analysis, as shown in Table 2 below. one.

Statistical analysis

Co-rank

R

Table 1 by Wilcoxon

R

Low dose toremifene placebo bonds 0.0003 * 0.0004 *

High dose toremifene placebo bond 0.0017 * 0.0071 * * Significance P <0.05

At week 33, at the time when all control animals developed tumors, 72% of the animals that received a low dose of toremifene, and 60% of the animals that received a high dose did not have tumors. Thus, treatment with toremifene, both at low and at high dosage, leads to a significant decrease in the incidence of tumors in the ventral lobe of the prostate gland in TBAMP mice. These results indicate that the incidence of prostate cancer is significantly reduced and the latency period increases.

As already mentioned, the administration of toremifene produces a significant chemoprophylactic effect against tumors in the ventral lobe of the prostate gland in TBAMR mice. Such a result gives hope for such a favorable effect on people in whom the prostate gland contains a segment corresponding to the ventral lobe of the rodent prostate gland.

Histological examination of prostate tissue

Tumors taken at the time of palpation in groups with placebo and received high and low dose of toremifene, histologically evaluated. FIG. 2A is a section H & E of the ventral lobe of the prostate in a 17 week-old healthy adult mouse. FIG. 2B shows a section of the ventral lobe of the prostate gland of a placebo-treated 16-week-old mouse of TAMPA, which, in contrast to the structure of the healthy prostate gland shown in FIG. 2A, the ventral lobe of the prostate gland of a mouse TBAMP is characterized by layers of undifferentiated anaplastic cells with a high mitotic index. In contrast, as seen in FIG. 2C, the prostate gland of toremifene-treated 30-week-old TAMPA mouse retains much of its normal granular structure and has tumors with a more differentiated structure, and the mitotic index is significantly lower than in animals treated with placebo. These results show that toremifene, even at low dosage, is able to inhibit prostate carcinogenesis on the TUMP model.

Western blot. Prostate tissue (dorsol-teral and ventral lobes) is harvested at the age of 10 weeks, quickly frozen in liquid ₂ and stored at -80 ° C. Tissue lysates are prepared using V1PA buffer (150 mM αΝίί. 1% 40, 0.5% deoxycholate, 0.1% 8Ό8, and 50 mM Tris, pH 7.5) containing a mixture of protease inhibitors (pefablock, aprotinin, bestainin , leupeptin and pepstein) and the phosphatase inhibitor №1 ₃ UO ₄ (10 mM). The homogenate is centrifuged at 14,000 / d at 4 ° C for 10 minutes, and the lysates are stored at -80 ° C.

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Determine protein concentrations using a Bradford protein assay (Βίο-Slave). Tissue lysates load 7.5% polyacrylamide gels, proteins (40 μg / lane) are separated by the method of 8Ό8ΡΆΟΕ and electrophoretically transferred to nitrocellulose membranes (0.2 μm, Βίο-Rab, Hercules, California) in a transfer buffer (192 mM glycine, 25 mM Tris-HC1 and 20% methanol). Prostate tumor tissue TRAMR is used as a positive control. Chemiluminescent markers Verse are used as molecular weight standards (Verse 8Alla, Snta Cruz, California). Blots are blocked overnight at 4 ° C in VYOTTO (6% non-fat dry milk in IX TB8) and incubated with primary antibodies to the large T-antigen (monoclonal mouse RaB 101, 1: 200, 8a1 Verse Vyu1es1odojoda) for 2 h at room temperature. The blots were washed with (3x) TTV8 (0.05% Tween 20, 50 mM Tris-HC1, 200 mM NaC1) and incubated with horseradish peroxidase (HPP) secondary antibodies (1: 5000) for 25 hours. WITH. Immunoreactive proteins are visualized on an autographic film using enhanced chemiluminescence (ES) (ARV, Piscataway, New Jersey). Actin protein expression is used to normalize the results of Thad. To this end, the above membranes are immersed in a removal buffer (100 mM 2-mercaptoethanol, 2% 8Ό8, 62.5 mM Tris-HCl, pH 6.7) and incubated at 50 ° C for 30 minutes, stirring occasionally. After blocking, the membrane is re-probed with primary antibodies to actin (1: 2500, Syetuop, Temecula, California) and then with secondary antibodies conjugated to HPP (1: 10,000). After the detection by the ECB method, the intensity of the bands is quantified using the Aboe RNoyUyur 5.0 data acquisition systems and the analysis of the first step of Oyap1 (Molesiup Oupatiuk).

Application of the chemoprophylactic action of toremifene against prostate cancer in the TRAM mouse model

This experiment confirms and demonstrates the chemoprophylactic efficacy of toremifene. This study focuses on the histological and molecular changes associated with the development of a prostate tumor in control animals and the mechanism of toremifene action on TRAMR animals, which are propagated, screened and treated with delayed-release granules of the drug. At predetermined times, animals are killed in groups of 5 animals and their prostate glands are removed for analysis. Prostate glands are assessed for the presence of a tumor histologically, dissecting the lymphoma and immunochemically with a large T-antigen. Placebo and toremifene are administered at 7, 10, 15, and 20 weeks to obtain data, and the results are discussed below.

Results. AMetoip! complete at 7, 10, 15 and 20 weeks for different groups of mice. An analysis of the U1I-type1 shows that in mice treated with placebo, prostate tumors develop at the age of 15-20 weeks, as in previous pilot studies. In addition, in animals treated with toremifene, there is a delay in the appearance of prostate cancer up to 20 weeks (Fig. 3). In addition to 20 weeks, in the group of animals treated with toremifene, there is an impressive delay in the appearance of a tumor up to 35 weeks (Fig. 4). These results confirm that even with a more sensitive assessment of oncogenicity, toremifene shows chemoprophylactic activity. For histological evaluation, tissue samples are fixed, processed and enclosed in paraffin.

Cuts are obtained (thickness 5 pm) and stained with the usual H & E method. Toremifene inhibits the development of the grooves and tissue differentiation (compared to week 17, prostate tumor of the mouse TRAMR and wild type (Fig. 4)); histology of the prostate gland treated with toremifine and placebo at week 15 (Fig. 5). Qualitatively, the immunohistochemistry of tissues treated with placebo and toremifene shows the presence of T-antigen in the ventral prostate gland. Thus, on the TRAMR model, it appears that the chemoprophylactic activity demonstrated by toremifene is not manifested by the suppression of the probazine (primus) promoter.

Findings. The ability of toremifene to prevent the occurrence of prostate cancer on the TRAM model is confirmed using more sensitive methods for assessing the formation of a tumor. It turns out that the mechanism of the chemoprophylactic action of toremifene does not constitute a loss of the transgene for a large T-antigen protein.

Toremifene induces regression of identified human prostate cancer on a model of nude mice

Currently, prostate cancer is the most commonly diagnosed cancer in Americans. However, there are questions of the etiology and treatment of the disease, especially in progressive forms. Hormone therapy remains the usual treatment for recurrent and progressive prostate cancer, despite the frequent development of a disease that cannot be treated with hormones. Therefore, new approaches to the prevention and treatment of prostate cancer are needed, which would help an increasing number of men diagnosed with this disease. The experiments and the results given below show that toremifene inhibits the growth of a hormone sensitive HChCaR tumor in athymic nude mice.

Materials and methods. One million BCSaP cells in matrigel are injected subcutaneously with atymic naked mice in each flank. In total, 40 mice are injected. After about 3-4 weeks

- 9 010263 visible tumors develop. After registering the size of the tumors in two dimensions, the mice are divided into placebo groups and treatments based on the equivalent tumor load. A separate granule (placebo versus 35 mg of Thorimifen) is implanted subcutaneously with each mouse between the shoulder blades. Register weekly observations of the size of the tumor. Calculate the tumor volume (tumor volume = 0.5 (L + ^) xbx ^ x0.5236, where b = tumor length and ^ = mass). The tumor volume at the time of the pellet implantation serves as a starting point for future comparison of tumor size changes. Weekly changes in the volume of each tumor are recorded as differences, as a percentage, with the initial measurement during the implantation of the granule.

Results. Of the 78 sites with injected tumors, 55 (70%) received tumors for evaluation, adequate in volume. Only 50 tumors are available for evaluation (24 animals with placebo and 26 animals treated with toremifene). The average tumor volumes at the time of implantation of the granules are similar for the groups treated with toremifene and placebo (1.90 mm ³ and 1.72 mm ^3, respectively). In the toremifene group, the average volume decreases to 1.68 mm ³ (-0.22 mm ³ ), while the average tumor volume in the placebo group increases to 2.33 mm ³ (+0.61 mm ³ ). The mean serum R8L level is higher in the placebo group (3.80 ng / ml) than in the toremifene group (2.80 ng / ml), but statistically this is not significant (p = 0.755). The total testosterone level is 2.18 ng / ml for the group with placebo (n = 17) and 2.96 ng / ml for the group with toremifene (n = 19).

Two mice died shortly after pellet implantation due to bites from other mice. One mouse treated with toremifene was excluded from the study due to hemorrhage in the tumor and the development of hematoma. All mice developed unilateral or bilateral tumors. Continued an independent study of each tumor. 24 tumors were treated with placebo and 28 tumors were treated with toremifene. The results are given in table. 2 and in FIG. 6 A and 6B.

table 2

Placebo group

Number of weeks Volume change relative to day 0 of treatment,%

3 eleven 9.44 four eight 115.27 five eight 271.71 6 eight 600.88

Toremifen

Number of weeks Change volume relative to day 0 of processing, 3 eleven -34,58 four 7 -61.01 five 7 -74,51 b five -61,72

Subsequent intervals for this population will be extended, and data for other animals are currently being collected.

Findings. Toremifene inhibits and induces regression of identified EI-R tumors. Although the mechanism by which toremifene exerts its action is unknown, the ability to exert such an effect is the argument for the use of toremifene as a method for treating prostate cancer and preventing regression of prostate cancer in risk patients with prostate cancer micrometastases.

Toremifene causes regression ΗΟΡΙΝ in clinical trials in humans with prostate cancer in phase 11a.

The chemopreventive effect of toremifene against prostate cancer is reproducibly demonstrated in this description on a generally accepted animal model of spontaneous prostate cancer. Toremifene is the first compound to demonstrate chemoprophylactic activity against prostate cancer.

In addition, a high degree of intraepithelial neoplasia (ΗΟΡΙΝ) and test time are established as a pathological change in the prostate cancer human precursor, also known as latent prostate cancer.

Then, ΗΟΡΙΝ is used as an intermediate target or a surrogate target in case of prostate cancer. Indeed, Νί.Ί is now recommended that ΡΙΝ should be used as an intermediate goal or a surrogate target in case of prostate cancer.

In phase 11a, a separate non-randomized study was conducted with open treatment with 17 patients. According to this scheme, patients with ΡΙΝ, proven by biopsy, are treated with toremifene 60 mg daily for 4 months. After 4 months, patients undergo a repeated biopsy (8 biopsies) and overestimate the status ΡΙΝ. Twenty-one patients entered the study, and completed the study pole

- 10 010263 eleven patients. The sum of pathological changes in prostate biopsy in these 16 patients shows that 12 patients have a regression of ΡΙΝ to benign or atrophic prostate tissue; thus, 12 out of 16 (75%) patients have a complete response. Of the remaining 4 patients, 3 have prostate cancer, but the number ΡΙΝ decreases, and 1 patient has a stable disease, but the epithelium in the area shows atrophic and degenerative changes.

An assessment of the pathology shows a complete decomposition ΡΙΝ with atrophic changes in the epithelium of the prostate gland. The patient has no acute or chronic toxicity when taking toremifene. Serum P8A, serum free testosterone, serum total testosterone and serum estradiol remain at normal intervals. The quality of life does not change, including the lack of influence on potency and libido. Therefore, the above results show the chemopreventive role of toremifene for the prostate gland.

The results show that toremifene weakens ΡΙΝ, which ultimately translates into a reduction in the incidence of the disease and prolongation of the prostate cancer latency and prevention of prostate carcinogenesis. Finally, toremifene was found to significantly induce TCE synthesis in human stromal fibroblasts.

Example 1. The use of tamoxifen citrate and raloxifene (8EKM) and faslodex (pure antiestrogen 1C1 182,780) in the prevention of prostate carcinogenesis.

Construction of the experiment. Chemotherapy with mice starts postnatally on day 30. Three groups of 50 male TCAMR hybrid mice each are treated with either anti-estrogens with tamoxifen citrate, or raloxifene (8EKM), or faslodex (pure antiestrogen 1C1 182,780). Drugs are obtained in the form of granules prepared for the delayed release of the drug (ΙηηννηΙίνε Keekaty o £ Atsps. Sarasota, Florida) and delivered as subcutaneous implants (see preliminary results). Control animals receive placebo implants without pharmacological activity. Animals (and = 10) are euthanized periodically at the age of 10, 15, 20, 25 and 30 weeks and evaluate the effectiveness of the treatment, leading either to the absence of the formation of a tumor, or a reduction in the size of the tumor, if any, comparing with control animals that received a placebo. Blood is collected to evaluate changes in serum levels of androgens and estrogens during each treatment. Prostate tissues are retained for a) morphometric studies; (B) for histological examination, tissue should be fixed in 10% buffered formalin, processed and embedded in paraffin; c) for molecular studies, the tissue is frozen in liquid nitrogen and stored at -70 ° C. Autopsy and survival data is also recorded.

The experimental results show the relative chemoprophylactic efficacy of various antiestrogens in delaying or preventing prostate cancer on the TCAMR model. Morphological studies show significant changes, if any, in the development of the size of the prostate gland and the ductal pattern as a result of each treatment. Paraffinized tissue sections are stained using the standard H & E method for histological changes, such as ΡΙΝ, which will be assessed by the appearance on the monitor of precancerous pathological changes as a precursor of prostate adenocarcinoma. The content of estradiol and total serum testosterone is measured for each age interval and the changes in the content of these hormones are assessed and whether or not they correlate with changes in, the content of peptide growth factors T6E, T6E 1, T6E 3 and ЬЕ6Е is measured quantitatively in the prostate gland samples taken in each interval. The corresponding peptide growth factor receptors E6EK and T6E КП and KP are also evaluated.

Table 3 The effect of selective estrogen receptor modulators (8 EKM) on the prevention of prostate cancer on the TCAMR model

8EKM Dose 20 weeks, 20 weeks, number of tumors (% tumors)

Placebo - 5/5 100% Toremifen 20 mg / kg / day 1/7 14.2% Tamoxifen 20 mg / kg / day 2/9 22% Raloxifene 20 mg / kg / day 3/10 thirty% * Faslodex * 10 mg / kg / day 8/11 72% (1C1 128,780)

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The animals are sacrificed at week 20, and the prostate glands are assessed by the method of analysis of the lymphoma and histologically.

Claims (18)

CLAIM 1. The use of raloxifene, or tamoxifen, or its derivative, isomer, or pharmaceutically acceptable salt, ester, β-oxide, or a mixture thereof, for the preparation of a pharmaceutical composition for the treatment of precancerous precursors of prostate adenocarcinoma. 2. The use of raloxifene, or tamoxifen, or a derivative thereof, an isomer, or a pharmaceutically acceptable salt, ester, Ν-oxide, or a mixture thereof, for the preparation of a pharmaceutical composition to reduce the number of precancerous precursors of prostate adenocarcinoma. 3. The use of raloxifene, or tamoxifen, or its derivative, isomer, or pharmaceutically acceptable salt, ester, Ν-oxide, or a mixture thereof, for the preparation of a pharmaceutical composition for the inhibition or suppression of precancerous precursors of prostate adenocarcinoma. 4. The use according to claims 1 to 3, in which the precursors of prostate adenocarcinoma are intraepithelial neoplasia of the prostate gland (ΡΙΝ). 5. The use of claims 1-3, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. 6. The use according to claim 5, in which the carrier is selected from the group consisting of resin, starch, sugar, cellulosic material and mixtures thereof. 7. The use according to claim 5, in which the pharmaceutical composition is made in the form of granules for subcutaneous implantation. 8. The use according to claim 5, in which the granules provide controlled release of raloxifene, or tamoxifen, or its derivative, isomer, or pharmaceutically acceptable salt, ester, Ν-oxide, or a mixture thereof for a certain period of time. 9. The use according to claim 5, in which the pharmaceutical composition is made in the form of a liquid for intravenous, intraarterial or intramuscular injection. 10. The use according to claim 5, in which the pharmaceutical composition is made in the form of a liquid or solid preparation for oral administration. 11. The use according to claim 5, in which the pharmaceutical composition is made in the form of a form suitable for topical application. 12. The use according to claim 5, in which the pharmaceutical composition is made in the form of granules, tablets, capsules, solution, suspension, emulsion, elixir, gel, cream or suppository. 13. The use of claim 12, wherein the suppository is a rectal or urethral suppository. 14. The use according to claim 5, in which the pharmaceutical composition is made in the form of a form suitable for parenteral administration. 15. The use of claim 14, wherein the parenteral form is liposomes comprising a complex of raloxifene or tamoxifen, or a derivative thereof, an isomer, or a pharmaceutically acceptable salt, ester, Ν-oxide, or a mixture thereof with a cyclodextrin compound. 16. The use according to claim 5, in which raloxifene, or tamoxifen, or its derivative, isomer or metabolite, or a pharmaceutically acceptable salt, ester, Ν-oxide, or a mixture thereof are made in a form that provides a dosage of from about 0.5 to approximately 80 mg per 1 kg of subject weight per day. 17. The use according to clause 16, in which raloxifene, or tamoxifen, or its derivative, isomer or metabolite, or a pharmaceutically acceptable salt, ester, β-oxide, or a mixture thereof are made in a form providing a dosage of from about 20 to about 60 mg per 1 kg of subject weight per day. 18. The use according to clause 16, in which raloxifene, or tamoxifen, or its derivative, isomer or metabolite, or a pharmaceutically acceptable salt, ester, Ν-oxide, or a mixture thereof are made in a form that provides a dosage of 20 mg per 1 kg of weight subject per day.